Advancing skeletal health and disease research with single-cell RNA sequencing.

Orthopedic conditions have emerged as global health concerns, impacting approximately 1.7 billion individuals worldwide. However, the limited understanding of the underlying pathological processes at the cellular and molecular level has hindered the development of comprehensive treatment options for these disorders. The advent of single-cell RNA sequencing (scRNA-seq) technology has revolutionized biomedical research by enabling detailed examination of cellular and molecular diversity. Nevertheless, investigating mechanisms at the single-cell level in highly mineralized skeletal tissue poses technical challenges. In this comprehensive review, we present a streamlined approach to obtaining high-quality single cells from skeletal tissue and provide an overview of existing scRNA-seq technologies employed in skeletal studies along with practical bioinformatic analysis pipelines. By utilizing these methodologies, crucial insights into the developmental dynamics, maintenance of homeostasis, and pathological processes involved in spine, joint, bone, muscle, and tendon disorders have been uncovered. Specifically focusing on the joint diseases of degenerative disc disease, osteoarthritis, and rheumatoid arthritis using scRNA-seq has provided novel insights and a more nuanced comprehension. These findings have paved the way for discovering novel therapeutic targets that offer potential benefits to patients suffering from diverse skeletal disorders.

The SIRT3-ATAD3A axis regulates MAM dynamics and mitochondrial calcium homeostasis in cardiac hypertrophy.

Mitochondria are energy-producing organelles that are mobile and harbor dynamic network structures. Although mitochondria and endoplasmic reticulum (ER) play distinct cellular roles, they are physically connected to maintain functional homeostasis. Abnormal changes in this interaction have been linked to pathological states, including cardiac hypertrophy. However, the exact regulatory molecules and mechanisms are yet to be elucidated. Here, we report that ATPase family AAA-domain containing protein 3A (ATAD3A) is an essential regulator of ER-mitochondria interplay within the mitochondria-associated membrane (MAM). ATAD3A prevents isoproterenol (ISO)-induced mitochondrial calcium accumulation, improving mitochondrial dysfunction and ER stress, which preserves cardiac function and attenuates cardiac hypertrophy. We also find that ATAD3A is a new substrate of NAD+-dependent deacetylase Sirtuin 3 (SIRT3). Notably, the heart mitochondria of SIRT3 knockout mice exhibited excessive formation of MAMs. Mechanistically, ATAD3A specifically undergoes acetylation, which reduces self-oligomerization and promotes cardiac hypertrophy. ATAD3A oligomerization is disrupted by acetylation at K134 site, and ATAD3A monomer closely interacts with the IP3R1-GRP75-VDAC1 complex, which leads to mitochondrial calcium overload and dysfunction. In summary, ATAD3A localizes to the MAMs, where it protects the homeostasis of ER-mitochondria contacts, quenching mitochondrial calcium overload and keeping mitochondrial bioenergetics unresponsive to ER stress. The SIRT3-ATAD3A axis represents a potential therapeutic target for cardiac hypertrophy.

Spatially multicellular variability of intervertebral disc degeneration by comparative single-cell analysis.

Previous studies have revealed cellular heterogeneity in intervertebral discs (IVDs). However, the cellular and molecular alteration patterns of cell populations during degenerative progression remain to be fully elucidated. To illustrate the cellular and molecular alteration of cell populations in intervertebral disc degeneration (IDD), we perform single cell RNA sequencing on cells from four anatomic sites of healthy and degenerative goat IVDs. EGLN3+ StressCs, TGFBR3+ HomCs and GPRC5A+ RegCs exhibit the characteristics associated with resistance to stress, maintaining homeostasis and repairing, respectively. The frequencies and signatures of these cell clusters fluctuate with IDD. Notably, the chondrogenic differentiation programme of PROCR+ progenitor cells is altered by IDD, while notochord cells turn to stemness exhaustion. In addition, we characterise CAV1+ endothelial cells that communicate with chondrocytes through multiple signalling pathways in degenerative IVDs. Our comprehensive analysis identifies the variability of key cell clusters and critical regulatory networks responding to IDD, which will facilitate in-depth investigation of therapeutic strategies for IDD.

