Global Analysis of Lysine Lactylation of Germinated Seeds in Wheat.

Protein lactylation is a newly discovered posttranslational modification (PTM) and is involved in multiple biological processes, both in mammalian cells and rice grains. However, the function of lysine lactylation remains unexplored in wheat. In this study, we performed the first comparative proteomes and lysine lactylomes during seed germination of wheat. In total, 8000 proteins and 927 lactylated sites in 394 proteins were identified at 0 and 12 h after imbibition (HAI). Functional enrichment analysis showed that glycolysis- and TCA-cycle-related proteins were significantly enriched, and more differentially lactylated proteins were enriched in up-regulated lactylated proteins at 12 HAI vs. 0 HAI through the KEGG pathway and protein domain enrichment analysis compared to down-regulated lactylated proteins. Meanwhile, ten particularly preferred amino acids near lactylation sites were found in the embryos of germinated seeds: AA*KlaT, A***KlaD********A, KlaA**T****K, K******A*Kla, K*Kla********K, KlaA******A, Kla*A, KD****Kla, K********Kla and KlaG. These results supplied a comprehensive profile of lysine lactylation of wheat and indicated that protein lysine lactylation played important functions in several biological processes.

Tetraspanin 7 promotes osteosarcoma cell invasion and metastasis by inducing EMT and activating the FAK-Src-Ras-ERK1/2 signaling pathway.

BACKGROUND: Tetraspanins are members of the 4-transmembrane protein superfamily (TM4SF) that function by recruiting many cell surface receptors and signaling proteins into tetraspanin-enriched microdomains (TEMs) that play vital roles in the regulation of key cellular processes including adhesion, motility, and proliferation. Tetraspanin7 (Tspan7) is a member of this superfamily that plays documented roles in hippocampal neurogenesis, synaptic transmission, and malignant transformation in certain tumor types. How Tspan7 influences the onset or progression of osteosarcoma (OS), however, remains to be defined. Herein, this study aimed to explore the relationship between Tspan7 and the malignant progression of OS, and its underlying mechanism of action. METHODS: In this study, the levels of Tspan7 expression in human OS cell lines were evaluated via qRT-PCR and western blotting. The effect of Tspan7 on proliferation was examined using CCK-8 and colony formation assays, while metastatic role of Tspan7 was assessed by functional assays both in vitro and in vivo. In addition, mass spectrometry and co-immunoprecipitation were performed to verify the interaction between Tspan7 and ß1 integrin, and western blotting was used to explore the mechanisms of Tspan7 in OS progresses. RESULTS: We found that Tspan7 is highly expressed in primary OS tumors and OS cell lines. Downregulation of Tspan7 significantly suppressed OS growth, metastasis, and attenuated epithelial-mesenchymal transition (EMT), while its overexpression had the opposite effects in vitro. Furthermore, it exhibited reduced OS pulmonary metastases in Tspan7-deleted mice comparing control mice in vivo. Additionally, we proved that Tspan7 interacted with ß1 integrin to facilitate OS metastasis through the activation of integrin-mediated downstream FAK-Src-Ras-ERK1/2 signaling pathway. CONCLUSION: In summary, this study demonstrates for the first time that Tspan7 promotes OS metastasis via interacting with ß1 integrin and activating the FAK-Src-Ras-ERK1/2 pathway, which could provide rationale for a new therapeutic strategy for OS.

Three-dimensional-printed titanium implants for severe acetabular bone defects in revision hip arthroplasty: short- and mid-term results.

PURPOSE: Severe acetabular bone defect is challenging in revision hip arthroplasty. In the present study, we aimed to present new treatment options with the 3D printing technique and analyze the clinical and radiographic outcomes of 3D-printed titanium implants for the treatment of severe acetabular bone defects in revision hip arthroplasty. METHODS: A total of 35 patients with Paprosky type 3 bone defect and pelvic discontinuity (PD), who underwent hip revisions using 3D-printed titanium implants between 2016 and 2019 at our institution, were retrospectively reviewed. Patient-specific 3D-printed titanium augments and shells (strategy A) were used in 22 type 3A and two type 3B patients. Custom 3D-printed flanged components (strategy B) were used in 11 type 3B patients, including five PD. The clinical outcomes were evaluated with the Harris hip score (HHS). In addition, radiographic results were analyzed by the hip centre of rotation (V-COR and H-COR), implant failure, and survivorship. RESULTS: The mean follow-up was 41.5 months (range, 16-62). The HHS was improved from 47.8 ± 8.2 pre-operatively to 78.1 ± 10.1 at one year follow-up and 86.4 ± 5.1 at the last follow-up (p < 0.01). Post-operative V-COR and H-COR of the operated side were 20.8 ± 2.0 mm and 30.2 ± 1.6 mm compared with 51.4 ± 4.1 mm and 33.9 ± 9.0 mm pre-operatively (p < 0.01). The complications included one dislocation and one partial palsy of the sciatic nerve. At the latest follow-up, no radiological component loosening or screw breakage was present. CONCLUSIONS: 3D-printed titanium implants showed satisfactory short- and mid-term clinical and radiographic outcomes. It was an effective therapeutic regimen with a low rate of complications, providing a patient-specific and reliable strategy for the severe acetabular bone defect in revision hip arthroplasty.

