Thus, the microenvironment provided by menstrual fluid, is likely responsible for the unique rapid and scar-free repair of this remarkable tissue. Insight gained from analysis of this fluid is likely to be of value not only for treating endometrial bleeding problems but also in providing potential new therapies for poorly repairing wounds such as those seen in the aged and in diabetics.Menstruation is a process whereby the outer functionalis layer of the endometrium is shed each month in response to falling progesterone and estrogen levels in a non-conception cycle. Simultaneously with the tissue breakdown, the surface is re-epithelialized, protecting the wound from infection. Once menstruation is complete and estrogen levels start to rise, regeneration progresses throughout the proliferative phase of the cycle, to fully restore endometrial thickness. Endometrial repair is unique compared to tissue repair elsewhere in the adult, in that it is rapid, scar-free and occurs around 400 times during each modern woman's reproductive life. The shedding tissue and that undergoing repair is bathed in menstrual fluid, which contains live cells, cellular debris, fragments of extracellular matrix, activated leukocytes and their products, soluble cellular components and extracellular vesicles. Proteomic and other analyses have revealed some detail of these components. Menstrual fluid, along with a number of individual proteins enhances epithelial cell migration to cover the wound. This is shown in endometrial epithelial and keratinocyte cell culture models, in an  The endometrium, the inner lining of the uterus, provides the maternal support for embryo development and undergoes remarkable remodeling on a cyclical basis. In humans and a few other species including old world primates, the endometrium develops more extensively during each menstrual cycle than in other placental mammals. In particular the process of decidualization, which provides the basis for the decidua of pregnancy, is initiated whether or not the cycle is one in which conception occurs. As this process is not reversible, the endometrium is shed in the process known as menstruation, then fully repaired and reconstructed during the subsequent cycle. Importantly, in contrast to most adult wounds, the endometrium repairs very rapidly (days) without scarring  and occurs over 400 times during a woman's reproductive lifespan. Most aspects of menstruation are discussed elsewhere in this volume. The focus of this article is the role of menstrual fluid (MF) in the scar-free repair of the endometrium.Understanding the cellular and molecular events of menstruation provides insights critical to understand the likely composition of menstrual fluid.in situ. re-epithelialization occurs rapidly but it is from the basalis that the full thickness endometrium subsequently regenerates  and uterine natural killer (NK) cells (CD56+/CD16−) but eosinophils, mast cells (both 3–5%) and T lymphocytes (1–2%) are less common  althougMacrophages, neutrophils, mast cells, and eosinophils, all degranulate upon activation, releasing their soluble contents. Importantly, active forms have been identified during menstruation by virtue of the extracellular immunostaining of granular contents –72. ThesWhile there is a paucity of information on immune cell products in menstrual fluid that may be relevant to endometrial repair, elsewhere, eosinophils produce a number of growth factors, including TGF-α and -β, fibroblast growth factor (FGF), EGF, PDGF, and VEGF, which participate in angiogenesis and myocardial repair . EosinopNeutrophils contribute to physiological tissue repair, and seem to be necessary for normal healing at least in part by promoting angiogenesis . FurtherMast cell actions are likewise realized through degranulation and secretion of the granules' content of cytokines or production of lipid mediators, depending on the nature of the stimuli received during activation. Relevant to wound repair, during cardiac tissue re-modeling their main function appears to be associated with regulation of fibrous tissue metabolism , and the+ leukocytes from menstrual fluid, endometrial stem/progenitor cells including clonogenic endometrial cells, sushi domain containing-2+ (SUSD2+) mesenchymal stem cells and N-cadherin+ (NCAD+) epithelial progenitor cells, have been isolated  previously termed exosomes, are released from all cells. They act as carriers of “cargo” of bioactive molecules including miRNA, proteins and lipids, which they deliver to specific target cells: the phospholipid membranes of the sEV protect the “cargo” from extracellular degradation. Importantly the proteomes of endometrial epithelial cell sEV depend upon the hormonal environment of the cells of their origin (estrogen or estrogen plus progesterone), but are substantially different from those of the cellular proteomes . Endometex vivo de-epidermized dermis (DED) pre-parations  will likely play roles in the initial repair of the vasculature and subsequent angiogenesis . RegrettSimultaneously with re-epithelialization, rapid repair of the open blood vessels must take place to stop the bleeding, some 5 days after menstruation starts. While experimental evidence for angiogenesis at this time of the cycle is lacking, circumstantial evidence indicates its likelihood. Menstrual fluid contains the potent angiogenic factor VEGF-A , 41), a, a41), aBy the time menses has ceased, the wounded surface is essentially “repaired”, covered by a new luminal epithelium with junctional complexes making a tight protective surface, that shields the underlying tissue from infection. During the next 10 or so days (cycle proliferative phase) and as estrogen levels rise, endometrial thickness and the full cohort of cellular structures, including glands, stroma, vascular structures and extracellular matrix  is regenerated through massive cellular proliferation in the growing functional layer. Current knowledge of this regeneration process has recently been detailed  and willRapid scar-free repair of the endometrium following menstruation is essential given that it occurs some 400 times during most women's reproductive lives and provides the basis for the subsequent regeneration and differentiation of the endometrium and its attainment of receptivity for embryo implantation in the new cycle. It is now indisputable that the microenvironment provided by menstrual fluid drives effective endometrial repair. Recent advances in analyses of menstrual fluid components has provided some insight but just which are the most important factors, soluble or those delivered to the damaged surface in extracellular vesicles, remains to be determined. Importantly menstrual fluid has the potential to provide factors for scar-free rapid wound repair, and to treat abnormal uterine bleeding .The author confirms being the sole contributor of this work and has approved it for publication.LS and her laboratory have been funded extensively by Fellowships, Program and Project grants from the Australian National Health and Medical Research Council. Research at the Hudson Institute of Medical Research is funded in part by a State Government of Victoria Infrastructure support program.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Stat1 signalling, rendering animals resistant to further tumour challenges. Our results indicate that caerin 1.1 and 1.9 treatment may enhance the efficacy of current immunotherapy modalities.Cancer immunotherapy has been enabled by immune checkpoint blockade, but is ineffective in many patients, mainly due to the high complexity of the immune-suppressive tumour microenvironment (TME). Here, with a murine transplantable tumour model, we demonstrate that host-defence caerin 1.1/1.9 peptides can alter the macrophages in the TME from immunosuppressive to immune active phenotypes, which appeared consistent with the macrophages observed at early-stage cervical cancer patients. The modulated macrophages by caerin 1.1/1.9 show significant activation of Type I interferons response and Macrophages are one of the essential components of the tumour microenvironment (TME) of many cancers and show complex heterogeneity and functions. More recent research has been focusing on the characterisation of tumour-associated macrophages (TAMs). Previously, our study demonstrated that caerin 1.1/1.9 peptides significantly improve the therapeutic efficacy of combined specific immunotherapy and immune checkpoint blockade in a murine transplantable tumour model (TC-1). In this study, the mice inoculated with TC-1 tumour were immunised differently. The TAMs were isolated using flow cytometry and characterised by cytokine ELISA. The survival rates of mice with different treatments containing caerin 1.1/19 were assessed comparatively, including those with/without macrophage depletion. The single-cell RNA sequencing (scRNA-seq) data of previous studies were integrated to further reveal the functions of TAMs with the treatments containing caerin 1.1/1.9. As a comparison, the TAMs of stage I and II cervical cancer patients were analysed using scRNA-seq analysis. We demonstrate that caerin induced tumour clearance is associated with infiltration of tumours by IL-12 secreting Ly6C+F4/80+ macrophages exhibiting enhanced IFN-α response signalling, renders animals resistant to further tumour challenge, which is lost after macrophage depletion. Our results indicate that caerin 1.1/1.9 treatment has great potential in improving current immunotherapy efficacy. Human papillomavirus (HPV) infection-related malignancy remains a severe public health problem worldwide, although HPV prophylactic vaccine has been introduced for more than a decade ,2. WorldClinically, the International Federation of Gynecology and Obstetrics (FIGO) Cervical cancer staging system is an effective prognostic factor in patients with cervical cancer and useful guidance for treatment . Stage IImmunotherapy is becoming a routine modality for the management of cancers, following the introduction of immune checkpoint inhibitors and the transfer of CAR-T cells for cancer management . HoweverIn both prophylactical and therapeutic settings, therapeutic vaccination with the simultaneous blocking of interleukin 10 (IL-10) increased antigen-specific CD8+ T cell responses, which consequently improved tumour growth inhibition . The effCaerin 1.1/1.9 peptides are able to inhibit the proliferation of TC-1 ,25, HeLaMacrophages are one of the essential components of the tumour microenvironment (TME) . The higIn this work, we revisited two scRNA-seq studies on the intratumoural injection  and topiSeven patients with pathological confirmed cervical cancer who underwent surgical removal of cervical cancer at the Department of Obstetrics and Gynecology at Foshan First People’s Hospital, between 2019 and 2021 were included in the current study, with three at AIII, one at CIN3 (referred to as CCI) and three patients at IIB (referred as CCII). The ethical approval code was L2016 (13).Experiments were approved by Animal Experimentation Ethics Committee . Specific pathogen-free (SPF) six to eight weeks old adult female C57BL/6 (H-2b) mice were ordered from the Animal Resource Centre of Guangdong Province China. Mice were kept under SPF conditions at the Animal Facility of the first affiliated hospital of Guangdong Pharmaceutical University. Five mice were kept in each cage at 22 °C and 75% humidity and provided with sterilised standard mouse food and water and a 12-h light/12-h dark cycle.Transformed with HPV16 E6/E7, a murine TC-1 cell line was purchased from Shanghai Institute for Cell Resources Centre of Chinese Academy of Sciences. Cell culture was following the procedure described previously .2), caerin 1.9 (GLFGVLGSIAKHVLPHVVPVIAEKL-NH2), and a control peptide P3 (GTELPSPPSVWFEAEFK-OH) were synthesised by Mimotopes Proprietary Limited , with a purity above 99% and the concentrations of lipopolysaccharide below 0.44 EU mL−1.Caerin 1.1  by Mimotopes Proprietary Limited .Rat anti-mouse anti-IL10 receptor (1B1.3), Anti-PD1 (J43) Hamster anti-mouse monoclonal antibody and IgG Isotype control antibody (LTF-2) were purchased from BioXcell, USA. Rat Anti-Mouse fluorescent conjugated antibodies were purchased from BD, Biolegend