STC1 competitively binding ßPIX enhances melanoma progression via YAP nuclear translocation and M2 macrophage recruitment through the YAP/CCL2/VEGFA/AKT feedback loop.

This study investigates the role of Stanniocalcin-1 (STC1) in melanoma progression, with a focus on its impact on metastasis, angiogenesis, and immune evasion. Systematic bioinformatics analysis revealed the potential influence of STC1 dysregulation on prognosis, immune cell infiltration, response to immune therapy, and cellular functions. In vitro assays were conducted to assess the proliferation, invasion, migration, and angiogenesis capabilities of A375 cells. In vivo experiments utilizing C57BL/6 J mice established a lung metastasis model using B16-F10 cells to evaluate macrophage infiltration and M2 polarization. A Transwell co-culture system was employed to explore the crosstalk between melanoma and macrophages. Molecular interactions among STC1, YAP, ßPIX, and CCL2 are investigated using mass spectrometry, Co-Immunoprecipitation, Dual-Luciferase Reporter Assay, and Chromatin Immunoprecipitation experiments. STC1 was found to enhance lung metastasis by promoting the recruitment and polarization of M2 macrophages, thereby fostering an immunosuppressive microenvironment. Mechanistically, STC1 competes with YAP for binding to ßPIX within the KER domain in melanoma cells, leading to YAP activation and subsequent CCL2 upregulation. CCL2-induced M2 macrophages secrete VEGFA, which enhances tumor vascularization and increases STC1 expression via the AKT signaling pathway in melanoma cells, establishing a pro-metastatic feedback loop. Notably, STC1-induced YAP activation increases PD-L1 expression, promoting immune evasion. Silencing STC1 enhances the efficacy of PD-1 immune checkpoint therapy in mice. This research elucidates STC1's role in melanoma metastasis and its complex interactions with tumor-associated macrophages, proposing STC1 as a potential therapeutic target for countering melanoma metastasis and augmenting the efficacy of PD-1 immunotherapy.

Bioinformatics methods in biomarkers of preeclampsia and associated potential drug applications.

BACKGROUND: Preeclampsia is a pregnancy-related condition that causes high blood pressure and proteinuria after 20 weeks of pregnancy. It is linked to increased maternal mortality, organ malfunction, and foetal development limitation. In this view, there is a need critical to identify biomarkers for the early detection of preeclampsia. The objective of this study is to discover critical genes and explore medications for preeclampsia treatment that may influence these genes. METHODS: Four datasets, including GSE10588, GSE25906, GSE48424 and GSE60438 were retrieved from the Gene Expression Omnibus database. The GSE10588, GSE25906, and GSE48424 datasets were then removed the batch effect using the "sva" R package and merged into a complete dataset. The differentially expressed genes (DEGs) were identified using the "limma" R package. The potential small-molecule agents for the treatment of PE was further screened using the Connective Map (CMAP) drug database based on the DEGs. Further, Weight gene Co-expression network (WGNCA) analysis was performed to identified gene module associated with preeclampsia, hub genes were then identified using the logistic regression analysis. Finally, the immune cell infiltration level of genes was evaluated through the single sample gene set enrichment analysis (ssGSEA). RESULTS: A total of 681 DEGs (376 down-regulated and 305 up-regulated genes) were identified between normal and preeclampsia samples. Then, Dexamethasone, Prednisone, Rimexolone, Piretanide, Trazodone, Buflomedil, Scoulerin, Irinotecan, and Camptothecin drugs were screened based on these DEGs through the CMAP database. Two modules including yellow and brown modules were the most associated with disease through the WGCNA analysis. KEGG analysis revealed that the chemokine signaling pathway, Th1 and Th2 cell differentiation, B cell receptor signalling pathway and oxytocin signalling pathway were significantly enriched in these modules. Moreover, two key genes, PLEK and LEP were evaluated using the univariate and multivariate logistic regression analysis from the hub modules. These two genes were further validated in the external validation cohort GSE60438 and qRT-PCR experiment. Finally, we evaluated the relationship between immune cell and two genes. CONCLUSION: In conclusion, the present study investigated key genes associated with PE pathogenesis that may contribute to identifying potential biomarkers, therapeutic agents and developing personalized treatment for PE.

