Cancer Discovery publication, Bianchi et al. unveil a crucial mechanism driving therapeutic resistance in pancreatic ductal adenocarcinoma (PDAC), orchestrated by cell-autonomous Cxcl1 and TNF from polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs). This insight provides fresh perspectives for surmounting such resistance in PDAC.1In a recent 2 Although significant advancements have been made in the field, numerous enigmas persist, particularly concerning the intricate interplay within the TME that fuels therapeutic resistance. A more profound comprehension of these underlying mechanisms is indispensable for devising enhanced treatment strategies and ultimately ameliorating patient prognoses. Bianchi’s groundbreaking research offers an enlightening vantage point on investigating the complex orchestration of interactions among tumor cells, immune cells, and stromal components within the TME.PDAC represents a highly aggressive malignancy exhibiting substantial therapeutic resistance, attributable to factors such as intractable genomic aberrations, immunosuppression mediated by myeloid cells, and pro-inflammatory activation of cancer-associated fibroblasts. For instance, spontaneous ferroptosis of PMN-MDSCs within the tumor microenvironment (TME), despite reducing their presence, indirectly suppresses T-cell activity and promotes tumor growth.KRAS-TP53 co-alterations specifically enriched pathways related to neutrophil function and CXCL1 expression. This finding was further validated in preclinical genetic models and patient-derived tumors. Utilizing single-cell RNA sequencing and imaging mass cytometry, the team ascertained that CXCR2, the receptor for CXCL1, was primarily expressed in intratumoral PMN-MDSCs. Furthermore, they observed strong spatial contiguity between CXCL1-expressing tumor islands and CXCR2+ PMN-MDSCs, with CD8+ T cells being conspicuously absent from these areas.Through analyzing transcriptomes of human PDAC cell lines, Bianchi et al. discovered that KRASG12D and TP53R175H increased CXCL1 production via CREB-dependent mechanisms. Inhibiting CREB led to a significant decrease in CXCL1 expression and secretion in both human and murine PDAC models. To investigate Cxcl1’s role in T-cell exclusion, the study employed CRISPR-Cas9 edited K-rasG12D/+;Trp53R172H/+;Pdx-1Cre/+ (KPC) tumor cells. The results revealed that Cxcl1 silencing contributed to reduced tumor growth and improved survival. Moreover, enriched immunoregulatory signaling and increased CD4+ and CD8+ T cells were observed in Cxcl1-silenced tumors, suggesting that Cxcl1 silencing overcomes T-cell exclusion and suppresses tumor growth through a CD8+ T-cell mechanism.The researchers also uncovered that the mutational cooperativity of cell-autonomous The authors then probed the impact of Cxcl1 silencing on MDSC trafficking dynamics and T-cell suppression using a novel in vivo adoptive transfer system. They found that reprogramming of PMN-MDSCs generated in Cxcl1-silenced tumor-bearing hosts disrupted their migratory potential and reduced immunosuppressive markers. This reprogramming of MDSC function could contribute to the improvement of T-cell exclusion and antitumor immunity in Cxcl1-silenced tumors. RNA sequencing identified enrichment in MAPK pathway signaling and Tnf as the top upstream regulator of differentially expressed MDSC transcriptomes. They established a signaling link between CXCR2 ligation, MAPK pathway activation, and Tnf production in PMN-MDSCs, with the dependency of MDSC-intrinsic TNF on CXCR2-MAPK signaling validated in vivo. The study exposed a previously unrecognized paradox where inhibiting Cxcl1-CXCR2 engagement invigorates T-cell activation while dampening tumor-wide TNF signaling, predominantly through disruption of MDSC-restricted TNF.Shifting focus to the impact of MDSC-derived TNF in the PDAC tumor microenvironment, the study unveiled its crucial role in immunoregulatory circuitries and stromal inflammation. The experiments demonstrated that transmembrane TNF-TNFR2 signaling upregulated the expression of Cxcl1 in tumor cells and cancer-associated fibroblasts (CAFs), leading to increased pro-inflammatory signaling. Etanercept treatment, a TNFR2 inhibitor, considerably decreased Cxcl1 expression and suppressed neutrophil trafficking dynamics in adoptive transfer experiments. Additionally, etanercept preconditioning rescued T-cell IFN-γ release suppression and enhanced intratumoral T-cell trafficking. This treatment also resulted in a significant decrease in PMN-MDSCs and M2-like macrophages. Inhibition of transmembrane TNF-TNFR2 impaired the IL-6/STAT3 signaling pathway in the tumor and diminished the iCAF:myCAF ratio, ultimately attenuating the inflammatory stromal components and fibrotic collagen deposition. Combining etanercept with gemcitabine + paclitaxel chemotherapy nearly doubled the median survival in orthotopic KPC models and reduced metastatic outgrowth compared to single treatments, without additional toxicity. These findings emphasize the potential of TNFR2 inhibition for mitigating stromal inflammation, reducing Cxcl1 expression, and enhancing PDAC’s chemosensitivity, which underscores its therapeutic value.KRAS-TP53 genomic co-alteration in PDAC, mediates T-cell restriction through interactions with CXCR2+ neutrophilic MDSCs, and that TNF signaling in cancer cell-neutrophil crosstalk drives stromal inflammation, immune tolerance, and therapeutic resistance (Fig. + T cells, remains nebulous. Additionally, the underpinning mechanism of CD8+ T-cell exclusion by PMN-MDSCs is yet to be elucidated. Furthermore, residual CAF-Il6 expression, notwithstanding transmembrane TNF-TNFR2 inhibition in MDSC-CAF co-cultures, suggests additional routes by which neutrophil-derived signals could influence iCAF polarization. Despite these limitations, the insights gained from this study offer valuable directions for future research and have important implications for the development of new therapeutic strategies.In summary, Bianchi and colleagues demonstrates that cell-autonomous Cxcl1, regulated by 3 emphasizing the need to determine if this discovery is exclusive to PDAC or a consistent phenomenon spanning a wide array of malignancies. In addition, certain cytokines in the TME were traditionally believed to be generated by specific cell types or stromal components, but it is important to note that other cell types, including previously unrecognized ones, may also produce these cytokines in response to TME-induced changes. Consequently, it is imperative to adopt a more comprehensive and meticulous approach to accurately determine the cellular sources of cytokines in the TME, which may lead to a deeper understanding of the mechanisms underlying cancer progression and inform the development of novel therapeutic strategies.The identification of neutrophils as a significant source of TNF in human and murine PDAC is a critical discovery that sheds new light on the intricate interplay between the immune system and cancer. Overexpression of TNF is a prevalent hallmark in various cancers,5 Additionally, TNFR2 has been reported to be highly expressed in some tumor cells, suggesting that targeting TNFR2 might also directly kill TNFR2-expressing tumor cells.5 However, the detailed role of TNFR2 in PDAC cells remains to be fully investigated. Bianchi et al. have reported that TNFR2 expression was induced in CAFs by co-culture with PMN-MDSCs, suggesting that targeting TNFR2 also alter the malignant behaviors of CAFs and further reshape the tumor microenvironment. The consequences of targeting TNFR2 on various TNFR2-expressing cell types within the TME merit additional exploration. These multiple potential benefits of TNFR2 targeting suggest that it might prove to be a promising approach for cancer therapy. Nevertheless, while preclinical studies have demonstrated efficacy, the efficacy and safety of TNFR2-targeting therapies need to be confirmed through human clinical trials.TNFR2 antagonism has been widely accepted as a strategy to eliminate tumor-residing Tregs and activate Teff expansion, making it a potentially attractive combination therapy.


COVID-19 presented with lung abnormalities on computed tomography (CT) scans in patient with false negative RT-PCR, which are helpful in diagnosis of this emerging global health emergency. It's a case report the young woman of 35-year-old patient with 2019-nCoV pneumonia confirmed with IgM-IgG serology underwent thin-section Chest CT.Our patient has the Chest CT with some lung abnormalities, the Ground-glass opacities, crazy paving pattern and smooth interlobular septal thickening. The clinical findings and with conspicuous ground grass opacity lesions in the peripheral and posterior lungs on CT are highly suspected of 2019-nCoV pneumonia. The 2019 novel coronavirus  is a viral respiratory illness, was firstly reported in Wuhan, Hubei Province, China, and rapidly spreads as a pandemicA 35 years old woman have just given birth by C-section, she was admitted to the emergency department, with fever, coughs and throat discomfort for 2 days. She had a history of good physical health and no underlying diseases. Physical examination showed fever, with a body temperature of 38.7°C, and the laboratory examination results showed abnormal leukocyte count, increased neutrophils, decreased lymphocytes, and elevated C-reactive protein.The first thin-section non contrast Chest CT scan from 2/4/2020 shows multiple ground-glass opacities and a crazy paving pattern in bilateral lung. The bilateratism of the peripheral lung opacities, without subpleural sparing, are common CT finding of the 2019 novel coronavirus pneumonia . The patIn the presence of the manifested clinical progression and the worsening symptoms, the second Thin-section none and with contrast Chest CT scan from 8/4/2020 (6 days after follow-up) shows progressive ground-glass opacities and multiple regions of patchy consolidation in two lungs with neither cardiopulmonary abnormalities such as pulmonary embolism . FollowiThe patient received a hydroxychloroquine with the azithromycin leading to increased benefit and good clinical course. 13 days after follow-up, CT showed regression of patchy consolidation and ground-glass opacities in both lungs with a right pleural effusion .In December 2019, the Novel coronavirus (2019 -nCOV) is identified in Wuhan, China, which causes a new type of pneumonia. Since then, the COVID- 19 is a respiratory pandemic disease The WHO has declared SARS-CoV-2 a pandemic, because the rapid rise in number of cases; uncontrolled and vast worldwide spread The clinical presentation of COVID-19 can from being asymptomatic to being in severe acute respiratory distress. The real-time polymerase chain reaction (RT-PCR) is method of diagnostic the COVID-19 to detect viral nucleotides from specimens obtained by oropharyngeal swab, nasopharyngeal swab, bronchoalveolar lavage, or tracheal aspirateTherefore, these false negatives caused repeat testing and critical resource constraintsSome studies have concluded that serologic assays are crucial method to identified the viral reservoir hosts and for epidemiological studies, as well as to reveal the forms of asymptomatic infections and to evaluate the morbidity and mortality.Generally, imaging is indicated in a patient with COVID-19 and worsening respiratory status, based essentially for Computed tomography (CT) which is key tools for pulmonary disease diagnosis and managementOn the one hand, Chest CT non-contrast has demonstrated about 56–98% sensitivity in differentiating COVID-19 from viral pneumonia, on the other hand it can be useful in early stages of disease course to detect this pathology with false negatives obtained from RT-PCRThe initial Thin-section non contrast Chest CT of COVID-19 reveals areas of “crazy-paving” patterns and ground-glass opacity (GGO) peripheral, developed in subpleural areas in bilateral lung. In Follow-up, the GGO it was increased of density and size in multiple lobes progressing to consolidation. CT signs gradually improve approximately 14 days post-symptom onset A diagnosis is made considering clinical findings, epidemiology, and laboratory testing to confirm SARS-CoV-2 infection. Chest CT can be a pivotal diagnostic role in the assessment of patients with COVID-19.


