Furthermore, gene enrichment analysis was carried out, and a regulatory gene network was constructed. Thus, the spectra of the common and unique effects of the PGP, PGPL, and Semax peptides under ischemia–reperfusion were distinguished.Glyprolines are Gly-Pro (GP)- or Pro-Gly (PG)-containing biogenic peptides. These peptides can act as neutrophil chemoattractants, or atheroprotective, anticoagulant, and neuroprotective agents. The Pro-Gly-Pro (PGP) tripeptide is an active factor of resistance to the biodegradation of peptide drugs. The synthetic Semax peptide, which includes Met-Glu-His-Phe (MEHF) fragments of adrenocorticotropic hormone and the C-terminal tripeptide PGP, serves as a neuroprotective drug for the treatment of ischemic stroke. Previously, we revealed that Semax mostly prevented the disruption of the gene expression pattern 24 h after a transient middle cerebral artery occlusion (tMCAO) in a rat brain model. The genes of this pattern were grouped into an inflammatory cluster (IC) and a neurotransmitter cluster (NC). Here, using real-time RT-PCR, the effect of other PGP-containing peptides, PGP and Pro-Gly-Pro-Leu (PGPL), on the expression of a number of genes in the IC and NC was studied 24 h after tMCAO. Both the PGP and PGPL peptides showed Semax-unlike effects, predominantly without changing gene expression 24 h after tMCAO. Moreover, there were IC genes ( Glyprolines are a class of oligopeptides that include an amino acid sequence consisting of Gly-Pro (GP) or Pro-Gly (PG) residues . GlyprolPreviously, we showed that both Semax and PGP modulated the expression of genes associated with processes of proliferation, differentiation, migration, survival, and cell death in a permanent middle cerebral artery occlusion (pMCAO) rat model. This model reflects the processes after acute stroke ,19. It wIn this study, we used PGP and another synthetic PGP-containing peptide, Pro-Gly-Pro-Leu (PGPL), in the rat tMCAO model. It should be noted that the PGP and PGPL peptides have numerous effects, which may be beneficial for the treatment of cerebrovascular accidents . The mulWhite two-month-old male rats of the Wistar line (weight: 200–250 g) were obtained from the AlCondi, Ltd., animal breeding house, Moscow, Russian Federation. The animals were divided into five groups: “sham operation” (SH), “ischemia–reperfusion” (IR), “ischemia–reperfusion + Semax” (IS), “ischemia–reperfusion + PGP” (IP), and “ischemia–reperfusion + PGPL” (IL). Each experimental group included at least eight animals.The transient middle cerebral artery occlusion (tMCAO) model was induced by endovascular occlusion of the right middle cerebral artery using a monofilament  for 90 min and was performed under magnetic resonance imaging (MRI), as described previously . The shaTo the rats of the IS, IP, and IL groups, the peptides Semax, PGP, and PGPl were administered intraperitoneally at doses of 100, 37.5, and 200 µg/kg rat weight, respectively. Animals of IR and SH groups were injected with saline. Peptides or saline were administered intraperitoneally after 1.5 h, as well as at 2.5 and 6.5 h after the surgical procedure. The used concentrations of substances and the time of peptide administration correspond to the data from the literature ,28,29,30The ipsilateral subcortical structures were obtained, and total RNA was isolated, as previously described . The RNA18 primers was synthesized, and RT-PCR was conducted as previously described [cDNA using oligo (dT)escribed . PCR-priGapdh was used to normalize the expression for the cDNA samples. The relative gene expression was calculated by the 2−ΔΔCt method [p < 0.05, as previously described [The reference gene t method . The datt method . At leasescribed .p-values, adjusted using the Benjamini–Hochberg procedure, lower than 0.05 (Padj < 0.05) were considered. Principal component analysis (PCA) of normalized read counts was conducted using R software. Hierarchical cluster analysis of DEGs was performed using Heatmapper  [The Database for Annotation, Visualization and Integrated Discovery (DAVID) v.6.8. , Gene Se Canada) . A volca Canada)  was usedWe detected the locations and volumes of ischemic foci in animals 24 h after tMCAO using the diffusion-weighted imaging (DWI) and T2-weighted imaging (T2 WI) modes of MRI. All ischemic rats from any of IR, IS, IP, and IL groups had ischemic injury in right hemisphere of the brain. A typical DWI with an ADC map and T2 WI scans of the formation of ischemic injury areas with a subcortical localization in the brains of rats 24 h after tMCAO are shown in iL1b, iL6, Ccl3, Socs3, Hspb1, and Fos) from an inflammatory cluster (IC) and six genes  from a neurotransmitter cluster (NC) associated with Semax’s action under IR conditions 24 h after tMCAO were selected for analysis based on our previous data [Padj < 0.05) between the IS and IR groups as determined by RNA-seq  and NC  genes under the influence of PGP and PGPL in the subcortical structures of the rat brain 24 h after tMCAO were studied.Using real-time RT-PCR, the expression changes for the IC  in IP versus IR , Chrm1 , and Gria3  genes under PGPL treatment 24 h after tMCAO was observed , iL1b , and iL6  genes under Semax versus PGP treatment 24 h after tMCAO was observed  from IC and three DEGs  from the NC related to PGPL treatment  and Ptk2b  and downregulation of Cplx2 , as well as Socs3 , Fos , iL6 , and Ccl3 , were observed in IL versus IS 24 h after tMCAO  predominantly translated signals of inflammatory and immune responses  involved in cluster functioning belonged to the IC. However, the majority of the genes (three) belonged to the NC. Thus, the Ptk2b gene was downregulated in IS versus IL, and Chrm1 and Gria3 were non-DEGs in both pairwise comparisons with the NC. Such genes were associated with neurosignaling pathways  that were included in the second pathway cluster . The peptides .Thus, a regulatory network illustrated the spectra of the effects of the Semax, PGP, and PGPL peptides on gene expression in the rat brain under IR conditions, against the background of the structures having a common PGP unit.In this study, we used three PGP-containing peptides  to differentiate their effects in a rat tMCAO model. The Semax peptide combines the ACTH4-7) and PGP structures and probably integrates the properties of each of the subunits. Semax is one of the most studied and practically significant peptides used in stroke therapy. The glyprolines PGP and PGPL can also be considered potential agents for the prevention and treatment of pathologies. They are able to exhibit neuroprotective, antihypertensive, anti-inflammatory, and antithrombotic effects  and PGP . The muliL1b, iL6, Ccl3, Socs3, Hspb1, and Fos) and six neurotransmitter  genes associated with Semax treatment under IR conditions were selected based on our previous data [Here, under MRI, a rat tMCAO model was implemented to test the peptides’ effects. Molecular genetics instruments were used. Six inflammatory  and Then, changes in the expression of the IC and NC genes under treatment with short PGP-containing peptides (PGP and PGPL) in the subcortical brain structures of rats 24 h after tMCAO were studied. Both the PGP and PGPL peptides showed effects, predominantly without changes in the gene expression under IR. This result emphasizes the importance of the structure of ACTH (MEHF) for Semax’s effects. In particular, the administration of the PGP and PGPL peptides did not have a statistically significant effect on the expression of the IC genes. It was shown that the PGP fragment structurally reproduces the key unit of the CXCL2 chemokine. CXCL2 is responsible for the activation of neutrophils. Moreover, PGP is able to bind to the CXCR2 receptor and partially recapitulate the chemoattractant function for neutrophils ,56.Gabra5 gene, encoding the α5 subunit of the γ-aminobutyric acid (GABA) receptor after PGP administration 24 h after tMCAO. The regulation of the activity of the GABA receptor is important in the formation of a protective reaction in response to the processes of excitotoxicity and microglia activation during ischemia [Cplx2, Chrm1, and Gria3) were increased in expression upon PGPL administration to ischemic rats. Such genes belong to the NC, and the proteins encoded by these genes are involved in the regulation of neurotransmitter processes. Moreover, the repression of the neurotransmitter system under conditions of cerebral ischemia has been noted previously [Among the NC genes, we found an increase in expression only for the ischemia ,57,58. Aeviously ,45,59,60iL1b, iL6, and Socs3) for PGP, as well as IC  and NC genes  for PGPL, that significantly changed in expression levels after peptide administration compared to Semax treatment under tMCAO conditions. Based on the differential expression results, a functional network was constructed. It linked genes from the IC and NC with 27 signaling pathways associated with Semax treatment during IR. At the same time, we showed that not all the Semax-related signaling pathways are equally characteristic of the effects of short PGP-containing PGP and PGPL peptides. The structures of PGP and PGPL are only partially similar to the structure of Semax, and the differences are great. Therefore, the network can specify the relationship between peptide structures and their functional effects.A search for DEGs associated with Semax treatment, compared to PGP and PGPL treatment, under tMCAO conditions was carried out. The results obtained made it possible to identify genes that characterize the overlapping and unique molecular signatures of the peptides’ effects under IR conditions. Both the PGP and PGPL peptides showed Semax-unlike effects, predominantly without changing gene expression 24 h after tMCAO. Moreover, there were IC genes  fragment, which is absent in the PGP and PGPL peptides. The manifestation of ACTH(4-7)’s action may be associated with the so-called synactonic mechanism. Synactone can be defined as a combination of the peptide itself and the products of its metabolism, acting in a certain sequence and by interaction [In the network, the signaling pathways formed three clusters depending on the effects of the peptides on gene expression. The first pathway cluster included 15 signaling pathways that were associated with genes that were differentially expressed after Semax treatment, versus both PGP and PGPL, under IR conditions. Such a pathway cluster can reflect Semax-unlike effects for both PGP and PGPL peptides. Interestingly, only the IC genes served as nodes in the first pathway cluster. In particular, the eraction ,62,63,64eraction ,62,63,64eraction . Moreoveeraction ,62,63,64Cplx2, Neurod6, and Ptk2b) from the NC related to PGPL treatment. Notably, the Fos and Ptk2b genes demonstrated a Semax-unlike expression pattern after PGPL administration. Such genes were associated with the inflammatory component of the second signaling cluster. The Fos gene encodes a transcriptional factor that is involved in the activation of the expression of a number of interleukin and chemokine genes during ischemia [The second pathway cluster included eight signaling pathways. They were associated with genes that were differentially expressed under IR conditions after Semax treatment versus PGPL, but not versus PGP administration. This cluster can reflect the Semax-unlike effect of PGPL, but not that of PGP peptides. Such a cluster included signaling pathways mainly related to the processes of neurosignaling and neuroreception. Indeed, PGP administration does not lead to statistically significant changes in the expression of NC genes relative to the level of expression of these genes in ischemic rats after Semax administration. Concomitantly, Semax treatment was associated with three DEGs  and encode acetylcholine, glutamate, and GABA receptors. The appearance of the third cluster of signaling pathways may be due to the presence of a common PGP structural unit in the structures of all the studied peptides. It is known that PGP is the main metabolite of Semax and PGPL and represents the common functional core of synactone for these peptides [The third pathway cluster included four signaling pathways that were associated only with non-DEGs after Semax treatment, versus both PGP and PGPL treatment, under IR conditions. The expression profiles of all the associated genes between Semax and PGP and PGPL treatment did not differ. Thus, the third pathway cluster can reflect Semax-like effects for both the PGPL and PGPL peptides. The genes included in the signaling pathways of this cluster belong to NC (peptides ,66,68. Mpeptides ,70 and apeptides ,63,68. Tpeptides ,63,68.Thus, our data provide insight into glyproline peptides’ activity through the modulation of the inflammatory and neurosignaling genetic responses following cerebral IR. The main limitation of our study is the lack of genome-wide expression data after the administration of the PGP and PGPL peptides under tMCAO conditions. A detailed study using RNA-seq will more accurately identify differences in expression patterns caused by synthetic peptides and their units. The results will also underlie the selection of targets for analysis of peptide functioning in protein levels in order to get even closer to understanding the phenomenon of their properties.In conclusion, the comparison of glyproline peptides allowed us to determine their general and individual effects on gene expression under cerebral IR. Furthermore, the results served as a first step in understanding the relationship between the chemical structure of the glyproline peptides and their possible effects at the genomic and functional levels. We believe that understanding the genesis of the relationship between the structure and function of peptides can lead to the creation of drugs that are even more effective than those currently available.


