The COVID-19 pandemic has unleashed unprecedented challenges upon the global population, demanding rapid and innovative solutions. Across various sectors, the incorporation of digital technologies, often prefixed with “tele-”, has played a pivotal role in reshaping societies and mitigating the impact of the virus . This SpThis Special Issue presents valuable insights from nine articles in the field under investigation . SpecifiThe COVID-19 pandemic has presented the world with an unprecedented opportunity to embrace digital health innovations . This Sp


The effects of COVID-19-associated restrictions on youth sexual and reproductive health (SRH) care during the pandemic remain unclear, particularly in sub-Saharan Africa. This study uses interrupted time series analyses to assess changes in SRH care utilisation  adolescent girls’ and young women’s  in eSwatini following COVID-19 lockdown beginning in March 2020. SRH utilisation data from 32 clinics in the Manzini region that remained open throughout the 2020 COVID-19 period were extracted from eSwatini’s electronic health record system. We tabulated and graphed monthly visits  by AGYW during the two-year period between January 2019 and December 2020. Despite the March to September 2020 lockdown, we did not detect significant changes in monthly visit trends from 2019 to 2020. Our findings suggest little change to AGYW’s SRH utilisation in eSwatini during the 2020 COVID-19 lockdown period. In the early months of the COVID-19 pandemic, researchers predicted that COVID-related lockdowns would cause disruptions to sexual and reproductive health (SRH) services, including human immunodeficiency virus (HIV) testing and treatment, family planning, and antenatal care, which are key elements of the Guttmacher-Lancet Commission’s essential package of SRH services . In thesDespite the growing literature documenting the impacts of COVID-19 on SRH and youth health care, findings remain mixed. Some studies identify substantial declines in SRH service utilisation in high-income countries (HICs) and LMICs, while other studies find modest disruptions . There aThis paper uses data from eSwatini’s electronic health record to explore the extent to which a six-month COVID-19-associated lockdown may have affected AGYW’s SRH service utilisation in primary care clinics in eSwatini. A secondary objective of the study was to assess whether there were differential changes in AGYW’s SRH service utilisation at clinics that had piloted enhanced youth-centred services prior to COVID-19. Findings may be useful in understanding the resiliency of health systems facing potential disruptions in care.tinkhundla . Over a third of the population is aged 10–24 years old. The current analysis builds on previous work that implemented and evaluated a strategic marketing campaign called Girl Champ ). After the lockdown, we observed a decrease in the monthly trend of total visits to Girl Champ clinics, relative to the pre-lockdown period   , Panel bWe also examined longitudinal change in utilisation for specific SRH services across all clinics (Our results suggest there were no major interruptions in AGYW’s SRH service utilisation in the first 10 months of the COVID-19 pandemic. We note that what had been an increasing trend in overall service utilisation pre-lockdown among the three Girl Champ clinics changed in the post-lockdown period; however, this was not statistically significant after applying the Bonferroni correction, and no similar effect was found among the other 29 clinics included in the study. Our study adds to the findings concerning adolescent SRH service utilisation and access in LMICs during COVID-19, which demonstrate that some services and settings were more resilient to disruptions than others ,16,17. AOur study should be interpreted in light of its limitations. First, our analyses focused on the first 10 months of the pandemic. While future trends in SRH service utilisation are not known, the study timeframe spanned a six-month COVID-19 lockdown, which was a period vulnerable to service disruption. Second, our sample of primary care clinics included those that stayed open to the general population throughout the study period. We do not quantify the loss of services in clinics that were transitioned to COVID-19 treatment facilities. In addition, we included clinics that were on the CMIS longer, and may have been more established organisations. We tried to limit potential bias by selecting a sample of clinics that ranged in size and geography, but recognise our sample may not be generalisable to all primary care centres in eSwatini. Finally, we acknowledge that there may have been other social and economic risk factors that influenced SRH service need and utilisation during COVID, such as increased rates of gender-based violence and unemployment . HoweverOur findings suggest minimal changes in AGYW’s use of SRH services in eSwatini during the COVID-19 lockdown in 2020. Further studies should identify and disseminate strategies that clinics employed to sustain service delivery and AGYW’s engagement in care.Click here for additional data file.


The efficacy of targeted albumin therapy in the management of decompensatory events in cirrhosis is unclear, with different reports showing conflicting results. It is possible that only certain subgroups of patients may benefit from targeted albumin administration. However, extensive conventional subgroup analyses have not yet identified these subgroups. Albumin is an important regulator of physiological networks and may interact with homeostatic mechanism differently in patients according to the integrity of their physiological network. In this study, we aimed to assess the value of network mapping in predicting response to targeted albumin therapy in patients with cirrhosis.This is a substudy of the ATTIRE trial, a multicenter randomized trial conducted to assess the effect of targeted albumin therapy in cirrhosis. Baseline serum bilirubin, albumin, sodium, creatinine, CRP, white cell count (WCC), international normalized ratio, heart rate, and blood pressure of 777 patients followed up for 6 months were used for network mapping using parenclitic analysis. Parenclitic network analysis involves measuring the deviation of each patient from the existing network of physiological interactions in a reference population.Overall network connectivity and deviations along the WCC-CRP axis predicted 6-month survival independent of age and model for end-stage liver disease in the standard care arm. Patients with lower deviation along the WCC-CRP axis showed lower survival in response to targeted albumin administration over a 6-month follow-up period. Likewise, patients with higher overall physiological connectivity survived significantly less than the standard care group after targeted albumin infusion.The parenclitic network mapping can predict the survival of patients with cirrhosis and identify patient subgroups that do not benefit from targeted albumin therapy.  There has been an epidemic of liver disease during the past 50 years in the United Kingdom, with a 4-fold increase in liver-related deaths . This isOther clinical trials of albumin infusion have shown conflicting results. For instance, a meta-analysis of albumin use in patients with spontaneous bacterial peritonitis reported significant benefit  that wasHealthy individuals show a high degree of functional connectivity between physiological organ systems. Disruption of organ system coupling is a hallmark of complex diseases, and recent studies showed that reduced network connectivity was linked with poor survival in patients with sepsis  and poorTargeted albumin infusion has the potential to challenge the physiological network through alteration of oncotic pressure, plasma volume, glomerular filtration rate (GFR), and transportation of various physiological molecules. Therefore, the assessment of physiological interaction as a network may allow for a more precise prediction of patients' response or outcome. The parenclitic network mapping allows network analysis of static, baseline clinical variables on the individual patient level . Our groThis is a substudy (and extension) of the ATTIRE trial, which was a randomized controlled trial of targeted albumin infusions vs standard care involving 777 patients hospitalized with decompensated cirrhosis from 35 hospitals across England, Wales, and Scotland 2016–2019)  for the definition of network indexes used in this study. In general, higher global parenclitic network topology indexes such as diameter, mean centrality, and shortest path length indicate lower connectivity among all components of the network  were computed and used to weigh the correlation network map of clinical variables. Furthermore, the parenclitic indexes, including deviations along variable pairs and a global network topology of all patients (treatment and standard care), were computed using an in-house code in MATLAB . For the measurement of global network topology, indexes such as network diameter, mean centrality, and shortest path length were calculated. Please see Supplementary Table S1 , and the remainder were used as the validation sample (n = 203). Survivors in the training group were used as a model for the calculation of the coefficients that were used for the calculation of parenclitic deviations in the validation group. Statistical analysis was performed to test whether the prognostic values persist following the random split in the validation sample. This is to confirm whether the result generated was independent of the population studied.P value of <0.05 was defined as statistical significance in all analyses. The Bonferroni correction was applied when multiple hypotheses