and eBioscienc, including FITC-CD45.2 (clone: 104), PE-F4/80 (clone:BM8), APC-Ly6C (clone: HK1.4), PerCP-Cyanine5.5-CD11b (clone: M1/70), BV421-MHCII (clone: M5/114.15.2), PE-Cy7-CD11c (clone: HL3), PE-Cy7-CD86 (clone: GL1), APC-Cy7-CD3 (clone: 145-2C11), PE-Cy7-PD-L1 (clone: 10F.9G2), and Fixable Viability Stain 510 (FVS510). They were stored at −80 °C until further use.5 cells/mouse in 0.2 mL of PBS were injected subcutaneously into the left flank. The average diameter of tumour size was assessed every 3 days using callipers, calculated as width × width × length. Mice were given 1% sodium pentobarbital by i.p. injection when treatment was performed. At the end of each experiment, mice were sacrificed when the tumour diameter reached 15 mm , by CO2 inhalation.TC-1 cells, at approximately 70% confluency, were harvested with 0.25% trypsin and washed repeatedly with PBS. About 5 × 10Three days post-TC-1 challenge, when tumours were palpable, mice were immunised intramuscularly (i.m.) with the vaccine on day 3, day 9 and 18, respectively. The vaccine contains 40 µg of four overlapping HPV16E7 peptides (EX) (10 µg of each peptide), 10 µg of monophosphoryl lipid A (MPLA) , with or without 300 µg of anti-IL10R antibodies, dissolved in 100 µL of PBS; 300 µg of anti-PD-1 was administered intraperitoneally (i.p.) on days 9 and 21 after the tumour challenge.Three days post-TC-1 challenge, the tumour diameters reached 3 to 5 mm and the mice were intratumourally injected with 30 µg of caerin peptides (caerin 1.1/1.9) in PBS daily for six consecutive days.Mice were injected intraperitoneally (i.p.) with 200 μL of Clodronate Liposomes  or Control Liposomes, and 24 h later injected subcutaneously (s.c.) with 5 × 105 TC-1 cells/mouse in 0.2 mL of PBS and into the right flank. Systemic and local macrophage depletion was achieved by injection of Clodronate Liposomes concomitantly intraperitoneally (200 μL) and subcutaneously near the tumour cell injection site (100 μL) per 4 days until the end of the experiment.To obtain single cells, TC-1 tumour tissues were isolated and homogenised with a tumour dissociation kit . Single cells were stained with FITC-CD45.2 (clone: 104), APC-Cy7-CD3 (clone: 145-2C11), PE-Cy7-CD8a (clone: 53–6.7), BV421- INFg (clone: XMG1.2), PE-B220 (clone: RA3-6B2) and FVS510. Viable cells were separated and analysed on a flow cytometer . Data were analysed with Flow Jo v10.0 software .Murine cytokine ELISA kits were purchased from BioLegend , and were performed following the instructions of the manufacturer. https://singlecell.broadinstitute.org/single_cell (accessed on the 13 May 2022), the accession numbers were SCP1371 and SCP1093.Previously, we performed single-cell RNA sequencing of CD45+ TC-1 tumour infiltrating cells and quantitative analysis of TC-1 tumours of application of caerin 1.1/1.9 and combination therapy of caerin 1.1/1.9 with ICB and therapeutic vaccination ,29. The For the scRNA-seq analysis of CC patients, the tumour tissues were collected with consent from the Human Ethnic Committee of the Foshan First People’s Hospital (No: L2016). Cells were washed once with ice-cold PBS containing 10% foetal bovine serum post-sorting and counted using a haemocytometer. The cells were then loaded onto a 10× chromium machine  and run through the library preparation procedures following guidance from the Chromium Single Cell 3′ Reagent Kits v2 . The data were analysed using the pipeline described elsewhere ,29.Interactions among the proteins encoded by significantly regulated genes were predicted using STRING . A requip-value < 0.05 using GSEA v4.1.0 [The genes differentially expressed between different treatments were analysed by Gene Set Enrichment Analysis (GSEA) with A v4.1.0 , to predThe transcription factor network inference was analysed using the SCENIC R package . Briefly® II Q RT SuperMix for qPCR Kit . The ChamQ SYBR qPCR Master Mix  was used for StepOnePlus™ Real-Time PCR System with Tower . The cycling conditions were: 90 s at 95 °C, 40 cycles at 95 °C for 5 s, 60 °C for 30 s and 72 °C for 20 s. β-actin (Actb) was used as an internal control. The relative level was calculated by the relative quantification 2-ΔΔCT method. The primer sequences were listed in The total RNA of tumour tissues was isolated with TRIzol reagent . First-strand cDNA was synthesised from 1 μg total RNA using a HiScriptPreviously, we demonstrated that intratumoural injection with caerin 1.1/1.9 (caerin) significantly increases the survival time of TC-1 tumour bearing mice treated with HPV16 peptides/monophosphoryl lipid A/anti-IL-10 and anti-PD-1 therapy . DT plus caerin significantly reduced M2 type MΦs but increased pro-inflammatory macrophages in the TME . Here, wTo test the hypothesis that tumour macrophages are necessary for caerin-mediated anti-tumour responses, TC-1 tumour bearing mice treated with DT with caerin or control, and untreated tumour bearing mice, were treated with clodronate liposomes (CL) to deplete MΦs A. ClodroArg1 in the TME and in tumour macrophages has been found to correlate with tumour growth, metastasis, and poor prognosis in many types of cancers [hiArg1 macrophage populations , to be more pro-inflammatory and immune response active, with respect to the untreated or control group [hiArg1 MΦs from the TC-1 tumour bearing mice treated with caerin. In the current study, the scRNA-seq data of this previous work, particularly the macrophages, were reanalysed.Aberrant expression of  cancers ,47. Prevol group . The INFhiEar2 MΦs from the caerin-treated animals when compared to TAMs from the control group, particularly Herc6, Parp9, Ripk2 and Ube2l6  with the survival of patients with CESC (Ccl4 (p = 0.041) and Gcnt2 (p = 0.021) with CESC survival time was identified, respectively (Coro7 (p = 0.04), Ccl4 (non-significant) and Gcnt2 (p = 0.011) in the caerin treated tumour bearing mice when compared to untreated mice  that might be regulating macrophage phenotypes, we examined the TF regulatory network of the six MΦ subtypes as mentioned above using SCENIC analysis. A total of 92 regulons were differentially detected among the MΦ subtypes . UpregulJun, Jund, Fos, Junb and Fosl2 were upregulated in the MΦs from the caerin-treated animals  (p < 0.001) (− and proinflammatory F4/80+Ly6C+ MΦs subpopulations accounted for over 80% of macrophages at day 7 and day 14. F4/80+Ly6C− and F4/80+Ly6C+ MΦs were sorted and stimulated with MPLA (100 ng/mL) overnight, and the supernatants were measured for IL-12, IL-10, IL-6 and TNF-α by ELISA  . In addi< 0.001) B. Tumour< 0.001) C. Tumourby ELISA D. IL-12 by ELISA D.Stat2, Isg15, Tap1, Psmb9, and Parp9, were more highly expressed in the CCI than CCII MΦs. IFN-α and IFN-γ signalling activation was observed in all subpopulations of MΦs in CCI subjects, and TNF-α signalling was observed in MΦ/DCs, resident- and M2-like MΦs , TAM (C1-TAM), M2-like (C2-M2), MΦ/DCs (C3-DC), and M1-like (C4-M1) MΦs, was assessed for four patients with early (CCI) and three patients with late (CCII) stage cervical cancer, using scRNA-seq analysis . This walike MΦs –E. IL6/Jlike MΦs . We examlike MΦs . NotablyIn this study, we demonstrated that the DT of ICB and therapeutic vaccination plus caerin 1.1/1.9 increased the proinflammatory cytokine IL12, while reducing IL10, IL6 and TNF-α secretion in tumour macrophages; the depletion of macrophages greatly attenuated the anti-tumour responses by DT boosted by caerin 1.1/1.9. By mining our previously published scRNA-seq data, we showed that tumour local administration  of caerin 1.1/1.9 significantly activated the IFN-α response pathway in different phenotypes of tumour macrophages, in general, similar to that exhibited by the tumour macrophages of early CC patients. Caerin 1.1/1.9 also promoted the expression of several TFs playing key roles in the regulation of genes that result in a proinflammatory response.Immunotherapy has become a routine modality for the treatment of cancers, especially with the introduction of ICB and CAR-T therapy, therapeutic vaccines in pre-clinical animal models have also demonstrated exciting efficacies. However, a great portion of cancer patients do not respond to ICB treatment. One of the most important caveats is that the immunosuppressive TME stunts the therapeutic effects of immunotherapy. Disturbing the TME is, therefore, a key issue to consider for immunotherapy. Given that caerin 1.1/1.9 can directly kill tumour cells and make the tumour cells release inflammatory cytokines we, therefore, set out to investigate whether caerin 1.1/1.9 can alter the TME and the efficacy of immunotherapy.The enhanced efficacy of immunotherapy mediated by tumour local treatment with caerin peptides was mediated by macrophage, most likely through the IFNα signalling pathway as demonstrated by the scRNA-seq analysis of the tumour infiltrating macrophages. scRNA-seq analysis of macrophages between early and later-stage cervical cancer tumour samples also indicate that the IFNα signalling pathway was more activated in the early-stage cervical cancer.Tyk2 and impairs Jak-STAT activation by IFN-α [Stat1 phosphorylation through a mechanism that involves the inhibition of tyrosine kinase 2 phosphorylation [Type I IFNs (IFN-α and IFN-β) constitute the first line of defence against both viruses and cancer cells, although it plays dual roles that can either prevent or promote tumour development . Virusesby IFN-α . Respirarylation ,52,53. Trylation ). These rylation .Cebpb, and IFN-α upregulates C/EBPβ by recruiting Stat1 and Stat5 to the novel 3′ distal enhancers of Cebpb [Cebpb was identified with significance in most MΦ phenotypes of the topical treatment containing the caerin peptides, which was also present as one of the differential TFs in the MΦs of the caerin + DT. The significant upregulation of Stat1 was confirmed by qPCR in this study and was previously detected in both the topical application and intratumoural injection treatments containing the caerin peptides [Cebpb and Gm8797 (ubiquitin B pseudogene) were detected as the marker genes of the MΦs of Vsir knockout mice, which showed an exacerbated inflammatory phenotype [Gm8797 was remarkably upregulated in the caerin + DT. This further suggested Cebpb may play an important role in the proinflammatory state induced by caerin 1.1/1.9.IFN-α has been used to treat cancers; however, the systemic administration of type I IFNs leads to significant side effects for patients . Recentlof Cebpb . The uprpeptides ,29. It wpeptides . Additiopeptides . Cebpb ahenotype . The expThe depletion of macrophages or avoidance of macrophage tumour infiltration in the TC-1 tumour model results in either a beneficial or detrimental anti-tumour response. The depletion of myeloid cells by injecting clodronate liposome increased the efficacy of the vaccination, in terms of reducing tumour size and prolonging the survival of tumour-bearing mice . A recenFos/Jun enhances inflammatory responses in macrophages [Jund was previously shown to be positively associated with inflammation, the knockdown of which resulted in significantly reduced macrophage activity and cytokine secretion in rat and human primary macrophages [Junb was found to play the key role in the full expression of Il1b and the genes involved in classical inflammation in macrophages treated with LPS and other immunostimulatory molecules [Fosl2 induced a systemic inflammatory phenotype with immune infiltrates in multiple organs in mice, by repressing the development of regulatory T cells [Usf1, Nfil3, and Mafb  involved. At the protein level, antibody-mediated signalling blockade or activation by agonists could be performed, with a particular focus on macrophage phenotypes. With respect to how caerin 1.1/1.9 activate IFN signalling of macrophages, the knockout of key regulators on the pathway, e.g., rophages ,82. It hrophages , which erophages . These wTaken together, for the first time, we unveil that caerin 1.1/1.9 are able to alter the tumour immune suppressive environment fundamentally to a more immune-active state, mainly through activating the type I type IFN signalling pathway of tumour MΦs, which significantly enhances the efficacy of ICB and warrants further investigation in more therapeutic vaccine-based immunotherapy.


