UBXN1 promotes liver tumorigenesis by regulating mitochondrial homeostasis.

BACKGROUND: The maintenance of mitochondrial homeostasis is critical for tumor initiation and malignant progression because it increases tumor cell survival and growth. The molecular events controlling mitochondrial integrity that facilitate the development of hepatocellular carcinoma (HCC) remain unclear. Here, we report that UBX domain-containing protein 1 (UBXN1) hyperactivation is essential for mitochondrial homeostasis and liver tumorigenesis. METHODS: Oncogene-induced mouse liver tumor models were generated with the Sleeping Beauty (SB) transposon delivery system. Assessment of HCC cell growth in vivo and in vitro, including tumour formation, colony formation, TUNEL and FACS assays, was conducted to determine the effects of UBXN1 on HCC cells, as well as the involvement of the UBXN1-prohibitin (PHB) interaction in mitochondrial function. Coimmunoprecipitation (Co-IP) was used to assess the interaction between UBXN1 and PHB. Liver hepatocellular carcinoma (LIHC) datasets and HCC patient samples were used to assess the expression of UBXN1. RESULTS: UBXN1 expression is commonly upregulated in human HCCs and mouse liver tumors and is associated with poor overall survival in HCC patients. UBXN1 facilitates the growth of human HCC cells and promotes mouse liver tumorigenesis driven by the NRas/c-Myc or c-Myc/shp53 combination. UBXN1 interacts with the inner mitochondrial membrane protein PHB and sustains PHB expression. UBXN1 inhibition triggers mitochondrial damage and liver tumor cell apoptosis. CONCLUSIONS: UBXN1 interacts with PHB and promotes mitochondrial homeostasis during liver tumorigenesis.

Prohibitin 2-mediated mitophagy attenuates renal tubular epithelial cells injury by regulating mitochondrial dysfunction and NLRP3 inflammasome activation.

Accumulating evidence demonstrates that mitochondrial dysfunction and inflammasome activation play a critical role in the pathogenesis of renal tubular injury through the production of reactive oxygen species and cytokines. Prohibitin 2 (PHB2) is a newly identified intracellular receptor of mitophagy (a type of autophagy) that mediates selective removal of damaged mitochondria, and it could possibly play a renoprotective role in kidney disease. In this study, we confirmed that autophagy is activated in tubular epithelial cells treated with angiotensin II and that inhibition of autophagy results in tubular cell injury. Strikingly, PHB2 knockdown reduced the level of mitophagy and augmented cell death, while overexpression of PHB2 provided protection against pyrin domain-containing protein 3 (NLRP3)-induced inflammatory pathways through amelioration of mitochondrial dysfunction. Our research is the first to experimentally demonstrate the role of PHB2 in renal proximal tubular cells and thereby to provide a better understanding of how autophagy modulates inflammation in renal tubules. These data highlight PHB2 as a therapeutic target in the future treatment of CKD.

Nicotine inhibits murine Leydig cell differentiation and maturation via regulating Hedgehog signal pathway.

Nicotine, the main toxic substance in cigarette smoke, significantly reduced the differentiation and maturation ratio of Leydig cell in murine testes. To investigate the underlying mechanism, C57BL/6J mice were divided into control (CT) and nicotine treated (NT) groups. Next generation RNA sequencing and bio-informatics analysis were carried out to analysis the effects of nicotine on the RNA profile of Leydig cells. Expression level of 7 pathways remarkably changed after nicotine treatment. As the positive regulating pathway of Leydig cell differentiation, Hedgehog signaling pathway was found among these pathways. PTCH1 and ß-TrCP were down-regulated in nicotine treated mice Leydig cells, while GSK3ß, Gli2 and Gli2 fragments increased significantly. Nicotine stimulated the destabilization of Gli2 via ß-TrCP induced ubiquitination and degradation. Gli2 was phosphorylated by up-expressed GSK3ß during this process. Destabilization of Gli2 reduced the activation rate of target genes of Hedgehog signaling pathway such as Ptch1. The differentiation of Leydig cell positively regulated by Hh pathway was thus inhibited by nicotine exposure. Consequently, the male reproduction process powered by Leydic cell-mediated androgen secretion was thus influenced. In conclusion, we find that nicotine inhibits murine Leydig cell differentiation and maturation via regulating Hedgehog signal pathway.

