A TaSnRK1α Modulates TaPAP6L-Mediated Wheat Cold Tolerance through Regulating Endogenous Jasmonic Acid.

Here, a sucrose non-fermenting-1-related protein kinase alpha subunit (TaSnRK1α-1A) is identified as associated with cold stress through integration of genome-wide association study, bulked segregant RNA sequencing, and virus-induced gene silencing. It is confirmed that TaSnRK1α positively regulates cold tolerance by transgenes and ethyl methanesulfonate (EMS) mutants. A plastid-lipid-associated protein 6, chloroplastic-like (TaPAP6L-2B) strongly interacting with TaSnRK1α-1A is screened. Molecular chaperone DJ-1 family protein (TaDJ-1-7B) possibly bridged the interaction of TaSnRK1α-1A and TaPAP6L-2B. It is further revealed that TaSnRK1α-1A phosphorylated TaPAP6L-2B. Subsequently, a superior haplotype TaPAP6L-2B30S /38S is identified and confirmed that both R30S and G38S are important phosphorylation sites that influence TaPAP6L-2B in cold tolerance. Overexpression (OE) and EMS-mutant lines verified TaPAP6L positively modulating cold tolerance. Furthermore, transcriptome sequencing revealed that TaPAP6L-2B-OE lines significantly increased jasmonic acid (JA) content, possibly by improving precursor α-linolenic acid contributing to JA synthesis and by repressing JAR1 degrading JA. Exogenous JA significantly improved the cold tolerance of wheat plants. In summary, TaSnRK1α profoundly regulated cold stress, possibly through phosphorylating TaPAP6L to increase endogenous JA content of wheat plants.

Functions of lysine 2-hydroxyisobutyrylation and future perspectives on plants.

Lysine 2-hydroxyisobutyrylation (Khib) is a novel protein post-translational modifications (PTMs) observed in both eukaryotes and prokaryotes. Recent studies suggested that this novel PTM has the potential to regulate different proteins in various pathways. Khib is regulated by lysine acyltransferases and deacylases. This novel PTM reveals interesting connections between modifications and protein physiological functions, including gene transcription, glycolysis and cell growth, enzymic activity, sperm motility, and aging. Here, we review the discovery and the current understanding of this PTM. Then, we outline the networks of complexity of interactions among PTMs in plants, and raise possible directions of this novel PTM for future investigations in plants.

Global crotonylatome and GWAS revealed a TaSRT1-TaPGK model regulating wheat cold tolerance through mediating pyruvate.

Here, we reported the complete profiling of the crotonylation proteome in common wheat. Through a combination of crotonylation and multi-omics analysis, we identified a TaPGK associated with wheat cold stress. Then, we confirmed the positive role of TaPGK-modulating wheat cold tolerance. Meanwhile, we found that cold stress induced lysine crotonylation of TaPGK. Moreover, we screened a lysine decrotonylase TaSRT1 interacting with TaPGK and found that TaSRT1 negatively regulated wheat cold tolerance. We subsequently demonstrated TaSRT1 inhibiting the accumulation of TaPGK protein, and this inhibition was possibly resulted from decrotonylation of TaPGK by TaSRT1. Transcriptome sequencing indicated that overexpression of TaPGK activated glycolytic key genes and thereby increased pyruvate content. Moreover, we found that exogenous application of pyruvate sharply enhanced wheat cold tolerance. These findings suggest that the TaSRT1-TaPGK model regulating wheat cold tolerance is possibly through mediating pyruvate. This study provided two valuable cold tolerance genes and dissected diverse mechanism of glycolytic pathway involving in wheat cold stress.

Proteomic Study on Multi-Organ Metastases of Human Ovarian Clear Cell Carcinoma Cell Line in a Xenograft Mouse Model Based on a Novel Sequence-Specific Analysis Strategy.