A Multicomponent Nucleic Acid Enzyme-Cleavable Quantum Dot Nanobeacon for Highly Sensitive Diagnosis of Tuberculosis with the Naked Eye.

Clinical tuberculosis (TB) screening and diagnosis are crucial for controlling the spread of this life-threatening infectious disease. In this work, a novel, rapid, and simple colorimetric detection platform for TB was developed based on a quantum dot-based nanobeacon (QD-NB) and multicomponent nucleic acid enzyme (MNAzyme). In the presence of target DNA (IS1081 gene fragment), the recombinase polymerase amplification (RPA) was performed and the amplicons were chemically DNA-denatured and then subjected to MNAzyme reaction. RNA-cleaving MNAzyme assembly included the recognition of target DNA and hybridization with a QD-NB fluorescence probe. Under the addition of Mg2+, the RNA-containing QD-NB as a cleavable substrate could be broken into two DNA fragments, leading to green fluorescence release due to their departure from a black hole quencher (BHQ2). The TB detection could be achieved with the naked eye under a portable and inexpensive UV flashlight. Our results demonstrated that QD-NB-based MNAzyme colorimetric assays improved the detection sensitivity by 1 order of magnitude compared with the detection using RPA. The limit of detection (LOD) of the visual reading was as low as 2 copies/µL (3.3 amol/L). Excellent specificity and reproducibility could also be achieved. Furthermore, the practical application of the colorimetric method for TB diagnosis was verified by 36 clinical TB patients and 20 healthy individuals. The developed QD-NB-based MNAzyme colorimetric assays provided a rapid, convenient, sensitive, and accurate alternative for clinical TB screening and diagnosis.

A novel and cost-efficient allele-specific PCR method for multiple SNP genotyping in a single run.

Cost-effective methods for DNA genotyping were needed because single nucleotide polymorphisms (SNPs) were essential biomarkers associated with many diseases. Allele-specific PCR (AS-PCR) has the advantages of mature instruments and high sensitivity. But conventional AS-PCR needs to multiply the number of reactions or primers for multiple targets, which complicates the operation and increases the cost. Herein, we proposed a novel AS-PCR method for multiple SNP genotyping in a single run. Wild-type allele-specific primer (WT primer) was designed for each target gene. The sample and WT primers only needed to undergo multiplexed AS-PCR once simultaneously. After AS-PCR, the concentration of remaining primers varied among the samples of each genotype combination, due to the different matching performance between template and WT primers. The remaining primers then triggered multiplexed molecular beacon-rolling circle amplification, and the molecular beacons labelled with different fluorescent dyes corresponded to different targets. The fluorescence ratios of the sample to the positive control were used as the genotyping indexes. This method was able to detect samples with concentrations as low as 10 fM. We successfully applied the method to the multiple genotyping of 23 hair root samples for ADH1B and ALDH2 genes, obtaining completely consistent results with sequencing. The reagent cost was 0.6 dollar for one sample, showing a good cost performance. This proposed approach had a great application prospect in simultaneously rapid and accurate genotyping of multi-SNPs, and provided a new method for personalized health management.

Fluorescence Biosensor for One-Step Simultaneous Detection of Mycobacterium tuberculosis Multidrug-Resistant Genes Using nanoCoTPyP and Double Quantum Dots.

The diagnosis of multidrug-resistant tuberculosis (MDR-TB) is crucial for the subsequent drug guidance to improve therapy and control the spread of this infectious disease. Herein, we developed a novel florescence biosensor for simultaneous detection of Mycobacterium tuberculosis (Mtb) multidrug-resistant genes (rpoB531 for rifampicin and katG315 for isoniazid) by using our synthesized nanocobalt 5,10,15,20-tetra(4-pyridyl)-21H,23H-porphine (nanoCoTPyP) and double quantum dots (QDs). Several nanoCoTPyPs with different charges and morphology were successfully prepared via the surfactant-assisted method and their quenching ability and restoring efficiency for DNA detection were systematically analyzed. It was found that spherical nanoCoTPyP with positive charge exhibited excellent quenching effect and sensing performance for the two DNAs' detection due to its affinity differences towards single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA). ssDNA attached on QDs (QDs-ssDNA) was specifically hybridized with targets to form QDs-dsDNA, resulting in fluorescence recovery due to the disruption of the interactions between nanoCoTPyP and ssDNA. Two drug-resistant genes could be simultaneously quantified in a single run and relatively low limits of detection (LODs) were obtained (24 pM for T1 and 20 pM for T2). Furthermore, the accuracy and reliability of our method were verified by testing clinical samples. This simple and low-cost approach had great potential to be applied in clinical diagnosis of MDR-TB.