were tested within a single analysis.Statistical analysis was performed using Stata statistical software  and SPSS Statistics 26 , with data presented as median and interquartile range (IQR) or mean ± SD. A 2-tailed U test to compare the median of computed parenclitic variables, including the deviations along physiological axes (denoted as δ in this report) and global network topology indexes, for the survivor and nonsurvivor groups. Significantly different parenclitic variables were then tested for prognostic value by computing univariate and then multivariate Cox regression controlling for MELD and age. Parenclitic variables with independent predictive values were combined with MELD to produce a combinatory prognostic index based on coefficients of bivariate Cox regressions as follows: composite index = β1 × MELD + β2 × δ, where β1 and β2 are, respectively, the regression coefficient of MELD and parenclitic indexes in the bivariate Cox model.Initially, we performed the Mann-Whitney Receiver operating curve (ROC) analyses were performed and the area under the curve (AUC) computed for individual and combinatory indexes to generate cutoffs. The specificity and sensitivity of resulting cutoffs were then used to generate the positive and negative predictive values based on Bayesian priors  .To test whether parenclitic indexes may differentiate between survivors and nonsurvivors at 6 months after targeted albumin treatment, we performed a multivariate Cox regression including patients' treatment arm  and each of the parenclitic indexes as interacting variables. The cutoff of parenclitic indexes that showed significant interaction with treatment in predicting survival was then used to categorize each patient into “1” if ≥cutoff and “0” if otherwise. The treatment arm of patients was then used to plot ROC curves to assess the 6-month survival grouped by the cutoffs of the significant parenclitic indexes.To assess possible improvement in prognostic performance of MELD due to the addition of parenclitic indexes, Brier scores, integrated discrimination improvement (IDI), and net reclassification indexes (NRIs) were computed on Stata statistical software. The Brier score provides the mean of the squared distance between observed and predicted risks of event  for the individual patient. Generally, the lower the Brier score, the better the predictive model . IDI andA total of 397 of 777 51%) patients received standard care and were subjected to parenclitic analysis. During the 6-month follow-up period, 119 (30%) of these patients died. Table % patientThe correlation network maps show that survivors had a significantly higher association between the baseline clinical variables compared with nonsurvivors Figure . OverallP < 0.001).According to our univariate Cox regression analysis, greater parenclitic deviations along WCC-Na, Bil-WCC, WCC-CRP, HR-Bil, and INR-Bil axes were all associated with an increased risk of 6-month mortality Table . LikewisMultivariate Cox regression was performed to assess whether parenclitic indexes that individually predicted 6-month mortality had a prognostic value independent of age and MELD. Parenclitic deviations along the WCC-CRP axis  and Bil-WCC axis  significantly predicted outcome independent of age and MELD of patients at trial entry (Table http://links.lww.com/CTG/A931), and parenclitic deviations of the validation sample (203 random patients) calculated from the training sample (194 random patients) demonstrated similar results to the original findings using the overall sample . Specifically, parenclitic deviation along the WCC-CRP axis as well as the mean shortest path length and diameter predicted 6-month survival independent of MELD and age of patients in this validation subset .The results of a parenclitic network analysis of a split sample (randomly selected patients) extracted from the study population showed similar correlation network maps of biomarkers  curves (AUC), cutoffs, and the sensitivity, specificity, positive and negative predictive values, and Brier scores of these cutoffs are presented in Table http://links.lww.com/CTG/A931). Furthermore, the Kaplan-Meier survival curves for patients grouped by the parenclitic indexes demonstrated significant differences when targeted albumin and standard care groups were compared , WCC-CRP parenclitic deviation, baseline serum albumin, and white cell count significantly interacted with targeted albumin treatment to predict survival .Potential competing interests: None to report.


Hematologic malignancies (HMs) pose a serious threat to patients’ health and life, and the five-year overall survival of HMs remains low. The lack of understanding of the pathogenesis and the complex clinical symptoms brings immense challenges to the diagnosis and treatment of HMs. Traditional therapeutic strategies for HMs include radiotherapy, chemotherapy, targeted therapy and hematopoietic stem cell transplantation. Although immunotherapy and cell therapy have made considerable progress in the last decade, nearly half of patients still relapse or suffer from drug resistance. Recently, studies have emerged that nanomaterials, nanotechnology and nanomedicine show great promise in cancer therapy by enhancing drug targeting, reducing toxicity and side effects and boosting the immune response to promote durable immunological memory. In this review, we summarized the strategies of recently developed nanomaterials, nanotechnology and nanomedicines against HMs and then proposed emerging strategies for the future designment of nanomedicines to treat HMs based on urgent clinical needs and technological progress. Hematopoietic malignancies (HMs) are originated from hematopoietic system, mainly including leukemia, lymphoma, multiple myeloma (MM) and myelodysplastic syndromes (MDS) Fig. , which iCurrent clinical strategies for treating HMs include radiotherapy, chemotherapy, targeted therapy and hematopoietic stem cell transplantation (HSCT) , 14. CheNanomedicine based on nanomaterials and nanotechnology show great advantages in cancer treatment and diagnosis , 23. NanNanomedicine refers to use of nanotechnology to make drugs into nanoparticles with particle size between 1 and 100 nm, or the combination of appropriate carrier materials with bulk drugs to form nanoparticles with nanometer scale and the final pharmaceutical preparation . CompareLiposome is the most widely used drug delivery system in biomedical research and clinical application –35.Liposomes are widely used and studied in the treatment of blood cancer , 37. CPX. reported a liposome modified with CD19 antibodies for the treatment of B cell leukemia [. reported a hyaluronic acid-modified liposome for the treatment of leukemia [Surface-modified liposomes are also widely studied. Antibody-modified liposomes can be used to specifically target leukemia cells . Myers eleukemia . Shao etleukemia . Combineleukemia .. constructed a liposome with dual targeting capability to remove leukemia stem cells and minimal drug residues [Shi et alresidues .Polymer-based nanoparticles are widely used in preparation of nanomedicine . Polymer. reported a PLGA-based nanodrug for the targeted treatment of AML [. constructed a PLGA nanodrug loaded with bendamustine, which can significantly enhance the toxicity to leukemia cells [. also constructed curcumin-loaded PLGA-based nanoparticles (Cur-PLGA NP) for the treatment of Jurkat leukemia cells [. constructed a erythrocyte membrane cloaked, ATO-loaded PLGA nanoparticle for APL treatment and to reduce the side effects of arsenic agents [. demonstrated the biosafety of PLGA as a delivery carrier [. demonstrated that PLGA nanoparticles encapsulated with Iguratimod could induce MM cell death, which could be used as a potential therapy for MM [Poly(lactic-co-glycolic acid) (PLGA)-based nanomaterials are widely used in drug carriers. Noureldien et alt of AML . Khan etia cells . In addiia cells . Leung eia cells . Su et ac agents . Venkatp carrier . Wang ety for MM .. reported a chitosan-based, self-assembled nanomedicine for the controlled release of 6-MP and treatment of AML [. showed that the preparation of zinc–chitosan nanoparticles (Zn-CSNPs) by linking zinc with chitosan could also enhance the toxicity against leukemia cells [. constructed a PEGylated, chitosan-based nanodelivery system for prolonging the blood circulation time of DOX [Chitosan nanoparticles are widely applied in drug delivery carriers. Gong et alt of AML . Derakhst of AML . Similart of AML . Sarangat of AML . Similaria cells . Termsare of DOX .. have constructed a light-induced polymer nanoparticle to enhance leukemia treatment [. have built a polymer-based self-assembly-based nanodrug to deliver leukemia treatment drugs [. reported a carrier-free catanionic drug-derivative nanodrug for the treatment of leukemia [Gigli’s study showed that simultaneous delivery of two polymer nanoparticles containing different drugs consistently down-regulates CML cancer development . Boto etreatment . To overnt drugs . Ma and nt drugs . Li et aleukemia .Carbon-based nanomaterials are widely studied as drug delivery carriers in anticancer research –84. Carb. reported a P-gp antibody (anti-P-gp) functionalized water-soluble single-walled carbon nanotubes (Ap-SWNTs) loaded with doxorubicin (Dox) to kill MDR human leukemia cells K562 [. reported the use of single-walled carbon nanotubes (SWNT) as antigen carriers to improve the immune response to weak immunogenic peptides [One-dimensional carbon nanotubes are widely used as drug delivery vehicles. Ruibin et allls K562 . Carlos peptides . In vitr. reported a RGO/Ag composite nanoparticle that has a strong anticancer activity against KG-1A cell line [. synthesized ND vectors capable of