The novel CMMRF fluid can simultaneously achieve Ra 0.32 nm (0.47 mm × 0.35 mm measurement area), Ra 0.22 nm (5 μm × 5 μm measurement area), and 1.91 × 10−2 mm3/min material removal efficiency. Comprehensive studies utilizing a scanning electron microscope and a magnetic rheometer show that the CMMRF fluid has a high mechanical removal effect due to the well-dispersed nanodiamond and nano-Fe3O4 particles. The results of Fourier transform infrared spectra and Young’s modulus test reveal the mechanism of the chemical reaction and the mechanical characteristics deterioration of the modified layer. Under co-enhanced chemical and mechanical effects, an ultra-smooth and highly efficient MRF technology for single-crystal silicon is realized.To improve the material removal efficiency and surface quality of single-crystal silicon after magnetorheological finishing, a novel green chemical-mechanical magnetorheological finishing (CMMRF) fluid was developed. The main components of the CMMRF fluid are nano-Fe An X-ray reflector is the essential part of the synchrotron radiation source ,2,3. ItsThe preferred substrate material for X-ray reflectors is single-crystal silicon, which has exceptional thermal characteristics, mechanical properties, and processability.Magnetorheological finishing (MRF) is a deterministic sub-aperture polishing technique based on a magnetically sensitive fluid that removes material via a shearing mechanism with minimal normal load ,11. DrivX-ray reflectors require the roughness of the single-crystal silicon substrate to be Ra 0.3 nm or lower. Current MRF technology is still far from achieving this level, so an additional ultra-smooth process needs to be introduced to improve the surface quality after MRF . In addiNumerous significant efforts have been devoted to enhancing the efficiency or surface quality of MRF. In terms of improving the efficiency of MRF, QED novel developments have been made with the fluid nozzle, magnetic field, and wheel geometry and wheel size to increase the removal rate. Custom nozzle shapes are used to create a very wide MRF ribbon resulting in a much wider removal function spot . The polChemical-mechanical polishing (CMP) is a process in which the material is removed by a chemical-mechanical synergetic mechanism, and it is regarded as the most effective method for achieving the sub-nano surface roughness ,34. Insp2 abrasive can react with the surface of single-crystal silicon to generate a soft layer, which is then removed by the abrasive to reveal a smooth substrate [2 is quite low. To achieve a high-efficiency and ultra-smooth CMMRF process for single-crystal silicon, it is important to find alternative chemical additives and abrasives.With the advancement of the semiconductor industry, the CMP technology of single-crystal silicon has reached a highly developed state. In an alkaline environment, the nano-SiOubstrate . However3O4, H2O2, CH3COOH, and deionized water. The novel polishing fluid can achieve a volume removal rate of 1.91 × 10−2 mm3/min and a surface roughness of Ra 0.22 nm (5 μm × 5 μm). The roughness is reduced by 51.2% and the efficiency has grown by 196.9% in comparison to the pure nanodiamond MRF fluid. These results demonstrate that the CMMRF fluid has an excellent performance in the high-efficiency process of ultra-smooth single-crystal silicon. In order to optimize and clarify the mechanism of the novel CMMRF fluid, a series of characterization experiments were carried out. The magnetic rheometer results demonstrate that nano-Fe3O4 increases the shear yield strength of MRF fluid, and the nanodiamond abrasives with the sharp cutting-edge lead to such a high material removal rate. A heterogeneous Fenton reagent composed of H2O2, nano-Fe3O4, and CH3COOH was characterized and optimized by spectrophotometer. The heterogeneous Fenton reagents generates a large number of hydroxyl radicals(·OH) which oxidize the surface and generate a silicon dioxide modified layer. The Young’s modulus test and Fourier transform infrared (FTIR) spectra indicate that the hydroxyl group in the solution can enhance the hydrolysis reaction of the modified layer, destroy its network structure, and degrade its mechanical properties. Compared to the mechanical MRF fluid, the novel CMMRF fluid considerably improves chemical and mechanical effects, enabling a high-efficiency ultra-smooth single-crystal silicon process.The objective of this study is to configure a novel eco-friendly CMMRF fluid to achieve efficient ultra-smooth processing of single-crystal silicon. The CMMRF fluid is composed of nanodiamond abrasives, carbonyl iron powder (CIP), nano-Fe3O4 with an average particle size of 20 nm, diamond polishing particles with an average particle size of 150 nm, deionized water, CH3COOH, PEG200, HCl, and NaOH. The details of different constituents used for the MRF are mentioned in The materials used to configure the MRF fluid in this paper include CIP with an average particle size of 2 μm, FeSingle-crystal silicon CZ-(111), with a diameter of 100 mm and a thickness of 10 mm, was utilized as the experiment sample for roughness and material removal efficiency. The initial surface roughness was approximately Ra 1 nm after fully pitched polishing, and the form error PV was less than 1 μm. As shown in The sample roughness results were measured by an atomic force microscope  and a white light interferometer . The measuring area of the AFM is 5 μm × 5 μm, and the tapping mode was used for surface microscopic topography imaging. The WLI has a magnification of 20 times, and the field of view is 0.47 mm × 0.35 mm.The content of ·OH in CMMRF fluid determines the oxidation effect of single-crystal silicon. Spectrophotometry was used to measure the content of ·OH in the CMMRF fluid. 2,3-dihydroxybenzoic acid and 2,5-dihydroxybenzoic acid produced by the reaction of salicylic acid with ·OH will have strong absorption peaks at 510 nm. We prepared a mixed solution of salicylic acid, ethanol, and deionized water as a catcher for ·OH, in which the concentration of salicylic acid was 1.8 mol/L, and the ratio of ethanol to water was 1:3. A UV-VIS-NIR 3600 spectrophotometer was used to measure the absorbance of mixed solutions containing different formulas of CMMRF fluid and catcher.3O4 was photographed using a scanning electron microscope . The shear yield strength of the MRF fluid was measured by Anton Paar MCR302 rheometer with an MRD180 module.MRF base fluid refers to the combination of the liquid components in the MRF fluid. In the CMMRF base fluid environment, the dispersion condition of diamond abrasive and CIP with nano-Fe4F, and deionized water was 1:10:50. The samples were immediately cleaned in a 270 kHz ultrasonic cleaner with deionized water for 5 min after the oxide layer had been removed. Transfer samples from which the oxide layer had been removed, was added to various CMMRF solutions and soaked for 24 h at 26 °C. We then took out the samples and cleaned them for 5 min in a 270 kHz ultrasonic cleaner with deionized water. The Young’s modulus of the modified layer was measured with a Bruker Dimension Icon AFM equipped with a PeakForce QNM module, and the probe model used for the measurement was DNISP-HS. The measurement area was 5 μm × 5 μm, and the average of the three measurements was regarded as the Young’s modulus of the modified layer. FTIR  was employed to detect the chemical composition of the modified layer.To characterize the chemical-mechanical mechanism of CMMRF fluid on single-crystal silicon, Young’s modulus and FTIR tests were conducted on single-crystal silicon CZ- (111) samples with a diameter of 10 mm and a thickness of 3 mm. After fully pitched polishing, the samples’ surface roughness was approximately Ra 1 nm. Using a plasma processing system (Ion wave 10), we conducted 1000 W power Ar gas plasma cleaning for 60 s to eliminate organic pollution from the sample surface. Then, the original oxide layer on the sample surface was removed by soaking it in buffered oxide etch (BOE) solution for 3 min. In the BOE solution, the volume ratio of HF, NHThe design idea of the CMMRF fluid is based on the uniform wet etching technique for single-crystal silicon. First, a powerful oxidant is chosen to uniformly oxidize the surface, followed by the removal of the oxide layer to obtain an ultra-smooth surface.3+ or Fe2+ in the homogenous Fenton reagent is too high for direct discharge without treatment.The Fenton reagent contains a large amount of ·OH, and the oxidation potential of ·OH has reached 2.80 V, which is suitable for oxidizing the single-crystal silicon. Meanwhile, the concentration of Fe2O2 reacts on the surface of the solid catalyst to generate ·OH, and the chemical reaction is as follows [2+ and ≡Fe3+ represent the iron ions on the solid catalyst surface, and Fe2+ and Fe3+ represent the iron ions in the solution. In an acidic environment, redox cycles occur on the catalyst surface and in the liquid. Ferrihydrite [3O4 [3O4 has exceptional catalytic and magnetic properties, and is widely utilized in MRF fluid and heterogeneous Fenton reagent. In this article, nano-Fe3O4 is used to develop a novel CMMRF fluid.In the heterogeneous Fenton reagent, H follows :(1)≡Fe3+ihydrite , goethitihydrite , and Fe3ite [3O4  are freq2O2, and concentration of the catalyst [2+ and ≡Fe3+ on the surface of the catalyst is the primary reason why pH influences oxidation activity. As the pH increases, the metal ions will precipitate, the oxidation potential of ·OH will decrease at the same time, and H2O2 is easily degraded into oxygen and water, which influences the generation of ·OH. For the concentration of H2O2, when H2O2 is inadequate, the amount of generated ·OH will be insufficient, whereas an excess of H2O2 will lead to the annihilation of ·OH.There are three factors affecting the oxidation effect of heterogeneous Fenton reagent, namely, pH value, concentration of Hcatalyst . The conWithin a certain concentration range, the active site increases with the concentration of the catalyst, but excessive catalyst will consume the generated ·OH. The reaction is shown in the following equation:To optimize these three factors, the absorbance of the mixture of CMMRF fluid and ·OH catcher was measured.3COOH and NaOH are utilized, and the range of pH is between 2.5 and 9. As shown in the As pH regulators, CH3COOH cannot be totally ionized since it is a monobasic weak acid. When pH = 2.5, the concentration of CH3COOH is 0.5 mol/L (1.5% vol), and when pH = 3 it is 0.05 mol/L (0.15% vol). The absorbance of the reagent is only 3.5 higher at pH = 2.5 than at pH = 3. Therefore, the pH of CMMRF fluid is set to 3, which saves CH3COOH, prevents the corrosion of processing equipment caused by high CH3COOH concentrations, and ensures the oxidation effect of CMMRF fluid.CH2O2 concentration on absorbance is shown in 2O2 rises, the absorbance of the reagent progressively increases, reaching a peak value at 5% vol H2O2, and then decreases as the concentration of H2O2 continues to rise.The effect of H3O4 on absorbance follows the same pattern as that of H2O2, with an initial increase followed by a decline. 