Determination of tuberculosis-related volatile organic biomarker methyl nicotinate in vapor using fluorescent assay based on quantum dots and cobalt-containing porphyrin nanosheets.

Methyl nicotinate (MN) is a representative and typical volatile organic marker of Mycobacterium tuberculosis, and the specific detection of MN in human breath facilitates non-invasive, rapid, and accurate epidemic screening of tuberculosis infection. Herein, we constructed a fluorescent assay consisted of CdTe quantum dots (QD) and cobalt-metalized tetrakis(4-carboxyphenyl) porphyrin (CoTCPP) nanosheets to determine methyl nicotinate (MN) in vapor samples. Red-emission QD (λex=370 nm, λem=658 nm) acts as signal switches whose fluorescence signals can be effectively quenched by CoTCPP nanosheets but restored in the presence of MN. The strategy relied on the distinct binding affinity of cobalt ion and MN. MN restored the fluorescence of QD quenched by CoTCPP in a concentration-dependent manner, which exhibited a well-linear relationship in the range 1-100 µM, and a limit of detection of 0.59 µM. The proposed platform showed sensitivity and selectivity to detect MN in vapor samples with satisfactory RSD below 3.33%. The method is cheap, simple, and relatively rapid (detected within 4 min), which suggests a potential in tuberculosis diagnosis in resource- and professional-lacked areas.

DNA functionalized double quantum dots-based fluorescence biosensor for one-step simultaneous detection of multiple microRNAs.

The disease diagnosis by detecting single microRNAs (miRNAs) can produce high false positive rate. Herein, a novel fluorescence biosensor method for one-step simultaneous detection of multiple miRNAs was proposed by using single-stranded DNA (ssDNA) functionalized double quantum dots (QDs) and black hole quencher (BHQ)-decorated magnetic nanobeads (MNs). MNs were linked with two black hole quenchers (BHQ1 and BHQ3) via a complementary DNA (cDNA). The ssDNA/cDNA hybridization contributed to the fluorescence quenching of double QDs due to the fluorescence resonance energy transfer (FRET) between double QDs and BHQ. In the presence of target miRNA-33 (miR-33) and miRNA-125b (miR-125b), the ssDNA1 and ssDNA2 were respectively hybridized with miR-33 and miR-125b to form more stable duplexes. Thus, the double QDs were released into supernatant after the magnetic separation, leading to the fluorescence signals recovery at 537 nm and 647 nm. A wide linear range (0.5 nM-320 nM for miR-33 and 0.1 nM-250 nM for miR-125b) and low limits of detection (0.09 nM for miR-33 and 0.02 nM for miR-125b) were achieved. Moreover, our approach has been demonstrated to simultaneously detect miR-33 and miR-125b in cell extracts. With advantages of high sensitivity, strong specificity, low background and low cost, the strategies show great potentials for the detection of various targets in bioanalysis and disease diagnosis.

Spatially defined single-cell transcriptional profiling characterizes diverse chondrocyte subtypes and nucleus pulposus progenitors in human intervertebral discs.

A comprehensive understanding of the cellular heterogeneity and molecular mechanisms underlying the development, homeostasis, and disease of human intervertebral disks (IVDs) remains challenging. Here, the transcriptomic landscape of 108 108 IVD cells was mapped using single-cell RNA sequencing of three main compartments from young and adult healthy IVDs, including the nucleus pulposus (NP), annulus fibrosus, and cartilage endplate (CEP). The chondrocyte subclusters were classified based on their potential regulatory, homeostatic, and effector functions in extracellular matrix (ECM) homeostasis. Notably, in the NP, a PROCR+ resident progenitor population showed enriched colony-forming unit-fibroblast (CFU-F) activity and trilineage differentiation capacity. Finally, intercellular crosstalk based on signaling network analysis uncovered that the PDGF and TGF-ß cascades are important cues in the NP microenvironment. In conclusion, a single-cell transcriptomic atlas that resolves spatially regulated cellular heterogeneity together with the critical signaling that underlies homeostasis will help to establish new therapeutic strategies for IVD degeneration in the clinic.