loading chemotherapeutics and gene delivery and applied them to the treatment of drug-resistant leukemia [Two-dimensional carbon nanomaterials such as graphene nanosheets also have been applied in targeted delivery of anticancer drugs . Roy et ell line . The incleukemia .. screened a variety of one- and two-dimensional carbon materials and unexpectedly found that GDYO showed a strong killing effect on DNMT3A-mutant AML cells [Inorganic nanoparticles are also widely used in the treatment of leukemia . In consML cells . By anal. reported that GDYO nanosheets can simultaneously kill lymphoma cancer stem cells and remodel tumor microenvironment, thus inhibiting the growth of lymphoma [GDYO also inhibits the growth of lymphoma. Lymphoma is a type of solid tumor originating in the lymphatic system. Li et allymphoma . Mechani. used MSN nanoparticles as the carrier to construct nanodrugs for active targeting of leukemia cells to deliver drugs [. prove that mesoporous silica microparticles can be used to enhance the toxicity of anticancer platinum drugs [Porous silica nanomaterials have great prospects in biomedicine as drug delivery carriers due to porous and good drug adsorption properties –98. Durfer drugs . Tao et um drugs .. prepared ATO into FA-HSA-ATO nanodrug to reduce the toxic and side effects of arsenic agents and improve the efficacy of targeted therapy [. reported that nanoencapsulating ATO can significantly improve the therapeutic efficacy of leukemia and reduce the toxic side effects on ovaries [Arsenic, as a traditional Chinese medicine, mainly includes arsenic and realgar. Arsenic trioxide, the active ingredient in arsenic, was first purified in the 1970s and used to treat APL, increasing the cure rate to more than 90%. However, the high toxicity and side effects limit the further anticancer application of arsenic agents. In view of this, Peng et al therapy . Similar ovaries . However ovaries , 104.4S4. Research evidence confirms that realgar has a similar therapeutic effect on APL as ATO, but it is insoluble in water and most organic reagents, resulting in poor bioavailability and limiting its clinical application. Inspired by nanodrugs, Wu et al. demonstrated that preparation of realgar into nanoparticles can increase bioavailability and enhance toxicity to cancer cells [. further confirmed the important role of caveolin-1 (Cav-1), a principal constituent protein of caveolae, in mediating the absorption of realgar nanoparticles by leukemia cells [Realgar is another important arsenic agent, mainly composed of Aser cells . Shi et ia cells . In addiia cells .. constructed nanomaterials based on gold nanoparticles to deliver small molecule inhibitors of FLT3 for the treatment of AML [. constructed gelatin-coated gold nanoparticles as carriers of FLT3 inhibitors for the treatment of acute myeloid leukemia [Gold nanoparticles are widely used as drug delivery carriers due to their good stability and adsorption ability. Molotkova and colleagues reported using gold nanoparticles as carriers to deliver Bosutinib, a TKI, for CML therapy . Simon et of AML . On thisleukemia .Silver nanoparticles have unique optical, electrical and catalytic properties and can be used in optical materials, battery electrodes and catalysts. In addition, nanosilver also shows certain application value in antifungal, anti-viral, anti-inflammatory and anti-thrombosis and antitumor and promoting wound healing . There a. found that PVP-coated nanosilver can induce apoptosis and necrosis of THP-1 cells in a dose- and time-dependent manner [. found that Ag NPs prepared from FA and CHA exhibited significant anticancer activity on K562 cells at a lower concentration due to ROS production and DNA fragmentation [. reported a simple, cost-effective and green method to synthesize Ag NPs nanomedicine using chitosan as a reducing agent and stabilizer [Foldbjerg et alt manner . This efentation . Hemmatiabilizer . These s. showed that the shape of Ag nanomaterials plays an important role in its anti-proliferative activity, and the activity of anti-proliferative silver nanomaterials highly depends on its nanostructure [Interestingly, the shape of the nanoparticles also affects their toxicity to tumor cells. Sakaguchi et altructure . The Ag Magnetic nanomaterials are an important category of nanomaterials, typically ranging in size from 1 to 100 nm. As a new type of functional material, magnetic nanoparticles have a broad application prospect in the field of magnetic materials and bioengineering . Due to . reported a Fe3O4-PLA nanocomposite material for drug delivery and explored the potential application of the daunorubicin (DNR) to the drug-resistant leukemia K562 cells [. linked magnetic Fe3O4 nanoparticles with Homoharringtonine (HHT), a natural cephalotaxine alkaloid, for tumor therapy, further demonstrating the potential of magnetic Fe3O4 nanoparticles as a carrier [Gang et al62 cells . The new carrier .. fabricated superparamagnetic anisotropic nanocomponents (SAN) and loaded vincristine (VCR) to form VCR-SAN nanoparticles [. reported the simple and effective combination of IONP nanoparticles with bile acid–cisplatin derivatives to use as antitumor drugs and demonstrated its cytotoxicity to T cell leukemia (Jurkat) cells [Anisotropic nanoparticles have a longer blood circulation lifespan than previous isotropic nanoparticles. Based on this strategy, Xiong et alarticles . VCR-SANarticles . Díez ett) cells .. have developed an iron oxide nanoformulation loaded with the anticancer drug Doxironide, which can be used as a selective drug carrier for different types of AML [. prepared a chitosan-coated magnetic nanoparticle (CS-SPION) and loaded the anticancer drug paclitaxel, FA-CS-PTX-SPION, through reverse microemulsion technology [Magnetic nanoparticles also show good properties in drug loading and delivery. El-Boubbou et als of AML . Musawi chnology . FA-CS-P. constructed nanoparticles using magnetic Fe3O4 nanoparticles as the carrier to deliver 2-methoxyestradiol for MDS treatment [In other blood cancer, Xia et alreatment .In addition to the commonly used metallic nanoparticles as delivery carriers, other metal nanoparticles such as titanium oxide, calcium and palladium have also been used in leukemia treatment research.2) is a nanomaterial widely used in medicine and life sciences. Song et al. reported the use of highly reactive TiO2 nanoparticles combined with daunorubicin to inhibit the MDR resistance of leukemia K562 cells [2. Regarding the toxicity of TiO2 nanoparticles to blood cells, Cui et al. found that TiO2 nanoparticles are not toxic to macrophages THP-1 within a concentration range as high as 220 µg/mL and showed that this is for any safe nanotechnology product [Nanotitanium dioxide  using the aqueous leaf extract of Geranium Geranium to kill leukemia cells [Geranium Geranium extract on human leukemia cell line (K562) showed a dose-dependent cytotoxicity. The green synthetic Pd NPs will bring new opportunities to the biomedical field. Kaur et al. developed a dual-functional nanocarrier using palladium [50 value indicated that it has the potential to be used as an anticancer agent.Palladium nanoparticles have gained attention in precious metal nanoparticles due to their wide application in materials science and medicine. Li et alia cells . The in alladium . The nan. reported a Targeted Alpha-particle Nano-generator Actinium-225 (225Ac)-lintuzumab (anti-CD33) nanomedicine for the treatment of acute myeloid leukemia (AML) [. used human monocytic leukemia cells (THP-1) as a model to evaluate the uptake and free radical scavenging activity of nanocerium oxide [As for other rare metal-based nanomedicines, Jurcic et alia (AML) . Cerium um oxide . The datum oxide .. constructed a nanoparticle using cadmium telluride quantum dots as nanocarriers to deliver DOX for MM treatment [. proved that zinc oxide nanoparticles could induce apoptosis of MM cells, which was a potential clinical agent for MM [For other blood cancer, briefly, Chen et alreatment . Li et at for MM .Thomas first utilized HSCT in leukemia treatment in the 1960s . HSCT isHowever, the lack of donor HSCs is an important factor limiting the effectiveness of HSCT, so increasing the number of HSCs is an effective strategy for improving HSCT. Ex vivo expansion of stem cells requires more complex conditions than that of mature cells. The use of nanomaterials to expand stem cells has been extensively investigated –138. SpeThe bone marrow microenvironment plays an important role in maintaining the homeostasis and expansion of HSCs –141. Mim. reported an amphiphilic hydrogel that can amplify HSCs [. constructed a fibrin scaffold for the expansion of cord blood hematopoietic stem cells in vitro [Hydrogel is commonly used for expanding stem cells ex vivo . Gvaramiify HSCs . Mechaniify HSCs , 147. Inify HSCs . Ferreirin vitro . Predictin vitro . Finallyin vitro –154.. reported that polyvinyl alcohol (PVA) can be used to amplify the number of HSCs [Nanoparticles can also promote HSCs amplification in vitro. PVA nanomaterials are widely used in LCD electronic screens, semiconductors and other applications. Wilkinson et al of HSCs . In mous of HSCs . The mec of HSCs , 158.. reported a method to facilitate T cell reconstruction and immune response after HSCT [After HSCT, the immune system cannot be rebuilt immediately, and thus, patients are susceptible to bacterial infection or external stimulation, finally leading to failure of transplantation. The efficiency of the reconstitution of the immune system after HSCT is an important factor affecting the prognosis of patients. Shah et alter HSCT . This stImmunotherapy is a major milestone in cancer treatment and is the most promising way to completely