Nicotine induces Nme2-mediated apoptosis in mouse testes.

In mouse testes, germ cell apoptosis can be caused by cigarette smoke and lead to declining quality of semen, but the exact molecular mechanisms remain unclear. To evaluate the effects of nicotine exposure on apoptosis during spermatogenesis, we first constructed a nicotine-treated mouse model and detected germ cell apoptosis activity in the testes using the TUNEL method. Then we analyzed the variation of telomere length and telomerase activity by real-time PCR and TRAP-real-time PCR, respectively. Further, we investigated a highly expressed gene, Nme2, in mouse testes after nicotine treatment from our previous results, which has close correlation with the apoptosis activity predicted by bioinformatics. We performed NME2 overexpression in Hela cells to confirm whether telomere length and telomerase activity were regulated by the Nme2 gene. Finally, we examined methylation of CpG islands in the Nme2 promoter with the Bisulfite Sequencing (BSP) method. The results showed that apoptosis had increased significantly, and then telomerase activity became weak. Further, telomere length was shortened in the germ cells among the nicotine-treated group. In Hela cells, both overexpression of the Nme2 gene and nicotine exposure can suppress the activity of telomerase activity and shorten telomere length. BSP results revealed that the Nme2 promoter appeared with low methylation in mouse testes after nicotine treatment. We assume that nicotine-induced apoptosis may be caused by telomerase activity decline, which is inhibited by the up expression of Nme2 because of its hypomethylation in mouse germ cells.

Nicotine Induced Murine Spermatozoa Apoptosis via Up-Regulation of Deubiquitinated RIP1 by Trim27 Promoter Hypomethylation.

Nicotine significantly promoted apoptosis in stages I, VII, VIII, and XI spermatogonia, stages I, VII, VIII, X, and XI spermatocytes, and stages I-V, VII, and VIII elongating spermatids. To explore the underlying molecular mechanisms, sperm mRNA next-generation sequencing of nicotine-treated mice was conducted. Out of the 86 genes related to apoptosis, Tnf (tumor necrosis factor alpha) was screened to be the most significant varied transcript, and the Onto-pathway analysis indicated that the TNF apoptotic pathway was especially activated by nicotine exposure. The TNF pathway was further studied at the gene and protein levels. The results showed that RIP1, the key component in the TNF apoptotic pathway, was up-expressed in its deubiquitinated form in nicotine-treated mice testis. TRIM27, an E3 ubiquitin ligase that activated TNF apoptotic pathway through up-regulating deubiquitinated RIP1, was also overexpressed in nicotine-treated spermatocytes; moreover, four consecutive CpG sites near the Trim27 transcription start site were less frequently methylated. Finally, in vitro experiments of Trim27 overexpression and RNA interference in GC-1 spermatogonial cells confirmed that the RIP1 deubiquitination and TRIM27 hyopmethylation were both positively correlated with spermatocyte apoptosis. In summary, our study suggests that nicotine may induce murine spermatozoal apoptosis via the TNF apoptotic pathway through up-regulation of deubiquitinated RIP1 by Trim27 promoter hypomethylation.

Protein profile screening: reduced expression of Sord in the mouse epididymis induced by nicotine inhibits tyrosine phosphorylation level in capacitated spermatozoa.