3O4 is 1%, the reagent produces the maximum concentration of ·OH.The effect of Fe2O2 5% vol, CH3COOH 0.15% vol, CIP 25% vol, nano-Fe3O4 1% vol, nanodiamond abrasive 3% vol, and PEG200 5.85% vol. Nanodiamond abrasive consists of carbon atoms, and possesses ultra-high hardness and stable chemical characteristics at room temperature. H2O2 is frequently used for disinfection and sterilization at low concentrations. CH3COOH is an organic acid that is the primary component of vinegar. A suitable amount of CIP can be used as a dietary additive to supplement the needs of the human body. PEG200 is polyethylene glycol with an average molecular weight of 200. It is a green organic solvent that can distribute particles in CMMRF fluid and adjust its viscosity. It is evident that the components of the CMMRF fluid are harmless to the environment and operators.The optimized CMMRF fluid composition is deionized water 60% vol, H−2 mm3/min, which increases by 196.9% compared with 0.97 × 10−2 mm3/min obtained by the mechanical MRF fluid. The peak removal rate (PRR) of CMMRF is 1.32 μm/min, which is 186.6% greater than the PRR of the mechanical MRF fluid (0.71 μm/min). Therefore, it is demonstrated that the novel green CMMRF fluid has superior performances when polishing single-crystal silicon.As a comparison of CMMRF fluid, the MRF fluid with pure mechanical material removal is configured as follows: deionized water 65% vol, CIP25% vol, nanodiamond 3%, PEG200 7% vol, and pH adjusted to 3 with HCl, which is consistent with the CMMRF fluid. Comparing the performance of these two kinds of fluid, 3O4 were photographed by a SEM in the CMMRF base fluid environment. It can be seen in 3O4 are spherical, and the regular spherical shape is beneficial for preventing surface defects during processing.To characterize the performance of the CMMRF fluid, the nanodiamond abrasive and CIP mixed with nano-Fe−1 when nano-Fe3O4 was added. Under the influence of an external directional magnetic field, the CIPs are arranged in a chain structure, and nano-Fe3O4 is embedded in the gap between the CIP and enhances the magnetic dipole–dipole force, which is reflected as an increase of shear yield strength. With an increase in shear yield strength, the capacity of MRF fluid to hold nanodiamond polishing particles is enhanced, which is beneficial for improving the removal efficiency of materials.3COOH.To investigate the reaction mechanism of CMMRF on the single-crystal silicon, various chemical substances were added to configure the different MRF fluids, and three samples with its original oxide layer removed were immersed in it. As shown in Young’s modulus is measured at three points on each sample, with a range of 5 μm × 5 μm at each point. The average of three points is regarded as the Young’s modulus of this sample. The measurement results are shown in −1 (Si-O), 778 cm−1 (Si-O), and 1113 cm−1 (Si-O-Si) [2, and the absorption peaks of Samples No. 2 and No. 3 are significantly stronger than that of Sample No. 1. In addition, compared with Sample No. 1 and No. 2, Sample No. 3 has a strong absorption band at 973 cm−1 (Si-OH). It can be seen from −1 (OH group), the absorption bands of Sample No. 1, Sample No. 2, and Sample No. 3 are intensified sequentially [From Si-O-Si) , which aentially .3O4 and H2O2 has a better oxidation performance and generates a thicker modified layer within the same treatment time.During the Young’s modulus test, the degradation of the sample mechanical properties will be aggravated by a thicker modified layer. It can be inferred that the thickness of the modified layer of Sample No. 1 is less than that of other samples, and the absorption peak intensity of Sample No. 2 and Sample No. 3 is higher than that of Sample No. 1, confirming this conclusion again. This is because the heterogeneous Fenton reagent composed of nano-Fe3COOH, which destroys the structure of the oxidation product SiO2, resulting in a further decrease in Young’s modulus [The modified layer of Sample No. 3 is attacked by the hydroxyl group in CH modulus ,45,46. A3) of abrasives, while the total penetration depth (δ2) is greater than that of mechanical MRF (δ1). The microscopic topography formed by abrasives will be more uniform when the substrate penetration depth is lower, and an increase in the total penetration depth will improve the material removal efficiency.As depicted in Furthermore, it should be noted that the chemical-mechanical balance and electrostatic force between the abrasive and modified layer also play vital roles in the high-efficiency ultra-smooth process. Further research is needed to fill the gaps in the CMMRF material removal mechanism.In this paper, a novel CMMRF fluid is developed and realizes high-efficiency ultra-smooth processing of single-crystal silicon. The novel fluid is harmless to the environment and the operators, and the solid components in the fluid can be recycled and reused.3O4 enhanced the shear yield strength of the fluid, and the employment of the hard nanodiamond abrasive was beneficial for improving the material removal efficiency. The heterogeneous Fenton reagent is composed of Fe3O4 and H2O2; the ·OH can rapidly oxidize single-crystal silicon. The addition of CH3COOH considerably enhanced the material removal rate by destroying the modified layer with the hydrolysis reaction. In addition, the modified layer reduces the substrate penetration depth of the abrasives, resulting in a significant improvement in surface roughness. This article could be a reference for the high-efficiency ultra-smooth processing of silicon. After the optimization of processing parameters, it can also be applied to a variety of materials as far as the modified layer can be generated on the surface.The polishing mechanism of CMMRF was revealed by comprehensive characterizations. The nano-Fe


Studies on the tumour microenvironment show that part of the body’s defence against neoplastic cells is carried out directly in the tumour through the organization of immune cellular aggregates, called tertiary lymphoid structures (TLSs). This study demonstrates the existence of TLS in canine mammary simple carcinomas, where some inflammatory infiltrates assume a follicle-like organization, especially in high-grade carcinomas and simulate the follicle of secondary lymphoid organs (SLOs). The role of TLS is controversial and it has been suggested that they contribute to an immunosuppressive environment or, on the contrary, have positive effects on neoplasm growth. Although canine mammary tumours have long been proposed as a model for studying breast cancer, there is still little research on the tumour stromal microenvironment in veterinary medicine and TLS evidence could allow for advances in understanding the process of tumour immunoediting.Neoplastic progression is influenced by the expression of tumour antigens that activate an anti-tumour immune response. Human medical studies show that this body defence is carried out in secondary lymphoid organs (SLOs) but also directly in the tumour through organized cellular aggregates that are called tertiary lymphoid structures (TLSs). However, their occurrence has different meanings in different tumour types. For example, the presence of TLSs in breast cancer is associated with the most aggressive subtypes. This paper aimed to study TLSs in canine mammary simple carcinomas. A morphological assessment of the inflammatory infiltrate was performed on H&E sections of fifty cases. Immunohistochemistry was then carried out to typify the inflammatory cells in the tumour microenvironment. Results showed that, sometimes, inflammatory infiltrates were organized in follicles close to high-grade carcinomas, simulating a lymphoid organization, as in breast cancer. Therefore, we can assume that even in canine mammary tumours, TLSs exist and they are entities to consider due to their presence in the most aggressive histotypes or tumours with a high degree of malignancy. Neoplastic immune surveillance is due to a series of genetic alterations, resulting in the expression of tumour antigens that activate an adaptive anti-tumour immune response committed to eliminating cancer cells. Typically, the adaptive immune response takes place in secondary lymphoid organs (SLOs), in which dendritic cells present molecule–peptide complexes to CD4+ T and CD8+ T cells. In SLOs, B lymphocytes, following antigen presentation, are activated in primary follicles, then secondary follicles, which eventually become germinal centres. This results in the proliferation and differentiation of T lymphocytes and memory B cells that migrate into the tumour and lead to the destruction of the neoplastic cells . HoweverThe density of TLSs correlates with the density of CD4+ T and CD8+ T cells in the tumour and the detection of tertiary structures has been associated with a favourable prognosis for many solid malignancies, probably related to their ability to induce a durable systemic anti-tumour response .Dendritic cells (DCs) present antigen to CD4+ T-cells, but studies in ovarian metastases of breast cancer have shown that DCs also appear to be involved in antigen presentation to germinal centre B-cells. Furthermore, locally produced anti-tumour immunoglobulin G increases the ability of dendritic cells to present antigen via IgG-specific receptors .In general, studies have shown that TLSs can generate a response from effector and memory B cells by producing antibodies against tumour antigen and protecting against metastasis . The linIn general, the presence of TLSs is associated with the most aggressive breast cancer subtypes, such as TNBC (triple-negative breast cancer), HER2 enriched and luminal B, while tumours without TLSs tend to be luminal A .In HER2- breast cancer, it has been seen that the presence of tumour cells inside the TLS is associated with lymphatic invasion and lymph node involvement . It coulAlthough canine mammary tumours have long been proposed as a model for studying breast cancer, there is still little research on the tumour stromal microenvironment in veterinary medicine compared with human medicine. There are already several studies investigating the role of various individual inflammatory populations, but no study has highlighted the presence of lymphoid tertiary structures in dog mammary tumours. The aim of this study was to investigate the immune cell infiltrates in simple carcinomas of the canine mammary gland and give evidence of TLS structures in order to allow for advances in understanding the processes of tumour immunoediting.Fifty cases of mammary gland lesions were retrieved from the archives of the Unit of Veterinary Pathology of the Department of Veterinary Sciences of Messina. All the cases were re-classified according to the most recent classification . We inclOn hematoxylin and eosin (H&E) sections, a careful morphological assessment of the inflammatory infiltrate was carried out. Immunohistochemistry was then performed to typify inflammatory cells and molecules of interest in the tumour microenvironment. Samples were fixed in formalin and embedded in paraffin. Then, 5 µm sections on poly-L-lysine-coated slides underwent antigen unmasking by the means of a microwave steaming in citrate buffer at pH 6 or EDTA at pH 8. Hydrogen peroxide was used to block the endogenous peroxidases and a blocking reagent to block non-specific protein reactions . The sections were then incubated with the selected primary antibodies , responsible for triggering the events that lead to the formation of TLSs ,29.In the present study, the overall prognostic role of TLSs can be only speculated because of the unavailability of the follow-up for the analysed samples. However, based on the higher representation of TLSs with a more defined organisation, in high-grade tumours close to the areas of infiltration and around neoplastic emboli, it can be suggested that dog tertiary lymphoid structures could also reflect a worse prognosis, as already highlighted in breast cancer by Sofopoulos et al. . It is cLooking more closely at the prognostic impact of tertiary structures in breast cancer, the presence of peritumoural TLSs is associated with grade 3 tumours . The pooSome studies suggest that inflammation-associated TLSs serve as niches for tumour progenitor cells and may lead, for example, to a recurrence of hepatocellular carcinoma. Peripheral TLS may, thus, contribute to an immunosuppressive environment, supporting tumour growth through negative effects on anti-tumour immunity or positive effects on the neoplasm itself .The tumour microenvironment is a dynamic entity characterised by the involvement of chemokines and immune cell populations that can be modulated by the tumour for its own survival and progression. In general, from what has been observed, we can, however, state that also in canine mammary tumours, we can speak of tertiary lymphoid structures with various degrees of organisation. The prognostic role of TLSs in dogs can, however, only be speculated on, as actual post-treatment follow-up would be required. Despite this, their greater representation in more malignant histotypes suggests that TLSs might also have a negative prognostic value in dogs. However, even though these are preliminary studies to be accompanied by further verification and research, they are fully in line with the concept of translational medicine, especially considering that the mammary tumour in dogs has long been considered for a model for female breast cancer. All this also opens the possibility of identifying new diagnostic and predictive markers and even new therapeutic targets.