Arterial pseudoaneurysms are rare vascular abnormalities that can occur as a complication of infections. Artery pseudoaneurysms associated with SARS-CoV-2 are a rare occurrence in COVID-19 patients, and their rupture can result in significant hemorrhage and sudden death. Few cases of SARS-CoV-2-associated artery pseudoaneurysms have been reported, and their underlying pathophysiological mechanisms remain unclear. This study presents the first reported case of a patient who developed both pulmonary and gallbladder artery pseudoaneurysms following SARS-CoV-2 infection. We investigate the potential pathogenesis of these pseudoaneurysms and aim to improve the understanding of this rare complication. This study presents a case of an 80-year-old male patient with chronic obstructive pulmonary disease (COPD) and type 2 diabetes mellitus who developed both pulmonary and gallbladder artery pseudoaneurysms one week after SARS-CoV-2 infection. The study aims to analyze the potential pathogenesis of these pseudoaneurysms to improve the understanding of this rare SARS-CoV-2-related complication.Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been known to cause damage to organs and tissues containing angiotensin-converting enzyme 2 (ACE2) receptors, resulting in various complications . Direct 2.The patient experienced symptoms such as fatigue, muscle soreness, chills, fever, and unsteady gait three days prior to admission. Upon admission, laboratory tests showed significantly elevated levels of cardiac enzymes, transaminases, bilirubin, creatinine, erythrocyte sedimentation rate and procalcitonin. High-resolution computed tomography (HRCT) of the lungs showed multiple pulmonary cysts , while aOn the seventh day after admission, the patient experienced four sudden and unexpected episodes of hemoptysis, with each episode producing approximately 30–50 ml of blood. Hemostatic medication, i.e., etamsylate, hemocoagulase bothrops atrox and tranexamic acid, was administered immediately, and a repeat HRCT scan of the lungs revealed the presence of a newly formed solid nodule in the posterior segment of the right lower lobe . FurtherAfter the cessation of hemoptysis, the patient developed severe right upper abdominal pain and recurrently vomited fresh blood. Emergency abdominal CT scan revealed acute cholecystitis with biliary bleeding . With thA multidisciplinary consultation was organized to discuss further treatment options. Considering the patient's current physical condition, which was no longer suitable for surgical intervention, the experts recommended an interventional approach with balloon occlusion of the pseudoaneurysm. However, the patient subsequently experienced massive gastrointestinal bleeding with hemorrhagic shock, and due to the critical condition of the patient, he was transferred to the intensive care unit for further treatment. Ultimately, surgical intervention was abandoned in favor of conservative treatment.3.The formation of arterial pseudoaneurysms is characterized by the destruction of the arterial wall, leading to an extravascular hematoma surrounded by a thin layer of connective tissue. Pseudoaneurysms typically occur due to the focal dilation of arterial branches, and they are commonly associated with infections caused by pathogens like staphylococci, streptococci, or Mycobacterium tuberculosis . AlthougThe specific mechanisms that lead to the formation of pseudoaneurysms following SARS-CoV-2 infection remain unclear. However, existing evidence suggests that the virus can attack endothelial cells in blood vessels, and cause multisystem inflammatory syndrome, which can exacerbate vascular injury , 9, but Research indicates that patients recovering from SARS-CoV-2 infection may develop iatrogenic and noniatrogenic pulmonary cysts, which can rupture and merge to form lung bullae or spontaneous pneumothorax . Slow aiIn addition, secondary mucormycosis infection following SARS-CoV-2 infection may also be an important mechanism for the formation of pseudoaneurysms , 8. AlthAlthough there have been no reports of SARS-CoV-2-related pseudoaneurysms in the gallbladder artery, the virus can damage organs that have ACE2 receptor , which m4.In conclusion, we present a case of pulmonary artery and gallbladder artery pseudoaneurysms that developed during SARS-CoV-2 infection and investigate the underlying mechanism of SARS-CoV-2-associated artery pseudoaneurysms. Elderly diabetic patients experiencing sudden hemoptysis or hematemesis during the recovery phase following SARS-CoV-2 infection should be evaluated for rare complications such as artery pseudoaneurysms. Early diagnosis and surgical intervention can significantly improve the long-term prognosis of patients with this condition.


Over-Expression by Degradation Rescue” (OEDR), provides an additional layer and potentially preferential tool for the control of RTKs expression in cancer, in addition to other mechanisms acting at the transcriptional and messenger transcript stabilization levels. We propose this newly defined phosphorylation/ubiquitination switch-dependent signal to bear wider unexploited relevance in cell biology and human pathophysiology. The recently identified mechanism underlying an OEDR-regulated RTK is discussed herein in the context of physiological endocrine circuits and cancer.Recently published work provide the first known evidence of a malignancy-associated regulatory mechanism, functionally connecting a phospho-regulated degron domain embedded in a receptor tyrosine kinase (RTK), with its ectopic expression in cancer, conditional to a specific autocrine growth factor signal. Mechanistically, the growth factor-triggered phosphorylation inhibits the degron domain present in the regulated RTK, blocking access to a specific degradation complex. This ultimately rescues the RTK from rapid ubiquitin-proteasome-system-mediated degradation and, most importantly, causes its cellular overexpression. This mechanism, which has been here assigned the new functional name “ This, depending on the respective localization of the hormone/growth factor producing cell and the target cell bearing its cellular receptor. Specifically, these three layers refer to (a) an endocrine mechanism, when the hormone/GF is produced by a cell/specialized tissue-organ distant from the targeted cell bearing the specific receptor; (b) a paracrine mechanism, when the target cell is in close proximity with the hormone/GF producing cell ; and (c) an autocrine mechanism, when the hormone/GF-secreting cell bears the activated receptor establishing a self-stimulatory loop, as observed in advanced cancer stages see . On a dy2bona fide degron motif-containing RTK degradation, as part of a negative feedback meant to down-modulate the RTK-signaling activity (pi)Degron embedded around the phosphorylated RTK region (piDegron recognition by an underlying ubiquitin E3 Ligase (UbE3L)-driven complex, ultimately blocking the RTK UPS-mediated degradation Degron domain, and (b) a UbE3L-driven complex docking the piDegron upon growth factor-signal deprivation causing piDegron dephosphorylation. Notably, this mechanism which we here define with the acronym “Over-Expression by Degradation Rescue” (OEDR) could offer a first mechanistic example of a positive feedback whereby autocrine GF-signals can acutely promote and maintain the overexpression of RTKs in cancer as well as in other RTK-dependent pathophysiological contexts. The realization of the existence of OEDR, we believe, can further intensify the search for unknown growth factor-regulated piDegrons within RTKs found expressed in the above contexts. OEDR also extends the number of post-transcriptional tools used by a cancer cell in order to maintain its malignant features beyond the known transcriptional, epigenetic and RNA transcript stabilization levels. In light of these evidence-based considerations, it is feasible to envision that specific targeting of autocrine growth factor circuits associated to RTKs’ OEDR may disclose a number of therapeutically exploitable anti-cancer effects. It is worth noticing that OEDR might similarly affect the expression of other signaling proteins beyond RTKs. However, our present perspective focuses on OEDR-regulated RTKs given the pathophysiological implications discussed herein.Control of protein stability by post-translational modifications has emerged as an important mechanism for controlling the expression and function of key cellular proteins , or phospho-inhibited Degrons (“piDegrons”). Indeed, phospho-regulated Degrons and their role in the regulation of cellular proteins expression has been recognized since a few decades . According to this proposed scenario, the UPS-degradation of RTKs bearing phospho-activating (pa) Degrons, such as those mediated by CBL reviewed in (OEDR regulation of an RTK through its phospho-inhibited (pi)Degron, such as that underlying the IGF-II/IR-EphB4 axis -associated UbE3Ls. While CBL has received various supporting evidences on its broad role in mediating degradation of RTKs  consistent with growth factors-activated negative feedback signals  Degrons versus OEDR-associated (phospho-inhibited) Degrons, under the same frame also UbE3Ls could be sub-classified as negative feedback-associated UbE3Ls versus positive feedback  is here introduced for the first time. It is proposed that OEDR-regulation of RTKs plays a constitutive and mechanistic role in previously described “hormone positive feedback loops”, under which a cellular self-stimuli provided by a growth factor promotes the upregulation and maintenance of a downstream directly phosphorylated RTK. The extent of OEDR-regulation of a cellular RTK by and autocrine growth factor, which bears the opposite function of a hormone/GF-induced negative feedback loop, is still to be clarified outside our published work. This, is partially due to the lack of optimized experimental methods for the identification and study of Degrons domains, which act as key sensors for the positive and negative feedback signals reported by the studies cited herein. Indeed, a novel experimental approach we set up for our discovery, namely a post-translational-modification-enhanced Degron bait pull-down assay (OEDR (Based upon extended revaluation of original findings contributed by the authors , 6, and wn assay , potentiwn assay  which weay (OEDR . We foreThe original contributions presented in the study are included in the article/supplementary material. Further inquiries can be directed to the corresponding author.PS: Conceptualization, Writing-original draft. SW: Writing-review & editing.