C-reactive protein (CRP) and calcitonin (PCT) The clinical value in the diagnosis of adult sepsis.  The aim of the study is to analyze the clinical characteristics, pathogen distribution, and drug sensitivity information of adult sepsis, and to provide reference for empirical clinical use; to explore the relationship between  We collected 455 cases of hospitalized patients with positive blood culture, including 352 cases with sepsis and 103 cases without sepsis; 1609 cases of hospitalized patients with suspected infection and negative blood culture, including 287 cases of sepsis, and 518 cases of non-infectious systemic inflammatory response syndrome (SIRS) and 804 cases of local infection. Age, gender, route of admission, admission status, CRP, PCT, and white blood cell (WBC) levels were collected from the patients. The differences between the factors were statistically analyzed, and the receiver operating characteristic curve (ROC curve) was plotted to obtain the optimal cut-off values of CRP and PCT and their area under the curve (AUC), and to compare the CRP, PCT and PCT, and the CRP + PCT tandem to diagnose sepsis sensitivity and specificity. Staphylococcus aureus was 87.5% resistant to penicillin and sensitive to vancomycin, milantropine, and teicoplanin; among Gram-negative bacteria, Escherichia coli was resistant to piracillin The resistance rate was 73.1%, fully susceptible to viraemia (100%) and resistant to imipenem, amikacin, and lacillin/tazole. (3) Among patients with positive blood cultures, CRP and PCT levels were higher in patients with sepsis than in those without sepsis. Pairwise comparison of ROC curves showed that the diagnostic value of PCT was greater than that of CRP (P=0.016).  (1) 387 pathogenic strains were isolated from blood cultures of patients with sepsis, 71.06% Gram-negative, 26.87% Gram-positive and 2.07% fungi. (2) Among Gram-positive bacteria,  CRP and PCT have a good reference value for diagnosis of sepsis patients and determination of the degree of infection in septic patients, especially PCT is more valuable for diagnosis of sepsis. Bacterial sepsis is an infectious disease that, if not controlled and treated in a timely manner, can develop into severe sepsis, posing a serious threat to the life and health of patients . In receC cells under physiological conditions; it contains a protein of 116 amino acids with a molecular mass of 14.5 kD, and PCT has a half-life of 25–30 h in the blood [PCT, first proposed in 1984, is a pro-peptide substance of calcitonin, which is mainly produced by thyroid he blood , 8. The he blood .Streptococcus pneumoniae, in the serum of patients with Streptococcus pneumoniae. Studies [CRP is a plasma protein produced by the liver and is considered to be a marker of inflammatory response. Studies , 12 firs Studies , 14 show Studies , 16 haveIn this study, we included patients with negative and positive blood cultures, analyzed clinical data from patients with sepsis, compared the differences in CRP and PCT levels between patients, and investigated the role of these two indicators in the diagnosis of sepsis. Optimal threshold values were obtained, and the sensitivity and specificity of the indicators for the diagnosis of sepsis at the threshold values and the relationship between CRP and PCT levels and the severity of sepsis were analyzed.Cases of blood culture-positive inpatients and suspected infectious, blood culture-negative inpatients admitted to our hospital from January 2015 to July 2021 were collected for retrospective analysis. The study population was divided into blood culture-positive sepsis, blood culture-positive non-septic, blood culture-negative sepsis, blood culture-negative non-infectious SIRS, and blood culture-negative localized infection , accordiHospitalized patients over 18 years old.The etiological data are complete.The disease course information recorded in medical records is complete.The inclusion criteria were as follows:Have a hospitalization record and have used antibiotics 2 weeks before admission.Patients who have been included in study cohort with repeated admissions.Died or discharged within 24 hours after admission.Emergency biochemical and procalcitonin examinations were not performed.The exclusion criteria were as follows:R). All operations were performed in strict accordance with the standard instructions. After identification of the samples, drug sensitivity analysis was performed using the Beidou Phoenix IM-100 fully automated microbiological analysis system; some drug sensitivity tests were performed using the K-B method or MIC broth dilution method, and the results were judged according to the American Clinical Laboratory Standardization Institute (CLSI) standards, and the specific instruments and manufacturers are shown in 5 mL of fasting elbow venous blood was collected from all patients within 24 h after admission, placed in EDTA-Na anticoagulated EP tubes, centrifuged at 3 500 r/min for 10 min, and the serum was separated and placed at −70°C for measurement. The PCT level was measured by electrochemiluminescence with a Roche e411; the CRP level was measured by immunoturbidimetric method, and platelet-related parameters including platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW), and large platelet ratio (P-LC) were measured by hematocrit analyzer in all patients after the onset of disease. Samples for CRP, PCT, and WBC testing were obtained from fresh or during hospitalization of patients. CRP was detected by automatic biochemical analyzer Cobas C501-2; PCT was detected by automatic electrochemiluminescence immunoassay analyzer Cobas e601; WBC was detected by Sysmex XN-3000 automatic blood analyzer. The specific instruments and manufacturers are shown in Escherichia coli, 79 strains (20.41%) of Klebsiella pneumoniae and 40 strains (10.34%) of Staphylococcus aureus. The distribution of pathogenic bacteria detected in blood cultures of 387 adult septic patients is shown in A total of 387 strains of pathogenic bacteria were isolated from blood culture of 352 patients, including 275 strains (71.06%) of gram-negative bacteria, 104 strains (26.87%) of gram-positive bacteria, and 8 strains of fungi (2.07%). The top three pathogenic bacteria with composition ratio are as follows: 134 strains (34.63%) of Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus cephalosporus, Staphylococcus humanus, and Staphylococcus saprophyticus, respectively. Among the Gram-positive bacteria, S. aureus was 87.5% resistant to penicillin and sensitive to vancomycin, mirantolimus, and teicoplanin; other types of staphylococci were more resistant to penicillin, benzocillin, and erythromycin and sensitive to rifampicin, linezolid, vancomycin, mirantolimus, and teicoplanin, as shown in E. coli had a resistance rate of 73.1% to piracillin, complete susceptibility (100%) to viraemia, and resistance to imipenem, amikacin, and laracillin/tazole. The susceptibility rate of bactam was above 95%; Klebsiella pneumoniae was 24.1% resistant to piroxicillin and relatively sensitive to amikacin with a sensitivity of 96.2%; Pseudomonas aeruginosa was resistant to amikacin, fully sensitive to gentamicin (100%) and had a relatively low antibiotic resistance rate; Acinetobacter baumannii had a relatively high antibiotic resistance rate and was 53.3% sensitive to amikacin, as During the study period, 40, 5, 4, 2, 2, 2, and 1 cases were infected with P > 0.05), but there was a statistical difference in admission route. Patients with toxemia had a higher proportion of emergency admissions and were relatively critically ill on admission, while nonseptic patients had relatively average admissions. In addition, levels of CRP  mg/L in sepsis were higher than nonseptic CRP  mg/L (P < 0.001), and levels of PCT in sepsis  ng/mL level was higher than PCT in non-septic  ng/mL (P < 0.001), WBC in κ toxemia  × 109/L and non-septic WBC  × 109/L There was a statistical difference between WBC  × 10°/L in toxemia (P < 0.001), and WBC was converted into a categorical variable for analysis, see A total of 455 hospitalized patients were included, including 352 patients with sepsis and 103 patients without sepsis. There was no difference in gender and age between sepsis and nonseptic patients (P=0.016). CRP and PCT levels were higher in patients with severe sepsis/septic shock. When CRP and PCT were 197.01 mg/L and 28.55 ng/mL, respectively, the AUC was 0.67  and 0.98 , and the diagnostic sensitivity and specificity of CRP was 50.5% and 79.7%, respectively, and that of PCT was 92.8% and 96.1%, respectively as shown in In patients with positive blood cultures, CRP and PCT levels were higher in sepsis than in non-sepsis. ROC curve analysis showed that the ACC for CRP, PCT and CRP + PCT tandem for the diagnosis of sepsis were 0.85 , 0.90  and 0.92 (95% CI 0.89∼0.94). When the critical value of CRP was 97.05 mg/L, the sensitivity and specificity were 77.8% and 80.7%, respectively; when the critical value of PCT was 1.34 ng/mL, the sensitivity and specificity were 86.5% and 79.5%, respectively; the sensitivity and specificity of CRP + PCT tandem diagnosis were 84.3% and 87.5%, respectively. Pairwise comparison of ROC curves indicated that the diagnostic value of PCT was greater than that of CRP (Sepsis is a relatively common infectious disease that poses a serious threat to the quality of daily life and the health of patients. If timely diagnosis can be made at an early stage in patients with sepsis and early effective treatment can be taken for them, the treatment effect can be significantly improved and the morbidity and mortality rate of patients with sepsis can be reduced . In the In this study, there were records in the hospital's electronic medical record system confirming that patients with pathogenic bacteria detected in clinical blood cultures were blood culture-positive patients. Studies , 20 haveIn this study, sepsis was divided into a sepsis group, a severe sepsis group, and a septic shock group according to the severity of sepsis. When CRP and PCT levels were compared, sepsis was found to be associated with severe sepsis and septic shock. A statistical difference was observed in septic shock: CRP and PCT levels were higher in severe sepsis/septic shock than in sepsis. A level of 28.55 ng/mL had a better diagnostic value. PCT performed better than CRP in the diagnosis of sepsis compared to the area under the ROC curve.In conclusion, the detection of serum PCT and CRP can effectively assess the development of sepsis patients, which has certain value for early diagnosis and prognosis, and the prognosis of patients is closely related to platelet count and related parameters, which has certain guiding significance for prognosis prediction. This study can provide some reference for the diagnosis of clinical sepsis, but there are some shortcomings in the study design. First, this part of the study was retrospective and failed to continuously monitor the dynamic changes of CRP and PCT and to follow up patients discharged from the hospital; therefore, the risk factors for sepsis-related mortality could not be analyzed. Second, because we did not have real access to the patients and the data came from information recorded in the electronic medical record system, data were missing for some patients, and a follow-up rigorous prospective study could be designed to compensate for the shortcomings of this study.