Parvimonas micra and Fusobacterium nucleatum, with survival in colorectal cancer patients. Our results suggest that patients with high tumoural levels of Parvimonas micra have decreased survival. In addition, we found associations for Parvimonas micra and Fusobacterium nucleatum with different clinicopathological and tumour molecular traits. A better understanding of the role of the gut microbiota in colorectal cancer may contribute to improved cancer care.The gut microbiota has been suggested to affect tumour development and progression in colorectal cancer. In this study, we investigated the associations of two colorectal cancer-related bacteria, Parvimonas micra and Fusobacterium nucleatum, was studied in relation to patient survival in a cohort of 257 CRC patients. Colonisation of P. micra and F. nucleatum was analysed in fresh frozen tumour tissue (n = 112) and in faeces (n = 250) by qPCR. When analysing tumour tissues, both P. micra and F. nucleatum were found to be associated with decreased five-year cancer-specific survival, an association that remained significant in multivariable analysis for P. micra. Furthermore, we found significant associations of high levels of P. micra and F. nucleatum with tumour molecular characteristics, i.e., tumours mutated in V600EBRAF, and tumours of the MSI subtype. The analysis of faecal samples showed weaker associations with prognosis and tumour molecular characteristics. In conclusion, our findings support a novel association of tumour colonisation of P. micra with decreased patient survival. A better understanding of the role of the gut microbiota in CRC might contribute to the advancement of prognostic tools and new targets for therapy.Increasing evidence suggests that the gut microbiota may impact colorectal cancer (CRC) development and progression. In this study, the tumour colonisation of two CRC-associated bacteria,  Colorectal cancer (CRC) is one of the most deadly cancers in both men and women worldwide with close to 1 million deaths per year, and the number is expected to rise due to an increasing incidence in younger adults and transitioning countries . A driveTogether, this makes the gut microbiota a potential and interesting target for future cancer therapy. Indeed, recent studies have suggested that the intestinal microecological composition is related to CRC prognosis  and specFusobacterium nucleatum [F. nucleatum has been suggested to contribute to colorectal tumour progression through various mechanisms, including expression of the virulence factor FadA on the cell surface, which induces E-cadherin mediated activation of Wnt/β-catenin signalling [F. nucleatum and decreased CRC survival [Parvimonas micra [P. micra and F. nucleatum to be more abundant in stool samples from CRC patients compared with healthy individuals [P. micra to tumour immunological traits [P. micra has a tumour-promoting effect through colonocyte proliferation and the alteration of the Th17 immune response [F. nucleatum, very little is known about the potential role of P. micra in CRC tumourgenesis and prognosis.Interestingly, many of the bacterial species suggested to be associated with CRC are often found colonising the oral microflora, one of the more well-studied being ucleatum ,10,11,12gnalling . Previousurvival ,19,20,21as micra ,23,24. Il traits . A recenresponse . Still, P. micra and F. nucleatum in tumour tissue and faeces of CRC patients affect cancer-specific survival. A potential link between microbial colonisation in CRC and patient prognosis would facilitate the development of new prognostic and predictive tools, as well as future targeted cancer therapies.In this study, we investigated if the levels of The study was performed on patient samples collected within the Uppsala-Umeå Comprehensive Cancer Consortium (U-CAN), which has been described previously . The proP. micra and F. nucleatum in fresh frozen tumour tissue and faeces was conducted using qPCR as described previously [® Soft Tissue Homogenizing CK14 Kit . The AllPrep DNA/RNA/miRNA Universal kit  was next used for DNA extraction. For faeces, the QIAamp PowerFecal DNA kit (Qiagen) was used to extract DNA from about 0.2 g stool. The Qubit dsDNA BR Assay Kit  was used to measure DNA concentration. The specific qPCR assays used to detect P. micra, F. nucleatum, and relevant reference genes were previously established [PGT assay, leaving 112 samples for further analyses. Additional exclusions included 2 samples for P. micra and 2 samples for F. nucleatum. For the faecal samples, 7 samples were excluded after the 16S rRNA gene analysis, leaving 250 samples for further analysis. For P. micra analysis, 4 additional samples were excluded and for F. nucleatum, 6 additional samples were excluded from analyses. The levels of P. micra and F. nucleatum were presented as a relative quantification using the 2−ΔCq method. The human gene PGT was used as reference for fresh frozen tissue and the 16S rRNA gene was used as reference for faeces.Detection of eviously ,25. In bablished ,25. The Molecular analyses were performed on DNA extracted from fresh frozen tumour tissues as described above. For patients lacking fresh frozen tumour tissue specimens, DNA was instead extracted from FFPE tumour tissues (five sections of 10 µm) using the AllPrep DNA/RNA FFPE kit (Qiagen) (n = 72). For determination of MSI status, the MSI Analysis System Version 1.2 (Promega) was used, based on the analyses of mononucleotide repeat markers BAT-25, BAT-26, NR-21, NR-24, MONO-27, as was previously described . The PeaV600EBRAF mutation, digital droplet PCR  was used, as described previously [KRAS mutation analysis, codon 12 and 13 were sequenced using Big Dye v.3.1 (Applied Biosystems). Primers and probes for BRAF and KRAS mutation analyses were described previously [For analyses of eviously . In brieeviously .p-values < 0.05 were considered statistically significant. Correlations between continuous variables were analysed using the Spearman’s rank correlation tests. For comparisons of categorical variables, Fisher’s exact test was used.IBM SPSS Statistics 28  was used for statistical analyses. P. micra or F. nucleatum as test variable and cancer-specific survival as state variable. Youden’s index was then used to calculate the optimal cut-off to identify samples with high or low levels of the markers.Included patients were followed from the date of surgery until the date of death or end of follow-up (October 2021). Cancer-specific survival was defined as death with disseminated or recurrent disease. Patients not undergoing resection of the primary tumour, and patients who died from post-operative complications within 90 days, were excluded from survival analyses. ROC curves were created using the relative levels of P. micra or F. nucleatum). Since the microbial variables included in the study were correlated, separate multivariable analyses were conducted for these.Kaplan–Meier plots were used to estimate cancer-specific survival over time and the log-rank test was used to calculate statistical differences in outcome between groups. Survival analyses were truncated at 5 years to fulfil the assumption of proportional hazards. Uni- and multivariable survival analyses were performed using the Cox Proportional Hazards model. Variables included in the multivariable analyses were selected using (1) Backward Stepwise Conditional model and (2) only including variables with a significant effect on survival in univariable analysis, including all available clinicopathological and molecular variables. Both methods resulted in the same variables being included in the multivariable analyses  and in faecal samples (n = 250) from a CRC patient cohort. For fresh frozen tumour tissues, relative quantification of P. micra and F. nucleatum was calculated using the human gene PGT as reference. P. micra was detected in tumour tissue from 30 out of 110 (27.3%) patients and F. nucleatum in 69 out of 110 patients (62.7%). For faecal samples, the relative levels of P. micra and F. nucleatum were calculated using the 16S rRNA gene as reference. P. micra and F. nucleatum were detected in faecal samples from 143 out of 246 (58.1%) and 185 out of 244 patients (75.8%), respectively.The levels of P. micra and F. nucleatum. The distribution of samples classified as high and low can be found in P. micra and 40 out of 110 patients (36.4%) with high levels of F. nucleatum in tumour tissue. In faeces, 108 out of 246 patients (43.9%) had high levels of P. micra and 114 out of 244 patients (46.7%) had high levels of F. nucleatum. Of the 24 patients with high levels of P. micra in tumour tissue, 20 patients also had high levels of P. micra in faeces .ROC curve analyses based on cancer-specific survival were used to calculate an optimal cut-off value for high or low levels of n faeces . For F. n faeces . FurtherP. micra also had high levels of F. nucleatum  and in faeces .Interestingly, 23 of the 24 tumour tissues with high levels of ucleatum . In faecucleatum . MoreoveP. micra and F. nucleatum to clinicopathological and molecular characteristics of the study patients were analysed using the cut-off for high/low levels  of CRC patients were included to increase sample size. The median follow-up time from date of cancer surgery was 93.2 (1.2–134) months. Kaplan–Meier curves truncated at 5 years were estimated for patients with high and low levels of P. micra or F. nucleatum in tumour tissue . For F. nucleatum, no significant association was found (p = 0.090).The levels of r tissue  and in fr tissue . High lesurvival . No signn faeces . When exP. micra in tumour tissue were also found to have a significant effect on survival in a multivariable analysis including tumour stage and grade (HR 3.82 (CI 1.12–13.0), p = 0.032), further strengthening the association between increased levels of p. micra in tumour tissue and decreased patient survival , p = 0.099)  .P. micra and F. nucleatum, in tumour tissue and in faeces of CRC patients in relation to prognosis. We found a novel association between high tumoural levels of P. micra and a decreased patient survival rate in CRC. High tumoural levels of F. nucleatum were also associated with decreased patient survival. We could further associate high levels of P. micra and/or F. nucleatum in tumour tissue with different patient clinicopathological and tumour molecular traits. In addition, we concluded that analyses of the tumour microbiota were more significant in terms of clinical and molecular associations than analyses of the faecal microflora.In this study, we investigated the presence of two CRC-associated microbes from the oral microflora, P. micra and F. nucleatum in tumour tissue in relation to tumour molecular features, high levels of both bacteria were found to be associated with BRAF-mutated tumours and tumours of the MSI subtype. Furthermore, F. nucleatum was significantly associated with right-sided tumours. The results are in line with earlier findings for F. nucleatum [P. micra and F. nucleatum in tumour tissue and tumours of CMS subgroup 1 were found [BRAF-mutated and right-sided, which is in line with the current findings [P. micra and F. nucleatum in tumour tissue were further associated both with age under 59 and age above 80. In a study by Bhem et al., an association between high age and high levels of F. nucleatum in tumour tissue was found in gastric cancer patients [F. nucleatum has also been associated with young onset CRC [P. micra, there was a significant association with the female sex, whereas for F. nucleatum there was a trend towards an association, although it was not significant. The association with the female sex has not previously been described and needs to be verified in other studies. However, right-sided tumours, tumours of the MSI subtype and BRAF-mutated tumours are known to be more frequent in older women [When analysing levels of ucleatum ,29. In are found ,25. CMS findings . High lepatients . Howevernset CRC . The nonnset CRC . In contnset CRC . For P. er women .P. micra and F. nucleatum in tumour tissue. High levels of P. micra or F. nucleatum in tumour tissue were significantly associated with decreased five-year survival in stage I-IV CRC. This association remained significant in multivariable Cox regression analysis for P. micra. When excluding stage IV CRC patients, the association with decreased survival was still significant for P. micra. To our knowledge, this