Tspan9 Induces EMT and Promotes Osteosarcoma Metastasis via Activating FAK-Ras-ERK1/2 Pathway.

Object: At present, there are few effective treatment options available to patients suffering from osteosarcoma (OS). Clarifying the signaling pathways that govern OS oncogenesis may highlight novel approaches to treating this deadly form of cancer. Recent experimental evidence suggests that the transmembrane protein tetraspanin-9 (Tspan9) plays a role in tumor development. This study was thus formulated to assess the molecular role of Tspan9 as a regulator of OS cell metastasis. Methods: Gene expression in OS cell lines was evaluated via qRT-PCR, while CCK-8, colony formation, Transwell, and wound healing assays were used to explore the in vitro proliferative, invasive, and migratory activities of OS cells. The relationship between Tspan9 and in vivo OS cell metastasis was assessed by injecting these cells into the tail vein of nude mice. Interactions between the Tspan9 and integrin ß1 proteins were explored through mass spectrometric and co-immunoprecipitation, and Western blotting to assess the functional mechanisms whereby Tspan9 shapes OS pathogenesis. Results: Both primary OS tumors and OS cell lines commonly exhibited Tspan9 upregulation, and the knockdown of this tetraspanin suppressed the migration, invasion, and epithelial-mesenchymal transition (EMT) activity in OS cells, whereas Tspan9 overexpression resulted in opposite phenotypes. Tumor lung metastasis were significantly impaired in mice implanted with HOS cells in which Tspan9 was downregulated as compared to mice implanted with control HOS cells. Tspan9 was also found to interact with ß1 integrin and to contribute to OS metastasis via the amplification of integrin-mediated downstream FAK/Ras/ERK1/2 signaling pathway. Conclusion: These data suggest that Tspan9 can serve as a promising therapeutic target in OS.

Real-time determination of flowering period for field wheat based on improved YOLOv5s model.

The flowering period is one of the important indexes of wheat breeding. The early or late flowering affects the final yield and character stability of wheat. In order to solve the problem that it is difficult to accurately and quickly detect the flowering period of a large number of wheat breeding materials, a determination method of flowering period for field wheat based on the improved You Only Look Once (YOLO) v5s model was proposed. Firstly, a feature fusion (FF) method combing RGB images and corresponding comprehensive color features was proposed to highlight more texture features and reduce the distortion caused by light on the extracted feature images. Second, the YOLOv5s model was selected as a base version of the improved model and the convolutional block attention model (CBAM) was adopted into the feature fusion layer of YOLOV5s model. Florets and spikelets were given greater weight along the channel and spatial dimensions to further refine their effective feature information. At the same time, an integrated Transformer small-target detection head (TSDH) was added to solve the high miss rate of small targets in wheat population images. The accurate and rapid detection of florets and spikelets was realized, and the flowering period was determined according to the proportion of florets and spikelets. The experimental results showed that the average computing time of the proposed method was 11.5ms, and the average recognition accuracy of florets and spikelets was 88.9% and 96.8%, respectively. The average difference between the estimated flowering rate and the actual flowering rate was within 5%, and the determination accuracy of the flowering period reached 100%, which met the basic requirements of the flowering period determination of wheat population in the field.

Self-Templating Synthesis of Hollow Co3O4 Nanoparticles Embedded in N,S-Dual-Doped Reduced Graphene Oxide for Lithium Ion Batteries.