In addition to melanocytic hyperfunction, changes are observed in the upper dermis of melasma, and fibroblasts play a central role in collagen synthesis and pigmentation induction.To explore the morphology, growth rate, and gene expression profile of fibroblasts from the skin with melasma in comparison to fibroblasts from the adjacent healthy skin.Ten women with facial melasma were biopsied , and the fragments were processed for fibroblast culture. Samples from five participants were seeded to evaluate growth  and senescence  curves. The samples from the other participants were submitted to real-time PCR to comparatively evaluation of the expression of 39 genes.WNT3A, EDN3, ESR2, PTG2, MMP1, and SOD2 genes were up-regulated, whereas the COL4A1, CSF2, DKK3, COL7A1, TIMP4, CCL2, and CDH11 genes were down-regulated in melasma skin fibroblasts when compared to the ones from adjacent healthy skin.Cultured fibroblasts from melasma skin were morphologically less fusiform in appearance and on average a 34% (95% CI 4%‒63%) greater proportion of cells labeled with SA-β-gal than the fibroblasts from the adjacent skin. The cell growth rate was lower for the melasma samples after eight days (p < 0.01). TheSmall sample size; absence of functional tests.Fibroblasts from the skin with melasma showed a lower growth rate, less fusiform morphology and greater accumulation of SA-β-gal than those from adjacent photo exposed skin. Moreover, their gene expression profile comprised factors that may contribute to upper dermis damage and sustained melanogenesis. Melasma is a highly prevalent acquired dyschromia, which results from an increase in the activity of the epidermal melanin unit.5Compared with the adjacent healthy skin (photoexposed), the skin with melasma has an increased amount of epidermal melanin, more mature melanosomes, hypertrophied melanocytes, and prominent solar elastosis. There is also an increase in the number of vessels and mast cells, as well as increased expression of inflammatory mediators, such as iNOS, NF-Kβ, endothelin, and growth factors, such as VEGF, and growth factors derived from hepatocytes,-stem cells, fibroblasts and nerves.Sun exposure is the main risk factor for the development of melasma. Chronic photoexposure, in addition to melanogenesis, induces oxidative stress, which promotes cellular senescence.10Fibroblasts play a central role in pigmentation and their transition to a senescent profile promotes a modification of their autocrine and paracrine activity.INK4A, which may justify the stimulus for sustained skin pigmentation.The senescent phenotype was identified in fibroblasts with melasma more intensely than in adjacent photo exposed skin, based on the immunoexpression of p16This study aimed to explore the morphology, growth rate and gene expression profile of fibroblasts from facial melasma, compared to adjacent photoexposed skin.This project was approved by the Research Ethics Council . and all participants consented to participate.Ten adult women with facial melasma underwent biopsy  of the melasma skin on the malar region and from the adjacent, clinically normal, photoexposed skin (<2 cm of distance), laterally. The participants were untreated for the dermatosis for at least 30 days, except for the use of sunscreen.3 in a 12-well cell culture plate.The skin fragments were sectioned and placed in fibroblast culture, maintained in medium 106 (Gibco™) containing growth factors . The fibroblasts were seeded (in triplicate) at a density of 5 × 104 in a 6-well cell culture plate, and the number of cells per cm2 was evaluated after two, five and eight days, based on the evaluation of 30 fields, and compared (between topographies) by a generalized linear mixed-effects model. Sample normality was evaluated by the Shapiro-Wilk test.Cultured fibroblasts, from healthy and damaged skin, from five participants, were seeded at a density of 1 × 10Subsequently, cell senescence was evaluated using senescence β-galactosidase  staining kit , following the manufacturer's instructions. After 24 hours of cell plating, the culture medium was removed, followed by washing with PBS and the addition of fixation solution for 15 minutes at room temperature. Then, it was washed with PBS twice, followed by overnight incubation with β-galactosidase solution  at 37 °C. After the solution was removed, 70% glycerin was added.A total of 300 cells were counted per sample. The morphology of the cultured fibroblasts and the percentage of cells labeled with cytoplasmic SA-β-gal were compared between the topographies .2 Profiler PCR Arrays, Qiagen) to assess the expression of 39 genes associated with growth factors and tissue repair, WNT pathway activation, neocollagenesis, metalloproteinase synthesis, estrogen receptors, and antioxidant mechanisms (2 First Strand Kit for RT-PCR (Qiagen), following the manufacturer's instructions.Fibroblasts from the primary cell culture of the other five participants were submitted to a real-time PCR array  was estimated by the fold change (melasma/adjacent skin) for each participant.The effect size for gene expression  than cultures from adjacent skin . MoreoveWNT3A, EDN3, ESR2, PTG2, MMP1, and SOD2 genes were up-regulated; COL4A1, CSF2, DKK3, COL7A1, TIMP4, CCL2, and CDH11 genes were down-regulated  is known to be the main trigger of age-related phenotypes, such as wrinkles and pigmentation.Senescent fibroblasts secrete inflammatory cytokines and stem cell factors, which promote collagen degradation through MMP activation and have reduced mitotic potential. There is increased stromal degradation and impaired tissue repair, as evidenced in the dermis of melasma. Therefore, a mosaic structure of tissue susceptibility to senescence can be theorized for the development of melasma in the skin under environmental stimuli such as UV radiation and sex hormones.COL4A1, COL7A1), and TIMP4 expression in fibroblasts isolated from melasma skin. During the senescence process, there is a progressive increase in metalloproteinase production and a decrease in the repair process.MMP9 showed a high fold change (>2), which, in association with the lower expression of TIMP1 and 4, suggests participation of melasma skin fibroblasts in the deficient repair of the upper dermis and angiogenesis.There was an increase in MMP1 gene expression and a decrease in collagen IV, VII  is part of the enzymatic antioxidant system and is the main enzyme responsible for scavenging oxygen free radicals through the conversion of superoxide anions into hydrogen peroxide and oxygen.37in vitro.The fibroblast is the skin cell that contains the most PTGS. This enzyme participates in the synthesis of prostaglandin E2 (PGE2) from arachidonic acid, and PGE2 inhibits collagen production by fibroblasts This study has limitations related to the small sample size and the lack of proteomic or functional assays to confirm the findings. However, the results were consistent and allowed the authors to explore, in a preliminary way, the hypothesis about the role of fibroblasts in the pathogenesis of melasma, proposing new investigations related to the pathogenesis and therapeutic strategies of melasma. The sample of patients submitted to the senescence and proliferation assay was different from the one submitted to the gene expression comparison, although the results were parallel. Finally, the investigation of fibroblasts from controls without melasma, matched by age, photoexposure pattern, photoaging, and phototype, could also elucidate the particular susceptibility of fibroblasts from individuals with melasma to senescence.Further studies should investigate the regulatory factors of these pathways and the cytokine network between keratinocytes, melanocytes, nerves, endothelium, and fibroblasts in melasma, preferably through studies with tissue cultures, for genomic, proteomic, and functional experiments.Fibroblasts from the skin with melasma showed a lower growth rate, less fusiform morphology, and greater accumulation of SA-β-gal than those from adjacent healthy photoexposed skin. Furthermore, their gene expression profile comprised pro-inflammatory, pro-melanogenic, and tissue repair deficit-related factors, which can induce damage to the upper dermis and support the focal pigmentary phenotype in melasma.FAPESP – number 2012/09233-5, 2012/05004-1; CNPq – number 401309/2016-9.Ana Cláudia Cavalcante Espósito: Design and planning of the study; collection, analysis, and interpretation of data; drafting and editing of the manuscript; Critical review of the literature; Approval of the final version of the manuscript.Gabrielli Brianezi: Design and planning of the study; collection, analysis, and interpretation of data; approval of the final version of the manuscript.Luciane Donida Bartoli Miot: Design and planning of the study; collection, analysis, and interpretation of data; effective participation in research orientation; approval of the final version of the manuscript.Hélio Amante Miot: Design and planning of the study; collection, analysis, and interpretation of data; statistical analysis; drafting and editing of the manuscript; critical review of the literature; critical review of the manuscript; approval of the final version of the manuscript.None declared.


