Photosensitive Hydrogel with Temperature-Controlled Reversible Nano-Apertures for Single-Cell Protein Analysis.

Single cell western blot (scWB) is one of the most important methods for cellular heterogeneity profiling. However, current scWB based on conventional photoactive polyacrylamide hydrogel material suffers from the tradeoff between in-gel probing and separation resolution. Here, a highly sensitive temperature-controlled single-cell western blotting (tc-scWB) method is introduced, which is based on a thermo/photo-dualistic-sensitive polyacrylamide hydrogel, namely acrylic acid-functionalized graphene oxide (AFGO) assisted, N-isopropylacrylamide modified polyacrylamide (ANP) hydrogel. The ANP hydrogel is contracted at high-temperature to constrain protein band diffusion during microchip electrophoretic separation, while the gel aperture is expanded under low-temperature for better antibody penetration into the hydrogel. The tc-scWB method enables the separation and profiling of small-molecule-weight proteins with highly crosslinked gel (12% T) in SDS-PAGE. The tc-scWB is demonstrated on three metabolic and ER stress-specific proteins (CHOP, MDH2 and FH) in four pancreatic cell subtypes, revealing the expression of key enzymes in the Krebs cycle is upregulated with enhanced ER stress. It is found that ER stress can regulate crucial enzyme (MDH2 and FH) activities of metabolic cascade in cancer cells, boosting aerobic respiration to attenuate the Warburg effect and promote cell apoptosis. The tc-scWB is a general toolbox for the analysis of low-abundance small-molecular functional proteins at the single-cell level.

Effects of ozone exposure on lipid metabolism in Huh-7 human hepatoma cells.

Ozone pollution is a major environmental concern. According to recent epidemiological studies, ozone exposure increases the risk of metabolic liver disease. However, studies on the mechanisms underlying the effects of ozone exposure on hepatic oxidative damage, lipid synthesis, and catabolism are limited. In this study, Huh-7 human hepatocellular carcinoma cells were randomly divided into five groups and exposed to 200 ppb O3 for 0, 1, 2, 4, and 8 h. We measured the levels of oxidative stress and analyzed the changes in molecules related to lipid metabolism. The levels of oxidative stress were found to be significantly elevated in Huh-7 hepatocellular carcinoma cells after O3 exposure. Moreover, the expression levels of intracellular lipid synthases, including SREBP1, FASN, SCD1, and ACC1, were enhanced. Lipolytic enzymes, including ATGL and HSL, and the mitochondrial fatty acid oxidase, CPT1α, were inhibited after O3 exposure. In addition, short O3 exposure enhanced the expression of the intracellular peroxisomal fatty acid ß-oxidase, ACOX1; however, its expression decreased adaptively with longer exposure times. Overall, O3 exposure induces an increase in intracellular oxidative stress and disrupts the normal metabolism of lipids in hepatocytes, leading to intracellular lipid accumulation.

Multifunctional 3D-printed bioceramic scaffolds: Recent strategies for osteosarcoma treatment.

Osteosarcoma is the most prevalent bone malignant tumor in children and teenagers. The bone defect, recurrence, and metastasis after surgery severely affect the life quality of patients. Clinically, bone grafts are implanted. Primary bioceramic scaffolds show a monomodal osteogenesis function. With the advances in three-dimensional printing technology and materials science, while maintaining the osteogenesis ability, scaffolds become more patient-specific and obtain additional anti-tumor ability with functional agents being loaded. Anti-tumor therapies include photothermal, magnetothermal, old and novel chemo-, gas, and photodynamic therapy. These strategies kill tumors through novel mechanisms to treat refractory osteosarcoma due to drug resistance, and some have shown the potential to reverse drug resistance and inhibit metastasis. Therefore, multifunctional three-dimensional printed bioceramic scaffolds hold excellent promise for osteosarcoma treatments. To better understand, we review the background of osteosarcoma, primary 3D-printed bioceramic scaffolds, and different therapies and have a prospect for the future.