BACKGROUND: To investigate the gene regulation of tumor cells in the process of different organ metastasis on a xenograft mouse model and screen the genes involved in the organ-target metastasis of tumor cells. METHODS: A multi-organ metastasis model was constructed with a human ovarian clear cell carcinoma cell line (ES-2) based on a severe immunodeficiency mouse strain (NCG). Differentially expressed tumor proteins among multi-organ metastases were successfully characterized by microliter liquid chromatography-high-resolution mass spectrometry, sequence-specific data analysis and multivariate statistical data analysis. Liver metastases were selected as typical for subsequent bioinformatic analysis. Selected liver metastasis-specific genes in ES-2 cells were validated by sequence-specific quantitation including high resolution-multiple reaction monitoring quantification at protein level and quantitative real-time polymerase chain reaction at mRNA level. RESULTS: From the mass spectrometry data, a total of 4503 human proteins were identified using the sequence-specific data analysis strategy. Of them, 158 proteins were selected as specifically regulated genes in liver metastases for subsequent bioinformatics studies. Based on Ingenuity Pathway Analysis (IPA) pathway analysis and sequence-specific quantitation, Ferritin light chain (FTL), lactate dehydrogenase A (LDHA) and long-chain-fatty-acid-CoA ligase 1 (ACSL1) were finally validated as specifically upregulated proteins in liver metastases. CONCLUSIONS: Our work provides a new approach to analyze gene regulation in tumor metastasis in xenograft mouse model. In presence of a large number of mouse protein interference, we validated the up-regulation of human ACSL1, FTL and LDHA in ES-2 liver metastases, which reflects the adaptive regulation of tumor cells to the liver microenvironment through metabolic reprogramming.

Value of growth arrest lines for predicting treatment effect on children with distal tibial epiphysis fractures.

Objective: This study aims to explore whether growth arrest lines can predict epiphyseal fracture healing. Method: The data of 234 children with distal tibial epiphysis fractures treated in our hospital from February 2014 to February 2022 were retrospectively analyzed. Imaging data were examined to record epiphyseal grade, fracture type, and the time to appearance of growth arrest lines. Follow-up data were retrieved to record treatment results (i.e., malunion, premature closure, or bone bridge formation). Results: There was a significant difference in the time to appearance of growth arrest lines between patients with epiphyseal grade 0-1 and grade 2-3 (P < 0.05) and between patients with normal healing and patients with a bone bridge (P < 0.05). Among patients with normal healing, there were no significant differences in the time to appearance of growth arrest lines between men and women and between patients with and without surgery (P > 0.05). There was a significant difference in the time to appearance of growth arrest lines between patients with different Salter-Harris fracture types (P < 0.05). Conclusion: For patients with epiphyseal grade 0-1, the time to appearance of growth arrest lines could be useful for predicting the treatment result of a distal tibial epiphyseal fracture.

The role of circRNA derived from RUNX2 in the serum of osteoarthritis and its clinical value.

BACKGROUND: Circular RNA (circRNA) has been shown to affect the pathological process of osteoarthritis (OA) and is expected to become a potential marker for disease diagnosis. This study aimed to investigate the association between circRNA derived from the gene of runt-related transcription factor 2 (RUNX2) and OA risk. METHODS: The expression profile of RUNX2-derived circRNAs in serum of OA patients was detected. Then, the cytological localization of screened differential circRNAs was studied. Luciferase (LUC) reporter assay was used to identify the microRNA (miRNA) sponge capacity of the circRNAs. Bioinformatics analysis was used to construct the functional pathway of this circRNA-miRNAs network. And then, the diagnostic value of RUNX2-derived circRNAs in OA was evaluated. RESULTS: RUNX2-derived hsa_circ_0005526 (circ_RUNX2) is significantly highly expressed in OA serum and mainly located in the cytoplasm within the cartilage cell by sponging multiple miRNAs (miR-498, miR-924, miR-361-3p, and miR-665). Bioinformatics analysis showed ECM-receptor interaction pathway ranked the most significant pathway of circ_RUNX2-miRNAs regulatory network in KEGG database. The ROC curve showed that there may be good diagnostic value of serum circ_RUNX2 in OA. CONCLUSION: RUNX2-derived circ_RUNX2 may be involved in OA development via ECM-receptor interaction pathways and may be used as potential clinical indicator of OA.