control cancer . Tumor iUtilizing nanotechnology to enhance immunotherapy holds great promise in cancer research . The appThe high recurrence rate is a key factor leading to the low five-year survival of leukemia . An effe. reported that a hydrogel-based AML vaccine can produce a long-lasting immune response in the body [Using biological materials is an effective way to improve the immune system’s recognition of antigens Fig. . Mooney the body . The vac. reported a therapeutic leukemia vaccine [Nanoparticle formulations also improve the immune system’s recognition of antigens. Ma et al vaccine . The vac. reported a nanomedicine that can enhance the immune response to lymphoma [. reported a BCMA peptide-engineered nanoparticle for the enhanced clinical treatment of multiple myeloma by enhancing the function of CD8 + cytotoxic T lymphocytes [Boosting the immune system is also an effective strategy for lymphoma and myeloma. Enhancing the body’s immune response and memory is an effective strategy to prevent recurrence. Islam et allymphoma . Regardiphocytes . Please . reported a method for preparing CAR-T cells in vivo by using nanoparticles [CAR-T therapy has made progress in the treatment of refractory and relapsed B cell leukemia and lymphoma. Due to the lack of suppressive cells in the TME, such as in solid tumors, CAR-T cells and immune cells can easily access cancer cells and then kill them. However, the preparation of CAR-T cells is complicated and costly and may cause serious CRS side effects, which seriously restricts their application. To simplify the preparation process, Smith et alarticles . This naThe clinical diagnosis of HMs currently mainly relies on blood, bone marrow, cytochemical staining and labeling of monoclonal antibodies to detect leukemia cell surface differentiation antigens and other biochemical detection methods. Currently, there is still lack of sensitive and specific molecular biology detection methods. Unclear detection of leukemia leads to undertreatment or overtreatment and side effects, seriously affecting the patient’s survival.The emergence of nanotechnology has made the diagnosis of tumors more accurate. With the help of the paramagnetic and fluorescent labels of nanoparticles, the location and progress of cancer cells can be accurately diagnosed. For example, a micro-nanodiagnostic instrument can be implanted in the human body to run with the blood, and real-time transmission of internal information to the external recording device. Nanotechnology can make the diagnosis of leukemia more rapid and accurate , thereby. reported a gold nanoparticle-modified glassy carbon electrode (GCE) used to distinguish different leukemia cancer cells [Electrochemical-based nanotechnology is widely used in diagnosis of leukemia –177. Hu er cells . By chaner cells .. reported that a fluorescently labeled antibody-conjugated nanotube can be used to quickly and label-free detect CD45 + microvesicles from leukemia cells in about 30 min [Antibody-modified nanoparticles can also be used to track tumor cells and improve diagnostic accuracy . Chaudhut 30 min . The det. have developed a monoclonal antibody (mAb)-conjugated magnetic nanobiological probes (FMBMNs) to detect and isolate a single type of tumor cells [Magnetic multifunctional probes have broad prospects in the fields of biomedical research and diagnosis. Song et alor cells . The pro. reported a gold nanorod that can induce aggregation at high temperature for rapid visual detection of lysozyme in serum. The gold nanorods are functionalized by connecting nucleic acid aptamers targeting lysozyme. Exposure of the nanoprobe to nanomolar level of lysozyme will cause specific structure aggregation. This research shows that the use of thermally induced gold nanorod aggregates has a broad prospect as a nanodiagnostic technology [Temperature responsive nanotechnology can also be used to detect leukemia cells. Gold nanorods are widely used in nano-biodiagnosis research. The high level of lysozyme expression is a characteristic sign of leukemia. Moghadam et alchnology .Principle of design nanomedicine is to specifically target and eliminate cancer cells while minimizing side effects. Based on tumor antigens which are highly expressed on the surface of cancer cells, modifying antibodies, peptides or aptamers that specifically recognize tumor antigens on the surface of nanoparticles is the most common strategy for nanomedicine design. According to recently reported HMs nanomedicines –192, we . reported a strategy utilizing hematopoietic stem cells (HSCs) to deliver drugs to bone marrow [The bone marrow is the main site for the pathogenesis and infiltration of leukemia and myeloma cells . Residuae marrow . In prine marrow . This ste marrow . The drue marrow .Fig. 5St. reported an MM nanomedicine for targeting the bone microenvironment [. reported a tumor microenvironment-targeted nanoparticle loaded with bortezomib and a ROCK inhibitor for the treatment of MM [. generated a nanoparticle that dually targets myeloma cells and cancer-associated fibroblasts for treatment of MM [. constructed a bone-targeting nanoparticle for co-delivery of decitabine and arsenic in the treatment of MDS [Bone is the place where myeloma cells survive and plays an important role in the occurrence and relapse of MM. Targeting bone to delivery drugs is an effective strategy. Swami et alironment . Similarnt of MM . Stromalnt of MM . Wu et at of MDS .. reported a nanomedicine that can target the lymph nodes and can considerably inhibit the growth of lymphoma [Lymph nodes (LNs) are the place where lymphocytes develop and mature, and also the place where lymphocytes transform into cancerous cells. Lymphoma has the characteristics of solid tumors, such as the presence of the immunosuppressive microenvironment and a high level of inflammation. Thus, targeted LNs delivery of drugs is an effective strategy for targeted elimination of lymphoma cancer cells. Schudel et allymphoma .Targeting the surface antigens of cancer cells is the most basic nanomedicine design strategy Fig. . Accordi. designed a high-affinity arsenic nanomedicine based on ferritin for specifically targeting leukemia cells [. delivered cytochrome C using ferritin as nanocarriers for the treatment of APL leukemia with high expression of CD71 on cell surface [The leukemia cells highly and stably express the transferrin receptor CD71 on the cell membranes due to the high demand for iron and abnormal iron metabolism. Based on this trait, Ma et alia cells . This na surface .. generated a nanodrug targeting CD123 molecules on the surface of LSCs [. reported a nanomedicine modified with CD123 antibody for the treatment of MDS [. reported a nanomedicine targeting the B220 antigen on the surface of AML stem cells [Cancer stem cells (CSCs) play a key role in the pathogenesis and relapse . Convent of LSCs . This nat of MDS . The nanem cells . This na. generated a nanomedicine for targeting leukemia stem cells in the bone marrow and delivered gene editing tools to silence oncogenes [Brian et al. developed a PDT therapy via targeting leukemia stem cells using CPSNP calcium phosphate nanoparticles loaded with ICG [Leukemia stem cells prefer to live in the bone marrow. Ho et alncogenes . The nanwith ICG . Among t. generated a nanodrug targeting folate receptors (FR) on the surface of lymphoma cells [. generated a biomimetic silver nanoparticle for targeting FA receptors on lymphoma cells [. reported an CD20 antibody-modified paclitaxel-loaded nanoparticle for the treatment of CD20 + B cell lymphoma [. reported a rituximab (anti-CD20)-modified AZD-2014 encapsulated nanoparticle for killing B lymphoma cells [. reported a BCR-targeted multimodal imaging-engineered nanoparticle for the therapeutic and diagnosis of B cell lymphoma [In lymphoma, construction nanomedicines for targeting lymphoma cells are also getting a lot of attention. Generating nanomedicine specifically targeting lymphoma cells is the basic strategy. The targets of lymphoma nanomedicines mainly include CD20, CD40, folate receptor (FR) and BCR. Qiu et alma cells . Similarma cells . Regardilymphoma . Tang etma cells . Torino lymphoma .. first performed an in vivo evaluation of use CD38 and CD138 as targets for nanoparticle-based drug delivery in MM [. reported a CD38-targeting nanoparticle for enhancing bortezomib activity and the specificity of proteasome inhibition in MM [. showed that modification of anti-CD38 on nanoparticles carrying STAT3 inhibitors can improve the therapeutic effect of myeloma [. generated a CD38-targeted daratumumab immunopolymer for chemotherapy of MM [. reported a VLA4-targeted nanoparticle for the treatment of MM that functioned by hijacking cell adhesion-mediated drug resistance [. generated bortezomib-loaded mesoporous silica nanoparticles (MSNs) targeting FA receptors to selectively alter metabolism and induce the death of myeloma cells [In other blood cancer, nanomedicine studies have also increased in recent years –224. Basry in MM . Puente on in MM . Huang e myeloma . Similarpy of MM . Regardisistance . Nigro ema cells .. reported a nanomedicine for targeted degradation of the BCR-ABL fusion gene in CML cells [. designed an Au nanoparticle-based AuNP@PEG@e14a2 for silencing BCR-ABL fusion gene overexpressed in CML [Abnormal activation of intracellular signaling is an important signature of cancer cells. In CML cells, the BCR-ABL fusion gene is continuously highly expressed. This gene encodes a tyrosine kinase, which plays a key role in maintaining the survival of leukemia cells. In this context, Liu et alML cells . In an ad in CML .. reported a nanomedicine for targeting and degradation of Bmi1 mRNA in leukemia cells. The drug uses PEI and has as