Knockdown of NLE1 inhibits development of malignant melanoma in vitro and in vivo NLE1 promotes development of malignant melanoma.

Melanoma, which originates from neural crest derived melanocytes, causes severe pain and even death to numerous patients. Previous studies reported that Notchless Homolog 1 (NLE1) plays an important role in cell proliferation, transcription and signal transduction. However, the clinical significance and biological behavior of NLE1 in melanoma remain a mystery. Thus, the role of NLE1 in melanoma was investigated in vitro and in vivo. The expression of NLE1 in melanoma was elevated and the expression level was positively correlated with lymphatic metastasis and tumor stage. In addition, NLE1 knockdown by shRNA specifically inhibited proliferation, enhanced the apoptotic sensitivity and hindered migration of melanoma cells in vitro. Mice xenograft model further showed that NLE1 knockdown could inhibit the tumor formation of melanoma in vivo. Additionally, the induction of apoptosis of melanoma cells by NLE1 knockdown required the participation of a series of apoptosis-related proteins. Besides, NLE1 can activate the PI3K/AKT signaling pathway. In summary, NLE1 was involved in the development and progression of melanoma, which may be a novel potential target for molecular therapy of melanoma.

Association of Fpr1 gene expression with osteogenesis and adipogenesis of adipose derived stem cells.

Formyl peptide receptors (Fprs) play fundamental roles in multiple cell functions including promotion of osteogenesis and bone fracture healing. In this study, the role of Fpr1 gene in osteogenic and adipogenic differentiation of adipose derived stem cells (ADSCs) was investigated. Primary ADSCs (mADSCs) from either Fpr1 knockout (KO) or wild type (WT) mice and human ADSCs (hADSCs) were treated by osteogenic (OM) or adipogenic (AM) medium, with basal medium as control. Osteogenesis and adipogenesis were measured by histological and biochemical methods. In both hADSCs and mADSCs, Fpr1 gene expression, osteogenic gene expression, as well as mineralization were increased after osteogenic induction. The osteogenic capacity of KO ADSCs was remarkably reduced compared to WT ADSCs, with decreased levels of expression of osteogenic markers, alkaline phosphatase activity, and mineralization. In contrast, the adipogenesis of KO ADSCs was remarkably enhanced compared with WT ADSCs, forming more lipid droplets, and increasing expression of adipogenic markers PPARγ and aP2. Expression of the nuclear transcription factor Forkhead box protein O1 (FoxO1) was decreased in KO ADSCs, while OM and AM caused increase and decrease in FoxO1 expression, respectively. The current study revealed a correlation of Fpr1 gene expression with osteogenesis and adipogenesis of mADSCs, underlying a mechanism involving FoxO1. Our present research suggests that targeting Fpr1 might be a novel strategy to enhance osteogenesis of ADSCs.

Increased first and second pulse harmonics in Tai Chi Chuan practitioners.

BACKGROUND: Tai Chi Chuan (TCC) is known to be a good calisthenics for people. This study examined the relationship between pulse harmonics and autonomic nervous modulation in TCC practitioners. METHODS: Power spectral measures of right pulse wave and heart rate variability (HRV) measures were compared between TCC practitioners and control subjects. Correlation analyses between pulse harmonics and HRV measures were performed using linear regression analysis. RESULTS: At baseline, the total power of pulse (TPp), powers of all individual pulse harmonics, normalized power of the 1(st) harmonics (nPh1) of TCC practitioners were greater, while the normalized power of the 4(th) pulse harmonics (nPh4) of TCC practitioners was smaller, than those of the controls. Similarly, the baseline standard deviation (SD(RR)), coefficient of variation (CV(RR)), and normalized high-frequency power (nHFP) of RR intervals were smaller, while the normalized very low-frequency power (nVLFP) and low-/high- frequency power ratio (LHR) were larger in the TCC practitioners. The TCC age correlated significantly and negatively with nPh1, and nearly significantly and negatively with nPh2 in the TCC practitioners. Thirty min after TCC exercise, the percentage changes in mRRI, SDRR, TP, VLFP were decreased, while the percentage changes in HR, ULFP, nLFP, and Ph2 were increased, relative to the controls. Correlation analysis shows that the %Ph2 correlates significantly and negatively with %mRRI and significantly and positively with %HR. CONCLUSION: The TCC practitioners had increased baseline total power of pulse and the 1(st) and 2(nd) pulse harmonics, and decreased power of the 4(th) pulse harmonics, along with decreased vagal modulation and increased sympathetic modulation. After TCC exercise, the power of the 2(nd) harmonics of TCC practitioners was increased which might be related to the increase in HR due to decreased vascular resistance after TCC exercise.