Objective. Dynamic cone-beam CT (CBCT) imaging is highly desired in image-guided radiation therapy to provide volumetric images with high spatial and temporal resolutions to enable applications including tumor motion tracking/prediction and intra-delivery dose calculation/accumulation. However, dynamic CBCT reconstruction is a substantially challenging spatiotemporal inverse problem, due to the extremely limited projection sample available for each CBCT reconstruction (one projection for one CBCT volume). Approach. We developed a simultaneous spatial and temporal implicit neural representation (STINR) method for dynamic CBCT reconstruction. STINR mapped the unknown image and the evolution of its motion into spatial and temporal multi-layer perceptrons (MLPs), and iteratively optimized the neuron weightings of the MLPs via acquired projections to represent the dynamic CBCT series. In addition to the MLPs, we also introduced prior knowledge, in the form of principal component analysis (PCA)-based patient-specific motion models, to reduce the complexity of the temporal mapping to address the ill-conditioned dynamic CBCT reconstruction problem. We used the extended-cardiac-torso (XCAT) phantom and a patient 4D-CBCT dataset to simulate different lung motion scenarios to evaluate STINR. The scenarios contain motion variations including motion baseline shifts, motion amplitude/frequency variations, and motion non-periodicity. The XCAT scenarios also contain inter-scan anatomical variations including tumor shrinkage and tumor position change. Main results. STINR shows consistently higher image reconstruction and motion tracking accuracy than a traditional PCA-based method and a polynomial-fitting-based neural representation method. STINR tracks the lung target to an average center-of-mass error of 1–2 mm, with corresponding relative errors of reconstructed dynamic CBCTs around 10%. Significance. STINR offers a general framework allowing accurate dynamic CBCT reconstruction for image-guided radiotherapy. It is a one-shot learning method that does not rely on pre-training and is not susceptible to generalizability issues. It also allows natural super-resolution. It can be readily applied to other imaging modalities as well. By STINR, we decoupled the complex spatiotemporal inverse problem of dynamic CBCT reconstruction into solving a spatial INR to represent a reference CBCT image, and several temporal INRs to represent the DVFs that characterize the time-resolved motion along different Cartesian directions. To reduce the complexity and leverage the inherent redundancy of DVFs, STINR combined PCA-based motion modeling with INR-based PCA coefficient learning to represent complex motion characteristics observed in each angle-varying projection. Compared to conventional machine/deep learning methods, STINR is a ‘one-shot’ learning technique which directly uses available cone-beam projection data to construct a patient-specific spatial and temporal imaging model that fits the specific projection set. In other words, the dynamic CBCT sequence is encoded by STINR as a neural network, which is solved on-the-fly in a self-supervised fashion with no prior training and no ‘ground-truth’ dynamic CBCTs required. Correspondingly, STINR does not suffer from the generalizability issues encountered by conventional pre-trained deep learning models  learning has gathered much interest in the artificial intelligence field , like t2.3.Minimizing the objective function in equation  solves tt figure  and recoequation ). At thiIn this study, we implemented the overall STINR framework based on the Pytorch backend (ver. 1.11.0). The optimization was performed automatically through the Pytorch framework. The initial learning rate was set to 0.002 for all the INRs. We used 500 iteration steps for stage 1, 500 iteration steps for stage 2, and 4000 iteration steps for stage 3.2.4.2.4.1.et alTo quantitatively evaluate the accuracy of the reconstructed dynamic CBCT volumes by STINR, in this study we used the 4D extended cardiac-torso (XCAT) phantom to simulate different breathing patterns and variations (Segars −1) used in clinical cone-beam projection acquisition scenarios , \documt figure , extractequation ). The obequation  was analon et al.et alIn addition to 2.4.3.et alTo quantitatively assess STINR and compare it against equation ). We alsequation ) and theon et al of the tIn equation , \documeEquation  defines 2.4.4.et alTo further assess the accuracy of STINR, we also performed a patient study in addition to the XCAT study. Compared with the XCAT phantom, the real patient images contained more detailed structures and degrading signals including scatter and noise. The patient imaging data were acquired under an IRB-approved protocol. A 4D-CBCT set was acquired for a lung cancer patient via an adaptive-speed slow-gantry rotation setting using 120 kVp, 80 mA, and 25 ms Lu et al. The proet al−1 and a frame rate of 11 frames s−1. Detailed cone-beam scan simulation parameters were summarized below in table To evaluate the accuracy and robustness of the STINR method by different breathing scenarios, we used a PCA-driven motion augmentation technique  direction was presented in figure v figure , as the 3.4.As shown in figures 4.4.1.Dynamic CBCT imaging generates volumetric images with superior temporal and spatial resolutions and is highly desired in clinical applications including radiotherapy targeting verification, tumor motion monitoring and prediction, treatment dose tracking and accumulation, and robust treatment planning. Our study developed a simultaneous spatial and temporal implicit neural representation learning (STINR) framework to address the extremely challenging problem of reconstructing dynamic CBCTs from singular x-ray projections. In comparison to traditional voxel-wise representations of volumetric images, the STINR method uses different multi-layer perceptrons to represent the spatial features and temporal evolutions of imaged anatomical structures. MLP is powerful in representing images or motions with complex, continuous, and differentiable functions, while needless to specify/constrain the detailed function forms in advance. Compared to conventional voxel-wise image representations, MLP maps the image/motion to neural networks and in theory can generate voxelized moving images of arbitrary spatial or temporal resolutions to achieve inherent super-resolution.Due to the difficulty of mapping the full spatiotemporal imaging series into a single MLP, we developed STINR to use individual MLPs to fit the spatial and temporal INRs separately, which allows the MLPs to be customized to tailor to the inherent complexity variations of different representation tasks (imaging/motion). We introduced patient-specific prior knowledge, the PCA-based motion models, into solving the temporal INRs to further reduce the complexity of the ill-conditioned spatiotemporal inverse problem. PCA-based motion modeling allows substantial dimension reductions to represent complex motion scenarios accurately and effectively.4.2.et alet alConventional PCA-based dynamic CBCT reconstruction methods solve the motion based on a fixed reference volume, and the reference volume can be reconstructed from onboard projections  to reconstruct dynamic CBCTs from singular x-ray projections acquired during a normal 60 s CBCT scan. The ‘one-shot’ learning STINR method uses the powerful representation capability of multi-layer perceptrons and adopts prior knowledge from PCA-based motion modeling. The combination successfully addresses the challenging spatiotemporal inverse problem of dynamic CBCT reconstruction. STINR can be easily adapted and implemented for other imaging modalities including magnetic resonance imaging. The spatially- and temporally-resolved images can benefit many clinical applications, including target motion tracking/prediction, intra-treatment dose calculation/accumulation, and robust planning/delivery. Future studies based on more real patient data with various motion/anatomy scenarios are warranted for a comprehensive evaluation and potential methodology fine-tuning.


Coronavirus disease 2019 (COVID-19) is a current pandemic caused by SARS-CoV-2 that has vastly affected the whole world. Although respiratory disease is the most common manifestation of COVID-19, the virus can affect multiple organs. Neurotropic aspects of the virus are increasingly unfolding, in so far as some respiratory failures are attributed to brainstem involvement. The neuro-ophthalmic manifestations of COVID-19 and the neuro-ophthalmic side effects of vaccination were reviewed. The major findings are that the SARS-CoV-2 infection commonly causes headaches and ocular pain. It can affect the afferent and efferent visual pathways by ischemic or inflammatory mechanisms. Optic nerve may be the origin of transient or permanent visual loss from papillophlebitis, idiopathic intracranial hypertension, or optic neuritis. Cerebrovascular strokes are not uncommon and may lead to cortical visual impairment or optic nerve infarction. SARS-CoV-2 may affect the pupillomotor pathways, resulting in tonic pupil (Adie's syndrome) or Horner's syndrome. Cranial neuropathies including third, fourth, sixth, and seventh nerve palsies have all been reported. Rhino-orbital mucormycosis superinfections in COVID-19 patients receiving steroids or other immunosuppressive therapies may result in unilateral or bilateral visual loss and ophthalmoplegia. Autoimmune conditions such as Guillain-Barré, Miller-Fisher syndrome, and ocular myasthenia have been reported. The disease presents in a widely variable range from asymptomatic carriers or mild infection to severe acute respiratory syndrome (SARS), multi-organ failure, and death. Besides respiratory distress syndrome, SARS-CoV-2 may affect many other organs and cause cardiac or renal injury, dermatologic conditions, gastrointestinal symptoms, coagulopathy, severe inflammatory reaction, and central or peripheral nervous system involvement. Up to one-third of patients with COVID-19 have neurological complications, and the incidence appears to be higher in patients with more severe infections. The most common severe neurologic manifestations of COVID-19 are acute cerebrovascular disease and altered level of consciousness. A retrospective study in Wuhan reported neurological symptoms in 36.4% of the hospitalized patients. Neurologic manifestations are mainly reported to be associated with severe cases of COVID-19.,5 Moreover, COVID-19-related cases of ischemic stroke involving visual pathways have resulted in prolonged hospitalizations and fatality.,7 Findings have suggested that ischemic strokes in patients with COVID-19 disease are more severe and disabling than strokes in the uninfected subjects. In contrast, isolated cranial neuropathies involving the eye are typically associated with mild to moderate COVID-19 disease that improves spontaneously or with local protocols.,10Coronavirus disease-2019 (COVID-19) is a novel β coronavirus of group 2B, first reported in December 2019 in Wuhan city, that rapidly disseminated all over the world, resulting in an immense pandemic. Based on an initial study in Wuhan, visual impairment was described in 3 out of 214 (1.4%) hospitalized patients with COVID-19.A recent review reported that over 4% of patients with COVID-19 had ophthalmic findings requiring clinical attention.Various neuro-ophthalmologic manifestations have been reported in patients with COVID-19 infection. In this article, we reviewed and categorized SARS-CoV-2-related neuro-ophthalmic presentations.,5 It is thought that direct viral invasion is mediated by viral fusion to the angiotensin-converting-enzyme 2 (ACE-2) receptor, which is expressed on the surface of pulmonary type II alveolar epithelial cells as well as neurons and glial cells. The SARS-COV-2 virus can affect nearly all neuro-ophthalmic pathways [Table 1]. COVID-19 can cause a wide range of pathologies within the afferent and efferent visual pathways. SARS-CoV-2 can cause central nervous system conditions such as seizures, anosmia or ageusia, altered level of consciousness, posterior reversible encephalopathy syndrome (PRES),,16 neuromyelitis optica (NMO) spectrum disorder,,17 myelin oligodendrocyte glycoprotein (MOG)-associated disease,,20,21,22 acute disseminated encephalomyelitis (ADEM), cerebral venous sinus thrombosis (CVST),,25 and cerebrovascular strokes.6,7]8],7[8]6,7],7[8]6,7] It can also affect the peripheral nervous system, leading to conditions such as Guillain-Barré, Miller Fisher syndrome,,28 polyneuritis cranialis, and myasthenia gravis.,30Several mechanisms have been postulated for neurologic involvement in COVID-19 disease. Direct viral invasion, hypoxia, hypercoagulable state, inflammatory reactions related to cytokine storm, delayed autoantibody formation, endothelial dysfunction, and retrograde axonal transport of the infection via cranial and peripheral nerves  are the proposed underlying mechanisms for neuro-ophthalmic manifestations of COVID-19.,31,32,33 or immune-mediated),4,17]21][[21][20][[[21][20] optic nerve infarction, papillophlebitis, and idiopathic intracranial hypertension.,37,38 The efferent visual pathways can be lesioned by COVID-19-related isolated cranial neuropathies,,39,40,41 nystagmus,,44,45,46 tonic pupil,,49,50,51 Horner's syndrome,,13,52 and opsoclonus-myoclonus-ataxia syndrome (OMAS).,54,55 Mucormycosis infections, which are of exponentially increasing incidence in COVID-19 patients, can occur as a superinfection and result in lesions of the afferent or efferent visual pathways or even both.,58SARS-CoV-2 is associated with pathologies of the afferent visual