nanocarriers and encapsulates si-Bmi1 for Bmi1 gene. In animal experiments, this nanomedicine substantially prolonged the survival of diseased mice [. reported a study using HSA as nanocarriers to deliver HDAC inhibitors for AML treatment [Mutations of genes involved in epigenetic pathways play an important role in the pathogenesis of leukemia. The Bim1 gene has been reported to maintain the stability of histones in the nucleus. Kushwaha et alsed mice . Similarreatment .. designed a Au nanomedicine to target the NCL/miR-221/NF-kB/DNMT1 pathway in leukemia cells [. generated a nanomedicine that can trigger apoptosis mediated by the ROS/TNF-α pathway in leukemia cells [Besides specific signaling molecules, abnormal activation of pathways also plays an important role in cancer cell survival. Deng et alia cells . In mousia cells .. generated a tissue factor-targeted “O2-Evolving” nanoparticle for photodynamic therapy of malignant lymphoma [. proved that the nanoparticle formulation of AZD2811 had stronger anti-B cell tumor effect [. have developed a nanodrug targeting BCL-2 for the treatment of B cell lymphoma [. generated an iron oxide nanoparticle that could remotely and magnetically control the autophagy process in mouse B lymphoma cells [. reported that preparing the adjuvant into the form of nanoparticles can enhance the killing effect of lymphoma cells [. demonstrated that asparagine-laminated gold nanoparticles (Asn-AuNP conjugates) can enhance toxicity to leukemia cells by targeted intracellular heat transfer [Besides cell surface antigens, intracellular signaling molecules and metabolic pathways, such as Ara-KB, hypoxia and autophagy pathways, can also be used as targets to design nanomedicines Fig. . Li et alymphoma . AZD2811r effect . Martucclymphoma . Lin et ma cells . Adjuvanma cells . Shadab transfer .Fig. 7TaDrug resistance in cancer cells is an important factor leading to poor chemotherapy outcome and cancer recurrence. Increasing the uptake of drugs in cancer cells is an effective strategy to improve the tumor killing effect. Nanoparticles loaded with chemotherapeutic drugs can considerably increase the uptake and accumulation of drugs in cells and enhance the therapeutic efficacy of free chemotherapy drugs , 246.. reported a strategy to use carbon nanotubes to overcome leukemia drug resistance [. designed a PLA polymer nanoparticle surface modified with P-glycoprotein antibody to overcome AML resistance and improve the therapeutic effect [. reported that modification of the chemotherapeutic drug DNR on the surface of TiO2 nanoparticles by electrostatic adsorption can drastically increase the accumulation of DNR in K562 cells and reduce P-glycoprotein-mediated drug resistance [Ion channel proteins on cell surface play a key role in mediating drug resistance. The P-glycoprotein plays an important role in the drug resistance of cancer cells. Li et alsistance . The druc effect . In addisistance .. reported that modifying DNRs on the surface of nanodiamonds can bypass ABC transporter-mediated drug resistance and increase the accumulation and toxicity of DNRs in cancer cells [The role of the ABC protein family in mediating drug resistance has been widely reported. Man et aler cells .. loaded the chemotherapeutic drug DNR and the photosensitive molecule NIR797 into PEG-PLL-PLGA polymer nanoparticles [. designed an albumin-based nanodrug system to increase drug load and achieve controlled release of Doxorubicin [. reported that used bovine serum albumin nanoparticles as delivery carriers and encapsulated Dox to prepare DOX-DBSA-NPs can enhance the killing effect of drugs on leukemia cells and reverse drug resistance [Organic polymer materials and biological macromolecules are also used as nanocarriers to load chemotherapeutics to overcome drug resistance. Guo et alarticles . In vitrorubicin . This drsistance .Enhancing the drug toxicity to cancer cells is also an effective strategy for enhancing the therapeutic effect. We selected representative nanomedicine to demonstrate their better pharmacokinetics and therapeutic effects compare to non-nanomedicine Table .Table 5C. proved that conjugation of photosensitizers sulfonated aluminum phthalocyanine with chemotherapy drugs DOX can significantly improve the efficacy of photodynamic therapy for leukemia [. demonstrated that nanoparticles encapsulating both bortezomib and DAC significantly enhanced the toxicity to MM cells [Qin et alleukemia . Kim proleukemia . Similarleukemia . For othMM cells .. reported a TCR-signaling responsive nanoparticle to improve the killing of CAR-T cells [Enhancing the function of CAR-T cells through nanotechnology is an extremely promising field in cancer treatment . Nanotec-T cells . Similar-T cells . Similar-T cells . Similar-T cells .. reported that using genetically programmable vesicles to improve the tumor microenvironment and enhance the killing function of CAR-T cells [In addition to cell surface modification nanotechnology, the combination of engineered nanoparticles also enhanced CAR-T therapy. Li et al-T cells .Moreover, the drawbacks and shortcomings of CAR-T therapy also need to be addressed. CAR-T therapy possesses serious side effects, which can produce serious CRS and neurotoxicity . In addiIn addition to adoptive T cell-based therapy, other immune cells can also be used in immunotherapy. Engineering NK cells into CAR-NK for cancer treatment also holds great promise , 269. MaRelapse is the most serious problem affecting the prognosis and survival of HMs patients. Most patients will face recurrence after first remission. Studies on relapsed AML patients after allogeneic transplantation have shown that reducing MHC-II levels in cancer cells is an important cause leading to immune escape and relapse .To achieve long-term tumor control, the immune system must be trained to form lasting memories . TherefoRegarding HSCT, using nanomaterials to promote the expansion of HSCs is of great help to improve the efficiency of HSCT. It is critical to understand and determine the key chemical and physical conditions that promote the expansion of HSCs  and mostAlthough nanomaterials show excellent properties and great prospects in drug delivery, the toxicity of various nanomaterials still need to be carefully studied , 277. ThTsai et al. showed that after treatment with gold nanoparticles (Au NPs), human chronic myeloid leukemia cells showed growth inhibition and apoptotic necrosis phenotype [In addition to being drug delivery carriers, metal nanoparticles themselves can also cause toxicity to cells. henotype . Mechani. showed that multi-walled carbon nanotube buckypaper can induce cell cycle arrest by regulating AKT and MAPK signaling pathways and raised the issue of biocompatibility and potential toxicity [. investigated the cytotoxicity of carbon nanotubes of different lengths to monocytes and found that longer nanotubes were more toxic because they were harder for macrophages to envelop [Carbon-based nanomaterials are also toxic to cells. Dinicola et altoxicity . Sato et envelop . Similar envelop .In the end, nanomedicine is gradually moving out of the laboratory and entering clinical trials, and there are also many problems to be faced . The bio


We present Multimodal Subspace Independent Vector Analysis (MSIVA), a methodology to capture both joint and unique vector sources across multiple data modalities by defining linked and modality-specific subspaces. In particular, MSIVA enables the estimation of independent subspaces of various sizes within modalities and their one-to-one linkage to corresponding subspaces across modalities. We compare MSIVA to a fully unimodal initialization baseline and a fully multimodal initialization baseline, and evaluate all three approaches with five distinct subspace structures on synthetic and neuroimaging datasets. We first demonstrate that MSIVA and the unimodal baseline can identify the correct ground-truth subspace structures from the incorrect ones in multiple synthetic datasets, while the multimodal baseline fails at detecting high-dimensional subspace structures. We then show that MSIVA can better capture the latent subspace structure with the minimum loss value in two large multimodal neuroimaging datasets compared to the unimodal baseline. Our results from subsequent per-subspace canonical correlation analysis (CCA) and brain-phenotype modeling demonstrate that the sources from the optimal subspace structure are strongly associated with phenotype measures, including age, sex and schizophrenia-related effects. Our proposed methodology MSIVA can be applied to capture linked and unique biomarkers from multimodal neuroimaging data. However, each neuroimaging modality has its own strengths and weaknesses, and only captures limited information of the brain. For example, structural MRI (sMRI) can show high-resolution anatomical structure of the brain but cannot capture temporal dynamics, while functional MRI (fMRI) can measure blood-oxygenation-level-dependent (BOLD) signals across time at the cost of lower spatial resolution. Therefore, it is important to take advantage of multimodal neuroimaging to effectively capture interpretable and multifaceted information about the brain.To jointly analyze multiple datasets or data modalities, a unified framework Multidataset Independent Subspace Analysis (MISA)  has beenMultimodal Subspace Independent Vector Analysis (MSIVA)   of all shttps://github.com/trendscenter/MSIVA.git.The analysis code is publicly available at 2.3.2.3.1.For each subspace structure, we generate a synthetic dataset  dataset . X[m] is2.3.2.We utilize two large multimodal neuroimaging datasets including two image modalities: T1-weighted sMRI and fMRI. The first dataset is from UK Biobank . 