Celiac disease is an immune-mediated, chronic, inflammatory, and systemic illness which could affect the eye. The aim of this study is to look for possible signs of retinal involvement in celiac disease that could be utilized as biomarkers for this disease. Sixty-six patients with celiac disease and sixty-six sex-matched healthy subjects were enrolled in this observational case–control study. A comprehensive ophthalmological evaluation, axial length measurements, and SD-OCT evaluation were performed. The thickness of the retinal layers at the circle centered on the fovea (1 mm in diameter) and the average of the foveal and parafoveal zones at 2 and 3 mm in diameter were evaluated, together with retinal volume and the peripapillary retinal nerve fiber layer (RNFL). Concerning the thicknesses of the retinal layers in each analyzed region, no statistically significant differences were found. The same results were obtained for the total volume. Regarding peripapillary RNFL, the celiac patients showed slightly thicker values than the healthy controls, except for temporal and nasal-inferior quadrants, with no statistically significant differences. All the analyzed parameters were similar for the celiac patients and the healthy individuals. This could be related either to the non-involvement of the retinal layers in celiac disease pathophysiology or to the gluten-free diet effect. Celiac disease is an immune-mediated, chronic, inflammatory, and systemic illness  characteClassic celiac disease presents malabsorption, failure to thrive, and diarrhea. At the same time, more subtle presentations such as latent, potential, oligosymptomatic, and extraintestinal signs related to otologic, dental, neurological, dermatological, and musculoskeletal symptoms may be less prevalent . HoweverAmong these extraintestinal findings, ocular manifestations due to celiac disease are of great concern because of the direct effect of visual function and ocular comfort on the quality of life ,7.The presence of circulating immune complexes or autoantibodies in ocular tissues, cross-reactivity of cell antigenic epitopes, vitamin deficiencies, and immunogenetic factors might all play a role in ocular involvement, especially for all the vascularized components of the eye .In fact, the choroid of celiac patients appears thicker than healthy controls ,9. In paTo the best of our knowledge, no studies have been published examining all the retinal layers concerning retinal involvement in celiac disease. Only a few studies evaluated the peripapillary retinal nerve fiber layer (RNFL), showing no consensus in children and adults ,12,13,14For these reasons, together with the disease’s autoimmune and inflammatory nature and the presence of the superficial and deep capillary plexuses among the retinal layers, this study aims to look for possible signs of retinal involvement, utilizing spectral-domain optical coherence tomography (SD-OCT), that could be utilized as biomarkers for this disease.Adult subjects with a diagnosis of celiac disease, evaluated at the Celiac Disease Center at the Department of Medicine, Surgery, and Dentistry of the University of Salerno between September 2019 and March 2020, and a control group of sex-matched healthy subjects were included in this observational case–control study.Diagnosis of celiac disease was confirmed by intestinal biopsy and serology, regardless of the time of diagnosis. Following the diagnosis, all the celiac patients were placed on a gluten-free diet. Regarding control subjects, they had at least one negative-specific serology for celiac disease and no diagnosis of any gastrointestinal diseases.Subjects younger than 18 years of age or with systemic and ocular diseases, or patients who underwent other ophthalmic surgical procedures which could affect the eye ,16,17,18According to the Declaration of Helsinki’s ethical principles, all participants were informed about the study’s purpose and written informed consent was acquired. Institutional Review Board approval was also obtained from the ComEtico Campania Sud (CECS), prot. n°16544. A comprehensive ophthalmological evaluation, including clinical history to identify possible exclusion criteria, slit-lamp examination, Snellen best-corrected visual acuity, axial length (AL) measurements with IOLMaster , and SD-OCT evaluation , was performed.All participants were examined between 2:00 p.m. and 3:00 p.m., without pupil dilation. For each participant, only the right eye was evaluated .A horizontal 30° volume OCT B-scan centered on the fovea was obtained for all examined eyes. Using the device’s built-in software , the segmentation of the retinal layers was obtained.Poor-quality images with a signal-to-noise score lower than 20 decibels were excluded.To study the 10 retinal layers, eleven optical interfaces were obtained  20]..20].In addition, utilizing the standard Early Treatment Diabetic Retinopathy Study (ETDRS) grid, the thickness of the retinal layers at the circle centered on the fovea (1 mm in diameter), the average of the 5 foveal and parafoveal zones (2 mm in diameter), and the average of the 9 foveal and parafoveal zones (3 mm in diameter) were evaluated .For all the analyzed regions , the values of the total thickness , photoreceptor (PHR) layer, retinal pigment epithelium (RPE), outer nuclear layer (ONL), outer plexiform layer (OPL), the inner retinal layer (IRL), and the GCC thickness were collected. IRL includes the sum of RNFL, GCL, IPL, and the inner nuclear layer (INL), while GCC is composed of RNFL, GCL, and IPL. However, the thickness value for all these layers was also evaluated individually in each studied region of the ETDRS grid.Moreover, the device’s built-in software automatically calculated the total volume at 3 mm diameter for each retinal layer.Concerning peripapillary RNFL, the optic nerve head protocol of the device generates an RNFL thickness map from which RNFL thickness is measured along a circle 3.45 mm in diameter centered on the optic disc. The average RNFL thickness of the seven quadrants  was measured for all patients.p > 0.05) for all data.All data were analyzed with GraphPad Prism 8 . Kolmogorov–Smirnov test was performed to assess normal distribution  . A diffeSixty-six patients with celiac disease  and sixty-six sex-matched healthy subjects were enrolled. The mean disease duration of the celiac patients was 9.1 ± 8.8 years (range: 0–41 years). None of the celiac patients included in this study presented previous ocular complications due to celiac disease.p = 0.75).The mean age of the celiac patients was 40.3 ± 11.6 years (range: 18–66 years), while the mean age of the healthy subjects was 39.9 ± 14.2 (range: 23–69 years), with no statistically significant difference between the two groups (p = 0.15).The mean AL of the celiac patients was 23.6 ± 1.0 mm (range: 21.7–26.1 mm), while the mean AL of the healthy subjects was 23.9 ± 1.2 mm (range: 20.7–27.5 mm), with no statistically significant difference between the two groups , at 2 mm , and at 3 mm .Considering the correlation between the years of gluten-free diet adherence and the total retinal thickness, no statistically significant correlation was found at 1 mm , and the inner plexiform layer (IPL) . The thiAccording to the assumptions by Karatepe Hashas et al.  and HazaThe present study’s results indicate that celiac disease’s inflammatory and autoimmune processes could not involve the retinal layers directly. However, this finding may also be explained by the gluten-free diet adherence of all analyzed celiac patients, possibly determining a remission of any retinal changes or a decrease in the inflammatory effects of the disease .The fact that the patients were on a gluten-free diet could represent a limitation of the present study. Further studies in naïve celiac patients, comparing the effects of a gluten-free diet versus a regular diet, would be needed to understand better the retinal baseline status of such subjects and its possible changes over time.In conclusion, retinal layer thicknesses, volumes, and peripapillary RNFL were similar in the celiac patients and the healthy individuals. The reason for these results could be due to either the non-involvement of the retinal layers in celiac disease or the gluten-free diet effect. However, the results of this study cannot omit a routine ophthalmological examination for these patients due to the association between celiac disease and other ocular disorders ,5,6,7.