pathways including optic neuritis , and OMAS were all postulated to have delayed-immune mediated mechanisms related to COVID-19. The delayed immune response may be from antibodies directed against SARS-CoV-2 proteins that cross-react with cellular proteins through a mechanism known as molecular mimicry. Alternatively, COVID-19 infection may impair immunologic self-tolerance.Neuro-ophthalmic manifestations of COVID-19 may present either concurring with systemic and pulmonary symptoms or days to several weeks after their resolution. Timeline latency between acute viral symptoms and neuro-ophthalmic manifestation, auto-antibody formation, and good response to steroid therapy favor an immune-mediated mechanism.,11,59 Headache was reported in 71% and ocular pain in 34% by healthcare workers in the Netherlands in patients with a positive SARS-CoV-2 polymerase chain reaction (PCR). In another study, eye pain was amongst the most common of the ocular symptoms and seen in 16% of patients. Photophobia and itchy eyes were seen at a similar frequency .Ocular pain and headache are common and may be the initial manifestation of COVID-19 infection. However, the exact mechanism is not clear, and viral encephalitis and meningitis should be considered.Headache characteristics in COVIDOverall, it seems that headache and periocular pain are the most prevalent neuro-ophthalmic presentations of COVID-19 but more comprehensive epidemiological studies are required to evaluate the prevalence of each neurological symptom.,31,32,33 COVID-19 has also been the harbinger to optic neuritis in NMO spectrum disorder,17 and MOG-related disease.,21,63,64 Optic neuritis in association with COVID-19 infection has also been reported in the pediatric age group even in the context of demyelinating conditions. COVID-19-related optic neuritis had a good response to intravenous (IV) corticosteroid therapy in all cases except four.,14,22,32Optic neuritis has been reported in association with COVID-19 infections in multiple cases where no other etiologies have been determined.Optic neuritis was reported in conjunction with panuveitis in one case of COVID-19, which resulted in optic atrophy despite corticosteroid treatment. However, the patient only received oral prednisone without high-dose IV corticosteroids. The authors postulated that ischemic insult due to the prothrombotic capacity of COVID-19 or an inflammatory reaction resulted in optic atrophy in this case.Novi et al reported an adult patient with COVID-19 disease and bilateral retrobulbar optic neuritis with enhancing lesions in optic nerves, brain, and thoracic spine, consistent with ADEM. Optic nerve infarction due to internal carotid artery occlusion was reported in a 50-year-old COVID-positive male by Tavakoli et al. Central retinal artery occlusion or ophthalmic artery occlusion due to internal carotid artery stroke have also been reported in COVID-19 disease.Cerebrovascular ischemia is a known complication of SARS-CoV-2 infection.There is one report of papillophlebitis in a 40-year-old male that occurred four weeks after the resolution of a typical COVID-19 infection with elevated D-dimer and fibrinogen levels. The authors proposed that the COVID-19 cytokine storm induced a hypercoagulable state and thrombotic microangiopathy, increasing the risk of papillophlebitis.,37,38 Two of these cases were associated with increased intracranial pressure inducing sixth nerve palsies in idiopathic intracranial hypertension due to COVID-19 multisystem inflammatory syndrome in children (MIS-C).,37Idiopathic intracranial hypertension can occur in adults with COVID-19 disease, but it is rare, and most of the cases have been reported in children. and bilateral cases. One patient with COVID-19-related bilateral tonic pupil also had inflammatory multifocal choroiditis. Additionally, Adie-Holmes syndrome  has been described in a patient with COVID-19 infection. Lastly, Ordás et al reported a case of tonic pupil with contralateral trochlear palsy in association with COVID-19 infection.Tonic (Adie's) pupil refers to parasympathetic denervation of the pupil sphincter, which causes poor constriction of the pupil to the light but the reaction to the accommodation remains relatively spared. Tonic pupil has been reported in association with COVID-19, including unilateral In each of the above cases of tonic pupil, there was a delay (ranging from two days to one month) between the onset of COVID-related respiratory symptoms and development of tonic pupil. Most of the authors hypothesized that tonic pupil in COVID-19 is secondary to a post-viral delayed immune-mediated injury rather than direct viral entry into the central nervous system. Moreover, a taper of prednisolone has been utilized for treatment with subsequent improvement in the tonic pupil.Tonic pupil is often idiopathic, but it has been associated with other infections , autoimmune processes, trauma, choroidal and orbital tumors, and surgery.,13,52 In each case, COVID-19 pneumonia involving the upper part of the lungs was the suggested etiology of Horner's syndrome, and no other causes were discovered with subsequent imaging studies.Horner's syndrome has also been rarely described in COVID-19 disease.,70 There are reports of ischemic stroke involving the visual pathways in association with SARS-CoV-2 infection, resulting in homonymous field defects or even cortical blindness from bilateral occipital infarcts. Bondira et al reported a patient with COVID-19 disease and bilateral occipital strokes, resulting in a partial right homonymous hemianopsia and difficulty with reading. Priftis et al presented a COVID-positive patient with a left occipito-temporal ischemic stroke, resulting in alexia without agraphia syndrome and right homonymous hemianopsia.COVID-19 disease can induce hypercoagulable and inflammatory states, increasing the risk of cerebrovascular stroke. or even hallucinatory palinopsia. COVID-associated CVST has also been described but is uncommon, and only a few cases have been associated with blurry vision and papilledema.,25 The disproportionally low frequency of reported visual involvement in COVID-associated CVST may be because of the severity of other symptoms or an altered level of consciousness.COVID-induced PRES has been reported, resulting in transient cortical visual loss,72In addition, there are reports of visual loss due to pituitary apoplexy in the context of acute COVID-19 infection.Damage to third, fourth, sixth, and seventh cranial nerves have all occurred in timeline relation to COVID-19 infection. Cranial nerve involvement may develop as a primary and isolated insult or in the context of a more generalized condition such as Miller Fisher syndrome, Guillain-Barré, myasthenia gravis, venous sinus thrombosis, and increased intracranial pressure.,40,73 Similarly, isolated abducens palsy has been described in COVID-19.,27 In these cases, there was no enhancement of the abducens nerve pathway on MRI, and patients had underlying systemic hypertension. Such findings raised the hypothesis that uncontrolled hypertension in the acute viral illness may cause abducens palsy. An alternative theory was that the abducens palsies occurred from the leptomeningeal invasion of SARS-CoV-2, given that two of these cases had optic nerve sheath enhancement on MRI. As previously mentioned, there is one case of unilateral trochlear nerve palsy with contralateral tonic pupil and another with bilateral trochlear nerve palsy in the setting of COVID-related cerebral vasculitis.Isolated oculomotor palsy with and without pupillary involvement has been reported in association with COVID-19.Different medications for COVID-19-related cranial neuropathies such as hydroxychloroquine, azithromycin, corticosteroids, and IV immunoglobulin have been used. However, it is not known whether the neuro-ophthalmic benefit was conferred. The deficits partially improved following IV immunoglobulin infusions. Gutiérrez-Ortiz et al detailed a similar case of Miller Fisher syndrome in a patient with right fascicular oculomotor palsy, right internuclear ophthalmoparesis, ataxia, areflexia, albuminocytologic dissociation, anosmia, ageusia, and positive reverse transcriptase-PCR for SARS-CoV-2. Most of the deficits were resolved following IV immunoglobulin infusions. These authors described a second COVID-positive case with bilateral abducens palsy, areflexia, and albuminocytologic dissociation without ataxia or limb weakness secondary to polyneuritis cranialis.Cranial neuropathy in relation to COVID-19 has also been seen in the context of Miller Fisher syndrome, Guillain-Barré, and polyneuritis cranialis. Dinkin et al described a case of presumed Miller Fisher syndrome in a COVID-positive patient with a partial left pupil involving oculomotor nerve palsy, bilateral abducens palsy, hyporeflexia, and gait ataxia.In a systematic review published by Sansone et al, of the 41 patients with SARS-CoV-2-related Guillain-Barré syndrome, 9 (22%) had diplopia, 7 (17%) had ocular palsy, and 13 (32%) had facial palsy. The average time between the onset of SARS-CoV-2 symptoms and the first neurological signs of Guillain-Barré syndrome was 10 days. Fatigability and other myasthenic symptoms started four to seven days after the initial COVID-19 manifestations.,30 Similarity between SARS-CoV-2 epitopes and neuromuscular junction components is postulated to induce molecular mimicry, resulting in auto-antibody formation and subsequently post-COVID-19 myasthenia gravis. Isolated peripheral facial nerve palsy has also been reported either during or at the onset of the SARS-CoV-2 clinical course. In one of the eight peripheral facial nerve palsy cases reported by Lima et al, ipsilateral abducens palsy was also present. Facial diplegia has also been reported as a parainfectious complication of COVID-19 disease and was considered a rare variant of Guillain-Barré syndrome in the setting of concomitant albuminocytologic dissociation.COVID-19 has also been the harbinger of new diagnoses of myasthenia gravis. Restivo et al detailed three patients who developed diplopia, ptosis, or dysphagia and tested positive for elevated serum acetylcholine receptor antibodies following the SARS-CoV-2 infection. and may be due to various underlying etiologies including acute labyrinthitis, benign paroxysmal positional vertigo, rhombencephalitis, and Bickerstaff encephalitis.Acquired nystagmus has been seen in association with COVID-19,,54,55 In all patients who were tested, work-up with a paraneoplastic panel was unrevealing for other causes of OMAS. A variety of infections besides SARS-CoV-2 including human immunodeficiency virus (HIV), West Nile virus, Epstein Barr virus, and enterovirus have also been associated with OMAS. Treatments used for COVID-associated OMAS include various combinations of the following: IV immunoglobulin, levetiracetam, sodium valproate, clonazepam, and corticosteroids. All cases were associated with a good therapeutic response and partial to complete recovery. Emamikhah et al stated that the dramatic effect of immunotherapy on recovery suggests an immune-mediated parainfectious mechanism for COVID-related OMAS.Several cases of OMAS have been described in COVID-positive patients. A retrospective, observational study was conducted in India involving 2826 patients with COVID-associated ROCM. The most common primary signs included periocular/facial edema (33%), loss of vision (21%), ptosis (12%), and proptosis (11%). Ocular movement restriction and diplopia were infrequent signs (3%). Most (87%) of the patients had received systemic corticosteroids prior to developing COVID-associated ROCM, and it was the most common risk factor for the condition followed by diabetes mellitus.There is strong evidence that infection with SARS-CoV-2 increases the risk of secondary fungal infections, notably rhino-orbital-cerebral mucormycosis (ROCM). The incidence of ROCM increased to epidemic proportions during the second wave of the COVID-19 pandemic in India.,78,79 These complications occurred after all types of vaccines. They include acute ischemic stroke, intracranial hemorrhages, and cerebral venous sinus thrombosis.,80 Other reported neuro-ophthalmic complications include: bilateral arteritic anterior ischemic optic neuropathy (AAION), post-vaccination cranial neuropathy, that is, third and sixth,83,84 nerve palsies, facial palsy,,85,86,87 multiple cranial nerve palsies, pupillary abnormalities such as Horner's syndrome, Holmes-Adie pupil, miosis and mydriasis, eight nerve involvement and benign paroxysmal positional vertigo, and post-vaccination optic neuritis.,91,92 In multiple sclerosis patients, BNT162b2 mRNA vaccine was reported to be associated with a transient increase of MS symptoms in 3.8% but none of them needed treatment.There are several reports of neuro-ophthalmic complications following COVID-19 vaccination.Coronavirus Disease 2019 and also all types of vaccines that have been made against it can affect the afferent and efferent visual pathways by ischemic or inflammatory mechanisms. The optic nerve may be the origin of transient or permanent visual loss from papillophlebitis, idiopathic intracranial hypertension, or optic neuritis. Cerebrovascular strokes are not uncommon and may lead to cortical visual impairment. The pupillomotor pathways and cranial nerve including the third, fourth, sixth, and seventh nerve may also be involved. Early detection of the neuro-ophthalmic features of COVID-19 disease/vaccine will assist in preventing vision loss and oculomotor deficits.This article is authors' own work and has no sources of support.The authors have no financial interest in the subject of this article.