Many studies have revealed the hazardous effects of cigarette smoking and nicotine exposure on male fertility, but the actual, underlying molecular mechanism remains relatively unclear. To evaluate the detrimental effects of nicotine exposure on the sperm maturation process, two-dimensional gel electrophoresis and mass spectrometry analyses were performed to screen and identify differentially expressed proteins from the epididymal tissue of mice exposed to nicotine. Data mining analysis indicated that 15 identified proteins were mainly involved in the molecular transportation process and the polyol pathway, indicating impaired epididymal secretory functions. Experiments in vitro confirmed that nicotine inhibited tyrosine phosphorylation levels in capacitated spermatozoa via the downregulated seminal fructose concentration. Sord, a key gene encoding sorbitol dehydrogenase, was further investigated to reveal that nicotine induced hyper-methylation of the promoter region of this gene. Nicotine-induced reduced expression of Sord could be involved in impaired secretory functions of the epididymis and thus prevent the sperm from undergoing proper maturation and capacitation, although further experiments are needed to confirm this hypothesis.

Alteration of sperm protein profile induced by cigarette smoking.

Cigarette smoking is associated with lower semen quality, but how cigarette smoking changes the semen quality remains unclear. The aim of this study was to screen the differentially expressed proteins in the sperm of mice with daily exposure to cigarette smoke. The 2D gel electrophoresis (2DE) and mass spectrometry (MS) analyses results showed that the mouse sperm protein profile was altered by cigarette smoking. And 22 of the most abundant proteins that correspond to differentially expressed spots in 2DE gels of the sperm samples were identified. These proteins were classified into different groups based on their functions, such as energy metabolism, reproduction, and structural molecules. Furthermore, the 2DE and MS results of five proteins (Aldoa, ATP5a1, Gpx4, Cs, and Spatc1) were validated by western blot analysis and reverse transcriptase-polymerase chain reaction. Results showed that except Spatc1 the other four proteins showed statistically significant different protein levels between the smoking group and the control group (P < 0.05). The expressions of three genes (Aldoa, Gpx4, and Spatc1) were significantly different (P < 0.05) at transcription level between the smoking group and the control group. In addition, five proteins (Aldoa, ATP5a1, Spatc1, Cs, and Gpx4) in human sperm samples from 30 male smokers and 30 non-smokers were detected by western blot analysis. Two proteins (Aldoa and Cs) that are associated with energy production were found to be significantly altered, suggesting that these proteins may be potential diagnostic markers for evaluation of smoking risk in sperm. Further study of these proteins may provide insight into the pathogenic mechanisms underlying infertility in smoking persons.

Estimated diversity of messenger RNAs in each murine spermatozoa and their potential function during early zygotic development.

To study the diversity of mRNAs in murine spermatozoa and their potential function during zygotic development, total RNAs in murine spermatozoa were sequenced via RNA-Seq and analyzed through bioinformatics techniques. The delivery and translation of sperm-borne mRNA in fertilized oocyte were detected using RT-PCR (reverse transcription-polymerase chain reaction), Western blot, and immunofluorescence. A total of 35 288 825 reads matching 33 039 transcripts, including 27 310 coding transcripts, were obtained. Based on our analyses, we hypothesized that the transcripts with RPKM (reads per kilobase of exon model per million mapped reads) higher than six may exist in each sperm cell as consistently retained transcripts. There were 4885 consistent transcripts in each sperm, and the remainder were randomly retained. If the baseline RPKM increased, the remaining coding transcripts were more likely related to reproduction and development. The sperm-borne transcripts Wnt4 and Foxg1 were delivered into fertilized oocytes on fertilization. Furthermore, Wnt4 was translated into protein in zygotes, whereas Foxg1 was not translated. In conclusion, approximately 4885 mRNAs were present in each murine spermatozoon, and the spermatozoal mRNAs related to reproduction and development were more likely retained. The sperm-borne mRNA Wnt4 was delivered into the fertilized oocyte and translated, evidence of a paternal effect on zygotic development.

Online advance respiration prediction model for percutaneous puncture robotics.