The design and synthesis of hollow-nanostructured transition metal oxide-based anodes is of great importance for long-term operation of lithium ion batteries. Herein, we report a two-step calcination strategy to fabricate hollow Co3O4 nanoparticles embedded in a N,S-co-doped reduced graphene oxide framework. In the first step, core-shell-like Co@Co3O4 embedded in N,S-co-doped reduced graphene oxide is synthesized by pyrolysis of a Co-based metal organic framework/graphene oxide precursor in an inert atmosphere at 800 °C. The designed hollow Co3O4 nanoparticles with an average particle size of 25 nm and wall thickness of about 4-5 nm are formed by a further calcination process in air at 250 °C via the nanoscale Kirkendall effect. Both micropores and mesopores are generated in the HoCo3O4/NS-RGO framework. Benefiting from the hierarchical porous structure of the hollow Co3O4 and the co-doping of nitrogen and sulfur atoms in reduced graphene oxide, the thus-assembled battery exhibits a high specific capacity of 1590 mAh g-1 after 600 charge-discharge cycles at 1 A g-1 and a promising rate performance from 0.2 to 10 A g-1.

Double Metal Diphosphide Pair Nanocages Coupled with P-Doped Carbon for Accelerated Oxygen and Hydrogen Evolution Kinetics.

Developing efficient and durable bifunctional transition metal phosphide (TMP) electrocatalysts is still a great challenge because of its relatively sluggish kinetics of oxygen evolution reaction (OER). Herein, we report a unique bimetallic diphosphide pair (FeP2-NiP2) forming spherical nanocages encapsulated in P-doped carbon layers (FeP2-NiP2@PC) as advanced bifunctional electrocatalyst synthesized by a very facile phosphorization approach. The obtained FeP2-NiP2@PC electrocatalyst exhibits an outstanding OER activity with an ultralow overpotential of 248 mV in 1 M KOH and a low overpotential of 117 mV for HER in 0.5 M H2SO4 (@10 mA·cm-2). Also it gives an exceptional long-term durability toward OER (60 h) and HER (20 h). Differently from the electrocatalysts as reported, after successive 3000 cycles CV acceleration, its overpotential decreases about 10 mV. Further investigation unveils that the electrochemical activation process boosts in situ phase transformation of oxides and phosphides to oxyhydroxides as the vital intermediates in FeP2-NiP2@PC during OER electrocatalysis. The direct observation of vital intermediates has been rarely reported on Fe/Ni-based phosphide electrocatalysts. Our exploration demonstrates an extraordinarily efficient and stable nonprecious TMP bifunctional electrocatalyst and provides a novel prospect to shed light on the intrinsic OER electrocatalytic behavior of Fe/Ni-based phosphide electrocatalysts.

POSS-Derived Synthesis and Full Life Structural Analysis of Si@C as Anode Material in Lithium Ion Battery.

Polyhedral oligomeric silsesquioxane (POSS)-derived Si@C anode material is prepared by the copolymerization of octavinyl-polyhedral oligomeric silsesquioxane (octavinyl-POSS) and styrene. Octavinyl-polyhedral oligomeric silsesquioxane has an inorganic core (-Si8O12) and an organic vinyl shell. Carbonization of the core-shell structured organic-inorganic hybrid precursor results in the formation of carbon protected Si-based anode material applicable for lithium ion battery. The initial discharge capacity of the battery based on the as-obtained Si@C material Si reaches 1500 mAh g-1. After 550 charge-discharge cycles, a high capacity of 1430 mAh g-1 was maintained. A combined XRD, XPS and TEM analysis was performed to investigate the variation of the discharge performance during the cycling experiments. The results show that the decrease in discharge capacity in the first few cycles is related to the formation of solid electrolyte interphase (SEI). The subsequent rise in the capacity can be ascribed to the gradual morphology evolution of the anode material and the loss of capacity after long-term cycles is due to the structural pulverization of silicon within the electrode. Our results not only show the high potential of the novel electrode material but also provide insight into the dynamic features of the material during battery cycling, which is useful for the future design of high-performance electrode material.

Cisplatin Enhances Hepatitis B Virus Replication and PGC-1α Expression through Endoplasmic Reticulum Stress.

Chronic hepatitis B infection remains a serious public health issue worldwide. Hepatitis B virus (HBV) reactivation is commonly reported in patients receiving anticancer therapy, immunosuppressive therapy, or organ and tissue transplantation. However, the precise mechanisms underlying chemotherapeutic agent-related HBV reactivation remain unclear. Here, we report that peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) plays a central role in cisplatin-induced HBV transcription and replication. First, cisplatin treatment upregulated the expression levels of PGC-1α and hepatocyte nuclear factor 4 alpha (HNF-4α) in both HBV-replicating cells and an HBV-transgenic mouse model. PGC-1α coactivates with HNF-4α, which interacts with a core promoter and enhancer II region of HBV genome, thereby promoting HBV production. In contrast, knockdown of PGC-1α and HNF-4α by RNA interference in hepatoma cells reversed HBV activation in response to cisplatin. Additionally, PGC-1α upregulation depended on cisplatin-mediated endoplasmic reticulum (ER) stress. We further observed that the recruitment of cyclic AMP-responsive element-binding protein plays a crucial role for PGC-1α transcriptional activation in cisplatin-treated cells. Finally, pharmacologic inhibition of ER stress impaired PGC-1α upregulation and HBV production induced by cisplatin treatment. These findings demonstrate novel molecular mechanisms indicating that ER stress-PGC1α signaling pathway plays a critical role in cisplatin-evoked HBV reactivation.