A Redox-induced Dual-mode Colorimetric and Fluorometric Method based on N-CDs and MnO2 for Determination of Isoniazid in Tablets and Plasma Samples.

We develop a simple hydrothermal method to prepare a novel nitrogen-doped carbon dots (N-CDs) originated from green carbon source Liu-bao tea and ethylene diamine. The N-CDs emits strong and stable blue fluorescence (Em = 440 nm) under the excitation wavelength of 350 nm with a quantum yield of 35%. And it is used as an excellent fluorescent output for the sensitive and visual dual-mode determination of isoniazid. The fluorescence of N-CDs is "turned off" first by manganese dioxide (MnO2) nanosheets due to inner filter effect, MnO2 nanosheets can also oxidize TMB (3,3',5,5'- tetramethylbenzidine) to blue oxTMB. Isoniazid, however, can reduce MnO2 nanosheets to Mn2+, turning on the fluorescence again. The color of the solution fades from blue to colorless because less TMB can be oxidized. Under the optimal conditions, the dual-mode method has a satisfying linear relationship ranging from 2.0 to 120.0 µM with a limit of detection of 0.7 µM (S/N = 3). And it has been applied successfully to colorimetric and fluorescent determination of isoniazid in tablets and clinical plasma samples, with recoveries ranging from 94.0% to 102.4%. The properties of N-CDs and MnO2 nanosheets were thoroughly characterized using TEM, FT-IR, XPS, AFM and fluorescence spectrophotometer, the quenching mechanism was also discussed.

High-Pressure Electrospray Ionization Yields Supercharged Protein Complexes from Native Solutions While Preserving Noncovalent Interactions.

Increasing charge state of protein complexes from native solutions while preserving noncovalent interactions in native mass spectrometry (MS) offers great opportunity to gain deeper insights into gas-phase protein structures. Several previous studies have disclosed the possibility of high pressure in supercharging small proteins, whereas its capability to supercharge large protein assemblies under native conditions and how it might affect protein structures remain open questions. Herein, we demonstrated that the high-pressure-induced supercharging strategy affords unique advantages of supercharging protein complexes with the highest charge state surpassing the Rayleigh limit (ZR) and concurrently preserving native-like topology. By examining 32 proteins and protein complexes with molecular weights (MWs) ranging from 8.58 to 801 kDa, we demonstrated that the increased average charge states of macromolecular ions have a strong dependence on the surface areas of native protein conformations and MWs. Factors that might contribute to the high-pressure-induced supercharging capability toward macromolecular ions were discussed. Furthermore, using collision cross section (CCS) variation as a function of charge state, we investigate the effects of gas pressure and charge states on gas-phase structures of proteins and protein complexes. Smaller proteins have the largest CCS variations once supercharged, while macromolecular protein complexes are less affected. The results revealed that both surface density of charge and charged surface basic residues contribute to the observed CCS-charge disciplines for all the macromolecules investigated. Taken together, the results presented here indicate that increasing gas pressure in the ion source affords a rapid, simple, and controlled supercharging method, offering the potency of facilitating further applications of native top-down MS analysis with improved transmission, fragmentation, and detection efficiency.

Rapid detection of five pesticide residues using complexes of gold nanoparticle and porphyrin combined with ultraviolet visible spectrum.