PURPOSE: Surgical robots have significant research value and clinical significance in the field of percutaneous punctures. There have been numerous studies on ultrasound-guided percutaneous surgical robots; however, addressing the respiratory compensation problem of deep punctures remains a significant obstacle. Herein we propose a robotic system for percutaneous puncture with respiratory compensation. METHODS: We proposed an online advance respiratory prediction model based on Bidirectional Gate Recurrent Unit (Bi-GRU) for the respiratory prediction requirements of surgical robot systems. By analyzing the main factors governing the accuracy of the respiratory motion prediction models, various parameters of the online advance prediction model were optimized. Subsequently, we integrated and developed ultrasound-guided percutaneous puncture robot software and a hardware platform to implement respiratory compensation, thus verifying the effectiveness and reliability of various key technologies in the system. RESULTS: The proposed respiratory prediction model has a significantly reduced update time, with an average root mean square error (RMSE) of less than 0.4 mm. This represents a reduction of ~ 20% compared to the online training long short-term memory(LSTM). By conducting puncture experiments based on a respiratory phantom, the average puncture error was 2.71 ± 0.65 mm and the average single-round puncture time was 65.00 ± 6.67 s. CONCLUSION: Herein we proposed and optimized an online training respiratory prediction network model based on Bi-GRU. The stability and reliability of this system are verified by conducting puncture experiments on a respiratory phantom.

Preparation of Ag@lignin nanotubes for the development of antimicrobial biodegradable films from corn straw.

Current environmental and energy issues have attracted considerable attention from industries, governments, and academia. Developing alternative diverse petrochemical-based plastics with biodegradable packaging materials from renewable resources is critical for ensuring both sustainability and safety. In this study, biodegradable films are fabricated from corn straw via a facile sol-gel process. Furthermore, these films are imbued with antimicrobial properties by coupling with silver@lignin nanotube hybrid antibacterial agents, formed via the in situ reduction of silver ions into elemental silver by lignin (mild reducing agent), followed by the self-assembly of lignin molecules into nanotubes assisted by an aqueous silver nitrate electrolyte solution. The developed antibacterial corn straw film exhibits strong mechanical and antibacterial properties, with a tensile strength and elongation at break of 68.7 MPa and 11.3 %, respectively, under optimum conditions and antibacterial activity against Escherichia coli and Staphylococcus aureus of 99.9 % and 97.2 %, respectively. The as-prepared corn straw films exhibit high hydrophobicity and ultraviolet resistance. The morphology, structure, and thermal properties of the corn straw films were characterized using Fourier transform infrared spectroscopy, scanning electron microscopy, energy-dispersive spectroscopy, X-ray diffraction, and thermogravimetric analysis. This study provides a straw-based biodegradable packaging film with antimicrobial properties.

Full-length transcriptome sequencing of pepper fruit during development and construction of a transcript variation database.

Chili pepper is an important spice and a model plant for fruit development studies. Large-scale omics information on chili pepper plant development continues to be gathered for understanding development as well as capsaicin biosynthesis. In this study, a full-spectrum transcriptome data of eight chili pepper tissues at five growth stages using the Oxford Nanopore long-read sequencing approach was generated. Of the 485 351 transcripts, 35 336 were recorded as reference transcripts (genes), while 450 015 were novel including coding, lnc, and other non-coding RNAs. These novel transcripts belonged to unknown/intergenic (347703), those retained introns (26336), and had multi-exons with at least one junction match (20333). In terms of alternative splicing, retained intron had the highest proportion (14795). The number of tissue-specific expressed transcripts ranged from 22 925 (stem) to 40 289 (flower). The expression changes during fruit and placenta development are discussed in detail. Integration of gene expression and capsaicin content quantification throughout the placental development clarifies that capsaicin biosynthesis in pepper is mainly derived from valine, leucin, and isoleucine degradation as well as citrate cycle and/or pyrimidine metabolism pathways. Most importantly, a user-friendly Pepper Full-Length Transcriptome Variation Database (PFTVD 1.0) (http://pepper-database.cn/) has been developed. PFTVD 1.0 provides transcriptomics and genomics information and allows users to analyse the data using various tools implemented. This work highlights the potential of long-read sequencing to discover novel genes and transcripts and their diversity in plant developmental biology.