Protein structure provides insight into how proteins interact with one another as well as their functions in living organisms. Protein backbone torsion angles (We present SAINT-Angle, a highly accurate method for predicting protein backbone torsion angles using a self-attention-based deep learning network called SAINT, which was previously developed for the protein secondary structure prediction. We extended and improved the existing SAINT architecture as well as used transfer learning to predict backbone angles. We compared the performance of SAINT-Angle with the state-of-the-art methods through an extensive evaluation study on a collection of benchmark datasets, namely, TEST2016, TEST2018, TEST2020-HQ, CAMEO and CASP. The experimental results suggest that our proposed self-attention-based network, together with transfer learning, has achieved notable improvements over the best alternate methods.https://github.com/bayzidlab/SAINT-Angle.SAINT-Angle is freely available as an open-source project at Bioinformatics Advances online. Proteins are responsible for various functions in cells and their functions are usually determined by their 3D structures. However, the experimental determination of protein structures using X-ray crystallography, cryogenic electron microscopy (cryo-EM) and nuclear magnetic resonance spectroscopy is costly and time- and labour-intensive (Given the growing availability of protein databases and rapid advances in machine learning (ML) methods , application of ML techniques to leverage the available data in accurate prediction of backbone angles has gained significant attention.Earlier ML-based methods used neural network , supportal-SPINE  leverageSeveral deep learning-based techniques have recently been developed that can predict backbone torsion angles with a reasonable accuracy. SPIDER2  to captuSPOT-1D , bidirecIn this study, we present SAINT-Angle, a highly accurate method for protein backbone torsion angle prediction, which is built on our previously proposed architecture of self-attention augmented inception-inside-inception network (SAINT) , CAMEO  ProtTrans features and (iii) Window features.SAINT-Angle takes a protein sequence feature vector Base feature class consists of a feature vector of length 57 for each residue. It contains features from PSSM profiles, HMM profiles and physicochemical properties (PCP) , polarizability, normalized van der Waals volume, hydrophobicity, isoelectric point, helix probability and sheet probability] were obtained from The Uniref90  databaseProtTrans features, generated by the pretrained language model for proteins developed by The Window features are generated by windowing the predicted contact information as was done in SPOT-1D and was subsequently used in SAINT. We used SPOT-Contact to generate the contact-maps. We varied the window lengths (the number of preceding or succeeding residues whose pairwise contact information were extracted for a target residue) to generate different dimensional features. We used three different window lengths The architecture of SAINT-Angle can be split into three separate discussions: (i) the architecture of SAINT  ProtTrans architecture and (iii) Residual architecture.We developed the following three architectures that we utilize in an ensemble network to create: (i) Basic architecture which is identical to the original SAINT architecture proposed for protein SS prediction  we have added residual connections  and predicted values (P) over all amino acid residues in a protein. The minimum value between N is the number of proteins, ith protein. jth amino acid residue in the ith protein, respectively. We performed Wilcoxon signed-rank test  are notable—indicating the successful utilization of ProtTrans-based transfer learning. Statistical tests suggests that these improvements of SAINT-Angle over other methods are statistically significant  is shown in We further compared the backbone torsion angles prediction performance of SAINT-Angle with the best existing methods on a newly introduced dataset TEST2020-HQ dataset, which was previously assembled and analyzed by SPOT-1D-LM . Given tThe performance comparison of SAINT-Angle with other methods is shown in Basic architecture with ESIDEN features SAINT-Angle-Single. We obtained the evolutionary features for the SPOT-1D training dataset and a collection of test datasets from the authors of ESIDEN. We also trained our ensemble network using the base features along with the novel ESIDEN features . The evolutionary features of ESIDEN were shown to be reasonably powerful . Note that these CASP datasets [analyzed in ESIDEN —performs notably well. Therefore, pretrained protein language model-based methods do not appear to require a certain level of homologous sequences for reasonably accurate prediction. On the other hand, MSA-based methods are less successful on ‘orphan proteins’—proteins with few or no homologs . Chowdhuvbad042_Supplementary_DataClick here for additional data file.


Among patients with OUD, those who inject or use high doses of opioids likely require higher doses of buprenorphine to maximize treatment efficacy. The objective of this analysis was to compare the efficacy and safety of 100-mg versus 300-mg maintenance doses of BUP-XR in OUD patients who inject opioids.BUP-XR  or placebo for 24 weeks. Abstinence was defined as opioid-negative urine drug screens combined with negative self-reports collected weekly. Each participant’s percentage abstinence was calculated after the first, second, and third maintenance doses in opioid-injecting and non-injecting participants. The proportion of participants achieving opioid abstinence in each group was also calculated weekly. Treatment retention rate following the first maintenance dose was estimated for opioid-injecting participants with Kaplan–Meier method. Risk-adjusted comparisons were made via inverse propensity weighting using propensity scores. Buprenorphine plasma concentration–time profiles were compared between injecting and non-injecting participants. The percentages of participants reporting treatment-emergent adverse events were compared between maintenance dose groups within injecting and non-injecting participants separately.BUP-XR 100-mg and 300-mg maintenance doses were equally effective in non-injecting participants. However, in opioid-injecting participants, the 300-mg maintenance dose delivered clinically meaningful improvements over the 100-mg maintenance dose for treatment retention and opioid abstinence. Exposure–response analyses confirmed that injecting participants would require higher buprenorphine plasma concentrations compared to non-injecting opioid participants to achieve similar efficacy in terms of opioid abstinence. Importantly, both 100- and 300-mg maintenance doses had comparable safety profiles, including hepatic safety events.These analyses show clear benefits of the 300-mg maintenance dose in injecting participants, while no additional benefit was observed in non-injecting participants relative to the 100-mg maintenance dose. This is an important finding as opioid-injecting participants represent a high-risk and difficult-to-treat population. Optimal buprenorphine dosing in this population might facilitate harm reduction by improving abstinence and treatment retention.Trial registration: ClinicalTrials.gov, NCT02357901.The online version contains supplementary material available at 10.1186/s12954-023-00906-7. The OI population was slightly younger; had higher percentages of males, Whites, Hispanics/Latinos, tobacco smokers, participants with a history of cocaine and polydrug use, and participants with psychiatric disorders; and had slightly longer lifetime opioid use and less employment at screening, compared to NIO (Table The distribution of participant risk factors (included in the propensity model) across the 2 maintenance doses was similar. After inverse probability weighting, the difference in covariates was reduced and within 5% of the standardized difference, indicating a good balance of baseline risk factors. Participants’ percentage of abstinence during the maintenance dose period is shown for both OI and NIO populations in Table Two possible factors might contribute to the higher mean percentage of abstinence in the 300-mg maintenance dose group for the opioid-injecting participants: (1) better treatment retention or 2) higher opioid abstinence while maintained on the 300-mg maintenance dose. Two additional risk-adjusted analyses were thus performed to evaluate potential differences on treatment retention and opioid abstinence for those who remained on treatment during the maintenance dose period. Consistent with their percentage abstinence, OI who received BUP-XR 300 mg during the maintenance dose period had higher retention rates than those who received BUP-XR 100 mg  relationship to characterize the effect of buprenorphine plasma concentration. Participants who injected opioids at baseline had a 3.6-fold higher buprenorphine EC50  compared to those who used opioids by non-injectable routes , suggesting that this subpopulation required higher buprenorphine concentrations to optimize buprenorphine efficacy. The current research not only consolidates those findings but extends this previous work by showing that in opioid-injecting participants, the efficacy of the higher maintenance dose results in improved treatment retention.The results of secondary analyses align with our previous exposure–response modeling work conducted on opioid abstinence using the same Phase 3 data . The promu-opioid receptor. Similarly, individuals achieving high opioid concentrations by intravenous injection are expected to require higher buprenorphine plasma concentrations to block the rewarding and reinforcing effects of opioids. Additional evidence of the benefit of high buprenorphine doses in higher-dose opioid participants also comes from other clinical studies. A retrospective study of individuals who use heroin on methadone maintenance treatment showed that their odds of using heroin decreased with every 1 mg increase in methadone maintenance dose [Findings also align with earlier literature demonstrating that higher doses of buprenorphine are needed to block the subjective effects of higher doses of a full agonist such as hydromorphone , 41–45. nce dose . In addince dose .From a safety perspective, both maintenance doses of BUP-XR were well tolerated, as previously reported . Interesmu-receptor signaling arising from rapid-onset, high-dose exposure associated with injection use. If OI are more opioid-tolerant than NIO, we hypothesize they would: (1) under controlled conditions, exhibit attenuated responses to a standard opioid challenge, as found in a preliminary study [One plausible explanation for the present findings is that, on average, OI are more opioid-tolerant than non-injecting participants, possibly due to changes in ry study , (2) in Overall, the current findings have implications for harm reduction approaches in the treatment of OUD across 3 key domains: public health, clinical practice, and patient-centeredness . InjectiA potential limitation of this study is that we did not compare BUP-XR 300 mg versus 100 mg during the maintenance dose period in the randomized intent-to-treat population. As a result, the treatment comparison might be subject to the bias introduced by participants’ discontinuation prior to receiving the maintenance dose. We attempted to limit such bias by performing the efficacy comparison via risk adjustment including both baseline and post-randomization variables. Despite these efforts, unmeasured confounders may have influenced the accuracy of efficacy comparison results. To overcome this limitation and confirm the benefit of BUP-XR 300-mg maintenance dose observed in the OI, a clinical study is currently ongoing to compare the efficacy, safety, and tolerability of the 100-mg versus 300-mg BUP-XR maintenance doses in participants with high opioid tolerance, including those who inject opioids and/or use high doses of opioids. In this study, treatment-seeking, high-risk opioid participants are randomized immediately prior to the first maintenance dose and receive twice-longer maintenance treatment than in the Phase 3 study used in the present analysis (8 vs. 4 maintenance doses).The 100-mg maintenance dose is well tolerated and may achieve sufficient efficacy outcomes in most NIO. In OI—a high-risk and difficult-to-treat population—the benefit of BUP-XR 300-mg maintenance dose is clinically relevant and may reduce harm by improving abstinence and treatment retention.Additional file 1.Supplemental Material. Supplemental Figure 1. Treatment Retention and Abstinence in Non-injecting Opioid Participants. Supplemental Figure 2. Mean (±SD) Buprenorphine Plasma Concentration-Time Profiles in Injecting vs. Non-injecting Opioid Participants for the Two BUP-XR Dosing Regimens.