Retrospective analysis of the effect of treatment of osteosarcoma complicated by pathological fracture by neoadjuvant chemotherapy combined with limb salvage surgery.

PURPOSE: To explore the efficacy of neoadjuvant chemotherapy combined with limb salvage surgery for the treatment of osteosarcoma complicated by pathological fracture. METHODS: 215 osteosarcoma patients who were admitted in our hospital from 2001 and 2012 were followed up for 5 years and were retrospectively analyzed among them. The patients were divided into 4 groups based on their condition and treatment method to observe the 5-year overall survival and tumor-free survival (TFS) in each group. Adverse reactions caused by chemotherapy were recorded and analyzed. In addition, the quality of life was compared in these 4 groups. RESULTS: No significant differences were observed in postoperative overall survival and TFS between patients who were subjected to limb salvage surgery (Group A) and amputation (Group B). Similarly, there was no difference between patients who underwent surgery for pathological fracture and those without fracture (Group D). However, the survival of non-preoperative chemotherapy group (group C) was significantly different from that of preoperative chemotherapy group (group C vs group A/B/D, p=0.008, p=0.042, p=0.010, respectively). Besides, the TFS of non-preoperative chemotherapy group was significantly lower than that of preoperative chemotherapy group (group C vs. group A/B/D, p=0.012, p=0.002, p=0.008, respectively). Vomiting was the main adverse effect in our research. In the comparison of quality of life, social function and physical limitations in the limb-salvage group were superior to the amputation group. CONCLUSIONS: Neoadjuvant chemotherapy combined with limb salvage surgery is effective for the patients with osteosarcoma complicated by pathological fractures.

The Ultrasonic Microsurgical Anatomical Comparative Study of the CHD Fetuses and Their Clinical Significance.

The aim of our study was to increase the detection rate of fetal cardiac malformations for congenital heart disease (CHD). The ultrasonic and microanatomical methods were combined to study the CHD cases firstly, which could provide the microsurgical anatomical basis to the prenatal ultrasonic diagnosis which was used in suspected CHD and help the sonographer to improve the quality of fetal cardiac diagnosis. We established the ultrasonic standard section of the 175 complex CHD cases and collected the fetal echocardiography image files. The induced/aborted fetuses were fixed by 4% paraformaldehyde and dissected by the ultrasonic microsurgical anatomy. This research could obtain the fetal cardiac anatomic cross-sectional images which was consistent with the ultrasonic standard section and could clearly show the internal structure of the vascular malformation that optimized the ultrasound examination individually. This method could directly display the variation of the CHD fetal heart clearly and comprehensively help us to understand the complex fetal cardiac malformation from the internal structure of the vascular malformation which was consolidated by the anatomical basis of the fetal heart. This study could improve the integrity and accuracy of the prenatal cardiac ultrasound examination tremendously.

The infection efficiency and replication ability of circularized HBV DNA optimized the linear HBV DNA in vitro and in vivo.

Studies on molecular mechanisms of the persist infection of hepatitis B virus have been hampered by a lack of a robust animal model. We successfully established a simple, versatile, and reproducible HBV persist infection model in vitro and in vivo with the circularized HBV DNA. The cells and mice were transfected or injected with circularized HBV DNA and pAAV/HBV1.2, respectively. At the indicated time, the cells, supernatants, serum samples, and liver tissues were collected for virological and serological detection. Both in vitro and in vivo, the circularized HBV DNA and pAAV/HBV1.2 could replicate and transcribe efficiently, but the infection effect of the former was superior to the latter (p < 0.05). The injection of circularized HBV genome DNA into the mice robustly supported HBV infection and approximately 80% of HBV infected mice established persistent infection for at least 10 weeks. This study demonstrated that the infection efficiency and replication ability of the circularized structure of HBV DNA overmatched that of the expression plasmid containing the linear structure of HBV DNA in vitro and in vivo. Meanwhile, this research results could provide useful tools and methodology for further study of pathogenic mechanisms and potential antiviral treatments of human chronic HBV infection in vitro and in vivo.