Cigarette smoking exposure alters pebp1 DNA methylation and protein profile involved in MAPK signaling pathway in mice testis.

Many studies have addressed the role of cigarette smoking on semen quality, but the exact mechanisms remain inconclusive. To evaluate the detrimental effects of smoking on the spermatogenesis process, we initially screened and investigated 31 differentially expressed proteins extracted from the testes of mice exposed daily to cigarette smoke using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry analysis. Data mining analysis showed that these 31 proteins were categorized into five functional clustering groups: metabolic process, cell growth and/or maintenance, RNA and protein processing, stress response, and spermatogenesis. Additionally, 23 of 31 proteins were involved in a main pathway network, including Pkc (s), ERK1/2, Akt, and NF-kappaB, which are known to be involved in cell communication, proliferation, and differentiation. Interestingly, among the 31 proteins, a spermatogenesis-associated protein, phosphatidylethanolamine-binding protein 1 (PEBP1), was especially expressed in serial sections of spermatids of spermiogenesis and interacted with ERKs. The bisulfite sequencing result showed four CpGs near the Pebp1 transcriptional start site were largely methylated in the treated group. A 5-aza-2'-deoxycytidine treatment on GC-1 spg cells reversed the hypermethylation status and elevated both Pebp1 mRNA and protein expression levels. ERK1/2 phosphorylation levels were also increased with upregulation of Pebp1 expression in GC-1 spg cells. In conclusion, protein profile in testes could be altered by cigarette smoking. Moreover, we also suggest that epigenetic Pebp1 inactivation may affect activation of ERK, and it could impair spermatogenesis of mice. Our data could provide further insight into the mechanisms of spermatogenesis.

Heparanase induced by advanced glycation end products (AGEs) promotes macrophage migration involving RAGE and PI3K/AKT pathway.

BACKGROUND: Advanced glycation end products (AGEs), inflammatory-associated macrophage migration and accumulation are crucial for initiation and progression of diabetic vascular complication. Enzymatic activity of heparanase (HPA) is implicated strongly in dissemination of metastatic tumor cells and cells of the immune system. In addition, HPA enhances the phosphorylation of selected signaling molecules including AKT pathway independent of enzymatic activity. However, virtually nothing is presently known the role of HPA during macrophage migration exposed to AGEs involving signal pathway. METHODS: These studies were carried out in Ana-1 macrophages. Macrophage viability was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays. HPA and AKT protein expression in macrophages are analysed by Western blotting and HPA mRNA expression by real time quantitative RT-PCR. Release of HPA was determined by ELISA. Macrophage migration was assessed by Transwell assays. RESULTS: HPA protein and mRNA were found to be increased significantly in AGEs-treated macrophages. Pretreatment with anti-HPA antibody which recognizes the nonenzymatic terminal of HPA prevented AGEs-induced AKT phosphorylation and macrophage migration. LY294002 (PI3k/AKT inhibitor) inhibited AGEs-induced macrophage migration. Furthermore, pretreatment with anti-receptor for advanced glycation end products (RAGE) antibody attenuated AGEs-induced HPA expression, AKT phosphorylation and macrophage migration. CONCLUSIONS: These data indicate that AGEs-induced macrophage migration is dependent on HPA involving RAGE-HPA-PI3K/AKT pathway. The nonenzymatic activity of HPA may play a key role in AGEs-induced macrophage migration associated with inflammation in diabetic vascular complication.

Outward-oriented sites within clustered CTCF boundaries are key for intra-TAD chromatin interactions and gene regulation.

CTCF plays an important role in 3D genome organization by adjusting the strength of chromatin insulation at TAD boundaries, where clustered CBS (CTCF-binding site) elements are often arranged in a tandem array with a complex divergent or convergent orientation. Here, using Pcdh and HOXD loci as a paradigm, we look into the clustered CTCF TAD boundaries and find that, counterintuitively, outward-oriented CBS elements are crucial for inward enhancer-promoter interactions as well as for gene regulation. Specifically, by combinatorial deletions of a series of putative enhancer elements in mice in vivo or CBS elements in cultured cells in vitro, in conjunction with chromosome conformation capture and RNA-seq analyses, we show that deletions of outward-oriented CBS elements weaken the strength of long-distance intra-TAD promoter-enhancer interactions and enhancer activation of target genes. Our data highlight the crucial role of outward-oriented CBS elements within the clustered CTCF TAD boundaries in developmental gene regulation and have interesting implications on the organization principles of clustered CTCF sites within TAD boundaries.