is the first study showing an association of P. micra in tumour tissue with decreased patient survival. However, one recent study has suggested a prognostic role of faecal P. micra in CRC [F. nucleatum, on the other hand, has been studied more extensively and has previously been associated with decreased survival [F. nucleatum were associated with decreased CRC-specific survival  [P. micra and F. nucleatum were associated with tumours of the MSI subtype, generally known to have a better prognosis [P. micra and F. nucleatum with BRAF mutations, which have been proposed to partially mitigate the positive prognostic effect of MSI [P. micra and/or F. nucleatum colonisation represent a subgroup of MSI tumours with a worse prognosis. To strengthen this hypothesis, further subgroup analyses are needed in larger patient cohorts. In a study by Wei Z et al., high levels of F. nucleatum in tumour tissue were associated with a decreased survival rate and increased expression of TNF-α, suggesting that dysbiosis might worsen the patient’s prognosis by upregulating gut inflammation [F. nucleatum has also been suggested to be associated with the development of tumour metastasis [F. nucleatum to promote CRC metastasis through the regulation of caspase activation and recruitment domain 3 , as well as activation of the nuclear factor-kappa B pathway resulting in migration of CRC cells [F. nucleatum has been associated with chemotherapy resistance through the regulation of autophagy pathways [Furthermore, we investigated associations between cancer-specific survival in colorectal cancer and levels of a in CRC . F. nuclsurvival ,19,20,21rognosis . Associat of MSI . It is tammation . F. nucltastasis , and hastastasis . Chen etRC cells ,39. In lRC cells . In addipathways . It is tP. micra were found to be strongly associated with the levels of F. nucleatum in both tumour tissue and faeces, which is in line with two of our previous studies, as well as a study by Jun Yu et al. [P. micra and F. nucleatum were shown to display synergistic effects in bacterial biofilm formation [P. micra and F. nucleatum [P. micra and F. nucleatum in CRC prognosis are needed to support these theories.The levels of u et al. ,23,25. Iormation . Drewes ucleatum , and rigucleatum . Thus, iucleatum . FurtherP. micra and F. nucleatum with clinical and molecular parameters were much weaker. P. micra and F. nucleatum were still associated with tumours of the MSI subtype. However, no association was found between P. micra or F. nucleatum and cancer-specific survival. One explanation for this could be that tumour tissue represents the local tumour microenvironment better than faecal samples do. In this study, we found significant associations between levels of P. micra and F. nucleatum in both tumour tissue and in faeces, respectively, suggesting that the faecal microflora still partly reflects the local tumour microflora. However, for more detailed analyses of the molecular role of P. micra and F. nucleatum in tumour progression and patient survival, tumour tissue appears to be the preferred choice.In faecal samples, the associations of One strength of this study is the inclusion of both tumour tissue and faecal samples originating from the same individuals, giving a better insight into the spatial presence of the examined species in association with studied traits. However, the lack of a validation cohort calls for further studies in order to confirm our results. Furthermore, our study does not include any mechanistic insights for the found associations. Thus, studies on the underlying mechanisms, including inflammatory and immunological processes, are needed in order to better understand the cause and effect relationships. Another area of interest is the association with dietary factors, which may directly and indirectly affect the constitution of the gut microbial content.P. micra and F. nucleatum in tumour tissue and decreased patient survival in CRC. We found that P. micra and F. nucleatum often colonise the same tumours, and that tumours colonised with P. micra and/or F. nucleatum more often are BRAF-mutated and of the MSI subtype. However, further studies on the role of the gut microbiota in CRC formation and progression, including underlying mechanisms, are needed. A better understanding of the role of gut microbial composition in CRC progression might contribute to future putative targets for therapy.Our study suggests associations between high levels of


Catalytic reactions on flexible sheets generate fluid flows that transform the shape of the sheet, which in turn modifies the flow. These complex interactions make computer models vital for designing and harnessing these feedback loops to create soft active matter that autonomously performs self-sustained mechanical work. Rather than requiring external electrical or mechanical power, the “nutrients” for operating SofAM can simply be carried in a small vial of chemicals.Chemically active matter commonly refers to nano- and micro-scale objects in a solution that consumes energy from catalytic reactions in the fluid to perform mechanical work. In converting reactants into products, these catalytic reactions generate local gradients in the concentration of chemicals and density of the fluid. The gradients propel the fluid to move and enable the system to mimic salient features of chemo-mechanical transduction in living organisms, which convert energy from nutrients into mechanical action . If the catalysts driving this biomimetic behavior are anchored to an immobile surface in a fluid-filled microchamber, then the system acts as a chemical “pump”, which drives the fluid to flow; the flow, however, does not alter the location of the immobilized catalysts. If the catalytic reaction occurs on hard, mobile particles , the chemically driven flow drives the particles’ motion, creating chemical “motors”, but does not deform the hard active matter. The situation is significantly more complicated if the catalysts are attached to soft active matter (SofAM), such as movable, flexible sheets. Now, the hydrodynamic interactions not only drive the sheets’ motion but also morph this elastic layer into three-dimensional shapes Fig.  or drive2 in 2014 that enzymatic reactions on immobile surfaces generate fluid flow, researchers have examined catalyst-coated, mobile surfaces, such as zero-dimensional (0D) rigid particles and one-dimensional (1D) flexible filaments3. To date, however, there have been few experiments on 2D or 3D soft active matter, which introduces additional degrees of freedom and complex dynamics. For example, computational models reveal that a soft catalyst-coated 2D layer can repeatedly bend and unbend to mimic the locomotion of inch worms3. Such distinctive dynamic behavior is due to a feedback loop  flow in a fluid-filled chamber; (2) advection and diffusion of chemicals in the solution and their reactions with deformable materials; (3) dynamic behavior of the nodes that make up the elastic material; and (4) fluid–structure interactions between these nodes and the solution -(4) interconnected processes can facilitate the design of synthetic SofAM that perform chemically driven, mechanical work, and thus serve as chemical gears or self-oscillating chemical clocks. More generally, the results reveal new modes of actuating and controlling the transformation of 2D biomimetic soft matter into 3D functional materials.9. The center of the sheet is pinned to restrict its horizontal motion but is free to move in the vertical direction. The entire sheet is coated with the enzyme catalase, with the nodes in black being heavier than the ones in green. The addition of hydrogen peroxide activates the catalyst to decompose H2O2 into the lighter products, water and oxygen, and thereby generate buoyancy-driven fluid flow. The generated flow drives the lighter, central region of the sheet to pop up out of the plane, morphing the 2D layer into a 3D shape. The sheet, in turn, exerts forces on the fluid that modify the flow; in particular, the distended spokes act as blades that “catch” the flow and rotate the 3D object. The computer models allowed us to capture and visualize the latter feedback between the fluid and sheet and design the optimal sheet architecture to maximize this fluid–structure interaction. The computer models thereby reveal new routes for actuating flexible material and achieving functionality that is not possible with chemically active matter that is immobile or mobile and inflexible.Figure With the addition of cogs to the outer rim, the active structure  centered at the patch. The combined hydrodynamic forces pull the sheets toward this center, while the excluded volume interactions keep the sheets apart. Through the inherently coupled interactions, the fluid flow affects the shape and movement of the flexible sheets, which in turn modify the fluid flow. This distinctive feedback, which combines hydrodynamic, steric, and fluid–structure interactions, causes the sheets to form self-regulating coupled oscillators, whose motion is synchronized in time and space. The phase dynamics of the coupled self-oscillators can be tuned by altering the extent of catalyst coverage on either the sheets and/or the surface patch. This breadth of dynamic behavior expands the functionality of the coupled oscillators, enabling soft robots to display a variety of self-sustained, self-regulating moves.The synchronization of self-oscillating systems is vital to various biological functions, from the coordinated contraction of heart muscle to the self-organization of slime molds. Through modeling, Balazs and co-authors designed bioinspired materials systems that spontaneously form shape-changing, self-oscillators, which communicate to synchronize both their temporal and spatial behavior3.Creating predictive models for controlling the actuation and motion of SofAM will require further developments in theory and simulation. While the above discussion focused on buoyancy-driven flow, two or more mechanisms for propelling fluids  can be triggered simultaneously by a single chemical stimulus. Models that capture these simultaneous processes can reveal new means to control the surface and bulk flows independently and thus, provide effective approaches to separately direct the self-organization of the immersed sheets and particles or autonomously sort different sized particles in microchannels14. Modeling this integrated behavior in SofAM to understand and ultimately direct their dynamic assembly could require simulations involving hundreds (or thousands) of actin filaments, myosin motors and cells. More coarse-grained modeling approaches are needed to simulate systems of that size scale in reasonable run times.As described above, the spatiotemporal behavior of SofAM sheets typically involves multiple coordinated, dynamic processes; capturing these processes within computationally feasible timescales introduces new challenges for modeling these systems in 3D. For example, the model of the spontaneous buckling of an acto-myosin sheet Fig.  encompasThe above studies focused on the conversion of chemicals into mechanical energy. Chemical reactions driven by light (or other stimuli) can involve multiple energy domains, and thus require models for photo-chemo-thermo-mechanical transduction. To capture this coupling of energy fields also requires longer simulation run times, as well as accurate models that describe the interaction among all these dynamic processes.Another challenge for modeling is to simulate cascade reactions that provide a programmable delay between individual chemical reactions. This delay is critical for directing particle motion in multi-step processes. For example, with regulation, a reaction yielding dense products can lead to outward motion , before the next reaction, which produces less dense products, would lead to inward motion. In other words, coordination between the chemical reactions will modulate directed transport (whereas simultaneous activation of both reactions would cancel out the fluid motion). Simulating cascades involving a number of reactions may require the development of new coarse-grained models, but meeting this goal would allow researchers to design multi-stage processes that perform autonomously in microchambers.All these challenges provide guidelines for new research directions, which can inspire the imagination of scientists for a considerable time.