Angiosarcoma is a rare subtype of soft tissue sarcoma with identifiable vascular differentiation. It can occur at any age and develop throughout the body, but it is most commonly found in skin, soft, and breast tissues. Primary retroperitoneal angiosarcoma is rarely reported in the relevant literature. This article reports a case of primary retroperitoneal angiosarcoma in a middle-aged man, with the relevant literature reviewed in detail. A 46-year-old male had experienced left waist pain for 2 months. An ultrasonic examination revealed a mass in the left retroperitoneum, and left retroperitoneal lesions were confirmed via computed tomography (CT) and magnetic resonance imaging (MRI). The tumor was removed surgically, and the CT scan revealed local tumor recurrence after 1 month when the first adjuvant therapy was performed. The patient died of a massive hemorrhage from a ruptured tumor. Angiosarcoma has high malignancy and a poor prognosis. Its early diagnosis and treatment significantly impact the long-term survival rate of patients. These tumors are rare, with an incidence rate of only 0.0027% . Among t2A 46-year-old male patient was admitted to The Second Affiliated Hospital of Fujian Medical University with the main complaint of left waist pain for 2 months. Previously, he had been in good health, with no underlying diseases and no tobacco or alcohol addiction. A physical examination revealed no bulges in either kidney region, the kidneys were not palpable, and the kidney areas were without percussion pain. Traveling along the bilateral ureteral area, there was no tenderness or palpable mass. Furthermore, there were no obvious abnormalities in the patient’s routine blood and urine tests and biochemical results, while the blood catecholamine, cortisol, and aldosterone readings were normal. A ultrasonic examination revealed no abnormality in the bilateral kidneys and adrenal glands. No obvious signs of metastasis were found in the lung and liver on computed tomography (CT) examination.The summary of the ultrasonic examination is as follows. The left posterior peritoneum, which was at the height of the left renal artery and abdominal aorta, had an irregularly shaped hypoechoic area (4.3 cm × 4.8 cm × 3.6 cm) with a non-uniform internal echo . The col-45, melan-A, podoplanin, CD117, discovered on gastrointestinal stromal tumor 1, thyroid transcription factor-1, chromogranin A, and smooth muscle actin). The antigen Ki-67 index was approximately 50% , and the tumor may rupture and hemorrhage . The CT This case supports the view of several studies that a tumor size of more than 5 cm is associated with a worse outcome . The labThe CT examination of angiosarcoma reveals the following: (i) soft oval tissue masses, some of which may be lobulated; (ii) an unclear boundary with a density much lower than that of muscle; and (iii) possible hemorrhage, necrosis, and calcification occurrence inside the mass.The MRI examination of angiosarcoma cases reveals the following: (i) T1 weighted imaging (T1WI) produces an uneven low signal, and (ii) T2 weighted imaging (T2WI) produces a high-signal imaging that shows facilitated water movement and a scattered low signal. However, the gold standard for the diagnosis of angiosarcoma is pathological immunohistochemistry, with the expression of platelet endothelial cell adhesion molecule-1 or recombinant cluster of differentiation 34 (CD34) and factor VIII-related antigen (+) characterizing the disease ,12. SincRetroperitoneal angiosarcoma needs to be differentiated from the following diseases: (i) pheochromocytoma ; (ii) retroperitoneal lipoma ; and (iii) neuroblastoma, which is more common in children; the echo of the mass is much lower than that of angiosarcoma.Surgical resection is the main method used in the traditional treatment of angiosarcoma. However, the median overall survival of patients with resection of the lesion alone is only 9 months, and the 5-year survival rate is only 20%. For patients supplemented with radiotherapy and chemotherapy, the median overall survival rate can reach 36 months, with a 5-year survival rate of up to 45%. The main causes of death with the disease are recurrence  and distant metastasis (rate: 34%). A multidisciplinary treatment approach is always warranted, but despite multimodality treatment, the recurrence remains high even for localized tumors . Recent In summary, angiosarcoma is a rare, fast-growing, and highly aggressive tumor with a very poor prognosis that can occur in any part of the body. The early diagnosis of retroperitoneal lesions is particularly difficult. Clinically, ultrasound, CT, MRI, and other imaging examinations should be integrated. An image-guided needle biopsy is particularly important since it can provide an early diagnosis. To improve the long-term survival rate of patients, a prompt surgical resection of the lesion should be combined with targeted radiotherapy and chemotherapy.4Angiosarcoma has high malignancy and a poor prognosis. Its early diagnosis and treatment significantly impact the long-term survival rate of patients. This study focused on the imaging manifestations of angiosarcoma; however, there remains a lack of understanding of the disease’s operation mode, radiotherapy, and chemotherapy. Therefore, additional similar cases must be identified and their characteristics summarized to inform further research.


The world has been dealing with Aids for forty years, covid-19 accentuated societal inequalities and promoted a rupture in care and prevention, including for people living with HIV. We compiled official HIV indicators, analyzed the impact of covid-19 in Brazil, at São Paulo State (SP), and compared it to the municipality of Santo André (in the state of São Paulo), which adopted linkage/retention strategies to mitigate the impact of covid-19. From 2019 to 2020, suppression/adhesion rates remained stable. The number of new treatments decreased both in Brazil (-19.75%) and São Paulo (-16.44%), but not in Santo André, where 80% of new patients started treatment within 30 days from their first TCD4 test (70% in São Paulo and 64% in Brazil). However, PrEP dispensing increased during this period. The distribution of 2,820 HIV self-tests in Santo André lead to only one documented new HIV diagnosis linked to care. Synergistic strategies to swiftly diagnose and connect new cases, ensuring retention as well as rescuing missing patients deserve priority in the fight against HIV, especially in times of covid-19. By March 29, 2021, the world had 196,553,009 confirmed cases, including 4,200, 412 deaths. Even though Brazil has 2.7% of the world population, it concentrated 13.3% of all covid-19 cases and 26.9% of all deaths at that time, with more than half a million deaths by July, 2021.The WHO. All over the world, health services have been disrupted, including those needed to support prevention, diagnosis, and treatment.Meanwhile, 40 years after the first documented Aids cases, its pandemic is far from reaching an end. In 2020 it was estimated that 37,700,000 people were living with HIV (PLWH), 1,500,000 new infections occurred during the year, 680,000 deaths related to Aids, with 73% of PLWH receiving antiretroviral therapy (ART). SARS-CoV-2 aggravated this scenario both directly, because of covid-19 disease, and indirectly, by disrupting HIV care. The covid-19 pandemic has accentuated existing inequalities and made the vulnerable population more vulnerable to hunger, violence, and diseases. The metaphor “we are not in the same boat, but in the same tempestuous sea” expresses the very unequal conditions in coping with covid-19 demands.The Brazilian Unified Health System (SUS) ensures access to health and services in a universal, equal, and integral way. Along with free ART for all PLWH and specific policies, Brazil had been a model for Aids response, a leadership that was further shaken in recent years as result of a crisis that originated from cuts and freezes in the budget for health, education, and social assistanceCoinciding with the beginning of the first covid-19 wave in Brazil (March 2020), the Santo André infectious diseases outpatient clinic (SA-IDC), the reference in the care of PLWH in the city, started a research project which aimed to test strategies to improve patient linkage/retention, an initiative that was adapted to the restrictions imposed by covid-19. The main strategies to mitigate the impact of covid-19 were: (i) screening at admission, managing users’ needs; ii) maintenance of new patients’ admission (without prior follow-up or transfer) as much as possible, and linking on the same day of admission; iii) maintenance of antiretroviral dispensing; iv) evaluation of complaints and need for laboratory exam collection at ART withdrawal; v) phone monitoring of newly admitted cases with recalls by the nursing staff until viral suppression was achieved; vi) contact with cases with delays in ART withdrawal for over ten days, first by WhatsApp message and, if unanswered, a home visit was made by the social worker; vii) provision of HIV rapid self-test for users seeking them, after discarding post-exposure prophylaxis (PEP) eligibility and; viii) psychological evaluations were maintained, whenever possible, online.. Number of T CD4 counts, HIV viral load tests, patients starting treatment, and antiretroviral dispensing were obtained from the covid-19 HIV panel. Clinical information for 2019 and 2020 on linked patients, late, and timely diagnosis, viralsuppression, and adherence to ART were evaluated...We conducted a descriptive study based on data on the Brazilian Ministry of Health official registry of HIV/Aids care and prevention indicators  to evaluate whether the strategies adopted in Santo André mitigated the difficulties imposed by the covid-19 pandemic. Information for 2019, 2020, and 2021  from Brazil, São Paulo State (SP), and Santo André (SA) were compared, moreover, ART is an fundamental tool to control the HIV epidemic, as the “undetectable=untransmittable” policy shows.The response to covid-19 included the cancellation of appointments and the routine collection of tests in all health units in mid-March 2020. The decrease in the number of ART dispensations as treatment shown inWhile pre-exposure prophylaxis (PrEP) dispensation increased , post-exposure prophylaxis (PEP) dispensation decreased  in 2020, when compared to 2019, in Brazil, São Paulo, and Santo André, respectively. At least in Santo André, where we had access to the details of PEP use, its decrease was due to the 37% reduction in the demand for accidents with biological material, from 155 in 2019 to 98 in 2020, whereas use for consented sexual exposure increased 6%, from 527 in 2019 to 557 in 2020. The increase coincided with a time when contact should have been restricted due to covid-19. PrEP dispensation increased overall due to efforts of national implementation, which was not so evident in Santo André, due to a reduction in the health care team that lead to the suspension of new PrEP program admissions.to contextualize the covid-19 pandemic phases. Covid-19 death rates, shown as deaths per 100,000 inhabitants, were estimated by dividing the number of deaths by the estimated population of 2021 (https://www.ibge.gov.br/) and multiplying the result by 100,000. Thus, we obtained 260 (Brazil), 298 (SP), and 335 (SA) deaths per 100,000 inhabitants. The higher documented death rate in Santo André suggests that the city was proportionally more affected by covid-19, with 12% and 29% more deaths/100,000 inhabitants than the state and country, respectively.. Albeit the important increase of self-tests dispensed, our observation may suggest that those who buy a test will more likely use it, whereas an individual who receives a test for