BACKGROUD: Pesticides are widely used to control insect infestation and weeds in agriculture. However, concerns about the pesticide residues in agricultural products have been raised in recent years because of public interest in health and food quality and safety. Thus, rapid, convenient, and accurate analytical methods for the detection and quantification of pesticides are urgently required. RESULTS: A nanohybrid system composed of gold nanoparticles (AuNPs) and tetrakis(N-methyl-4-pyridiniumyl) porphyrin (TMPyP) was used as an optical probe for the detection and quantification of five pesticides (Paraquat, Dipterex, Dursban, methyl thiophanate and Cartap). The method is based on the aggregation effect of pesticides on the carboxyl group modified by AuNPs. Subsequently, with the help of particle swarm optimization-optimized sample weighted least squares-support vector machine (PSO-OSWLS-SVM), all the pesticides could be successfully quantified. In addition, partial least squares discriminant analysis (PLS-DA) was applied and the five pesticides were satisfactorily recognized based on data array obtained from the ultraviolet visible (UV-visible) spectra of AuNP-TMPyP complex. Furthermore, the quantitative and qualitative analysis of the five pesticides could be also achieved in the complex real samples, in which all the relative standard deviations (RSDs) were less than 0.3‰ and all the linear absolute correlation coefficients were more than 0.9990. Furthermore, recognition rate of the training set and the prediction set based on multiplicative scatter correction (MSC), or second-order derivative (2nd derivative) UV-visible spectra in PLS-DA model could reach 100%. CONCLUSION: This method was successfully applied for the rapid and accurate determination of multicomponent pesticide residues in real food samples. © 2020 Society of Chemical Industry.

Fluorescence paper-based sensor for visual detection of carbamate pesticides in food based on CdTe quantum dot and nano ZnTPyP.

The needing of rapid and sensitive detection method for pesticides is increasing, to facilitate its detection without complicated instruments. Herein, a novel paper-based senor was developed for the visual detection of three carbamate pesticides (metolcarb, carbofuran, and carbaryl) based on CdTe quantum dots (QDs) and nano zinc 5, 10, 15, 20-tetra(4-pyridyl)-21H-23H-porphine (nano ZnTPyP) with a "turn-off-on" mode. This fluorescence sensing model could be applied in the highly selective and sensitive detection of carbamate pesticides both by fluorescence spectrometry or paper-based sensors. Based on the extracted RGB color values of paper, the partial least squares regression (PLSR) was used to accurately quantify the concentrations of carbamate pesticides in different food matrices (apple, cabbage and tea water). This method featured in high speed, low price and high accuracy, and provided a new strategy for the detection of food safety.

Double quantum dots-nanoporphyrin fluorescence-visualized paper-based sensors for detecting organophosphorus pesticides.

A high-sensitivity fluorescence visualization paper-based sensor is developed and used to achieve specific detection and analysis of three organophosphorus pesticides (OPPs: dimethoate, dichlorvos, and demeton) in a "Turn-off-on" detection mode. The fluorescence visualization paper-based sensor is established through combining double quantum dots (QDs) with high-activity nanoporphyrins (QDs-nanoporphyrin), realizing double nanometer signal amplification and producing different color change responses to these three OPPs. In particular, this approach is based on Partial least squares discriminant analysis (PLSDA) for fingerprint spectrum analysis of three kinds of organophosphorus pesticides in complex matrix (apple and cabbage). Therefore, different concentrations of pesticide residues can be quickly identified at the same time with 100% accuracy for both training set and prediction set. In summary, this method has high selectivity and stability, and thus provides a new approach for the identification of organophosphorus residues in complex systems.

Simultaneous Recognition of Species, Quality Grades, and Multivariate Calibration of Antioxidant Activities for 12 Famous Green Teas Using Mid- and Near-Infrared Spectroscopy Coupled with Chemometrics.

In this paper, mid- and near-infrared spectroscopy fingerprints were combined to simultaneously discriminate 12 famous green teas and quantitatively characterize their antioxidant activities using chemometrics. A supervised pattern recognition method based on partial least square discriminant analysis (PLSDA) was adopted to classify the 12 famous green teas with different species and quality grades, and then optimized sample-weighted least-squares support vector machine (OSWLS-SVM) based on particle swarm optimization was employed to investigate the quantitative relationship between their antioxidant activities and the spectral fingerprints. As a result, 12 famous green teas can be discriminated with a recognition rate of 100% by MIR or NIR data. However, compared with individual instrumental data, data fusion was more adequate for modeling the antioxidant activities of samples with RMSEP of 0.0065. Finally, the performance of the proposed method was evaluated and validated by some statistical parameters and the elliptical joint confidence region (EJCR) test. The results indicate that fusion of mid- and near-infrared spectroscopy suggests a new avenue to discriminate the species and grades of green teas. Moreover, the proposed method also implies other promising applications with more accurate multivariate calibration of antioxidant activities.