The use of heart rate variability measures as indicators of autonomic nervous modulation must be careful in patients after orthotopic heart transplantation.

The precise relation between heart rate variability (HRV) and autonomic re-innervation has not been established explicitly in patients after orthotopic heart transplantation (OHT), but can be inferred from the fact that the HRV is reduced immediately after OHT and may increase gradually with time. The aim of this study was to investigate the residual HRV in patients about 1-2 years after OHT, as compared with patients after coronary artery bypass graft (CABG) surgery. Thirteen patients who had received OHT and 14 patients who had received CABG surgery were recruited. HRV analysis was performed and the HRV measures in supine position were compared between these two groups of patients. We found that the mean (mRRI), standard deviation and coefficient of variation of RR intervals, total power, very low frequency power (VLFP), low frequency power, high frequency power (HFP), normalized VLFP (nVLFP) and low-/high-frequency power ratio in the OHT group were all significantly decreased, while the heart rate (HR) and normalized HFP (nHFP) were significantly increased, as compared with the CABG group. The decrease in HRV was more severe in the VLFP region. A smaller nVLFP and a greater nHFP were associated with a smaller mRRI and a larger HR in the OHT patients. The slope of the power law relation of HRV became positive in OHT patients, instead of negative in CABG patients. We conclude that patients after OHT have residual HRV which were characterized by severely depressed time and frequency domain HRV, increased HR and nHFP, decreased nVLFP, and positive slope of the power-law relation of HRV. The use of nHFP as the indicator of vagal modulation and the use of nVLFP as the indicator of renin-angiotensin modulation, thermoregulation and vagal withdrawal must be careful in the OHT patients.

CXCL13 promotes the effect of bone marrow mesenchymal stem cells (MSCs) on tendon-bone healing in rats and in C3HIOT1/2 cells.

OBJECTIVES: Mesenchymal stem cells (MSCs) are potential effective therapy for tissue repair and bone regeneration. In present study, the effects of CXC chemokine ligand-13 (CXCL13) were evaluated on tendon-bone healing of rats. METHODS: Tendon bone healing of the rat model was established and biomechanical testing was performed at 2, 4, 8 weeks after surgery. Murine mesenchymal cell line (C3HIOT1/2 cells) was cultured. The expression of miRNA-23a was detected by real-time PCR. The protein expression of ERK1/2, JNK and p38 was detected by western blotting. MiR-23a mimic and inhibitor were used to overexpress or silence the expression of miR-23a. RESULTS: MSCs significantly elevated the levels of ultimate load to failure, stiffness and stress in specimens of rats, the effects of which were enhanced by CXCL13. The expression of miR-23a was down-regulated and the protein of ERK1/2 level was up-regulated by CXCL13 treatment in both in vivo and in vitro experiments. ERK1/2 expression was elevated by overexpression of miR-23a and reduced by miR-23a inhibitor. CONCLUSIONS: These findings revealed that CXCL13 promoted the tendon-bone healing in rats with MSCs treatment, and implied that the activation of ERK1/2 via miR-23a was involved in the process of MSCs treated bone regeneration.

Breathing frequency-independent effect of Tai Chi Chuan on autonomic modulation.