2907 su2.4.For each of five subspace structures ce (ISI) , 18, a mWe then perform the same experiments on two multimodal neuroimaging datasets using these five candidate subspace structures, and identify the optimal structure yielding the lowest final MISA loss value. Separate follow-up CCA of each subspace recovered projections with maximum cross-modal correlation.2.5., 1]) on the training set, then train the model using all training subjects and evaluate it on the hold-out test set. The age regression performance is measured by mean absolute error (MAE) and the classification performance is measured by accuracy.To further evaluate that association between the post-CCA sources and phenotype measures, we perform an age prediction task and a sex classification task for the UKB dataset, as well as an age prediction task and a binary SZ classification task (controls vs SZ patients) for the patient dataset. Specifically, we train a ridge regression model to predict the age, and a linear support vector machine to classify sex groups or SZ patients. For the UKB dataset, 2907 subjects are divided into a training set of 2000 subjects and a hold-out test set of 907 subjects. For the patient dataset, 999 subjects are divided into a training set of 699 subjects and a hold-out test set of 300 subjects in the age prediction task; 875 controls and SZ patients are grouped into a training set of 612 subjects and a test set of 263 subjects in the SZ diagnosis classification task. We perform 10-fold cross-validation to choose the best hyperparameters , to capture linked cross-modal sources as well as modality-specific sources. We first show that both MSIVA and the unimodal baseline can correctly identify the ground-truth subspace structures from the incorrect ones, and verify that the correct subspace structures result in the lowest loss values from multiple synthetic data experiments. We next apply both approaches on two large multimodal neuroimaging datasets and demonstrate that MSIVA can better reveal the latent subspace structure across two imaging modalities with lower loss values. Among all cases, MSIVA findings .Furthermore, MSIVA can be viewed as an extension of MMIVA except that their initialization methods  and subspace structures (MSIVA: flexible subspace structures; MMIVA: rces see . Thus, tIt is important to acknowledge limitations of our current work. The first limitation is the subspace structure used in MSIVA. MSIVA selects the subspace structure that best fits the data from a predefined set, according to the loss value. However, it is not efficient to exhaustively evaluate potential subspace structures and corresponding performance metrics. Additionally, we make two assumptions on the subspace structure that the linked subspaces share the same size across modalities, and that the modality-specific subspaces are one-dimensional. However, it is possible that these assumptions may not represent the underlying data structure. Ideally, we want to learn the underlying subspace structure from the data in an unsupervised manner. The second limitation is the linear mixing assumption in MSIVA. MSIVA assumes that the data of each modality can be transformed to linearly mixed sources, but the true mixing process can be nonlinear. Also, individual differences, site effects and preprocessing-related effects may introduce nonlinear artifacts, which are not taken into account in MSIVA. Future work will include applying data-driven subspace structures such as NeuroMark template , and dev5.Our proposed approach MSIVA can capture linked and modality-specific sources on both synthetic and neuroimaging datasets and yield a lower loss value compared to the unimodal baseline. The sources in the linked subspaces are strongly associated with age, sex and psychosis according to brain-phenotype modeling. Our results support that MSIVA can be applied to capture linked and unique imaging biomarkers from large multimodal neuroimaging datasets.


Hyperspectral stimulated Raman scattering (SRS) microscopyis arobust imaging tool for the analysis of biological systems. Here,we present a unique perspective, a label-free spatiotemporal map ofmitosis, by integrating hyperspectral SRS microscopy with advancedchemometrics to assess the intrinsic biomolecular properties of anessential process of mammalian life. The application of spectral phasoranalysis to multiwavelength SRS images in the high-wavenumber (HWN)region of the Raman spectrum enabled the segmentation of subcellularorganelles based on innate SRS spectra. Traditional imaging of DNAis primarily reliant on using fluorescent probes or stains which canaffect the biophysical properties of the cell. Here, we demonstratethe label-free visualization of nuclear dynamics during mitosis coupledwith an evaluation of its spectral profile in a rapid and reproduciblemanner. These results provide a snapshot of the cell division cycleand chemical variability between intracellular compartments in single-cellmodels, which is central to understanding the molecular foundationsof these fundamental biological processes. The evaluation of HWN imagesby phasor analysis also facilitated the differentiation between cellsin separate phases of the cell cycle based solely on their nuclearSRS spectral signal, which offers an interesting label-free approachin combination with flow cytometry. Therefore, this study demonstratesthat SRS microscopy combined with spectral phasor analysis is a valuablemethod for detailed optical fingerprinting at the subcellular level. In addition, the coordinated divisionof subcellular organelles during mitosis supports their inheritanceand cellular homeostasis.2 Mitosis comprisesfive sequential stages known as prophase, prometaphase, metaphase,anaphase, and telophase/cytokinesis, where mechanisms such as chromatincondensation, nuclear envelope remodeling, modulation of the spindleapparatus, and chromosome alignment/separation facilitate equal distributionto the daughter cells with elegant precision.2 The functional dissection of mitotic biochemistry is still an activearea of investigation due to the multifaceted nature of cell cycledynamics and is of great importance to researchers, as its dysregulationhas catastrophic consequences, including cancer.4 Thedevelopment of approaches sensitive enough to evaluate the molecularprofiles of individual cells provides an opportunity to gain a newperspective on some of the most fundamental processes of life.The mammalian cell cycle isregulated by a well-studied but complex biochemical reaction system,which has fascinated microscopists for over a century.5 however, labelingtarget molecules is a key element to its execution. Labeling probes,such as fluorescent proteins or organic dyes, may perturb the intrinsicbiochemical properties of the cell, and their application can oftenimpair or modify its biological functions, coupled with potentialimaging artifacts from nonspecific targeting.7 Inaddition, photobleaching, phototoxicity, and low multiplexing levelsresulting from broad fluorescent spectra are also limitations of thismethod.8 Thus, label-free imaging modalitiespresent a distinct advantage over the use of imaging dyes or fluorophoresfor visualizing cellular processes, i.e., the preservation of chemicalspecificity. Vibrational techniques like infrared (IR) spectroscopycan be used in histological analysis without the requirement of labels;10 however, IR radiation is strongly absorbed in aqueous specimens,and the resolution is restricted to whole-cell analysis.11 When imaging cellular models, Raman spectroscopyis a preferred optical tool because it can provide label-free characterizationand distribution of biomolecules under biocompatible imaging conditions.This noninvasive technique has been widely applied to the study ofcellular processes pertinent to biomedicine12 including cancer diagnostics,13 cardiovasculardysfunction,14 endothelial pathologies,15 and pharmacokinetics;16 however, long image acquisition rates can affect its biologicalsampling potential.Optical microscopy methods are often used to characterize cellularprocesses because they offer information on the distribution and dynamicsof biomolecules at the single-cell level. Fluorescence microscopyis classically used for visualizing the specific structures and functionsof biological systems;17 SRS has enabled the biochemicalprofiles of lipids, proteins, and nucleic acids within numerous biologicalmodels to be visualized including cells,19 tissues,21 and animals.23 Considering its high molecular specificity, sensitivity, and imagingrates, SRS imaging has emerged as a unique tool for pharmaceuticaldiscovery.24 Drug uptake and distributioncan be observed in high resolution through the use of endogenous chemicalbonds or small bio-orthogonal tags,25 permittingintracellular pharmacodynamics to be monitored in real time.26 To enhance the chemical sensitivity of the SRSdata obtained, a hyperspectral imaging approach can be applied,27 such that each pixel of an acquired image containsan intensity value as a function wavenumber in the Raman spectrumby automatic retuning of the pump wavelength in between image frames.To delineate these often rich hyperspectral data sets, multivariateanalysis techniques have previously been employed, including principalcomponent analysis (PCA), independent component analysis (ICA), andmultivariate curve resolution (MCR), to extract underlying biochemicalinformation from cellular constituents .30 Recently, the application of k-means cluster analysis(KMCA) to multiwavelength SRS images in the high-wavenumber regiongenerated a visual interpretation of endogenous LD composition inprostate cell models following drug perturbation.31 An alternative chemometric analysis tool for Raman spectraldata is spectral phasor analysis, which has proven advantageous forcellular and tissue segmentation,33 cytometry,34 exploring drug–cell interactions,35 and chemoresistance mechanisms.36 SRS microscopy coupled with advanced chemometrics providesa label-free means to explore critical aspects of cell biology.The development of stimulated Raman scattering(SRS) microscopyis credited with improvements in the spatial resolution, temporalanalysis, and 3D imaging capabilities for bioanalysis using Ramanspectroscopy.