Resveratrol attenuates oxidative injury in human umbilical vein endothelial cells through regulating mitochondrial fusion via TyrRS-PARP1 pathway.

BACKGROUND/AIMS: Oxidative stress-induced damage in endothelial cells is a crucial initiator of atherosclerosis (AS), which is highly related to excessive reactive oxygen species (ROS) and mitochondrial dynamics. Resveratrol (RSV) exerts beneficial effects against endothelial oxidative injury, while the underlying mechanisms have not been fully elucidated. Thus, we aimed to explore the role of mitochondria dynamics during the anti-oxidative activities of RSV in palmitic acid (PA)-stimulated human umbilical vein endothelial cells (HUVECs) and to verify whether tyrosyl transfer- RNA synthetase (TyrRS) and poly (ADP-ribose) polymerase 1 (PARP1) are targeted during this process. METHODS: HUVECs were exposed to 200 µM of PA for 16 h before treated with 10 µM of RSV for 8 h. Cell viability was detected using Cell counting kit-8 (CCK-8) assay. The intracellular ROS level and mitochondria membrane potential (MMP) were measured using microplate reader and flow cytometry. The malondialdehyde and superoxide dismutase were measured using the microplate reader. The mitochondrial morphology and fusion process was observed under transmission electron microscopy and confocal microscopy. TyrRS and PARP1 were knocked down with the specific small interference RNAs (siRNA), and the protein expressions of TyrRS, PARP1, and mitochondrial fusion proteins (MFN1, MFN2, and OPA1) were measured by western blot. RESULTS: RSV treatment suppressed the PA-induced injuries in HUVECs, including the damage to cell viability, oxidative stress, and loss of MMP. Additionally, RSV improved the protein levels of MFN1, MFN2, and OPA1 as well as inhibited the PA-induced fragmentation of mitochondria. However, the effects of RSV on oxidative stress and mitochondrial fusion were abolished by the pretreatment of siRNAs of TyrRS and PARP1, indicating that these effects of RSV were dependent on the TyrRS-PARP1 pathway. CONCLUSIONS: RSV attenuated endothelial oxidative injury by regulating mitochondrial fusion via TyrRS-PARP1 signaling pathway.

UPLC-Q-TOF/MS-based untargeted metabolomics coupled with chemometrics approach for Tieguanyin tea with seasonal and year variations.

The taste and aroma quality of Tieguanyin tea fluctuate seasonally and yearly. However, the compounds responsible for the seasonal and year variations of metabolic pattern and its sensory quality are far from clear. 60 Tieguanyin tea samples harvested in different years and seasons were analyzed by ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) and chemometrics. Principal component analysis (PCA) explained 33.2% of the total variance, while orthogonal projection to latent structures discriminate analysis (OPLS-DA) can obtain potential metabolites with better discrimination, and with R2X and Q2 of cross-validation as 0.974 and 0.937, respectively. Subsequently, heat map analysis (HCA) visualized relationships between Tieguanyin teas with these significantly different potential metabolites by Mann-Whitney U test (p < 0.05). Furthermore, the best discriminate metabolites contributing to different sensory qualities were revealed by stepwise liner discrimination analysis (SLDA) with 100% accuracy rate. The present strategy also exhibited great potential for untargeted metabolomics of other foods.

Dihydromyricetin Ameliorates Nonalcoholic Fatty Liver Disease by Improving Mitochondrial Respiratory Capacity and Redox Homeostasis Through Modulation of SIRT3 Signaling.