OBJECTIVE: This study investigates the breathing frequency (BF)-independent effect of Tai Chi Chuan (TCC) on autonomic nervous modulation in TCC practitioners. METHODS: Twenty-five TCC practitioners and 25 sedentary normal controls were recruited. The stationary heart rate variability (HRV) measures of TCC practitioners and controls were compared. The same HRV measures in TCC practitioners and among the controls, TCC practitioners before TCC and TCC practitioners 30 min after TCC were compared. RESULTS: In TCC practitioners, the BF, normalized high-frequency power (nHFP), and normalized very low-frequency power were significantly increased, while the normalized low-frequency power (nLFP) was significantly decreased 30 min after TCC. The BF correlated significantly and negatively with heart rate (HR), nHFP and nLFP, and correlated significantly and positively with mean RR interval (MnRR) before TCC in TCC practitioners. A slower BF is associated with a higher HR, a greater vagal modulation, and a greater combined sympatho-vagal modulation before TCC. To remove the effect of BF on HRV measures, new indices such as HR*BF, nHFP*BF, nLFP*BF, and MnRR/BF were introduced for comparison among the controls, TCC practitioners before TCC, and TCC practitioners 30 min after TCC. Thirty minutes after TCC, the MnRR/BF of TCC practitioner was smaller whereas HR*BF and nHFP*BF were greater than those before TCC. INTERPRETATION: The BF-independent effects of TCC on the autonomic nervous modulation of TCC practitioners are an increase in vagal modulation and HR, and a decrease in mean RR interval. The mechanism underlying the parallel increase in HR and vagal modulation in TCC practitioners is not understood yet at present.

[Study on Ficolin-A against infection of Plasmodium berghei in mouse model].

OBJECTIVE: To evaluate the effect of Ficolin-A, a lectin complement against Plasmodium berghei in mice model. METHODS: The Mr 19,000 fragment of merozoite surface protein-1 of P. berghei (MSP1(19)) was cloned and then subcloned into the vector pGEX-KG. The recombinants of pGEX-KG-Ficolin-A and pGEX-KG-MSP1(19) were transformed into Escherichia coli BL21, and followed by expression of the protein induced by 1 mmol/L IPTG. The fusion protein was purified by affinity chromatography using Glutathione Sepharose 4B, and then identified by SDS-PAGE and Western-blotting. Five mouse model groups were treated with PBS, GST, Ficolin-A, MSP1(19), or Ficolin-A+MSP1(19), respectively. Each group had eight mice. Mice in Ficolin-A or MSP1(19) groups were injected with 20 microg Ficolin-A or MSP1(19) protein each time, respectively. Mice in Ficolin-A+MSP1(19) group were injected with 20 microg Ficolin-A and 20 microg MSP1(19) each time. Mice in control groups were injected with 200 microl PBS or 20 microg GST, respectively. All the mice received four immunizations at 2-week intervals. Two weeks after the last immunization, all the mice were inoculated with 300 microl Plasmodium berghei-infected red blood cells. On day 2, 4, 6, 8, and 10 post-infection, blood samples were collected from three mice of each group, and the Giemsa stained-blood films were microscopically examined. Density of malaria parasites was calculated. The survival rate was evaluated on day 20 post-infection. RESULTS: The recombinant vectors of pGEX-KG-Ficolin-A and pGEX-KG-MSP1(19) were constructed. Purified fusion proteins, Ficolin-A-GST and MSP1(19)-GST, were obtained. Western blotting analysis indicated that the relative molecular mass of fusion proteins Ficolin-A-GST and MSP1(19)-GST was about Mr 69,000 and Mr 41,000. Animal experiments showed that on day 10 after infection, the parasite density in Ficolin-A+MSP1(19) group [(22.2 +/- 1.7)%] was slightly lower than that of the groups MSP1(19) [(33.4 +/- 2.7)%], Ficolin-A [(36.2 3.1)%], GST [(43.8 +/- 4.8)%] and PBS [(45.3 +/- 3.6)%], but the difference was not statistically significant (P > 0.05). No mouse survived in PBS group on day 20 after infection. There was no significant difference in number of survival mice between Ficolin-A group (3 mice) and GST group (2 mice). Six mice survived in Ficolin-A+MSP1(19) group, which was significantly more than that of GST group (P < 0.05). CONCLUSION: Ficolin-A cannot significantly suppress parasite density. However, Ficolin-A+MSP1(19) can increase the survival rate of Plasmodium berghei-infected mice.