free, may not do it properly. This needs further evaluation as the effectiveness of this and other modalities of diagnostic incentive are crucial to inform prevention policy.The Santo André public health system prioritizes the diagnosis of HIV infections via HIV rapid tests (RT) performed on SUS health services. The amount of RT performed for diagnosis in 2019, 2020, and in the first half of 2021 was provided by the Santo André health department. Out of 30,910 tests in 2019, 324 (1.1%) were reactive. Out of 28,414 in 2020, 308 (1.1%) were reactive, an 8% decrease in testing. Until June 2021, only 11,997  tests were performed. On the other hand, there was a marked increase in self-tests dispensed, from only six in 2019 to 2,820 in 2020. Those were almost entirely dispensed in the SA-IDC, and 635/2,820 were provided directly in saunas and bars attended by the more vulnerable population. However, among 115 cases admitted without prior follow-up in 2020, only one reported having sought the service due to the reactive result from the self-test obtained free of charge from SUS. 1/2,820, (0.04%) with six additional cases referring a paid self-test acquired in a pharmacy. In the first semester of 2021, only 274 self-tests were dispensed and in-service testing resumed. HIV testing is a cornerstone for prevention and treatment, needed to achieve suppression and break the transmission cycle.New cases linked to the service in the year are estimated by the number of people who started ART.. With covid-19, we tried to start treatment on the same day, consistent with the fact that in the presence of exposure risk, we prescribe PEP to mitigate the risk of a new infection. Therefore, we could use the same logic for treating quickly to prevent new infections. Worst-case estimates by North American researchers suggest that disruptions in HIV prevention and care during the pandemic may result in a 9% increase in new HIV infections over the next few years. Monitoring laboratory tools suffered a major temporary interruption during the initial response to the pandemic. For a while, there was no evidence for a decrease in sustained suppression. However, longer follow-up data are needed to assume the durability of ART suppression.The proportion of individuals who start ART within a month after collecting their first TCD4 count test (TimeCD4/ART) may be a proxy for how quickly linking to care occurs. Santo André showed faster linking and maintained more than 80% of its patients in 2020, compared with 70% in São Paulo and 64% in Brazil . The fa. According to data from 37 countries, HIV infection is independently associated with an increased 30% risk of death among patients hospitalized with suspected or confirmed SARS-CoV-2 infection. These inequalities in confronting covid-19 should not repeat what happened with Aids. Similar imbalances are emerging and repeating mistakes will be unforgivable. Vaccine disparities are the clearest evidence of this gap. The richest countries must act now to ensure that no one is left behind in the coronavirus vaccine rollout.Disparities in the social determinants of health and comorbidities likely have a greater influence on the susceptibility of PLWH to covid-19. PLWH bear a disproportionate burden of alcohol, drug use, and mental health disorders, as well as other structural vulnerabilities, which might increase their risk to both viral infections, essential for a robust health system. Priorities cannot be neglected, and managers need to use science creatively, adapting the response to local challenges.Although official Brazilian data show stability in retention and viral suppression of PLWH, careful monitoring in the coming years will be necessary, due to the impoverishment the country is experiencing amid a disruption of social public policies


The detection of volatile organic compounds (VOCs) exhaled by human body fluids is a recent and promising method to reveal tumor formations. In this feasibility study, a patented device, based on nanostructured chemoresistive gas sensors, was employed to explore the gaseous exhalations of tumoral, immortalized, and healthy cell lines, with the aim of distinguishing their VOC patterns. The analysis of the device output to the cell VOCs, emanated at different incubation times and initial plating concentrations, was performed to evaluate the device suitability to identify the cell types and to monitor their growth. The sensors ST25 (based on tin and titanium oxides), STN , and TiTaV  used here, gave progressively increasing responses upon the cell density increase and incubation time; the sensor W11 (based on tungsten oxide) gave instead unreliable responses to all cell lines. All sensors (except for W11) gave large and consistent responses to RKO and HEK293 cells, while they were less responsive to CHO, A549, and CACO-2 ones. The encouraging results presented here, although preliminary, foresee the development of sensor arrays capable of identifying tumor presence and its type. Tumor detection and monitoring are fundamental topics in medicine worldwide. According to American Cancer Society statistics, one out of three people will be diagnosed with cancer in their lifetime ,2. For tTumor gaseous biomarkers are volatile organic compounds (VOCs) produced by cancer cells mainly as a result of cell metabolic process alterations  and cell2 molecules that are quickly released into the atmosphere , entirely designed, produced, and assembled in the “Laboratorio Sensori” of the University of Ferrara, Italy. The working principle of these sensors is based on the chemoresistivity, i.e., the capability of varying the film conductivity as a consequence of the chemical reactions occurring between the surface of the grains and the gaseous molecules of the surrounding atmosphere. The chemoresistivity process requires the thermal activation of the sensor that consists in maximizing the number of electrons (in conduction band) able to overcome the potential barrier  in water or alcohol solution , during which several changes in the gel structure and properties occur as polycondensation, lead to an increase in thickness of the colloidal particles and to the reduction of their porosity.The last step is the drying and calcination: the GEL is dried first (at about 100 °C), and then calcinated at higher temperatures (at about 400–800 °C) to completely remove the residuals of solvents and of other chemical additives. In this process, the temperature and the relative humidity greatly influence the quality of the final MOX powder.To optimize the performances of a chemoresistive MOX gas sensor, it is primarily necessary to increase the surface–volume ratio of its sensitive porous material as much as possible, to maximize its interaction with the surrounding gases. With this aim, the sol-gel chemical process was employed to synthesize networks of MOX nanometric spherical grains, precisely interconnected ,43,44,45The MOX nanopowder, synthesized through the sol–gel technique ,42,43, wST25, based on tin oxides and titanium (25%) and gold nanoparticles (1%);W11, based on tungsten oxide;STN, based on tin, titanium, and niobium oxides;TiTaV, based on titanium, tantalum and vanadium oxides.The four MOX sensors hosted in SCENT B1, listed below, were selected on the basis of previous laboratory tests carried out on pure standard gases, on VOCs exhaled by blood, feces, biopsies, and preliminary tests on cultured cells ,32,33,34All sensors were thermally treated with a firing temperature of 650 °C and were heated to the working temperature of 450 °C by the platinum coil ,32,33,34uctivity ,32. The In order to assess the reliability of the sensor responses on the cell samples presented in the following, four different STN sensors were tested, as an example, in both wet , but the human skin fibroblasts were:A549, explant culture of lung carcinomatous tissue;CACO-2, human colorectal adenocarcinoma cell line;CHO, epithelial cell line derived from the ovary of the Chinese hamster;HEK293, specific immortalized cell line originally derived from human embryonic kidney;RKO, human colorectal cancer cells.The cell types employed here, purchased from the American Type Culture Collection  taken out from the incubator were quickly cooled to room temperature on a copper plate. Room temperature was monitored and kept approximately constant at 22.0 ± 1.5 °C.4, 2.5 × 105, 5 × 105, 1 × 106 cells per dish (3.5 cm of diameter); all dishes were kept in the same incubator at 37 °C in a humidified atmosphere containing 5% of CO2.To maximize the sample exhaling surface, in order to improve the signal-to-noise ratio, three petri dishes were simultaneously measured by placing them in the sample chamber stacked in an holding tray . Each roSince the cells were kept and grown in media rich in organic compounds, it was necessary to measure 2O at 37 °C  upon the cell density increase and withdrawal time from incubator .However, all sensors poorly discriminated among the 80, 250, and 500 K cells/dish plated at 24 h, except for the 1 M one. To check that the responses to the first three cell concentrations were small, they were averaged  and are shown in Sensor responses to CHO cells, an epithelial line derived from the ovary of the Chinese hamster, and A549 cells, an explant culture of lung carcinomatous tissue, were negligible after 24 h of plating, regardless of the plating concentration. In particular, the A549 responses were smaller than one, i.e., they were smaller than the sensor response to culture medium only. Both these cell types grew quite fast, and at a concentration of 500 K, cells/dish were already confluent after 48 h of plating , insert;Lastly, the cell lines CACO-2 and human skin fibroblasts gave negligible responses . This waIn summary, it is possible that the VOCs emitted by CHO, A549, and CACO-2 cell lines, although they are expected to be quite sustained since these cells are fast growing and therefore have a high metabolism, are poorly detected by the sensors. If this is the case, it may become advantageous, since the sensors here employed, giving different response patterns with different cancer cells, could be used in the future not only to detect them (hopefully to an early stage), but also to identify their type. To further explore this idea, as shown in The same sensor gave different response amplitudes to different cell lines a, or difThe sensors ST25, STN, and TiTaV used in this work gave progressively increasing responses upon increasing the cell number, no matter if it was attained by the initial seeding concentration or by the cell growth. The sensor W11 gave instead unreliable responses to all the cell lines here employed, and therefore, they were discarded in the final analysis. The sensors gave large responses to the RKO and HEK-293 cell lines, while they were progressively less responsive to the CHO, A549, and CACO-2 cell lines. This suggests that a particular cell type could be identified by properly combining the response to its VOCs via a different sensor, allowing for the future development of sensor sets that are able to identify the presence of a tumor and even its type. The novelty of this approach, in respect to the current methods in development for tumor identification, is the employment of a portable, compact, and fast (the outcome is attained within 10–15 min) device, not requiring expensive consumables or complex and expensive analysis of the samples (that may require days), and that can be operated by untrained people.Malagù, C.; Gherardi, S.; Zonta, G.; Landini, N.; Giberti, A.; Fabbri, B.; Gaiardo, A.; Anania, G.; Rispoli, G.; Scagliarini, L. Combinazione di materiali semiconduttori nanoparticolati per uso nel distinguere cellule normali da cellule tumorali (2015), National #:102015000057717 .