Dorsal Hexagon Local Flap Without Skin Graft for Web Reconstruction of Congenital Syndactyly.

PURPOSE: The ideal web reconstruction for syndactyly requires both satisfactory function and aesthetic appearance. In this study, we report a dorsal hexagon local flap with adequate size and that leaves most of the scars in the interdigital space. METHODS: Between June 2015 and June 2017, 16 patients (10 males and 6 females) with 22 syndactylies underwent surgical reconstruction using the dorsal hexagon local flap technique. All patients had simple syndactyly. The postoperative evaluation included the quality of scarring, the extent of flexion and extension deformity, web creep, lateral flexion deformity, and rotational deformity of the digit. RESULTS: All flaps survived and there were no postoperative complications. All patients achieved satisfactory interdigital commissure depth. During 12 to 34 months of follow-up, no case of flexion contracture or web creep after reconstruction was noted. CONCLUSIONS: The dorsal hexagon local flap is an alternative method for syndactyly reconstruction. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV.

Tetrahydrohyperforin prevents articular cartilage degeneration and affects autophagy in rats with osteoarthritis.

Osteoarthritis (OA) is a highly prevalent disease, which is associated with extracellular matrix degradation and cell death in articular cartilage. The aim of the present study was to identify whether tetrahydrohyperforin (IDN5706) ameliorates the degeneration of articular cartilage and affects autophagy in OA. The rat model of experimental OA was induced by intra-articular injection of collagenase solution. IDN5706 was administered intragastrically to rats for 6 weeks. Histopathological changes in articular cartilage were examined using hematoxylin and eosin (H&E) and safranin O staining, and Mankin scoring systems. The effect of IDN5706 on autophagy was examined using western blotting. ELISA was performed to detect cartilage inflammation. H&E and safranin O staining, Mankin scores, and electron microscopy indicated that IDN5706 could lessen the degeneration of articular cartilage in OA rats. In addition, western blotting revealed that IDN5706 treatment may activate the suppressed autophagy in OA rats. In conclusion, the present study demonstrated that IDN5706 was able to reduce the severity of experimental OA, alleviate the degeneration of articular cartilage, and affect autophagy in OA model rats.

Anti-Osteoarthritic and Anti-Inflammatory Activities of Diazine: In Vitro and In Vivo Studies.

BACKGROUND The present study evaluated the effects of diazine (DZN) on collagenase-induced osteoarthritis (OA) in rats. MATERIAL AND METHODS OA was produced via intra-articular injections of collagenase type II into the knee joint. The rats were then treated with DZN (25, 50, or 100 mg/kg, p.o.) for three weeks. At the end of the protocol, all rats were evaluated for paw latency, paw edema, and knee swelling. Additionally, serum concentrations of glycosaminoglycan (GAG), alkaline phosphatase (ALP), and C-reactive protein (CRP) were determined. X-rays were performed to estimate radiological and histopathological changes in the knee joint. The expressions of antioxidant enzymes, matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases (TIMPs) were estimated in the synovial tissues. RESULTS DZN treatment attenuated inflammation in osteoarthritic rats, as evidenced by decreases in paw edema and knee swelling and enhanced paw latency compared to the negative control group. Additionally, there were significant decreases in the serum levels of CRP and GAG and increases in ALP in the DZN-treated groups compared to the negative control group. The radiological and histopathological results showed that DZN protected against cartilage damage in the knee joint. Additionally, MMP levels decreased and there were significant reductions in the expressions of antioxidant enzymes and TIPMs in the DZN-treated groups compared to the negative control group. CONCLUSIONS The present findings demonstrated the chondroprotective effects of DZN via its modulation of the expressions of TIMP-1 and MMPs in the synovial tissues of osteoarthritic rats.

A predictive model to estimate the pretest probability of metastasis in patients with osteosarcoma.