Chemical probes can be used to understand the complex biological nature of diseases. Due to the diversity of cancer types and dynamic regulatory pathways involved in the disease, there is a need to identify signaling pathways and associated proteins or enzymes that are traceable or detectable in tests for cancer diagnosis and treatment. Currently, fluorogenic chemical probes are widely used to detect cancer-associated proteins and their binding partners. These probes are also applicable in photodynamic therapy to determine drug efficacy and monitor regulating factors. In this review, we discuss the synthesis of chemical probes for different cancer types from 2016 to the present time and their application in monitoring the activity of transferases, hydrolases, deacetylases, oxidoreductases, and immune cells. Moreover, we elaborate on their potential roles in photodynamic therapy. For example, Yang et al. synthesized a C–Se-based photo-crosslinker that can cleave and capture peptide pools from chaperones . A . A 2 gen cancers . Another cancers . Similarer cells . Moreovet oxygen . Yuan eter cells .50; selectivity should be 30-fold higher than other competitor portion and should have reasonable cell penetrant with proper on target selectivity [https://pubchem.ncbi.nlm.nih.gov, accessed on 4 May 2022), ChEMBL , Probe miner , SGC chemical probe collection , Guide to Pharmacology , and Chemical Probe Portal . Chemical probes have improved the current PDT approaches for studying tumor incidence and progression as well as the therapeutic inventions in cancer. These novel probes aid researchers in understanding the disease-causing mechanisms in more sophisticated and precise ways; however, considerably better probe selection needs to be a focus for further development in this field. To analyze hypertoxicity, a series of new probes with better physical properties and reduced levels of immunotoxicity are needed. It is expected that improvements in the overall applicability of probes will contribute to the scientific community’s pursuit of better clinical treatment options and therapies against cancer.Chemical probe tools offer highly reliable analyses of the biological complexity of cancer through monitoring and understanding the key regulatory factors with a dynamic range of fluorogenic properties. Another broad application of probe proteomics is in the identification of cancer biomarkers. In this approach, AfBP facilitates target identification by phenotypic screening rather than by observing and monitoring activities. For example, in the cellular system, ANXA2, PDIA3/4, FLAD1, and NOS2 targets show hindering properties. A series of tetrazole-based probes have been used to identify these targets by phenotypic screening and affinity-based proteome profiling . In thisectivity . In addi


R0, and the permanence of a system with at least one tumour-present periodic solution is investigated. Furthermore, the effects of the death rate, effector cell injection dosage, therapeutic period, and effector cell activation rate on indicator R0 are determined through numerical simulations, and the results indicate that radioimmunotherapy is more effective than either radiotherapy or immunotherapy alone.In this paper, an antitumour model for characterising radiotherapy and immunotherapy processes at different fixed times is proposed. The global attractiveness of the positive periodic solution for each corresponding subsystem is proved with the integral inequality technique. Then, based on the differentiability of the solutions with respect to the initial values, the eigenvalues of the Jacobian matrix at a fixed point corresponding to the tumour-free periodic solution are determined, resulting in a sufficient condition for local stability. The solutions to the ordinary differential equations are compared, the threshold condition for the global attractiveness of the tumour-free periodic solution is provided in terms of an indicator  Cancer is a major public health issue and the leading cause of death worldwide. According to the World Health Organization (WHO), there were nearly 10 million cancer-related deaths in 2020 . Host ceα, are soluble proteins that mediate cell-to-cell communication  or ((n − 1)T + lT, t] givesIntegrating  from 23) thatN1 > 0 such that when n ≥ N1, it holds thatThen, according to  and 17)17), thert ∈ ((n − 1)T, nT], where n − 1 ≥ N1, it follows from , a, at ∈ ((ows from  that(27ε(1) > 0 is arbitrary, we conclude that Since This completes the proof.Similarly, we arrive at the following conclusion.y(t), z(t), x(t)) of system  denote the solution of the first three equations of T, as follows:Let tions of  for initI1, I2 : R3⟶R3 as follows:Additionally, we can define the mappings R3⟶R3 asand the map Ψ : (i)y∗(t), z∗(t), 0) of system y∗(t), z∗(t), 0) of system (The tumour-free periodic solution (f system  is globaX0) at point X0∗ = (y∗(0+), z∗(0+), 0) arey∗(t), z∗(t), 0) is locally stable , where n ≥ N5, we obtainLet egrating  on ((n −x(((N5 − 1) + lT + nT)+) ≥ x(((N5 − 1)T + lT)+))n⟶+∞ as n⟶∞, which contradicts the boundedness of x(t). Thus, there exists a t6 > t5 > 0 such that x(t6) ≥ m0.Then, based on  and 74)74), we hx(t) ≥ m0 for all t ≥ t6, then our goal is obtained. Otherwise, x(t) < m0 for some t > t6. We set t∗ = inf{t|t > t6, x(t) < m0}; then, we have t∗ ≥ t6 > t5 > 0, which implies that (45) holds on the interval  and x(t∗+) ≤ m0.According to the definition of the infimum n2, n3 ∈ Z+ such thatWe choose t7 ∈ T) such that x(t7) > m0. Otherwise, we can assume that x(t) ≤ m0 is valid for t ∈ T]. Then, similar to t ∈ T], it holds thatWe claim that there exists a ution of . Thus, wt ∈ T], it follows from T]. Similar to , , a, at ∈  ≤ m0 and y(t) < ME hold for t ∈ , a, a93), at∗∗ = inf{t|t > t∗, x(t) > m0}; then, t∗∗ ∈  and x(t∗) = m0. Suppose that t∗ ∈ ((n6 − 1)T + lT, n6T + lT), where n6 ∈ Z+.It is clear that t ∈  such that x(t) > m0.There exists some t∗∗∗ = inf{t|t > t∗, x(t) > m0}; then, t∗∗∗ ∈ [t∗, n6T + lT). For t ∈ , it follows that x(t) ≤ m0; thus, x(t∗∗∗) = m0. Similar to , we haveLet milar to , when t t > t∗∗∗, the same arguments can be continued since x(t∗∗∗) = m0.For t ∈ , x(t) ≤ m0.For all t∗∗∗∗ = inf{t|t > t∗, x(t) > m0} ; then, t∗∗∗∗ ∈ [n6T + lT, (n6T + lT) + (n2 + n3)T) and x(t∗∗∗∗+) = m0. Similar to Cases t ∈ , it holds thatmT < m1 < m2 < m0.Let  + n2 + nT) and x(x(t) ≥ mT for t ≥ t6.We repeat the above procedure to prove that Furthermore, according to Schauder's fixed point theorem, there exists a tumour-present periodic solution for system .This completes the proof.pE, pH, and pT, the dosage of infusing the ECs (σ1), the therapeutic period T, and the activation rate of the ECs (ρ)) affect the threshold value R0 defined in (+0Ty∗(t)dt is independent of pT, R0 decreases monotonically with respect to pT, indicating that the strong tumour cell killing effect of radiotherapy can increase tumour cell death.We are interested in how the key factors  decreases to the threshold value TpH = 0.1440 (or TpE = 0.0377) of the tumour-free periodic solution, R0 monotonically decreases to 0. In addition, when TpH < pE, pH < 0.2, R0(pH) is much smaller than R0(pE), where R0(pE) > 1. Therefore, in this case, the optimal control strategy is achieved when pH and pE are sufficiently small, and, compared to parameter pE, a smaller parameter pH is more beneficial for tumour control. Similarly, Figures R0 monotonically decreases as σ1 increases or T decreases, indicating that a higher dosage of infusing the ECs or more frequent radioimmunotherapy can accelerate the eradication of tumour cells. In addition, ρ increases to the threshold value Tρ = 0.2003 of the tumour-free periodic solution, R0 rapidly decreases to 0. Thus, strong activation of the ECs by the HTCs is beneficial for tumour control.σ1 = 0 andMoreover, when T is smaller than the threshold value Ta = 2.0076, the global attractiveness of the tumour-free periodic solution can be validated  the parameters pE, pH, and T  are, the smaller the indicator R0 is. In particular, decreases in pH are more beneficial for tumour control than decreases in pE, and radioimmunotherapy is more effective than either radiotherapy or immunotherapy alone.Our results demonstrate that the effectiveness of radioimmunotherapy, the therapeutic period, and the activation rate of the R0, as shown in T on the threshold value R0. The tumour-free periodic solution is locally stable for Ta < T < Tp, whereas system  note that system 3) in  includes in  inclSimilar to , 29, the


BackgroundSocial media has been more widely used by urology residency programs since the COVID-19 pandemic. There are no studies on the relationship between Doximity residency ranking and social media usage in urology.ObjectivesIn this study, we aim to investigate the relationship between social media usage and the academic prestige of urology residency programs.MethodsResidency reputation data was acquired from the Doximity Residency Navigator website. Twitter and Instagram activity in 2019 and 2021 was analyzed by collecting data on the total number of posts and followers. Data on residency virtual recruitment was obtained from Twitter and UroResidency website.ResultsBy the end of 2021, 122/139 (87.8%) urology residency programs had a Twitter account and 61/139 (43.9%) had an Instagram account. A significant linear regression was found between Doximity ranking and the number of Twitter followers (p<0.001), Twitter posts (p=0.005), and Instagram followers (p=0.026). Virtual recruitment events were held by 107/139 (77%) programs in 2021. There was a significant linear regression between Doximity rankings and the number of virtual events (p<0.006).ConclusionsSocial media use by urology residency programs has increased since the COVID-19 pandemic. A program’s higher Doximity ranking was correlated with the presence of Twitter and Instagram accounts as well as the number of Twitter followers, Twitter posts, and Instagram followers. There was a significant relationship between Doximity rankings and the number of hosted virtual events. Programs should consider increasing social media visibility to potentially improve their Doximity rankings. Social media use has become more prevalent and important in the field of academic urology. In 