Mechanisms of the effect of fertility policies on the labor-capital income gap.

This paper investigates the impact mechanism by which an incentive-based fertility policy may reduce the labor income share. First, the specific paths through which this impact mechanism is realized are analyzed using the production function. It is found that an incentive-based fertility policy triggers high savings, which implies more, cheaper, and more readily available capital to be invested in production. A distribution system that earns income based on factor contributions results in more gains for capital than labor, i.e., a lower share of labor income and a wider income gap between labor and capital. Second, the impact mechanism includes three theoretical hypotheses. They are that an encouraging fertility policy is negatively related to labor income share; this relationship is valid provided that the study subject is in a closed economy; and that capital intensification is a mediator variable of fertility policy affecting labor income share. Finally, to further corroborate the impact mechanism in this paper, a Hansen threshold panel model is applied to verify that the effect of fertility policy on labor income share has a threshold effect. This indicates that the effect of the former on the latter changes significantly before and after the change in fertility policy, confirming the existence of an impact mechanism. The established literature has paid little attention to the impact of incentivised fertility policies on the labour income gap. Using capital intensification as the mediating variable, this paper demonstrates the existence of the former effect on the latter. In view of this, under the encouraged fertility policy, this paper proposes specific measures to enhance the labor income share in order to narrow the income gap between labor and capital.

Determinants of hand pulse wave velocity and hand pulse transit time in healthy adults.

Arterial pulse wave velocity (PWV) is recognized as a convenient method to assess peripheral vascular stiffness. This study explored the clinical characteristics of hand PWV (hPWV) and hand pulse transit time (hPTT) in healthy adults (sixty males = 42.4 ± 13.9 yrs; sixty-four females = 42.8 ± 13.9 yrs) voluntarily participated in this study. The arterial pulse waveform and the anatomical distance from the radial styloid process to the tip of the middle finger of both hands were recorded in the sitting position. The hPWV was calculated as the traversed distance divided by hPTT between those two points. Male subjects showed significantly greater hPWV, systolic blood pressure, and pulse pressure than age-matched female subjects, while the hPTT was not significantly different between genders. Multiple linear regression analysis showed that gender is a common determinant of hPWV and hPTT, and that age and heart rate (HR) were negatively correlated with hPWV and hPTT, respectively. We conclude that male subjects have greater hPWV than female subjects. Ageing is associated with decreased hPWV, while increased HR is associated with a smaller hPTT. The hPWV and hPTT might be used as non-invasive indices to characterise the ageing and arterial stiffness of peripheral blood vessels.

Agreement of ultra-short-term heart rate variability measure after different repeated bouts of sprint ability tests.