–1) probed biomoleculesat each phase of mitosis in the SK-BR-3 cell model. Cellular segmentationby multivariate analysis enabled the biochemical distribution of lipids,protein, and DNA to be considered specific to endogenous featuresin a robust and reproducible manner. This approach also allowed cellsin the same field of view to be characterized in separate phases ofmitosis without labeling. These results demonstrate the potentialof hyperspectral SRS imaging and spectral phasor analysis for investigatingfundamental processes of cell biology.Herein, we report the use of spectral phasor analysis to assesssubcellular features associated with mitosis and its associated spectralprofile by SRS microscopy. A label-free assessment of the high-wavenumberregion (2800–3050 cm–1) across the range 2800–3050cm–1 to collect a hyperspectral stack of 40 SRSimages. A spectral phasor algorithm32 wasapplied to this data set, clustering each data point based on its SRS spectral similarity within the stack of images togenerate a 2D chemical map of the cell model. Spectral phasor usesthe Fourier transform to depict the spectrum of every pixel in a 3Dhyperspectral SRS image stack as a point on the 2D phasor plane, providinga global overview of the ensemble of pixels. As such, spectral phasoranalysis can simplify the interpretation of hyperspectral data setsto visualize the biochemical differences between cellular populations.Segmentation of the phasor plot enabled the identification of intracellularregions of interest (ROIs), and the corresponding SRS spectrum associatedwith the nuclear region could be used to probe the changes in cellularbiomolecules during cell division.The aim of this research wasto capture the dynamic biologicalphases of mitosis and evaluate the molecular contributions which facilitatecell division via a label-free approach. Herein, we report the firstapplication of hyperspectral SRS imaging, supplemented with spectralphasor analysis, to investigate the distribution of cellular DNA,protein, and lipids throughout mitosis. With readily identifiableand compartmentalized morphological structures, the SK-BR-3 cell modelwas considered an excellent candidate for phenotypic assessment. Toprobe its cellular biochemistry, a wavelength scanning experimentwas performed by tuning the pump laser wavelength at increments of0.4 nm , protein , and lipids  in theHWN region can be observed which are key C–H stretch assignmentspreviously described in the DNA imaging of live cells.18 Lipids are predominately distributed throughoutthe cell cytoplasm and as LDs which serve a plethora of intracellularfunctions.37 In contrast, the total proteincontent of the cell is largest in the nucleus through its intrinsicassociation with DNA, where histones and many other nuclear proteinsare localized.2 Interestingly, despitecomparable spectral profiles (–1) in combination with phasor analysis offers a platformto explore the biochemical transformation of a cell during mitosis.Differentiation between cellular protein contentis demonstratedin profiles 1C, the nprofiles 1A, which2 To capture this short biological process (<1h), we performed a double-thymidine block to synchronize a populationof SK-BR-3 cells in culture and preserved the dividing cells in paraformaldehyde(PFA) at key time points during mitotic entry.38 Mitosis is a highly dynamic process, and under the microscope,live cells may become stressed; therefore, to minimize changes incell biochemistry during the analysis, PFA fixation was chosen. Threebiological replicates were imaged at each of the five phases of mitosis,and an interphase model was used for comparison. SRS images were acquiredacross a range of wavenumbers (2800–3050 cm–1), and from this stack, an average intensity projection was generated.Spectral phasor plots were then produced for each mitotic phase . To corroborate our label-free interpretation of the mitotic phasesby SRS, we adopted a multimodal approach by labeling the cells witha DNA fluorescent dye (DAPI), which allowed sequential SRS microscopyand fluorescence imaging via a Leica microscope system. Parity betweenSRS imaging via phasor segmentation of the nucleus and the DNA contrastagent, DAPI, is evident, demonstrating the density and distributionof the condensed chromatin within the cell at each of the phases ofmitosis and prophase b (proB).39 A phasor approachreadily segments the nuclear domain  and delivers subtle molecular interactions specific to the nucleolusin proA and proB , which mayultimately be overlooked during whole cell analysis. In the late prophasecell (proB), a noticeable change in SRS signal attained from the nucleolarsegment is observed ), whilethe ratio at 3020/2930 cm–1 associated with thelocalized RNA/protein levels40 declines), concomitant with theordered release of nucleolar complexes and repression of RNA transcription.39 These data suggest it is possible to identifychemical contrast within the mammalian nucleus at different cell cyclestages. In addition, multiple labels would be required to resolvethese features by fluorescence imaging, whereas label-free SRS cancharacterize these endogenous structures efficiently and effectively.In early prophase, the homogeneously distributedchromatin of interphaseform visible thread-like structures  in eukaryotic cells provide a physicalbarrier to separate DNA replication and transcription from other cytoplasmicevents.modeling 3A. The C nucleus 3B.2 Once aligned at the equator of the spindle inmetaphase, the chromatin adopts its most compacted state during mitosis(45 The percentage change in SRS intensity in thenucleus (∼12.5%) between prometaphase and metaphase at 2970cm–1 (DNA) may be indicative of this process (46 This may explainthe difference in the intensity ratio (DNA:CH3) valuesobserved between mitotic and interphase cells  that link the spindle microtubulesfacilitate elongation between the poles, prizing the homologous chromosomesapart toward opposite ends of the cell .This finding led us to probe the LD distribution using phasor segmentation,cholesterol esters , and saturatedlipids .49 An increase in saturated lipids was observed at early prophase andprometaphase in both LD spectra . Due to the essential role lipid unsaturation plays in the biophysicalproperties of membranes ,55 we analyzed the ratio at 3015/2851 cm–1 .56 Overall, we observed higher rates of lipid unsaturation during mitosisin the cytoplasm , cell boundary, and nuclear boundary , with the highest rates of unsaturationin membrane-associated regions . The highest degree of unsaturation associated with membrane fluiditywas found during anaphase , and where both the expansion of the plasma membrane aids correctspindle alignment for symmetric division and the nuclear membranereassembly transpires.57 In addition, NEstability is influenced by the degree of saturation of lipids in ERmembranes which could regulate the enzyme activity of lipids to controllipid content and abundance crucial to its reformation.59 The lowest measurements of unsaturation were recorded in telophase,where a decrease in membrane turnover is typically observed at thisfinal stage of the cell cycle.60 Nuclearreassembly and chromatin decondensation re-establish the cells’interphase structure, as the nucleolus reforms and transcription ofRNA begins. In contrast to anaphase and metaphase, the ratio of DNAto protein (2970/2930 cm–1) decreased during telophase,since the distribution of DNA has already become relatively diffuseat this late stage of mitosis (3) indicatesextensive redistribution in both nuclear and cytoplasmic compartments,where abscission takes place  as a functionof its composition.35During our investigationwe discovered that a spectral phasor approachcan also be utilized to detect cells in different phases of division,as shown in the segmented SRS images presented in –1 5B, whichWe have reported the use of hyperspectralSRS imaging to visualizethe intrinsic vibrational signals from single cells to measure theirbiochemistry throughout mitosis. Label-free SRS imaging of this fundamentalprocess of eukaryotic life captured the gradual spectral changes inbiomolecules at various subcellular compartments. A spectral phasorapproach to image analysis demonstrated chromatin distribution duringmitosis, which paralleled the fluorescent DNA contrast agent withoutdisturbing cellular composition. Our results also demonstrate theuse of chemometric-based image analysis for cell cycle differentiation,which in tandem with flow cytometry could be a transformative platformfor label-free cytometry. Potential future research may include thedevelopment of organelle-specific probes  that facilitate the precise imaging of division structures,which would enhance the spatial resolution of these dynamic nanoscalesubcellular components during the cell cycle. In addition, automaticsegmentation of the spectral phasor plot using machine learning methodologythat is based on a priori spectral features  could increase the throughput of the technique, thereproducibility of the phasor segmentation, and the end-user accessibility.To conclude, SRS microscopy coupled with spectral phasor analysisoffers a versatile modality to visualize the role of molecules atthe subcellular level.