Osteosarcomas (OSs) represent a huge challenge to improve the overall survival, especially in metastatic patients. Increasing evidence indicates that both tumor-associated elements but also on host-associated elements are under a remarkable effect on the prognosis of cancer patients, especially systemic inflammatory response. By analyzing a series prognosis of factors, including age, gender, primary tumor size, tumor location, tumor grade, and histological classification, monocyte ratio, and NLR ratio, a clinical predictive model was established by using stepwise logistic regression involved circulating leukocyte to compute the estimated probabilities of metastases for OS patients. The clinical predictive model was described by the following equations: probability of developing metastases = ex/(1 + ex), x = -2.150 +  (1.680 × monocyte ratio) + (1.533 × NLR ratio), where is the base of the natural logarithm, the assignment to each of the 2 variables is 1 if the ratio >1 (otherwise 0). The calculated AUC of the receiver-operating characteristic curve as 0.793 revealed well accuracy of this model (95% CI, 0.740-0.845). The predicted probabilities that we generated with the cross-validation procedure had a similar AUC (0.743; 95% CI, 0.684-0.803). The present model could be used to improve the outcomes of the metastases by developing a predictive model considering circulating leukocyte influence to estimate the pretest probability of developing metastases in patients with OS.

Facile solid-phase synthesis of the diammoniate of diborane and its thermal decomposition behavior.

The recent mechanistic finding of the hydrogen release pathways from ammonia borane (AB) has sparked new interest in the chemistry and properties of the diammoniate of diborane (DADB), an ionic isomer of AB. We herein report a facile one-step solid-phase synthesis route of DADB using inexpensive starting materials. Our study found that mechanically milling a 1 : 1 NaBH(4)/NH(4)F powder mixture causes the formation of crystalline DADB via a NH(4)BH(4) intermediate. The produced DADB can be readily separated from the sodium fluoride (NaF) by-product by a purification procedure using liquid ammonia at -78 °C. The thermal decomposition behavior of DADB was studied using synchronous thermal analyses, particularly in comparison with AB. It was found that the decomposition steps and products of DADB are similar to those of AB. But meanwhile, DADB exhibits a series of advantages over AB that merit its potential hydrogen storage application, such as lower dehydrogenation temperature, free of foaming and lack of an induction period in the thermal decomposition process. Our study further found that the volatile non-hydrogen products from DADB can be effectively suppressed by milling with MgH(2).

Observation of mass analyzed threshold ionization using synchrotron radiation on a new-style time of flight mass spectrometer.

We have developed an efficient and applicable apparatus that combines mass-analyzed threshold ionization (MATI) with continuous molecular-beam mass spectrometry using tunable vacuum ultraviolet synchrotron radiation at National Synchrotron Radiation Laboratory. The new design, in which the spoiling field and the pulsed ionization field are perpendicular to each other, can obtain efficiently the ionic spectra of molecule. The MATI spectra of Ar and N(2) have been recorded in the energy region between 15.5 and 17.5 eV to illustrate the feasibility of this scheme. With its unique features, the important experiment considerations are potentially a powerful tool for study of information of ionization energies and ionic states of complex organic compounds.

Performance of the atomic and molecular physics beamline at the National Synchrotron Radiation Laboratory.

At the National Synchrotron Radiation Laboratory, The University of Science and Technology of China, an atomic and molecular physics beamline with an energy range of 7.5-124 eV has been constructed for studying the spectroscopy and dynamics of atoms, molecules and clusters. The undulator-based beamline, with a high-resolution spherical-grating monochromator (SGM), is connected to the atomic and molecular physics end-station. This end-station includes a main experimental chamber for photoionization studies and an additional multi-stage photoionization chamber for photoabsorption spectroscopy. A mid-photon flux of 5 x 10(12) photons s(-1) and a high resolving power is provided by this SGM beamline in the energy range 7.5-124 eV. The size of the synchrotron radiation beam spot at the sample is about 0.5 mm in the vertical direction and 1.0 mm in the horizontal direction. Some experimental results of photoionization efficiency spectroscopy and photoabsorption spectroscopy of atoms and molecules are also reported.