This study aims to explore: (1) the validity of post-exercise ultra-short-term heart rate variability (HRVust) after two different bouts of repeated sprint ability test (RSA), and (2) the relationship between HRVust measure and RSA performance. Twenty adolescent male futsal players voluntarily participated in this study (age: 17.65 ± 1.81 years, body height: 170.88 ± 4.98 cm, body weight: 61.78 ± 4.67 kg). The participants performed a standard RSA test (RSAstandard) and an RSA test with a 10% decrement of the best sprint time test (RSA10%decrement) on two separate occasions within a week. On both occasions, a 5-min resting electrocardiography was administered pre- and post-RSA exercise protocols. The first 30-s (HRVust30s), 60-s (HRVust60s), and 60-120-s (HRVust1-2min) were extracted and used to compare with the standard of 5-min HRV recording (HRVcriterion). The natural logarithm (ln) of the standard deviation of normal-to-normal intervals (SDNN) and root mean square of successive normal-to-normal interval differences (RMSSD) HRV indices were utilised to establish intraclass correlation coefficient (ICC2,1), coefficient of variation (%CV), and Pearson product-moment correlation (r). Results revealed the ICC values of HRVust lnSDNN (RSAstandard = 0.77-0.88; RSA10%decrement = 0.41-0.71) and lnRMSSD (RSAstandard = 0.81-0.86; RSA10%decrement = 0.57-0.82). Furthermore, significantly positive correlations between best sprint time and post-exercise HRVust indices were found in lnSDNN (r = 0.47-0.62; p < 0.05) and lnRMSSD (r = 0.45; p < 0.05). Additionally, a large CV of lnSDNN (RSAstandard = 32%-45%; RSA10%decrement = 29%-39%), lnRMSSD (RSAstandard = 50%-66%; RSA10%decrement = 48%-52%), and ratio (RSAstandard = 45%-126%; RSA10%decrement = 27%-45%) was found after the RSA protocols. In conclusion, the number of bouts of RSA exercise potentially influences the agreement of post-exercise time-domain HRVust indices to standard HRV measure.

Knockout of formyl peptide receptor 1 reduces osteogenesis and bone healing.

AIMS: Formyl peptide receptor 1 (FPR1), from a G-protein coupled receptor family, was previously well-characterized in immune cells. But the function of FPR1 in osteogenesis and fracture healing was rarely reported. This study, using the FPR1 knockout (KO) mouse, is one of the first studies that try to investigate FPR1 function to osteogenic differentiation of bone marrow-derived stem cells (BMSCs) in vitro and bone fracture healing in vivo. MATERIALS AND METHODS: Primary BMSCs were isolated from both FPR1 KO and wild type (WT) mice. Cloned mouse BMSCs (D1 cells) were used to examine role of FoxO1 in FPR1 regulation of osteogenesis. A closed, transverse fracture at the femoral midshaft was created to compare bone healing between KO and WT mice. Biomechanical and structural properties of femur were compared between healthy WT and KO mice. KEY FINDINGS: FPR1 expression increased significantly during osteogenesis of both primary and cloned BMSCs. Compared to BMSCs from FPR1 KO mice, WT BMSCs displayed considerably higher levels of osteogenic markers as well as mineralization. Osteogenesis by D1 cells was inhibited by either an FPR1 antagonist cFLFLF or a specific inhibitor of FoxO1, AS1842856. In addition, the femur from WT mice had better biomechanical properties than FPR1 KO mice. Furthermore, bone healing in WT mice was remarkably improved compared to FPR1 KO mice analyzed by X-ray and micro-CT. SIGNIFICANCE: These findings indicated that FPR1 played a vital role in osteogenic differentiation and regenerative capacity of fractured bone, probably through the activation of FoxO1 related signaling pathways.

pH-Responsive ß-Glucans-Complexed mRNA in LNPs as an Oral Vaccine for Enhancing Cancer Immunotherapy.

mRNA vaccines for cancer immunotherapy are commonly delivered using lipid nanoparticles (LNPs), which, when administered intravenously, may accumulate in the liver, potentially limiting their therapeutic efficacy. To overcome this challenge, the study introduces an oral mRNA vaccine formulation tailored for efficient uptake by immune cells in the gastrointestinal (GI) tract, known for its high concentration of immune cells, including dendritic cells (DCs). This formulation comprises mRNA complexed with ß-glucans (ßGlus), a potential adjuvant for vaccines, encapsulated within LNPs (ßGlus/mRNA@LNPs). The ßGlus/mRNA complexes within the small compartments of LNPs demonstrate a distinctive ability to partially dissociate and reassociate, responding to pH changes, effectively shielding mRNA from degradation in the harsh GI environment. Upon oral administration to tumor-bearing mice, ßGlus/mRNA@LNPs are effectively taken up by intestinal DCs and local nonimmune cells, bypassing potential liver accumulation. This initiates antigen-specific immune responses through successful mRNA translation, followed by drainage into the mesenteric lymph nodes to stimulate T cells and trigger specific adaptive immune responses, ultimately enhancing antitumor effects. Importantly, the vaccine demonstrates safety, with no significant inflammatory reactions observed. In conclusion, the potential of oral ßGlus/mRNA@LNPs delivery presents a promising avenue in cancer immunotherapy, offering needle-free and user-friendly administration for widespread adoption and self-administration.