Cleiothyridina and 405 Crinisarina specimens, 18 and 39 individuals were malformed, respectively; an abnormality ratio of nearly 10%. Considering the preservation status and buried environment of the abnormal specimens, and analysis of trace elements and rare earth elements from whole-rock and brachiopod shells, we conclude that the appearance of malformed athyrids is likely related to epi/endoparasites, or less likely the slightly higher content of heavy metal in the sea.Although malformations are found in both extant organisms and the fossil record, they are more rarely reported in the fossil record than in living organisms, and the environmental factors causing the malformations are much more difficult to identify for the fossil record. Two athyrid brachiopod taxa from the Upper Devonian Hongguleleng Formation in western Junggar  show distinctive shell malformation. Of 198  However, malformations are known from the fossil record too, and have been reported from individuals of different fossil groups, including foraminifera , trilobiCleiothyridina and Crinisarina) specimens from the Upper Devonian Hongguleleng Formation in the Buninuer section, of which nearly ten percent of individuals were malformed. The aims of current study are to explore the biotic or abiotic stressors of these malformed brachiopods, and the effect of change in environmental factors on the brachiopod shells.Although many malformed fossils have been described, abnormal fossils  are generally rare, with sometimes only one or two specimens known. Therefore, previous studies have generally been limited to description of malformed specimens and simple classification of the cause(s) of malformation, only in a few cases the relationship between malformed specimens and their habitat has been discussed . AlthougThe material studied in this paper was collected from the Upper Devonian Hongguleleng Formation in western Junggar, Xinjiang. The Hongguleleng Formation is a widely distributed marine unit near the Devonian–Carboniferous boundary in western Junggar. The formation is divided into three members: the Lower Member is composed of thin bioclastic limestones, muddy limestones and shales; the Middle Member is mainly made up of fine pyroclastic rocks with a few sandy and muddy limestones; and the Upper Member consists of calcareous clastic rocks with a small amount of bioclastic limestones . The forCrinisarina and Cleiothyridina) are non-flattened specimens with well-preserved dorsal and ventral valves. Although athyrids occur in other beds of the Hongguleleng Formation, no malformed specimens were found in those levels.Brachiopods occur in all three members of the Hongguleleng Formation. Brachiopod abundance and diversity is highest in the Lower Member, with the groups present including Productida, Orthida, Rhynchonellida, Athyridida and Spiriferida . AthyridThe 603 specimens include a wide range of size and may include individuals representing different growth stages . We divianom is equal to lg[3Cen/(2Lan + Ndn)], and Cen, Lan and Ndn were NASC-normalized of Ce, La and Nd, respectively.To explore whether athyrid malformations were caused by environmental factors, trace and rare earth elements of whole-rock samples from specific levels within the Hongguleleng Formation were measured. Samples BL–1 and BL–2 were obtained from the lower part of the Lower Member of the Hongguleleng Formation, which yielded abundant non-malformed athyrids. Samples BL–3 and BL–4 came from the upper part of the Lower Member, form where the malformed fossils described in this paper were obtained. Samples BL–5, BL–6 and BL–7 were collected from the Middle Member of the Hongguleleng Formation; only a few athyrids occurred at the bottom of this member, and the group almost disappeared above that level. Sample B9b–1 was from the Upper Member, which yielded a small number of athyrids. The trace elements and rare earth elements of four athyrid shells were measured. Samples CL69 and CR178 are non-malformed shells, while CLJ13 and CRJ29 are malformed shells. All samples were ground into powder and analyzed by ALS Minerals/ALS Chemex Co. Ltd . Rare earth and trace elements were fused with lithium borate, and quantitatively analyzed by ICP-MS with Elan 9000 Perkin Elmer that was made in America. The CeOf the 603 athyrid fossils, macroscopic abnormalities were detected in 57 specimens. The most common teratomorphy is obvious asymmetry on the left and right sides of the shells , 1G–1J, Cleiothyridina and 405 of Crinisarina, 18 (9.1%) Cleiothyridina and 39 (9.63%) Crinisarina were malformed. The overall malformation ratio was 9.45%, nearly one-tenth of all specimens , 8.5% (15 mm ≤ H < 20 mm), 13.7% (20 mm ≤ H < 25 mm) and 11.1% (25 mm ≤ H < 30 mm); those of Cleiothyridina are 9.68% (10 mm ≤ H < 15 mm), 9.3% (20 mm ≤ H < 25 mm) and 14.8% (25 mm ≤ H < 30 mm). Thus, shell-malformation occurs at different athyrid growth stages, and the probability of abnormality is higher in larger specimens. That indicates a higher probability of malformation during advanced ontogenetic stages of the studied brachiopod taxa.Of the 198 specimens of pecimens . In all ne shell . Moreovene shell , 2C. Thee.g., trilobites, crinoids, and corals) that are well-preserved in three dimensions , which are all non-malformed in the same layer. In addition, the fossils preserved in the upper part of the Lower Member of the Hongguleleng Formation are relatively complete, and there is no evidence of strong bottom currents, so the influence of bottom currents can be excluded as a teratogenic factor. Pentlandina loveni, from the Sliurian Högklint Formation in Gotland (Sweden), are commonly markedly asymmetric, and some groups of shells occur in tight clusters, each apparently attached to other shells of the same species. He interpreted that these asymmetrical shells resulted from the limited space for growth, i.e., overcrowded conditions. However, all specimens from western Junggar are isolated, instead of grouped in tight clusters or attached to other shells. Furthermore, if they are living in overcrowded space, the distribution of malformation should be random or almost uniform on both sides of the shells, but the malformations commonly occur on the right side of athyrids shells  from western Junggar  in the layer with the malformed athyrid shells are relatively high compared to the levels in the lower part of the Lower Member, particularly cadmium and lead. The levels of cadmium are 0.02 and 0.03 ppm in the lower part of the Lower Member, and are 0.03 and 0.04 ppm in the horizon that yielded the malformed fossils. The lead contents are 1.3 and 2.2 ppm in the lower part, but 5.1 and 5.2 ppm in the upper part. In the Middle Member of the Hongguleleng Formation, where athyrids almost disappeared, the heavy metal levels are even higher. In the Upper Member, where athyrids reappear, the heavy-metal content decreases again  did not change significantly in the sediments from the non-malformed athyrids-bearing layers to the layers containing malformed specimens, and the contents of arsenic and silver even decreased in the layers containing malformed athyrids  epi/endoparasites, or a low probability that slightly high heavy-metal  in the sea, rather than eutrophication, bottom current activity, overcrowded conditions, hypoxia or other factors.Some specimens of two athyrid genera, 10.7717/peerj.13447/supp-1Supplemental Information 1Click here for additional data file.10.7717/peerj.13447/supp-2Supplemental Information 2Click here for additional data file.10.7717/peerj.13447/supp-3Supplemental Information 3Click here for additional data file.


The occurrence and development of malignancies are closely related to abnormal cell cycle regulation. Myeloid leukemia factor 1 (MLF1) is a small nucleocytoplasmic shuttling protein associated with cell cycle exit, apoptosis, and certain immune functions. Therefore, it is pertinent to explore the role of MLF1 in health and diseases. Studies to date have suggested that MLF1 could act as a double-edged sword, regulating biochemical activities directly or indirectly. In hematopoietic cells, it serves as a protective factor for the development of lineages, and in malignancies, it serves as an oncogenesis factor. The diversity of its functions depends on the binding partners, including tumor inhibitors, scaffolding molecules, mitochondrial membrane proteins, and transcription factors. Emerging evidence indicates that MLF1 influences immune responses as well. This paper reviews the structure, biological function, and research progress on MLF1 in health and diseases to provide new insights for future research. Myeloid leukemia factor (MLF) is a poorly characterized family of conserved proteins which earliest member, myeloid leukemia factor 1 (MLF1), is associated with hemopoietic lineage commitment and malignancies. MLF1 has so far been shown to be a double-edged sword, acting as either a tumor suppressor or an oncogene, depending on the context of the cell. MLF1 has been initially described in the leukemic fusion protein NPM-MLF1, which is generated by a rare t3;5)q25.1;q34) chromosomal translocation in patients with acute myeloid leukemia (AML) , which leads to the downregulation of COP1; therefore, the cell cycle of hematopoietic cells becomes arrested because the bonding accelerates the accumulation of wild-type p53 in the nucleus . MLF controls LZ activity and prevents its degradation, which is critical to control crystal cell number in the fly  is a key transcription factor regulating myeloid differentiation in normal hematopoiesis and is frequently dysregulated in AML   . Furthere system . However protein , and its protein , 59. Thi protein . However3. MLF1 in antitumor protection. MLF1 protein is directly associated with the deubiquitinase ubiquitin-specific peptidase 11 (USP11), which is a promising therapeutic target. Additionally, USP11 has promoted the accumulation of MLF2 in all tested cells  has been confirmed by genome-wide DNA methylation analysis, and aberrant methylation is associated with inverse changes in mRNA levels  is more likely to occur in M6 patients than in patients with other leukemia types . A preclrelevant . The prerelevant . An exteferation . Recent feration . Howeverxtensive , as indixtensive . MLF1 isc cancer , 73. A crization . Shuang arcinoma . Defectsarcinoma . HypermeA levels . MarcelaA levels .In conclusion, MLF1 is a small shuttling protein playing a critical role in biological and pathological processes. Currently, research regarding MLF1 has mainly focused on cancer development, which is still an obscure and disputed topic. In general, although there is more evidence supporting the point that MLF1 contributes to tumor suppression, a few studies have confirmed the tumorigenesis of MLF1 in solid and hematologic tumors, which cannot be neglected. Additionally, the functions of MLF1 in immune response still need further investigations despite some already reported studies. Given the complexity and variety of involved proteins, we may draw a conclusion that MLF1 might be a double-edged factor in the regulation of cell cycle, immunity, stem cell development, and cancer. However, studies about MLF1 are still inadequate; therefore, expanding the research on MLF1 is significant and may enrich the knowledge of MLF1 in the above-mentioned conditions. On the other hand, exploring regulations of MLF1 shuttling will provide a better understanding of MLF1, which helps develop novel specific MLF1-aimed drugs that might provide a promising strategy for cancer treatment, as well as other pathologies, such as neurological diseases. Therefore, analysis of the partner protein, localization, and shuttling mechanism might provide new insights for future research.All authors contributed to the article and approved the submitted version.