The most serious problem for people suffering from severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection is liver damage. The liver is a frequently affected organ due to the metabolizing and detoxifying functions of several endogenous and exogenous molecules. In COVID-19-affected individuals, even moderate loss of hepatic function could dramatically affect the therapeutic efficacy of antiviral drugs metabolized in the liver. The clear mechanism of hepatocellular damage from SARS-CoV-2 infection is not fully understood. The main objective of this review is to identify potential mechanisms of SARS-2 induced liver damage, treatment outcomes in SARS-CoV-2-infected patients, and future direction. Electronic databases including Web of Science, Google Scholar, MEDLINE, Scopus, and Cochrane library were used to systematically search without limitation of publication date and status. Observational, retrospective cohort, prospective case-control, cohort studies, cross-sectional studies, or clinical trials were included. Liver damage in coronavirus patients is characterized by histopathological changes and abnormal elevation of some liver function tests. These abnormalities include elevation of Alanine aminotransferase, Aspartate aminotransferase, Gamma-glutamyl transferase, Alkaline phosphatase, and Serum bilirubin levels. Histopathological changes of the liver might consist of complete or partial thrombosis of the portal and sinusoidal vessels, portal tract fibrosis, and focally markedly enlarged and fibrotic hepatocytes. Understanding the fundamental molecular and immunological processes of COVID-19-related liver injury is essential for the selection of appropriate drugs and the logical development of successful treatment. Even though most COVID-19 patients suffer from pulmonary symptoms, approximately 5% of individuals suffer from multi-organ dysfunction, which increases of morbidity and mortality from the disease. SARS-CoV-2 is an enveloped virus that can access the cell via angiotensin-converting enzyme 2 (ACE2), the main receptor for this virus. The attachment of SARS-CoV-2 to hepatocyte cells may be mediated by the interaction of the spike (S) protein and host receptor called ACE2. Following attachment of the virus to the host receptor, the S protein is cleaved by the transmembrane serine protease 2, which allows entry of the virus into the host cell.,4In Wuhan, People’s Republic of China, pneumonia of unknown origin was first noticed and reported to the World Health Organization in December 2019. Severe acute respiratory syndrome corona virus-2 (SARS-CoV-2) was the designation given to the illness after it was later determined to be caused by a new coronavirus.,6 Previous studies have also suggested that the S glycoprotein of SARS-CoV-2 may use another transmembrane protein known as CD147 receptor which is highly expressed in hepatocyte cells.–9The direct effect of SARS-CoV-2 on the liver may not be fully achieved through ACE2 alone. According to some studies, dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin and liver-specific intercellular adhesion molecule-3-grabbing non-integrin (L-SIGN) are the alternative receptors that promote ACE2-mediated virus infection of hepatic cells.,11 Viral replication also occurs in double-membrane vesicles. Additionally, one of the nonspecific proteins called nsp-6 mediated inhibition of auto phagosome and lysosome expansion to favor viral replication. Then after, newly synthesized viral components assemble to form the nucleocapsid and viral envelope at the Endoplasmic reticulum–Golgi intermediate compartment of the hepatocyte. Finally, mature virions are then released through the exploitation of the host vesicular system.,13Once entered into the host cell, the viral components are translated by the host ribosome into polyproteins (pp1a and pp1b) that further cleave into 16 nonstructural proteins.Electronic databases including Web of Science, Google Scholar, MEDLINE, Scopus, and Cochrane library were used to systematically search without limitation of publication date and status. Observational, retrospective cohort, prospective case-control, cohort studies, cross-sectional studies, and clinical trials were included.The protocol of the review does not require ethical approval because it does not involve humans. This article will be published in a peer-reviewed journal and presented at relevant conferences.–16The most serious problem for people suffering from SARS-CoV-2 infection is liver damage. Liver is frequently injured due to the metabolizing and detoxifying functions of several endogenous and exogenous molecules. A COVID-19-associated liver injury can be explained as any liver injury that occurs during the course of the disease or treatment in patients with COVID-19. Liver damage in COVID-19 patient is usually characterized by abnormal liver function tests. Moreover, affected liver cells could impair the therapeutic efficacy of antiviral drugs.,18 Some laboratory findings following SARS-CoV-2 induced liver damage are elevated liver enzymes like Alanine aminotransferase (ALT), aspartate aminotransferase (AST), Gamma-glutamyl transferase, Alkaline phosphatase (ALP), and Serum total bilirubin levels. Additionally, histopathological changes of the liver might encompass complete or partial thrombosis of portal and sinusoidal vessels, portal tract fibrosis, and fibrotic hepatocytes.Additionally, infection with SARS-CoV-2 impairs the function of cholangiocellular tight junction proteins which have a protective role in a normal liver. Morphological impairment of the liver can cause the leakage of toxic bile component into the liver parenchyma and cause liver injury.,20 Understanding the basic molecular mechanism and processes of COVID-19-related liver injury is essential for the selection of appropriate drug and rational development of successful treatment.Even though the clear mechanism of liver damage in SARS-CoV-2-infected patients is not fully understood, some proposed mechanisms were due to the direct cytopathogenic effects of the SARS virus, cytokine storms, hypoxic from respiratory failure, and certain drugs used for the management of COVID-19. Single-cell RNA-sequencing analysis found that there is a significant ACE2 expression in cholangiocytes.,23 ACE2 receptors in the liver cells are expressed mainly in cholangiocytes. However, this receptor is minimally expressed in hepatocytes, and almost absent in Kupffer cells.,25 Following the invasion of liver cell by SARS-CoV-2, at early phase the expression of ACE2 in hepatocytes will increase. Systemic inflammation signals such as type I interferon or Interleukin-6 (IL-6),27 also enhance the expression of ACE2 receptors of hepatocyte cells. This mechanism facilitates the easy accessibility of the virus to liver cells. SARS-CoV-2 invades hepatocytes and cholangiocytes through various classical receptors, including Toll-like receptors, Nod-like receptors, and RIG-I-like receptors. Active replication and virus release by infected cells can lead to proptosis, which is responsible for releasing damage associated molecular patterns. When hepatocytes recognize damage associated molecules, proinflammatory cytokines and chemokines, including IL-6, are released. This process switch-on cellular signaling and cytokine molecules attract professional leukocytes like monocytes, macrophages, and T cells to the site, increasing inflammation. Therefore, IL-6 can act to intensify cytokine response syndrome activation. Furthermore, SARS-CoV-2 will downregulate ACE2 as the disease advances. As a result, angiotensin II accumulates and acts as a proinflammatory cytokine via the AT1R-metalloprotease 17 axis to intensify hepatocyte damage.Many recent studies are conducted to know the exact mechanism of liver damage by SARS-CoV-2 virus among COVID-19 patients. Some of the recent findings propose that SARS-CoV-2 enters alveolar epithelial cells and hepatocytes via ACE2 receptors.–32Overactivation of immune cells and proinflammatory cytokines causes a cytokine storm that exacerbates further hepatocyte damage. In the liver, hepatocellular damage caused by proinflammatory cytokines can trigger Kupffer cell activation. Activated Kupffer cells release proinflammatory cytokines and reactive oxygen species, which can continue hepatocyte injury. Nitric oxide generated by activated neutrophils can lead to peroxynitrite formation, cause mitochondrial damage, and induce liver cell necrosis.–36One of the acute complications in COVID-19 patients is severe respiratory distress that can cause tissue hypoxia. When the oxygen delivery to the liver is compromised, liver ischemic injury will develop. Insufficient oxygen supply to the liver will end up with metabolic disturbances resulting from abnormal lipid accumulation, and adinosine triphosphate depletion that leads to hepatocyte cell death. In reperfusion injury, damaged liver cells release damage associated molecular patterns, which activate the complement cascade, and mitochondrial reactive oxygen species production that causes ultimate reperfusion injury in the liver. Additionally, hepatic mitochondrial swelling and narrowed glycogen granules were observed in the SARS-CoV-2-infected liver cell, which indicated hepatic injury caused by direct cytotoxicity of SARS-CoV-2.SARS-CoV-2 can also promote intracellular cytotoxicity to hepatocytes through membrane destruction and mitochondrial dysfunction. The virus might also activate rapamycin signaling, which inhibits autophagy and facilitates viral escape from the immune system of the host cell. Several findings have been proposing that the virus potentially causes damage to bile duct epithelial cells. There is evidence that typical coronavirus particles were identified in the cytoplasm of hepatocytes. Many infected patients were treated with antipyretic agents containing acetaminophen, a drug that can cause significant liver damage. It is known that acute ingestion of > 7.5 to 10 g of acetaminophen in adults or 150 to 200 mg/kg in children is likely to cause hepatotoxicity. Due to the hepatotoxicity of these drugs, the US Food and Drug Administration Committee has proposed reducing the maximum daily dosage of acetaminophen from 4 to 3 g to minimize drug-related liver damage in SARS-CoV-2 patients.–42Another set of histological findings from liver biopsies of patients with SARS suggested that SARS-CoV-2 may induce apoptosis of hepatocytes.–45 SARS-CoV-2 may damage the biliary tract which increases indirect and total bilirubin, gamma-glutamyl transferase (GGT), and ALP levels. Another study proposed that there are variable levels of increments of GGT, ALP, and ALT observed in 58% to 78% of patients.,46From the above mechanisms of liver damage, it is evident that individuals presenting with COVID-19 have a hepatic injury and abnormal liver function tests. The histopathological changes of the liver might include complete or partial thrombosis of the portal and sinusoidal vessels, portal tract fibrosis, and focally markedly enlarged and fibrotic hepatocytes. Studies have shown an increase in some liver biomarkers, particularly the elevation of ALT and AST levels due to hepatocellular damage. Mild increases in AST and ALT levels have been found in 14% to 53% of severe SARS-CoV-2 infection cases.An observational cohort study done in China, in Wuhan, with 2912 COVID-19 patients, found that 1414 patients (48.6%) had abnormal LFTs, with ALT, AST, total bilirubin, ALP, and GGT elevation in 662 (22.7%), 221 (7.6%), 52 (1.8%), 135 (4.6%), and 536 (18.5%), respectively. According to this study, abnormal liver function tests are an independent risk factor for mortality, intensive care unit admission, and mechanical ventilation among COVID-19 patients.Liver damage is the most serious issue for those with SARS-CoV-2 infection. Any liver damage sustained by COVID-19 patients throughout their illness or treatment is referred to as a COVID-19-associated liver injury. Abnormal liver function tests are typically used to identify liver disease in COVID-19 patients. Moreover, damaged liver cells may reduce the therapeutic effectiveness of antiviral medications. Although the precise cause of liver damage in SARS-CoV-2-infected patients is still unknown, several theories have been put forth, including those involving the direct cytopathogenic effects of the SARS virus, cytokine storms, hypoxia brought on by respiratory failure, and specific medications used to treat COVID-19.Proinflammatory cytokines and immune cell overactivity result in a cytokine storm that exacerbates further hepatocyte injury. Severe respiratory distress, which may result in tissue hypoxia, is 1 acute consequence of COVID-19 in patients. A liver ischemia injury develops if the liver’s oxygen delivery is impaired. Inadequate oxygen delivery to the liver will result in aberrant lipid accumulation, metabolic abnormalities, and adinosine triphosphate depletion, which kills hepatocyte cells.Elevated liver enzymes including ALT, AST, GGT, ALP, and Serum total bilirubin levels are some of the laboratory results after SARS-CoV-2 induced liver injury.In addition, portal tract fibrosis, sinusoidal artery thrombosis, and fibrotic hepatocytes are histological abnormalities in the liver. Even though viral fragments appeared in SARS-CoV-2-infected liver cells, more proof is required.Conceptualization: Fikadu Seyoum.Validation: Fikadu Seyoum.Visualization: Fikadu Seyoum.Writing – original draft: Fikadu Seyoum.Writing – review & editing: Fikadu Seyoum.