COX5A as a potential biomarker for disease activity and organ damage in lupus.

Systemic Lupus Erythematosus (SLE) is a complex autoimmune disease with limited therapeutic targets or clinical outcome predictors. This study aimed to gain more insights into the underlying immunological pathways and prognostic biomarkers of SLE. Integrated analyses of RNA-seq data from 64 SLE and 62 healthy controls, examining 27 immune cell types to explore the key pathways and driver genes in SLE pathogenesis. Single-cell RNA sequencing data from the skin and kidney were used to determine the association of COX5A expression with organ damage. The associations of COX5A with SLE phenotypes were further evaluated in two independent cohorts, and receiver operating characteristic (ROC) curves were constructed to assess the value of COX5A as a biomarker for disease activity and organ damage in SLE. We found that oxidative phosphorylation (OXPHOS) is the most significantly altered metabolic pathway in SLE, especially in effector T cells. Notably, we identified an OXPHOS-related enzyme, COX5A, whose expression was significantly higher in effector T cells than in naïve T cells and showed associations with disease activity, organ damage, and steroid treatment of SLE. Furthermore, ROC curves showed that COX5A is a robust biomarker for disease activity, kidney involvement, and new-onset skin lesions, with the area under the curve (AUC) values of 0.880, 0.801, and 0.805, respectively. Our results identified the OXPHOS signature as a prominent feature in SLE T cells, and COX5A as a potential candidate biomarker for disease activity and organ damage in SLE.

P2X7R Mediates the Synergistic Effect of ATP and MSU Crystals to Induce Acute Gouty Arthritis.

Activation of the nod-like receptor protein 3 (NLRP3) inflammasome by monosodium urate (MSU) crystals has been identified as the molecular basis for the acute inflammatory response in gouty arthritis. However, MSU crystals alone are not sufficient to induce acute gouty arthritis (AGA). Adenosine triphosphate (ATP) is an endogenous signaling molecule involved in the NLRP3 inflammasome activation. We aimed to explore the role of ATP in MSU crystal-induced AGA development. In peripheral blood mononuclear cell-derived macrophages obtained from gout patients, we observed a synergistic effect of ATP on MSU crystal-induced IL-1ß release. Furthermore, in a rat model of spontaneous gout, we demonstrated that a synergistic effect of ATP and MSU crystals, but not MSU crystals alone, is essential for triggering AGA. Mechanistically, this synergistic effect is achieved through the purinergic receptor P2X7 (P2X7R). Blockade of P2X7R prevented AGA induction in rats after local injection of MSU crystals, and carrying the mutant hP2X7R gene contributed to the inhibition of NLRP3 inflammasome activation induced by costimulation of MSU crystals and ATP in vitro. Taken together, these results support the synergistic effect of ATP on MSU crystal-induced NLRP3 inflammasome activation facilitating inflammatory episodes in AGA. In this process, P2X7R plays a key regulatory role, suggesting targeting P2X7R to be an attractive therapeutic strategy for the treatment of AGA.

Chitosan targets PI3K/Akt/FoxO3a axis to up-regulate FAM172A and suppress MAPK/ERK pathway to exert anti-tumor effect in osteosarcoma.