Glycolipids improve the stability of liposomes: The perspective of bilayer membrane structure.

The storage and thermal stability of liposomes, which are amphiphilic carriers, cause very large challenges. However, glycolipid modification may be a potential method to improve the stability of liposomes. In this study, the mechanism by which tilapia head glycolipids improve the stability of liposomes was studied. The head groups of glycolipids and liposomes have a strong interaction (Ka = 633.650 M-1), mainly due to hydrogen bonds, which promote the formation of microstructure domains between glycolipids and liposomes. In addition, glycolipids caused the bilayer structure of liposomes to rearrange, resulting in an increase in the phase transition temperature, tight arrangement of membrane molecules, and increase in membrane thickness (from 2.4 nm to 3.5 nm). Novelty, the formation of microstructure domains helped prevent the liposomes membrane structure from being disrupted during storage and heat. Therefore, glycolipid modification improved the stability of liposomes. This study can provide new insights into the development of high-stability liposomes.

Selection of candidate genes for differences in fat metabolism between cattle subcutaneous and perirenal adipose tissue based on RNA-seq.

The site of fat deposition plays an important role in meat quality and body health. Biologically, the perirenal visceral fat (PF) and back subcutaneous fat (BF) are distinct. Angus and Simmental cattle (Bos taurus) were used as models. HE staining, triglyceride assay kit and RNA-seq were used to analyze the differences in tissue morphology and lipid accumulation, co-genes, and differentially expressed genes (DEGs) between the two tissues. According to the findings, BF has a smaller cell area and greater lipid deposition ability than PF. RNA-seq generated approximately 10.99 Gb of data in each library, and 23,472 genes were identified. The genes FABP4, ADIRF, and SCD that are related to adipose deposition were highly expressed in four tissues. There were 1678 DEGs and 1955 DEGs between BF and PF in Angus and Simmental cattle respectively. Gene Ontology function analysis identified several DEGs involved in metabolism. KEGG pathway analysis showed that four pathways related to fat metabolism were enriched. In the BF, seven genes (COL1A1, COL1A2, COL3A1, COL2A1, RXRA, C1QTNF7, and MOGAT2) were up-regulated. Five genes (ADRB3, ABHD5, CPT1B, CD36, LPIN1) were down-regulated. This study identified candidate genes that led to differences in fat metabolism, which could be useful in cattle breeding.

The bHLH transcription factor regulated gene OsWIH2 is a positive regulator of drought tolerance in rice.

Drought is a major abiotic stress limiting crop growth and yield. In this study, we characterized a novel drought tolerance induced WIH gene in rice, OsWIH2. Overexpression of OsWIH2 in rice resulted in significantly higher drought tolerance, probably due to the decreased water loss rate and reactive oxygen species (ROS) accumulation under drought stress. We identified a long-chain fatty acid HOTHEAD (HTH) that interacted with OsWIH2 using yeast two-hybrid screening. OsWIH2 is an enzyme which is involved in fatty acid synthesis. We further demonstrated that the drought-inducible bHLH transcription factor OsbHLH130 could activate the expression of OsWIH2. Overall, our results suggest that drought stress may induce OsbHLH130 accumulation, which in turn activates OsWIH2 expression, and the latter improves rice drought tolerance by participating in cuticular wax biosynthesis and reducing the water loss rate as well as ROS accumulation. This research provides new genes for crop improvement.

Relation of kidney function and homocysteine in patients with hypothyroidism.