Blinatumomab is a bispecific CD19-directed CD3 T-cell engager that has proven efficacy in children with relapsed or refractory B-cell acute lymphoblastic leukemia (ALL). Despite its efficacy, it has also been associated with the development of potentially serious adverse events such as the cytokine release syndrome (CRS) and neurologic events. The present meta-analysis aimed to assess the safety profile of blinatumomab in terms of serious adverse events, CRS, and neurologic events  in pediatric patients with B-cell ALL.A systematic review was conducted in Pubmed up to December 10, 2021 to retain pediatric clinical trials on blinatumomab. A random effect meta-analysis approach was used. This study followed the PRISMA statement. Four out of the 255 initial references were selected, of which 2 were phase 1/2 clinical trials and 2 phase 3 clinical trials. Blinatumomab was associated with a lower risk of serious adverse events , febrile neutropenia , infection , and grade ≥ 3 adverse events  compared to chemotherapy. No difference in the risk of CRS  and seizure  was observed between groups, while for encephalopathy a higher risk was associated with blinatumomab compared to chemotherapy .Our data support the good safety profile of bliantumomab in treating pediatric patients with B-ALL. Relapse in vivo . Moreove in vivo  and high in vivo , 12. Des in vivo . Moreove in vivo . Howeverpost-hoc analysis, etc.) were also excluded.The Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guideline were used to perform a standardized data search, extraction, reporting, and presentation. A systematic literature search of clinical trials of blinatumomab in pediatric patients with ALL published up to December 10, 2021 was performed in PubMed. The search strategy is shown in supplementary file 1 and it was performed by two investigators (AM and MM) who screened the scientific literature. One investigator identified selection criteria and started the review of titles and abstracts of all retrieved studies (MM). Another investigator validated these criteria and independently screened the selected papers (AM). In case of discrepancies between the two investigators, a third investigator was consulted (FR) to get a final decision. A population/participant, intervention, comparator/control, outcome, study design, known as PICOS, framework was developed. Population: pediatric patients with acute lymphoblastic leukemia; Intervention: blinatumomab; Comparator: standard chemotherapy; Outcome: number of adverse events, serious adverse events, adverse events graded ≥ 3, cytokine release syndrome (CRS), encephalopathy, seizure, neurologic events, infection, or febrile neutropenia; Study design: clinical trial. We included articles that met the PICOS criteria. Articles in languages other than English were excluded. Reviews, letters, commentary, case reports/series, editorials, preclinical studies, guidelines, observational studies, or other type of studies .2 test statistics used to evaluate the heterogeneity of the effect across studies. A Q statistic with a p-value < 0.10 was considered significant. The percentage of total variation across studies due to heterogeneity rather than chance was assessed with I2 statistics, which values <25% were used for low heterogeneity, between 25 and 75% for moderate heterogeneity, or > 75% for high heterogeneity. Risk ratios (RRs) and 95% confidence intervals (95% CIs) were computed for dichotomous outcomes. Results were also displayed as fixed effect model. For phase 1/2 and 3 clinical trials data were pooled to estimate the incidence rate of CRS and neurologic events. Incidence rates were computed by dividing the number of events with the number of pediatric patients exposed.For phase 3 clinical trials, the endpoint was to assess the risk ratio of adverse events, serious adverse events, adverse events graded ≥ 3, cytokine release syndrome (CRS), encephalopathy, seizure, and neurologic events for blinatumomab compared with comparator arm. A random effect meta-analysis was applied, with the QStatistical analyses were computed with software R version 3.6 .N = 124), case report/series (N = 35), observational studies (N = 24), letters/commentary (N = 20), post-hoc analysis (N = 15), preclinical studies (N = 5), cost-effectiveness analyses (N = 4), genetic studies (N = 2), and qualitative researches (N = 2). Observational studies were primarily excluded because they do not fall within our aim of assessing the safety profile of blinatumomab by using pediatric clinical trials. Moreover, because they investigated other therapies rather than blinatumomab (N = 8), they were conducted in adults (N = 5), they were not in English (N = 1), or they were small studies without a comparison group (N = 6). Then, a total of 24 clinical trials was identified, of which 20 were excluded because they were conducted in adult patients (N = 16), they were related to biological samples (N = 3) or were sub-analyses of already included clinical trials (N = 1). Finally, 3 (19%) of aforementioned clinical trials conducted in adult patients evaluated the efficacy of chimeric antigen receptor (CAR) T-cell therapy rather than blinatumomab. As a result, a total of 4 clinical trials was identified, of which 2 were phase 1/2 clinical trials . Among safety outcomes both studies evaluated neurologic events and CRS. One open-label clinical trial enrolled 9 Japanese children with B-ALL, who received blinatumomab for 4 weeks (at a dose of 5 μg/m2/day for week 1 and followed by a dose of 15 μg/m2/day for weeks 2–4), followed by a treatment-free interval of 2-week. In this study a total of 7 neurologic events and 5 CRS were identified .One randomized phase 3 clinical trial included children aged > 28 days and < 18 years with B-ALL at high-risk of first-relapse with morphologic complete remission  or M2 marrow (blasts ≥ 5 and <25%) at time of randomization. These patients were previously treated with induction therapy and 2 blocks of consolidation therapy and then randomized to receive a third consolidation course with 1 cycle of continuous intravenous infusion of blinatumomab for 4 weeks (dose: 15 μg/m2/day) or chemotherapy .p = 0.67). For the number of serious adverse events, 13 events for blinatumomab and 22 for chemotherapy were observed. Blinatumomab was associated with a lower risk of serious adverse events compared to chemotherapy . For CRS, 24 events were found in the blinatumomab group and 1 event in the comparator group. No difference in the risk of CRS was observed between groups . For encephalopathy a total of 12 events were observed with blinatumomab, while no event was observed with the control group. Blinatumomab showed a higher risk of encephalopathy compared to chemotherapy . Finally, 6 events of seizure were observed only in the blinatumomab group. No difference in the risk of seizure was observed between the two groups . Among the events potentially associated with life-threatening complications, 9 events of febrile neutropenia occurred with blinatumomab and 69 with the comparator group. Blinatumomab showed a lower risk of febrile neutropenia then the comparator arm . The number of infection were 31 for the blinatumomab group and 72 for the comparator group, with a lower risk for blinatumomab than the comparison .The other randomized phase 3 clinical trial included patients aged 1 to 30 years with B-ALL at high- and intermediate-risk of first relapse. The trials enrolled a total of 208 patients, of which 33 (15.9%) were aged 18–30 years (15 of blinatumomab group and 18 of comparator group). All patients were treated with a course of reinduction chemotherapy for 4-weeks, followed by 2 cycles of blinatumomab or multiagent chemotherapy based on randomization . Based oThe present meta-analysis is the first to our knowledge that evaluated the safety profile of blinatumomab with a focus on the risk of encephalopathy, seizure, and CRS in the pediatric population. A previous meta-analysis on blinatumomab included both adult and pediatric patients and reported data published up to 2017, thus not including pediatric phase 3 clinical trials . TherefoWe observed an incidence rate of 0.21 (95% CIs 0.16–0.27) for neurologic events and 0.16 (95% CIs 0.11–0.21) for CRS. The previous meta-analysis found instead a lower occurrence rate of CRS  and neurologic events , but we should consider that these rates were estimated only for grade 3 events and by considering a mixed population (adult and pediatric) . For theFor the risk of neurologic events, blinatumomab showed a higher risk of encephalopathy compared to chemotherapy, while no difference was observed for the risk of seizure. This is in accordance with studies that reported among the most frequent neurologic events with blinatumomab: tremor, dizziness, confusional state, and encephalopathy , 31–33. Although blinatumomab has shown a significant survival benefit compared to chemotherapy, it has a particular toxicity profile that requires a scrupulous surveillance by all healthcare professionals , both duThe use of a single repository such as PubMed is an important limitation in our meta-analysis. Indeed, the meta-analysis was performed on a limited number of clinical trials, which also have differences in the characteristics of population and treatment schedules. Furthermore, in the trial by Brown et al. was not possible to select only events that occurred in the pediatric cohort, because it included patients aged up to 30 years. All this could have affected our results. Finally, conclusions could be susceptible to study's limitations, publication bias and the formulation of eligibility criteria. These limitations may create bias and alter the analyses. Therefore, results should be interpreted accordingly.In conclusion, blinatumomab showed a good safety profile in treating pediatric patients with B-ALL. The risk of serious adverse event was indeed higher with chemotherapy than blinatumomab. The occurrence of CRS and neurologic events was very low among clinical trials. Moreover, CRS and neurologic events have showed a generally good response to drug withdrawal and interventional therapies in the analyzed clinical trials. Moreover, an important role in limiting the seriousness of these events is recovered by the appropriate management of patients that has to consider the patients' history and the use of prophylaxis therapies. Limited progress in treating relapsed disease have been done in the last 20 years, with few new drugs that have been developed for ALL. CAR T cells demonstrated significant results in determining sustained and durable responses in patients not only with bone marrow relapse but also with relapse in the central nervous system. CAR T cells thus have the potential to be an alternative to HSCT as cure of relapsed disease in spite of the remaining concern represented by CRS, since complications of HSCT should not be underestimated as well. Current trials are investigating instead the use of inotuzumab ozogamicin in multidrug regimens, including a phase I study evaluating the safety of this drug in combination with prednisone, cyclophosphamide, and vincristine in patients affected by refractory CD22 + acute leukemia. However, further studies are needed to better evaluate blinatumomab safety profile and to allow the execution of more reliable meta-analytic approach.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation. And I did not detect any particular expressions.AM, MM, and FR performed the screening of the literature. AM and MM performed the analysis and wrote the paper. AM, MM, GM, GM, EP, and FR drafted the work and revised it for important intellectual content. FR developed the concept and designed the study. All authors approved the final version of the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.