Mesenchymal stromal cells (MSCs) show considerable promise in regenerative medicine with superior anti-fibrotic, immunomodulatory, and angiogenic functions. More recently, discovered with the tumor tropism, MSCs have been exploited as the basis of targeted cancer therapy. In this scenario, MSCs can directly home to tumor tissues and play anti-tumor properties. In addition, MSCs, MSC-derived exosomes and MSC-derived membranes are often developed as carriers for precisely delivering cytotoxic agents to cancer sites, including chemotherapeutic drugs, therapeutic genes, or oncolytic viruses. However, it has revealed the tumorigenic risk of MSCs as an important component within the tumor microenvironment, hampering the translation of MSC-based cancer therapies into clinical settings. Therefore, in this review, we introduce the specific tumor-tropic ability of MSCs and underlying mechanisms. We also summarize the current application of MSC-based therapeutic approaches in treating gynecologic cancers, mainly including cervical, ovarian, and endometrial cancers. Moreover, we discuss the main challenges that the current MSC-based cancer therapies are facing. Mesenchymal stromal cells (MSCs) are a bulk population of multipotent stem cells, progenitors and differentiated cells , 2. MSCsGynecologic cancers are still the prevalent and fatal diseases among women worldwide, mainly including cervical, ovarian, and endometrial cancers . SurgicaCurrently, cell-based therapies have gained increasingly clinical attraction in cancer treatment, such as MSCs , 20, redNumerous studies that employ MSCs or MSCs’ derivatives for treating gynecologic cancers have been reported with mixed results over recent years –41, encoThe strong tropism toward cancer cells has predisposed MSCs to oncologic therapies in recent years , 28. ImpNumerous efforts have been made on developing MSC-based therapies for gynecologic cancer treatment. In the following section, we summarize therapeutic strategies involved in the use of MSCs, MSC-derived exosomes, and MSCMs as a powerful tool for killing cancer cells Figs.  and 3.FiDespite few studies directly utilizing MSCs for tumor therapy, it is important to mention that tumor-suppressive roles of MSCs have been verified in several preclinical cervical and ovarian cancer models Fig. .in vitro transwell co-culture system was established by loading En-MSC and cervical cancer HeLa cells in the upper and lower chamber, respectively. And the co-culture results revealed that En-MSC blocked the cell cycle at G0/G1 phase, and suppressed the proliferation and invasion in HeLa cells. In vivo, a HeLa-bearing tumor xenograft mouse model was generated by subcutaneously co-injecting HeLa cells with En-MSCs at the density of 1:1. It was found that the average volume and weight of xenografted tumors were markedly decreased by En-MSC. Mechanistic experiments further revealed that En-MSC secreted TGF-β1 and activated the JNK/P21 signaling pathway in HeLa cells to exert anticancer effects. In another study, umbilical cord-derived MSC (UC-MSC) were co-cultured in direct contact with HeLa cells at a density ratio of 2:1. Experimental results found that UC-MSC could time-dependently inhibit the growth and invasion of HeLa cells through decreasing the activities of AKT/PI3K/STAT3/mTOR pathway and epithelial-mesenchymal transition (EMT), but did not alter the apoptosis and cell cycle [The direct anticancer effects of MSCs have been studied on cervical cancer cells. For example, Liu et al.  isolatedll cycle .Recently, therapies based on cellular secretome have emerged as promising treatments for human diseases . ConditiAziz et al.  also emplow CB-MSC significantly suppressed tumor growth and increase the survival time of mice, and the anticancer effects were further enhanced after the combination of immune activator VIC-008. Notably, this was mediated by the activation of anti-tumoral CD4+ and CD8+ T cells and the reduction of Treg cells within the tumor microenvironment (TME). Collectively, these studies demonstrate the direct anticancer functions of MSCs in gynecologic cancers, which are partially through the paracrine signaling mechanisms.Similarly, the CM from hyperthermia-treated (HT) adipose tissue-derived MSC (AT-MSC) and AF-MSC (HT-MSC-CM) also exerted tumor-suppressive effects on human ovarian cancer SKOV3 cells. The reduced viability and cell cycle arrest were observed after HS-MSC-CM treatment in SKOV3 cells by the morphologic, cell proliferation and Sub-G1 analyses . GauthamMounting data have developed MSCs as vehicles for delivering therapeutic agents in a targeted way, including chemotherapy drugs, tumor-suppressive genes, and oncolytic viruses. Therefore, we will describe experimental evidence to summarize the applications of these engineered MSCs in gynecologic cancer therapy Fig. .in vitro toxicities in human ovarian cystadenocarcinoma MA148 cells and lung cancer A549 cells. Although the in vivo anticancer effects of nano-engineered MSCs on ovarian cancer were not investigated, this study revealed that nano-engineered MSCs were selectively accumulated and retained in the orthotopic lung tumors.Poor availability into tumors and catastrophic side effects are major challenges that limit the efficacy of conventional chemotherapy drugs. Owing to the tumor-homing property, approaches that use MSCs have been designed to specifically enhance drug delivery and improve the therapeutic efficacy. To solve this issue, Sadhukha et al.  preparedAnother study by Borghese et al.  determinHSVtk) was constructed by transduction with the lentiviral vector carrying HSVtk gene. It is noted that TC-1 cell lines, derived from primary lung epithelial cells of C57BL/6 mice and immortalized with human papilloma virus (HPV)-16 E6/E7 oncogenes, are often used to establish cervical cancer mouse model [HSVtk, followed by intraperitoneally administration of GCV after 14 days. Results demonstrated that AT-MSCHSVtk/GCV led to a significant decrease in tumor size in a common TC-1 cells-grafted cervical cancer model, which might be mediated by enhanced anticancer activities of natural killer (NK) cells and cytotoxic T cells. The inhibitory effects of AT-MSCHSVtk were also investigated in highly metastatic ovarian cancer models. Toro et al. [HSVtk in female nude mice, following by intraperitoneally injection of 5-FC after 14 days. A significantly decreased tumor volumes and prolonged survival time were observed in A2780 tumor-bearing mice after AT-MSCHSVtk/5-FC treatment.Despite few studies on the direct delivery of chemotherapy drugs by MSCs, researches using MSCs as vectors for gene therapy in treating gynecologic cancers are widely conducted. Herpes simplex virus thymidine kinase (HSVtk) is known as one of the most frequently used suicide gene therapy tools. Indeed, HSVtk could convert non-toxic ganciclovir (GCV) monophosphate or 5-fluorocytosine (5-FC) into toxic GCV or 5-fluorouracil (5-FU) to disrupt DNA synthesis . In studse model . After to et al.  subcutanTRAIL) and AT-MSC (AT-MSCTRAIL) were successfully generated by retrovirus- and lentivirus-mediated transductions, respectively [TRAIL and AT-MSCTRAIL could target HeLa cells and caused a cell ratio-dependent apoptosis in vitro. In vivo studies in HeLa-bearing nude mice found that AT-MSCTRAIL localized into tumors via either intravenous or subcutaneous administration and induced apoptosis without apparent toxicities to normal tissues [Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a transmembrane pro-apoptotic ligand that causes apoptosis of various cancer cells . Althougectively , 75. FurIFN−α) and intravenously injected them into a mouse model with HeLa cell xenografts. The tumor tropism of AF-MSCIFN−α was maintained and tumor growth was significantly restricted by AF-MSCIFN−α, which was mediated by cell proliferation suppression, angiogenesis inhibition, and apoptosis induction in cancer cells.Interferon-α (IFN-α), known as an important antiviral cytokine, plays also a tumor-suppressive role by inducing cell apoptosis . Zhou etE7+HSP27). In addition, immunized with BMSCE7+HSP27-based vaccination significantly inhibited the tumor growth through boosting the E7-specific T-cell responses in TC-1-grafted cervical cancer mouse models. Taken together, these results provide evidence that MSCs could serve as good carriers for targeted gene transfer in treating cervical and ovarian cancers.HPV E7 and E6 oncoproteins have emerged as specific immunotherapeutic targets in cervical cancer . PreviouOncolytic viruses have emerged as promising therapeutics for treatment of human cancers in the last decade, whereby oncolytic viruses can be preferentially replicated within cancer cells and destroy them via direct lysis . Among vSeveral studies have investigated the possibility of using MSCs as OAdV carriers owing to the documented tumor tropism. For instance, Komarova et al.  and DembAnother study by Alfano et al.  preparedExosomes are an important type of extracellular vesicles (EVs) secreted by living cells and mediate the cell-to-cell communication. Exosomes often carry specific bioactive cargoes like proteins, lipids, and nucleic acids to play various biological functions . MSC-derTwo important oncogenic markers for cervical cancer were identified in another two studies, namely the centrosomal protein, 55 Kd (CEP55)  and mateThe exosomes released from MSCs have also been revealed to possess great therapeutic effects in cervical cancer via the delivery of chemotherapy drugs. In a most recent study, PTX was loaded into the exosomes derived from human UC-MSC (PTX-UC-MSC) by electroporation and then incubated with HeLa cells in vitro . The PTXLi et al.  demonstr3+,Er3+ upconversion nanoparticles (UCNPs@SiO2) with human BMSCMs (SUCNPs@mSiO2). Experimental studies showed that SUCNPs@mSiO2 displayed MSC-mimicking long blood circulation time and tumor-targeting property with large accumulation at the tumor site. Further PDT experiments were performed under irradiation with 980 nm laser at 0.35 W/cm2 in HeLa cells and HeLa-bearing nude mouse models. In vitro co-culture or tail intravenous administration of SUCNPs@mSiO2 showed stronger suppressive efficacy in tumor growth than pure UCNPs@SiO2, providing the evidence that artificial modification of anticancer agents with natural MSCMs holds considerable potential for cervical cancer treatment by systemic administration. More studies using the biomimetic platform to treat ovarian and endometrial cancers should be carried