Robotic system for accurate percutaneous puncture guided by 3D-2D ultrasound.

PURPOSE: Ultrasound (US)-guided robotic systems can reduce the reliance on the experience and skills of surgeons and enable automatic and accurate percutaneous puncture. Two-dimensional (2D) and three-dimensional (3D) US guidance have various advantages and disadvantages. The planned puncture path in the US data directly affects the puncture quality and tissue injury risk. It is difficult to define the optimal path in 2D US images and achieve accurate and safe puncture under the guidance of 3D US volume. This study aims to propose a robotic system guided by 3D-2D US to realize accurate and safe percutaneous puncture. METHODS: We proposed a 3D-2D US-guided percutaneous puncture robotic system by integrating a 3D US scanning robotic system and a 2D US-guided puncture robotic system. The optimal spatial puncture path that targets the lesion and avoids other important tissues was determined in the 3D US volume reconstructed through robotic US scanning. Thereafter, the puncture robot was placed at the puncture site determined according to the planned path. The optimal path was mapped to the 2D US image taken at the puncture site. Finally, the 2D US image and puncture path were used to guide the robot in performing an accurate and safe percutaneous puncture. RESULTS: The proposed robotic system based on the guidance of 3D-2D US exhibits the advantages of both 3D US and 2D US to improve the accuracy of percutaneous puncture and reduce the risk of tissue injury. The experimental results of phantom puncture demonstrate that the mean puncture accuracy of the system is 1.09 ± 0.35 mm, and the puncture success rate with single needle insertion is 100%. CONCLUSION: A percutaneous puncture robotic system based on 3D-2D US guidance was proposed and tested successfully. The experimental results demonstrate the feasibility of the proposed system for achieving accurate and safe robotic percutaneous puncture.

Method for Carotid Artery 3-D Ultrasound Image Segmentation Based on CSWin Transformer.

Precise segmentation of carotid artery (CA) structure is an important prerequisite for the medical assessment and detection of carotid plaques. For automatic segmentation of the media-adventitia boundary (MAB) and lumen-intima boundary (LIB) in 3-D ultrasound images of the CA, a U-shaped CSWin transformer (U-CSWT) is proposed. Both the encoder and decoder of the U-CSWT are composed of hierarchical CSWT modules, which can capture rich global context information in the 3-D image. Experiments were performed on a 3-D ultrasound image data set of the CA, and the results indicate that the U-CSWT performs better than other convolutional neural network (CNN)-based and CNN-transformer hybrid methods. The model yields Dice coefficients of 94.6 ± 3.0% and 90.8 ± 5.1% for the MAB and LIB in the common carotid artery (CCA) and 92.9 ± 4.9% and 89.6 ± 6.2% for MAB and LIB in the bifurcation, respectively. Our U-CSWT is expected to become an effective method for automatic segmentation of 3-D ultrasound images of CA.

Preparation of Bio-Foam Material from Steam-Exploded Corn Straw by In Situ Esterification Modification.

In this work, we engineered a corn-straw-based bio-foam material under the inspiration of the intrinsic morphology of the corn stem. The explosion pretreatment was applied to obtain a fibrillated cellulose starting material rich in lignin. The in situ esterification of cellulose was adopted to improve the cross-linking network of the as-developed foam bio-material. The esterification of lignin was observed in the same procedure, which provides a better cross-linking interaction. The esterified corn-straw-derived bio-foam material showed excellent elastic resilience performance with an elastic recovery ratio of 83% and an elastic modulus of 20 kPa. Meanwhile, with surface modification by hexachlorocyclotriphosphazene-functionalized lignin as the flame retardant (Lig-HCCP), the as-obtained bio-foam material demonstrated quite a good flame retardancy (with 27.3% of the LOI), as well as a heat insulation property. The corn-straw-derived bio-foam material is prospected to be a potential substitution packaging material for widely used petroleum-derived products. This work provides a new value-added application of the abundant agricultural straw biomass resources.