Aims: Our previous clinical trial indicated that the flavonoid dihydromyricetin (DHM) could improve hepatic steatosis in patients with nonalcoholic fatty liver disease (NAFLD), altough the potential mechanisms of these effects remained elusive. Here, we investigated the hepatoprotective role of DHM on high-fat diet (HFD)-induced NAFLD. Results: DHM supplementation could effectively ameliorate the development of NAFLD by inhibiting hepatic lipid accumulation both in HFD-fed wild-type mice and in palmitic acid-induced hepatocytes. We reveal for the first time that mitochondrial dysfunction characterized by ATP depletion and augmented oxidative stress could be reversed by DHM treatment. Moreover, DHM enhanced the mitochondrial respiratory capacity by increasing the expression and enzymatic activities of mitochondrial complexes and increased mitochondrial reactive oxygen species scavenging by restoring manganese superoxide dismutase (SOD2) activity. Interestingly, the benefits of DHM were abrogated in SIRT3 knockout (SIRT3KO) mice and in hepatocytes transfected with SIRT3 siRNA or treated with an SIRT3-specific inhibitor. We further showed that DHM could increase SIRT3 expression by activating the adenosine monophosphate-activated protein kinase (AMPK)-peroxisome proliferator-activated receptor-γ coactivator-1 alpha (PGC1α)/estrogen-related receptor-α (ERRα) signaling pathway. Innovation: Our work indicates that SIRT3 plays a critical role in the DHM-mediated beneficial effects that include ameliorating mitochondrial dysfunction and oxidative stress in a nutritional NAFLD model both in vivo and in vitro.Conclusion: Our results suggest that DHM prevents NAFLD by improving mitochondrial respiratory capacity and redox homeostasis in hepatocytes through a SIRT3-dependent mechanism. These results could provide a foundation to identify new DHM-based preventive and therapeutic strategies for NAFLD.

Fusion of near-infrared and fluorescence spectroscopy for untargeted fraud detection of Chinese tea seed oil using chemometric methods.

BACKGROUND: This paper investigated the feasibility of data fusion of near-infrared (NIR) and fluorescence spectroscopy for rapid analysis of cheap vegetable oils in Chinese Camellia oleifera Abel. (COA) oil. Because practical frauds usually involve adulterations of multiple known and unknown cheap oils, traditional analytical methods aimed at detecting one or more known adulterants are insufficient to identify adulterated COA oil. Therefore, untargeted analysis was performed by developing class models of pure COA oil using robust one-class partial least squares (OCPLS). RESULTS: The most accurate OCPLS model was obtained with fusion of standard normal variate (SNV)-NIR and SNV-fluorescence spectra with sensitivity of 0.954 and specificity of 0.91. Robust OCPLS could detect adulterations with 2% (w/w) or more cheap oils, including rapeseed oil, sunflower seed oil, corn oil and peanut oil. CONCLUSION: Fusion of NIR and fluorescence data and chemometrics provided enhanced capacity for rapid and untargeted analysis of multiple adulterations in Chinese COA oils. © 2018 Society of Chemical Industry.

"Turn-off" fluorescent sensor based on double quantum dots coupled with chemometrics for highly sensitive and specific recognition of 53 famous green teas.

Fluorescent "turn-off" sensors based on double quantum dots (QDs) has attracted increasing attention in the detection of many materials due to their properties such as more useful information, higher fluorescence efficiency and stability compared with the fluorescent "turn-off" sensors based on single QDs. In this work, highly sensitive and specific method for recognition of 53 different famous green teas was developed based on the fluorescent "turn-off" model with water-soluble ZnCdSe-CdTe double QDs. The fluorescence of the two QDs can be quenched by different teas with varying degrees, which results in the differences in positions and intensities of two peaks. By the combination of classic partial least square discriminant analysis (PLSDA), all the green teas can be discriminated with high sensitivity, specificity and a satisfactory recognition rate of 100% for training set and 100% for prediction set, respectively. The fluorescent "turn-off" sensors based on the single QDs (either ZnCdSe QDs or CdTe QDs) coupled with PLSDA were also employed to recognize the 53 famous green teas with unsatisfactory results. Therefore, the fluorescent "turn-off" sensors based on the double QDs is more appropriate for the large-class-number classification (LCNC) of green teas. Herein, we have demonstrated, for the first time, that so many kinds of famous green teas can be discriminated by the "turn-off" model of double QDs combined with chemometrics, which has largely extended the capability of traditional fluorescence and chemometrics, as well as exhibits great potential to perform LCNC in other practical applications.