OBJECTIVE: It has been found that both serum homocysteine (Hcy) and serum creatinine levels were increased in hypothyroidism patients. The aim of this study was to investigate the correlation between serum Hcy and kidney function in patients with subclinical hypothyroidism or hypothyroidism. METHODS: A total of 448 subjects were enrolled and divided into three groups: hypothyroidism (n = 129), subclinical hypothyroidism (n = 141), and control group (n = 168). Anthropometric information, metabolic parameters, serum Hcy and creatinine levels, and estimated glomerular filtration rate (eGFR) were analyzed. RESULTS: Compared with healthy subjects, patients with subclinical hypothyroidism or hypothyroidism had significantly higher serum Hcy and creatinine levels and lower eGFR level (all P < 0.001). Serum Hcy was negatively correlated with eGFR in subclinical hypothyroidism patients (r = -0.220, P = 0.009), and in hypothyroidism patients (r = -0.422, P < 0.001). After adjusting for age, sex and BMI, eGFR was still significantly correlated with serum Hcy in subclinical hypothyroidism or hypothyroidism patients (both P < 0.05). Levothyroxine treatment resulted in significantly decreased Hcy and increased eGFR in hypothyroidism patients (both P < 0.001). The decrease in Hcy was correlated with the increased eGFR after treatment (P = 0.001). CONCLUSION: Serum Hcy was negatively correlated with eGFR in subclinical hypothyroidism or hypothyroidism patients. After levothyroxine treatment, a correlation was found between the decrease in serum Hcy and the increase in eGFR in hypothyroidism patients.

Copper-catalyzed alkylarylation of vinylarenes with cycloalkylsilyl peroxides and boronic acids.

A mild copper-catalyzed alkylarylation of vinylarenes with cycloalkylsilyl peroxides and boronic acids is described. This three-component protocol provides a straightforward approach to the remote keto-functionalized 1,1-diarylmethane derivatives. A radical pathway initiated by C-C bond cleavage is proposed for this tandem reaction.

A calcium-dependent lipid binding protein, OsANN10, is a negative regulator of osmotic stress tolerance in rice.

Annexin, a multi-gene family in plants, is essential for plant growth and stress responses. Recent studies demonstrated a positive effect of annexin in abiotic stress responses. Interestingly, we found OsANN10, a putative annexin gene in rice, negatively regulated plant responses to osmotic stress. Knocking down OsANN10 significantly decreased the content of H2O2 by increasing Peroxidase (POD) and Catalase (CAT) activities, further reducing oxidative damage in rice leaves, suggesting a negative regulation of OsANN10 in protecting cell membrane against oxidative damage via scavenging ROS under osmotic stress.

Mechanisms of TET protein-mediated DNA demethylation and its role in the regulation of mouse development.

TET (ten-eleven translocation) protein family includes three members TET1, TET2 and TET3, which belong to alpha-ketoglutaric acid ( α-KG )- and Fe(2+)-dependent dioxygenase superfamily, and have the capacity to convert 5-methylcytosine (5 mC) to 5-hydroxymethylcytosine (5 hmC), 5-formylcytosine (5 fC) and 5-carboxylcytosine (5 caC). At present, growing lines of evidence indicate that TET proteins are involved in the control of active or passive DNA demethylation via different mechanisms; moreover, their activities may be regulated by some cellular factors. TET proteins play vital roles in modulating mammal development, including primordial germ cell formation, embryonic development, stem cells pluripotency, nerve and brain development, etc. The identification of biological roles of TET proteins will open a new field in epigenetic research, and these studies on TET proteins are of great significance to life science research. Here, we review TET proteins from their structure, molecular mechanisms of DNA demethylation and function in the regulation of mouse development, which may provide the basis for understanding the functions of TET proteins.

Homogeneous and sensitive detection of microRNA with ligase chain reaction and lambda exonuclease-assisted cationic conjugated polymer biosensing.

A simple and homogeneous microRNA assay is developed by integration of ligase chain reaction (LCR) and lambda exonuclease-assisted cationic conjugated polymer (CCP) biosensing. LCR is utilized for exponential amplification of microRNA, and lambda exonuclease is introduced to degrade excess fluorescein-labeled probes in LCR for eliminating background signal. After addition of CCP, efficient fluorescence resonance energy transfer from CCP to fluorescein in LCR products occurs. The method is sensitive enough to detect 0.1 fM target microRNA and specific to discriminate one-base difference of microRNAs, which paves a new way for homogeneous microRNA detection and molecular diagnosis.