2014, the American Urological Association (AUA) reported that 71% of its members used some form of social media . InstagrThere have been a few studies analyzing the use of social media with the AUA Match outcomes ,6, but tWith travel restrictions and away rotation limitations due to COVID-19 making it more challenging for prospective residents to explore residency programs , programA comprehensive list of urology residency programs in the United States was acquired and ranked 1 to 145 based on the Doximity Residency Navigator, with 1 being the highest ranked . ProgramTwo independent reviewers used the Google search engine with the search term “name of institution + urology residency” + “social media account”  urology residency Twitter) to determine the presence of a social media account. Reviewers also used the Association of American Medical Colleges’ Electronic Residency Application Service and official urology departmental websites to identify social media accounts ,15. All If a program had multiple accounts identified within the same social media platform, the account with more followers was used in the statistical analysis. Finally, for the 2021-2022 application cycle, the presence and number of virtual residency recruitment events  were collected using Twitter and UroResidency, a urology residency application advisement website, and analyzed against Doximity ranking.Since the variable of interest in social media usage was the COVID-19 pandemic, the years 2019 and 2021 were chosen as the proxy for “before the pandemic” and “after the pandemic,” respectively. The year 2021, rather than 2020, was chosen for comparison because COVID-19 was not declared a pandemic until March 11, 2020, and the first stay-at-home order in the United States was not ordered until March 19, 2020 ,17.The Shapiro-Wilk test was run to assess for normality of all study distributions, and the appropriate statistical analyses were then performed. Significance was set at p<0.05. All statistical analyses were performed on Statistical Package for the Social Sciences (SPSS) version 28.0 . This study was deemed Exempt Category 4 by the Institutional Review Board (IRB).Social media among urology residency programsA total of 145 urology programs were available on the Doximity Urology Residency Navigator, of which 139 programs were included. Six programs were excluded from the analysis; five were military programs, and one was not recruiting for the 2021-2022 cycle. Overall, 122 (87.8%) programs had Twitter accounts and 61 (43.9%) had Instagram accounts by the end of 2021, an increase from 88 (63.3%) and 12 (8.6%) in 2019, respectively  but normally distributed for Instagram (p=0.456). Both Twitter and Instagram posts were not normally distributed in 2021 . Of the programs that owned Twitter accounts in 2019 versus 2021, the median (interquartile range (IQR)) number of Tweets made by each program was 27 (44) versus 23 (51), respectively  had a median Doximity ranking of 67, whereas programs without Twitter (n=17) had a median Doximity ranking of 133 (Table 22 had a Urology residency programs with Instagram n=61) had a median Doximity ranking of 66, whereas programs without Instagram (n=78) had a median Doximity ranking of 84  urology programs hosted virtual residency recruitment events. For the programs that hosted virtual events, the median  number of hosted events was 2 . A significant linear regression relationship was found between a program’s Doximity ranking and the number of virtual events (p<0.006)  of urology residency programs hosted virtual recruitment events during the 2021-2022 AUA Match cycle, compared to 75% of orthopedic surgery , 59% of 7% of uroThe main limitation of our study is the relative paucity of metrics to rank urology residency programs. Our study relied on the Doximity reputation rankings, which are based on three components: satisfaction data via current resident and alumni surveys, reputation data via board-certified physician surveys, and objective data via research output and a “proprietary Doximity database” . While iA final comment is that our study does not imply that social media is the only factor in rankings. With urology’s highly competitive environment in both education and practice, many variables determine academic prestige, and social media may be one of many. The goal of this study is primarily to voice our recommendations by providing additional data for urology programs to make informed decisions on their own social media activity.More research is needed to investigate the relationship between social media usage by urology residency programs and their academic prestige. A future direction of interest would be to analyze the content of Twitter and Instagram posts, particularly how many of these posts were used for residency recruitment specifically. Although Tweet categories  have been examined in prior studies, it was not done so in relation to a program’s ranking . AnalyziThe use of social media by both urology residency programs and applicants has been largely increasing as a result of the COVID-19 pandemic requiring residency recruitment and interviews to become primarily virtual. There was a significant relationship between a program’s Doximity ranking and the presence of Twitter or Instagram, as well as the number of Twitter followers, Twitter posts, Instagram followers, and virtual recruitment events. Although Twitter appears to be more widely utilized and a better indicator than Instagram regarding academic prestige on Doximity, both platforms have been used by the majority of urology residency programs.


Giant cell tumour is a benign lesion classified as a fibrocystic tumour whose localization in Hoffa's fat pad is very rare. Clinical symptoms are insidious and non-specific causing a frequent confusion and delay in diagnosis therefore it should be distinguished radiologically from other similar conditions such as Hoffa´s disease and lipomas. We report a case of a 37-year-old patient, with no relevant history, who complained of a right knee pain for 5 years. Magnetic resonance imaging showed a small nodular mass in Hoffa's pad which was excised through a direct approach. Histologic examination of the specimen revealed a giant cell tenosynovial tumour. One year after surgery, the patient was asymptomatic with no local recurrence. The surgical removal of the tumour is the ideal treatment. The choice between open surgery and endoscopy depends on the site, size, and extent of the tumour. Tenosynovial giant cell tumour may arise almost anywhere in the body, Hoffa´s fat pad localization is rarely seen . ClinicaPatient information: we report a case of a 37-year-old patient, with no relevant history, who complained of right knee pain for 5 years. It was discontinuous and aggravated by strenuous activity. The patient had no sensation of the knee giving way when walking, nor did he experience any locking of his knee.Clinical findings: the tof motion was limited from 0 to 110° of flexion with a moderate pain on further flexion. Neither swelling nor effusion was visible and there was no palpable mass across the joint line. There was no history of general symptoms such as night sweats, fever, loss of appetite, or weight loss.Diagnostic assessment: the patient´s blood parameters were normal. Antero-posterior and lateral X-ray views showed no abnormality (ormality . MRI shoormality . The bonTherapeutic interventions: due to the localization of the tumour, we opted for open resection of the mass through a medial parapatellar approach. A 1.5 cm diameter encapsulated tumour was found. The outer surface was focally fat-laden, and the surface was yellow-brown. Therefore the mass was totally excised. Histopathological examination showed mononuclear stromal cells, osteoclastic giant cells, few lymphocytes, and macrophages. No cellular atypia or other evidence of malignancy was seen. These features allowed us to confirm the diagnosis of a giant cell tumour.Follow-up and outcome of interventions: at 2 years of follow-up, the patient was asymptomatic and was able to return to his previous level of daily activities. No local recurrence of the tumour was noted.Patient perspective: the patient was happy with the successful outcome of the surgery.Informed consent: written informed consent was obtained from the patient for participation in our study.Giant cell tumour was firstly reported by Chassaignac in 1852 as a pigmented villonodular synovitis and tenosynovitis, but recently the term of giant cell tumour is being accepted as a more accurate expression [et al. estimated the incidence for the local giant cell tumour to be 1.8 cases for each million [Meyers  million . Other i million . However million ,5-8 (TabLipomas are seen on MRI as a typical discrete lipomatous mass with hyperintensity on both T1- and T2- weighted images with the loss of signal on appropriate fat-suppressed sequences. Hoffa´s disease is an intrinsic disease of Hoffa´s fat pad firstly described in 1904 by Albert Hoffa as an acute or chronic inflammation of the infrapatellar fat pad [In the case of a giant cell tumour, MRI shows T1 hypointense and a discrete T2 hyperintense signal with gadolinium homogeneous enhancement. This lesion is characterized by hypointense punctuations best seen on fast field echo sequence MRI images suggesting the presence of hemosiderin deposits that are typical of this condition. This sequence was not used to establish the diagnosis in our case and even in the majority of reported cases. The suitable treatment of the local form is controversial due to the lack of evidence and the high risk of recurrence. The local Hoffa´s fat extra-articular form of giant cell tumour can be removed by open surgery -8, endosAlthough tenosynovial giant cell tumour has a rare occurrence in Hoffa's fat pad, it should be considered as a potential cause of non-traumatic knee pain. The presence of hemosiderin deposits in MRI has best seen, especially on fast field echo sequence is strongly suggestive of the diagnosis and permits to eliminate of differential diagnoses such as lipomas and Hoffa´s disease. The diagnosis is only confirmed by histopathological examination. Open surgical resection is preferred in order to ensure complete resection and to avoid recurrences.