Osteosarcoma (OS) is a serve and the most frequent primary malignant tumor of bone. Chitosan was reported to have anti-tumor effect on human cancers including OS. However, the molecular mechanism by which chitosan suppresses tumor growth is not fully illustrated. In this study, human OS cell lines, including both Saos-2 and U2OS cells, were used to dissect the underlying mechanisms. RNA sequencing results show that a candidate biomarker family with sequence similarity 172 member A (FAM172A) was up-regulated in both of the two cell lines treated with chitosan. We observed that the mitogen-activated protein kinase (MAPK) signaling pathway could be inactivated by chitosan, and the MAPK inhibition caused by chitosan was reversed by FAM172A knockdown. Moreover, we uncovered a direct interaction between C-terminal domain of FAM172A (311-415) and mitogen-activated protein kinase kinase 1 (MEK1) (270-307) by immunoprecipitation assay. Finally, we also found that chitosan could bind with subunit p85 of PI3K to further inactivate the PI3K/Akt pathway. Taken together, our study demonstrates that chitosan binds with PI3K p85 subunit to suppress the activity of PI3K/Akt pathway to up-regulate the expression of FAM172A, and which exerts its function by suppressing phosphorylation of MEK1/2 and blocking the activity of MAPK/ERK signaling pathway. Taken together, our study deepens the understanding of the molecular mechanism of MAPK/ERK pathway inhibition induced by chitosan, and provides insights into the development of new targets to enhance the pharmacological effect of chitosan against OS.

Role of the tumor microenvironment in malignant melanoma organoids during the development and metastasis of tumors.

Malignant melanoma (MM) is the most metastatic and aggressive form of skin cancer, and carries a high risk of death. Immune-checkpoint inhibitor therapy and molecular-targeted therapy can prolong the survival of patients with advanced MM significantly. However, the low response rate and inevitable drug resistance prevent further improvements in efficacy, which is closely related to the tumor microenvironment (TME). The TME refers to the tumor stroma, including fibroblasts, keratinocytes, immune cells, soluble molecules, and extracellular matrix (ECM). The dynamic interaction between the TME and tumor cells is very important for the growth, local invasion, and metastatic spread of tumor cells. A patient-derived organoid (PDO) model involves isolation of tumor tissue from patients with MM and culturing it in vitro in a three-dimensional pattern. Compared with traditional cultivation methods, the PDO model preserves the heterogeneity of the tissue structure of MM and demonstrates the interaction between MM cells and the TME. It can reproduce the characteristics of proliferation, migration, and invasion of MM cells, and better simulate the structural function of MM in vivo. This review explores the role of each TME component in development of the PDO model. This review will provide a reference for research on the drug screening and targeted treatment using PDOs, particularly for the immunotherapy of MM.

Silicone rubber sealed channel induced self-healing of large bone defects: Where is the limit of self-healing of bone?

Background: Large defects of long tubular bones due to severe trauma, bone tumor resection, or osteomyelitis debridement are challenging in orthopedics. Bone non-union and other complications often lead to serious consequences. At present, autologous bone graft is still the gold standard for the treatment of large bone defects. However, autologous bone graft sources are limited. Silicon rubber (SR) materials are widely used in biomedical fields, due to their safety and biocompatibility, and even shown to induce nerve regeneration. Materials and methods: We extracted rat bone marrow mesenchymal stem cells (BMMSCs) in vitro and verified the biocompatibility of silicone rubber through cell experiments. Then we designed a rabbit radius critical sized bone defect model to verify the effect of silicone rubber sealed channel inducing bone repair in vivo. Results: SR sealed channel could prevent the fibrous tissue from entering the fracture end and forming bone nonunion, thereby inducing self-healing of long tubular bone through endochondral osteogenesis. The hematoma tissue formed in the early stage was rich in osteogenesis and angiogenesis related proteins, and gradually turned into vascularization and endochondral osteogenesis, and finally realized bone regeneration. Conclusions: In summary, our study proved that SR sealed channel could prevent the fibrous tissue from entering the fracture end and induce self-healing of long tubular bone through endochondral osteogenesis. In this process, the sealed environment provided by the SR channel was key, and this might indicate that the limit of self-healing of bone exceeded the previously thought. The translational potential of this article: This study investigated a new concept to induce the self-healing of large bone defects. It could avoid trauma caused by autologous bone extraction and possible rejection reactions caused by bone graft materials. Further research based on this study, including the innovation of induction materials, might invent a new type of bone inducing production, which could bring convenience to patients. We believed that this study had significant meaning for the treatment of large bone defects in clinical practice.