LXRα gene expression, genetic variation and association analysis between novel SNPs and growth traits in Chinese native cattle.

Liver X receptor α (LXRα) has emerged as an important regulator of lipid and energy metabolism. In this study, to better understand the effects of LXRα gene on growth traits in cattle, the mRNA tissue expression patterns and the polymorphisms of some exons of LXRα were revealed. The expression profile of the bovine LXRα gene was detected by quantitative real-time polymerase chain reaction (qRT-PCR) in 11 different Jiaxian cattle tissues and was found mainly expressed in spleen, liver, fat tissue, kidney, muscle, and lung. Meanwhile, it showed that four single nucleotide polymorphisms (SNPs), named g.1028 T>C, g.1514 T>C, g.2929G>A, and g.3493 T>C, were detected and 12 different haplotypes were constructed. Haplotype with CCGT was dominant with frequency of 40.8 %. There was a strong link between g.1028 T>C and g.1514 T>C (r (2) = 0.374). Association analysis of SNPs with growth- and body-related traits was carried out in 445 Chinese native cattle. The results displayed that the heterozygous genotypes of g.1028 T>C and g.1514 T>C showed a molecular heterosis on four performance traits related to body size: height at withers, body length, hipbone width, and hip width (P < 0.05). The multiple effects of four sites showed that the height at withers, body length, hipbone width, and hip width of individuals of TC-TC-GG-TT combined genotypes were significantly higher than other genotypes (P < 0.05). The effects of the four loci genotype combination on conformation traits were consistent with the effects of g.1028 T>C and g.1514 T>C loci. The SNPs of g.1028 T>C and g.1514 T>C of the bovine LXRα gene could be potential genetic markers for growth traits in cattle. These results suggest that LXRα gene is expressed in many tissues and may provide primary molecular information for further studies on body size traits in Chinese indigenous cattle.

Molecular cloning of bovine FABGL gene and its effects on bovine bioeconomic traits.

The complete CDS sequence of the bovine FABGL gene was determined by homology cloning approach combined with RT-PCR and 3'- and 5'-RACE. The results of sequence analysis and bioinformatics study showed that this cDNA contained 994 nucleotides, with a 780 bp open reading frame (ORF) flanked by a 16 bp 5'-UTR (incompletely) and a 198 bp 3'-UTR. The deduced amino acid sequence (260 AA) shows 88% identity with the corresponding sequence in humans. Two single nucleotide substitutions, one located in intron 5 (I5) at position 1 065 bp (Y = C/T) (GenBank: DQ409814) and the other in intron 8 (I8) at position 1 792 bp (R = A/G), were detected using the PCR-SSCP method. Analysis of the allele frequencies of the two polymorphic sites in three different cattle breeds (Angus, Hereford, and Simmental) with different genotypes showed large differences: in locus I8, cattle with the GG genotype showed higher beef performance index (BPI) (4.283 +/- 0.475 kg/cm) in comparison with cattle with the AA genotype (4.008 +/- 0.465 kg/cm) (P = 0.01). Regarding the ribeye area, cattle with the GG genotype showed significantly higher ribeye area (73.380 +/- 13.005 cm(2)) compared with cattle with the AA genotype (67.744 +/- 12.777 cm(2)) (p = 0.05). In locus I5, some associations for the average daily gain (ADG) were found at the significance level of 0.01 between three different genotypes (CC, CT, TT): cattle with the TT genotype showed the highest ADG (0.652 +/- 0.330 kg/d), whereas cattle with the CC genotype showed the lowest ADG value (0.421 +/- 0.178 kg/d).