It can be difficult to diagnose coronary artery disease in patients with acute coronary syndrome if coronary angiography does not identify stenosis. Coronary inflammation, which can contribute to the pathogenesis of coronary artery disease and acute coronary syndrome, can be quantified using the perivascular fat attenuation index. Furthermore, the perivascular fat attenuation index is a marker for all-cause mortality, cardiac-related mortality and impaired global coronary flow reserve.Here we report a case of a patient presenting with symptoms of acute coronary syndrome. The patient had hypokinesis of the lateral-posterior wall of the left ventricle, decreased myocardial perfusion in the posterior wall myocardium and elevated myocardial troponin-T and creatine phosphokinase levels. However, coronary computed tomography angiography did not identify arterial stenosis. The patient did have an increased perivascular fat attenuation index, indicating coronary inflammation. Moreover, the fat attenuation index was higher around the left circumflex artery than around the right coronary artery or left anterior descending artery. Intravascular ultrasonography identified an intramural haematoma, leading to a diagnosis of type 3 spontaneous coronary artery dissection in the left circumflex artery.Perivascular fat attenuation index may be a useful tool to help identify and localise disease-causing lesions, and to direct further testing to confirm a diagnosis of spontaneous coronary artery dissection in acute coronary syndrome patients without significant arterial stenosis. The diagnosis of coronary artery disease (CAD) is often difficult to make in patients with symptoms of acute coronary syndrome (ACS) if coronary angiography does not show severe stenosis of the coronary arteries. In such cases, measurement of the perivascular fat attenuation index (FAI) may help identify coronary inflammation and direct further diagnostic testing to identify CAD.Epicardial adipose tissue, including the perivascular fat around coronary arteries, can secrete pro-inflammatory adipokines , 2. InflPerivascular FAI is a validated non-invasive marker for coronary inflammation , 4. It iHere, we present a case of a patient with signs of ACS but with no significant coronary artery stenosis. We used CCTA images to identify increased perivascular FAI and intravascular ultrasonography to diagnose SCAD in the LCX of this patient.4, we traced a 3-mm width of the coronary adventitia along the total length of the three main coronary arteries in the CCTA images. Our measurements differed from those of the original report because we traced the proximal 40-mm segments of all three major epicardial coronary vessels. The total length of the coronary artery was approximately 155 mm for all three branches. We defined perivascular fat as adipose tissue surrounding the vessel wall within a distance equal to the diameter of the vessel [A 49-year-old woman with the clinical characteristics of ACS was referred to our hospital. She had a 4-day history of chest pain. A 12-lead electrocardiogram (ECG) showed negative T waves in leads II, III and aVF. Transthoracic echocardiography (TTE) showed hypokinesis of the lateral-posterior wall of the left ventricle. The patient’s myocardial troponin-T and creatine phosphokinase (CPK) isoenzyme levels were elevated . Coronary computed tomography angiography (CCTA) images showed decreased myocardial perfusion in the posterior wall myocardium; however, they did not show significant coronary artery stenosis. Therefore, we measured the perivascular FAI for all three coronary arteries to quantify coronary inflammation. Using a previously described and validated methodWe found significantly higher FAI around the LCX  than around the LAD and RCA  study mapped perivascular fat attenuation in the proximal RCA, the LAD and the LCX in two cohorts of patients from Germany and the US . High peIn contrast, the link between inflammation, perivascular FAI and SCAD remains unclear. Perivascular fat attenuation has been reported in two SCAD patients . ConversHere, we present a case of an ACS patient who did not show significant coronary arterial stenosis, but who did have a higher FAI in the LCX than that in the RCA or LAD. The high FAI helped us to direct further imaging to the LCX and to diagnose SCAD in this vessel. The data presented in this report is from a single case and, therefore, further studies are required to confirm these findings.This case shows that using CCTA to measure perivascular FAI in the coronary arteries may be a useful tool to direct further testing and confirm a diagnosis of SCAD in ACS patients, even if no obvious atherosclerotic lesion is found in the coronary arteries.


We present an intriguing and rare case of a 71-year-old male who presented with a non-ST elevation myocardial infarction (NSTEMI). Initial coronary angiography revealed severe and unusual systolic extrinsic compression of the left main coronary artery (LM), warranting further advanced imaging investigations. Computed tomography angiography (CTA) and transesophageal echocardiography (TEE) were employed to determine the underlying cause, which was identified as a contained aortic rupture leading to the formation of a pseudoaneurysm in the left coronary sinus of Valsalva and aortic root. This condition was found to be a sequela of previously undiagnosed endocarditis, likely secondary to lower extremity osteomyelitis and bacteremia, for which the patient received prolonged intravenous (IV) antibiotic therapy. This case highlights the critical role advanced imaging techniques play in accurately diagnosing and characterizing complex cardiovascular abnormalities, enabling early intervention and optimizing patient outcomes. Healthcare providers should remain vigilant for such atypical presentations to ensure timely and appropriate management. Myocardial ischemia and infarction resulting from extrinsic compression of the left coronary artery are exceedingly rare and have only been documented in a limited number of case reports -6. This Enterococcus faecium. A transthoracic echocardiogram (TTE) performed during that admission failed to show any valvular vegetation, valvular structural damage, or valvular malfunction. He was on a regimen of multiple medications, including amiodarone, metoprolol tartrate, isosorbide mononitrate, warfarin, aspirin, atorvastatin, hydralazine, torsemide, and amlodipine. The patient completed 12 weeks of tailored antibiotic therapy. The patient's electrocardiogram (ECG) displayed ST depressions in the inferior and anterolateral leads, accompanied by elevated aVR  two years prior with coronary artery bypass grafts. Grafts were attached from the left internal mammary artery (LIMA) to the left anterior descending coronary artery (LAD), along with a saphenous vein graft (SVG) to the right coronary artery (RCA) and diagonal 1 (D1). He also had atrial fibrillation, arterial hypertension, type 2 diabetes mellitus, chronic kidney disease stage 4, and chronic warfarin use. The patient had a history of prolonged hospitalization and was completing intravenous (IV) antibiotics at a skilled nursing facility for osteomyelitis with bacteremia caused by vancomycin-resistant High-sensitivity troponins were consistent with acute coronary syndrome (ACS). Due to the presence of acute chest pain consistent with angina, elevated cardiac markers, and ECG changes suggestive of ischemia, the patient underwent urgent coronary and graft angiography. Angiography revealed that the grafts were patent, but it also exposed severe systolic compression of the mid- to distal sections of the LM coronary artery Figures ; Video 1In light of the patient's renal disease, a transesophageal echocardiogram (TEE) was done initially, which suggested the presence of a pseudoaneurysm in the aortic root with two entry points from the left coronary sinus of Valsalva Figures ; Video 2Further evaluation with gated cardiac computerized tomographic angiography (cCTA) revealed an extraluminal contrast-filled cavity adjacent to the left-sided aortic root, measuring 2.4 cm × 3.8 cm × 3.6 cm, and multiple peri-, supra-, and infra-valvular perforations Figures -4F. Based on the imaging findings, the patient was suspected to have a contained rupture of the aortic root, which encased the LM coronary artery and resulted in its collapse during systole, leading to angina and myocardial ischemia. The case was thoroughly discussed by the heart team and evaluated by cardiothoracic surgeons to determine the most appropriate course of action. After careful consideration, the patient underwent proximal aortic root repair along with homograft aortic valve repair. This surgical intervention aimed to address the aortic root rupture and restore normal blood flow to the compressed left main coronary artery. Post-operative management involved close monitoring in the intensive care unit, with a focus on optimizing cardiac function and preventing complications. Additionally, the patient's complex medical history required a comprehensive care plan involving collaboration among various specialties to ensure a successful recovery.Aneurysms of the sinus of Valsalva can be classified as either congenital anomalies or acquired conditions. Congenital sinus of Valsalva aneurysms are rare and occur due to the absence of muscular and elastic tissue in the aortic wall of the sinus of Valsalva ,5. ConveOur case presents an exceptional occurrence of non-ST elevation myocardial infarction (NSTEMI) caused by extrinsic compression of the LM coronary artery due to a contained aortic root perforation and a left coronary sinus pseudoaneurysm. This exceedingly rare condition emerged as a late consequence of previously undiagnosed prosthetic aortic valve endocarditis. Notably, there have been no reported cases of such extrinsic compression of the LM coronary artery to date. The unique etiology highlighted in this report underscores the significance of considering rare conditions in patients presenting with acute coronary syndromes, necessitating tailored management approaches for uncommon scenarios. Ultimately, this case report emphasizes the value of interdisciplinary collaboration and careful evaluation of atypical presentations to enhance patient care in complex cardiovascular cases. By sharing and analyzing such cases, we can deepen our understanding of rare cardiovascular complications and improve patient outcomes.This case highlights the vital role of advanced imaging techniques in accurately diagnosing and characterizing complex cardiovascular abnormalities, enabling early intervention for better patient outcomes. The unique etiology discussed emphasizes the importance of considering rare conditions in patients presenting with acute coronary syndromes, requiring tailored management approaches for such uncommon scenarios. Overall, this case report underscores the value of interdisciplinary collaboration and careful evaluation of atypical presentations to enhance patient care in complex cardiovascular cases.