out in the future.It should be noted that roles of MSCMs for drug encapsulation in the treatment of gynecologic cancers have not been fully studied. To date, there have been only one study reported in using MSCM-camouflaged nanosystems to treat cervical cancer . The upcAlthough the anticancer effects of MSCs have been well documented, MSC-based therapies are also challenged by oncogenic functions of MSCs as an exogenous regulators of cancer cells. In addition, although MSCs derived from distinct tissues have similar morphology and immunophenotypes, the homing ability of diverse MSCs may be discrepant and tissue-specific –95. TherThe pro-tumorigenic roles of exogenous MSCs that do not initially exist within TME have been described in some co-culture studies.For example, Castro-Oropeza et al.  performeOb, but not SC-AT-MSCOb, SC-AT-MSCLe, or O-AT-MSCLe, induced more infiltration of inflammatory CD3+ T cells and F4/80+ macrophages to support tumor growth after intraperitoneal co-injection with mouse ovarian cancer ID8 cells in C57BL/6 mice.Researchers have found that AT-MSC in adipose tissue can engraft into neighboring tumors to form supportive tumor stroma. Zhang et al.  isolatednu nude mice bearing HEC-1-A xenografts in the upper flank, O-AT-MSC was found to be more efficiently recruited to the tumors. Moreover, O-AT-MSC produced an accelerated growth and vascularization of endometrial tumors while SC-AT-MSC did not. In addition, Li et al. [Klopp et al.  collectei et al.  first isi et al.  further Exogenous MSCs can also aggravate malignant traits of gynecologic cancer cells through contact-independent or contact-dependent paracrine mechanisms. For contact-independent way, Ding et al.  collecteThe exact mechanism underlying the cancer tropism of MSCs has not been fully elucidated. Transplanted MSCs with higher tumor tropism are more prone to reach lesion sites and exert more effective therapeutic functions , 107. SiClinicalTrials. Gov database (NCT02530047 and NCT02068794). However, no data have been published from such trials. Moreover, the trials concerning the therapeutic value of MSCs on cervical or endometrial cancers are still lacking. By solving these challenges, we believe that MSC-based therapeutic strategies could more efficiently and safely kill cancer cells in human gynecologic cancers.The specific tumor-homing properties of MSCs have attracted increased attention by researchers to use MSCs to treat human gynecologic cancers. MSCs can possess the direct anti-tumoral functions toward cancer cells, but in most cases, they are functionally or genetically engineered to deliver chemotherapeutics, suicide genes, or oncolytic viruses in gynecologic cancer therapy. Furthermore, MSC-derived exosomes and MSCMs can be extracted as a bioactive carrier of anticancer drugs, miRNAs, or multifunctional composite nanoparticles. However, several questions or challenges need to be addressed before translating the MSC-based cancer therapies into clinical settings. First, MSCs possess pro-tumorigenic and anti-tumorigenic roles in cancer development. However, the reported discrepancies with respect to the capacity of MSCs to promote or inhibit tumorigenesis are based on the differences of experimental settings including animal models, cell lines, doses, and duration times of treatment. Therefore, whether MSCs should be considered as anticancer agents or a therapeutic target in cancer treatment is still a matter of debate. How to reduce the extrinsic tumorigenicity of exogenous MSCs such as functional modification will be another topic we need to focus on. Second, different tropism abilities require further delineation about the homing map of each type of MSCs in various cancers, which will guide the selection of proper MSCs in cancer therapy. Third, it is necessary to develop advanced extraction methods for scalably producing MSC-derived exosomes or MSCMs in the future. Lastly, up to date, there are only two clinical trials accessing the MSC-based therapies for the treatment of ovarian cancer that have been registered on


MGMT promoter status. The authors concluded that well-powered prospective study on the clinical efficacy of TMZ for patients with IDH-wildtype anaplastic gliomas (meeting criteria for glioblastoma) is warranted. Given that the prognostic and predictive role of MGMT status for grade 2–3 gliomas is unresolved, we determined the effect of MGMT status on OS in patients with 1p19q-intact gliomas in the National Cancer Database (NCDB).Standard-of-care for 1p19q-intact anaplastic gliomas is defined by the international randomized phase III CATNON trial, which found an overall survival (OS) benefit for adjuvant temozolomide (TMZ) when added to radiotherapy. Paradoxically, TMZ did not appear to benefit patients with IDH-wildtype gliomas, regardless of MGMT with OS.We queried the NCDB from 2018–2019 for patients with IDH-wildtype or -mutant astrocytomas who received chemotherapy with follow-up through 2022. The Kaplan-Meier method and Cox proportional hazards regressions models were used to determine the association of We identified 1,514 patients who were newly diagnosed with IDH-wildtype  or - mutant astrocytomas  and received chemotherapy during initial management. An unmethylated promoter was associated with poorer survival in patients with IDH-wildtype  but not IDH-mutant astrocytomas .MGMT status. Determining the optimal strategy for diffuse gliomas that are IDH-wildtype will be particularly important. MGMT promoter methylation should be considered as a stratification factor in future clinical trials for these patients.This ancillary analysis supports adjuvant TMZ as standard-of-care for anaplastic astrocytomas (IDH-mutant and 1p19q-intact), irrespective of  No benefit was observed in a subset of 159 patients with tumors that met the WHO 2021 molecular criteria for glioblastoma, IDH-wildtype.4Standard-of-care for 1p19q-intact anaplastic gliomas is defined by the international randomized phase III CATNON trial, which found an overall survival (OS) benefit for adjuvant temozolomide (TMZ) when added to radiotherapy.6-methylguanine-DNA methyltransferase (MGMT) promoter methylation was included as a stratification factor in the CATNON trial, given its predictive role for benefit from TMZ in glioblastoma. Promoter methylation was prognostic in patients with IDH-wildtype but not IDH-mutant tumors.5 However, to the surprise of the investigators, MGMT status did not predict benefit from TMZ in patients with IDH-wildtype tumors. They concluded that a well-powered prospective study on the clinical efficacy of TMZ for patients with anaplastic gliomas who meet the contemporary definition of glioblastoma is warranted. Consistent with CATNON, we also found that MGMT status is prognostic in IDH-wildtype but not IDH-mutant, 1p19q-intact gliomas in a pooled analysis of three prospective cohorts.6MGMT status was not an independent predictor of OS on multivariable analysis in either study, though sample sizes in both our study and the CATNON study were limited. Therefore, the prognostic and predictive role of MGMT status for grade 2–3 gliomas, particularly IDH-wildtype tumors, is unresolved.OMGMT status on OS in patients with 1p19q-intact gliomas in the National Cancer Database (NCDB) from 2018–2019 with follow-up through 2022. The NCDB accounts for approximately 70% of invasive cancers in the United States, so it is unlikely that inadequate power would lead to ambiguous results.7To address this uncertainty, we determined the effect of MGMT promoter methylation status.15We queried the NCDB (2022 submission) to identify patients with diffuse or anaplastic astrocytoma -O-3 codes 9400 or 9401), IDH-mutant (Brain Molecular Markers 1 or 2) or IDH-wildtype (2 or 4) newly diagnosed between January 1, 2010 and December 31 2019 with follow-up through December 31, 2022. Patients were included if they received chemotherapy. Patients were excluded if they had less than one month of follow-up or missing data for age, sex, race, Charlson-Deyo Comorbidity Index, extent of resection, receipt of radiotherapy, or MGMT status with OS. Variables that were statistically significant on univariable analysis were included in the multivariable model. Schoenfeld’s test of weighted residuals was utilized to assess proportional hazard assumption in the Cox model. All analyses were performed at the .05 significance level based on two-sided statistical testing.All statistical analyses were conducted using the RStudio software Version 1.4.1106 . The Kaplan-Meier method with the log-rank test and adjusted Cox proportional hazards regressions models were used to determine the association of MGMT status (44.9%) or other missing data (1.6%). There were 1,514 patients (802 IDH-wildtype and 712 IDH-mutant) included in our final analysis. MGMT promoter methylation was observed in 33% and 48% of patients with IDH-wildtype and -mutant astrocytomas, respectively  will be particularly important. The largest randomized trials , paradoxically, have not demonstrated an improvement in OS with alkylating chemotherapy for patients with IDH-wildtype grade 2–3 gliomas, though these were post-hoc analyses with modest sample sizes.18 Based on our results, MGMT promoter methylation should be considered as a stratification factor in future clinical trials of patients with IDH-wildtype gliomas.This ancillary analysis supports adjuvant TMZ as standard-of-care for anaplastic astrocytomas (IDH-mutant and 1p19q-intact), irrespective of MGMT promoter methylation may be a prognostic or predictive biomarker in IDH-mutant tumors, longer follow-up is required in both our dataset and the CATNON trial, as approximately 80% and 66% of patients were still alive at the database lock. However, we are not aware of any prospective studies that support a role for MGMT status in IDH-mutant astrocytomas. In contrast, emerging data suggest a role for MGMT as a biomarker in 1p19q-codeleted oligodendrogliomas.7 The method of testing for MGMT promoter methylation is not reported in the NCDB, and results and cutoff levels may vary depending on the specific assay. Considering these limitations, our study is the largest on the subject and our results are consistent with other literature. Collectively, these data support a role for MGMT testing in IDH-wildtype gliomas and IDH-mutant/1p19q-codeleted oligodendrogliomas but not IDH-mutant/1p19-intact astrocytomas.Limitations of our study include retrospective analysis with relatively short follow-up. To clearly determine if