Astragalus membranaceus inhibits inflammation via phospho-P38 mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-κB pathways in advanced glycation end product-stimulated macrophages.

Advanced glycation end products (AGEs) and inflammation contribute to the development of diabetic complications. Astragalus membranaceus has properties of immunological regulation in many diseases. The aim of this study was to determine the function of A. membranaceus extract (AME) on the AGE-induced inflammatory response in Ana-1 macrophages. The viability of cells treated with AME or AGEs was evaluated with the MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] method. The secretion and mRNA levels of IL-1ß and TNF-α were measured by ELISA and RT-PCR, respectively. The activity of NF-κB was assayed by EMSA. The phosphorylation of p38 MAPK was assessed by western blotting. The results showed that AME was not toxic to macrophages. The treatment of macrophages with AME effectively inhibited AGE-induced IL-1ß and TNF-α secretion and mRNA expression in macrophages. These effects may be mediated by p38 MAPK and the NF-κB pathway. The results suggest that AME can inhibit AGE-induced inflammatory cytokine production to down-regulate macrophage-mediated inflammation via p38 MAPK and NF-κB signaling pathways and indicate that AME could be an immunoregulatory agent against AGE-induced inflammation in diabetes.

Astragalus embranaceus extract activates immune response in macrophages via heparanase.

Astragalus membranaceus (AM), a traditional Chinese medicinal herb, has immunoregulatory properties in many diseases. We investigated the effects and mechanism of Astragalus membranaceus extract (AME) in the macrophage migration and immune response mediator release. The viability of Ana-1 macrophages treated with AME was evaluated by the MTT method. The secretion and mRNA levels of IL-1ß and TNF-a were measured by ELISA and RT-PCR, respectively. Macrophage migration was assayed by transwell assay. The activity of heparanase (HPA) was determined by a heparin-degrading enzyme assay. Our results didn't show any toxicity of AME in macrophages. AME increased the activity of HPA, cell migration, mRNA levels and secretion of IL-1ß and TNF-a in macrophages. Pretreatment with anti-HPA antibody reduced cell migration, secretion of IL-1ß and TNF-a did not change the mRNA levels of IL-1ß and TNF-a significantly in AME-treated macrophages. This suggests that AME may increase the release of immune response mediator and cell migration via HPA to activate immune response in macrophages.

Design and implementation of a surgical planning system for robotic assisted mandible reconstruction with fibula free flap.

PURPOSE: Free fibula flap is the gold standard for the treatment of mandibular defects. However, the existing preoperative planning protocol is cumbersome to execute, costly to learn, and poorly collaborative with the robot-assisted cutting of the fibular osteotomy plane. METHODS: A surgical planning system for robotic assisted mandibular reconstruction with fibula free flap is proposed in this study. A fibular osteotomy planning algorithm is presented so that the virtual surgical planning of the fibular osteotomy segments can be obtained automatically with selected mandibular anatomical landmarks. The planned osteotomy planes are then converted into the motion path of the robotic arm, and the automatic fibula osteotomy is completed under optical navigation. RESULTS: Surgical planning was performed on 35 patients to verify the feasibility of our system's virtual surgical planning module, with an average time of 13 min. Phantom experiments were performed to evaluate the reliability and stability of this system. The average distance and angular deviations of the osteotomy planes are 1.04 ± 0.68 mm and 1.56 ±1.10°, respectively. CONCLUSIONS: Our system can achieve not only precise and convenient preoperative planning, but also safe and reliable osteotomy trajectory. The clinical applications of our system for mandibular reconstruction surgery are expected soon.