Prostate cancer is known to lose the capability to absorb and secrete zinc compared to normal prostate tissue, suggesting that the evaluation of zinc in prostate secretion can be a tool to identify the risk of developing cancer. In our study, we observed that the average amount of zinc detectable in urine after a prostatic massage is lower in patients with prostate cancer than in healthy subjects. Moreover, there is an inverse correlation between the concentration of urinary zinc and the tumor stage. This evidence suggests that the evaluation of urinary zinc may be a parameter for better diagnosis and prognosis of prostate cancer.Prostate Cancer (PCa) is one of the most common malignancies in men worldwide, with 1.4 million diagnoses and 310,000 deaths in 2020. Currently, there is an intense debate regarding the serum prostatic specific antigen (PSA) test as a diagnostic tool in PCa due to the lack of specificity and high prevalence of over-diagnosis and over-treatments. One of the most consistent characteristics of PCa is the marked decrease in zinc; hence the lost ability to accumulate and secrete zinc represents a potential parameter for early detection of the disease. We quantified zinc levels in urine samples collected after a standardized prostatic massage from 633 male subjects that received an indication for prostate biopsy from 2015 and 2019 at AOU Città della Salute e della Scienza di Torino Hospital. We observed that the mean zinc levels were lower in the urine of cancer patients than in healthy subjects, with a decreasing trend in correlation with the progression of the disease. The combination of zinc with standard parameters, such as PSA, age, digital rectal exploration results, and magnetic resonance findings, displayed high diagnostic performance. These results suggest that urinary zinc may represent an early and non-invasive diagnostic biomarker for prostate cancer. The diagnostic process to detect prostate cancer (PCa) includes measuring serum prostatic specific antigen (PSA) levels and a digital rectal examination (DRE). When PCa is diagnosed early, the treatment can be effective and with minimal morbidity , underlyCurrently, there is an intense debate regarding the serum PSA test as a screening tool for PCa due to the lack of specificity and high prevalence of over-diagnosis and over-treatments . It is, In particular, the controversial PSA screening sustains the need for alternative biomarkers with better diagnostic and predictive potential, capable of distinguishing between aggressive and indolent cancers. In the last years, several biomarkers have been discovered, suggesting better performances compared to PSA. Among these, we cite the Prostate Health Index (PHI) , 4K scorHowever, all these new approaches led only to limited benefits in the diagnostic-therapeutic pathway of PCa, mainly because a unique gene signature cannot be universally applicable to all patients with an oncologic disease. A shared aspect that characterizes neoplastic transformation is a profound change at both phenotypic and functional levels. One of the physiological functions of the prostate is to produce part of the fluid that forms semen. The majority of the prostate is composed of glandular cells; hence the most common type of cancer that affects this organ  can impair the composition of the prostate fluid. The amount of several products of the prostate, such as PSA is decreased in advanced cancer ,9, and tTogether with the mammary and pancreatic tissues, the prostate is one of the organs that physiologically accumulate most of the zinc circulating in our body . Zinc inProstate cells actively accumulate zinc. Several studies have shown that damage to the functionality of the cells, for example, due to a neoplastic transformation, leads to a loss of the ability to absorb zinc and therefore, to a lower expression in the tissue ,15,16.In this study, the level of zinc present in the urine of men who received an indication to undergo a prostate biopsy for a suspected tumor was quantified. The amount of zinc was found to be lower in the urine of patients with cancer than those with negative biopsy. Also, the average urinary zinc level gradually decreases as the disease progresses. These results are in line with what has already been observed in the tissue. However, here we reported for the first time that urinary zinc can be used for the diagnosis and prognosis of PCa, alone and in combination with standard parameters such as PSA or multiparametric Magnetic Resonance (mpMRI).Men scheduled for prostate biopsy at the urology unit in the AOU Città della Salute e della Scienza di Torino  from June 2015 to May 2019 (four years) were invited to participate in the study. The indication for prostate biopsy was defined according to routine clinical parameters, such as PSA levels, digital rectal exploration (DRE), and MRI imaging. Before the biopsy, we collected a urine sample from each subject after a standardized prostatic massage. Participants (n = 633) were divided into Training Cohort 1 (n = 411) and Validation Cohort 2 (n = 212). Subjects with PSA levels above 25 ng/mL (n = 38) were excluded from the analysis. Subjects that received a diagnosis of prostate cancer (PCa patients) were stratified according to PSA, Gleason Score (GS), and tumor staging  into lowA urine sample was collected from 78 men with an indication of prostate biopsy without prostate massage prior to collection.Urine samples were collected and processed as previously described . A standardized prostate massage was performed to extract prostatic secretions through three digital compressions in each lobe, starting at the base, moving down to the center and apex in a 30-s time lapse.Approximately 30 mL of first voided urine were collected after gentle agitation, a 15 mL aliquot was stored in plastic tubes at −80 °C within 5 min of collection.Urinary zinc measurement was performed in the Atomic Absorption Laboratory , AOU Città della salute e della Scienza di Torino, Turin, Italy.Samples were first acidified with 100 μL 18% HCl and then diluted 1: 5 with CsLaCl 1 g/L (1 mL sample + 4 mL CsLaCl 1 g/L). They were then analyzed by the flame atomic absorption technique. In addition, four calibration curve standards were used for the analysis , and two controls (Seronorm Urine L-1 and Seronorm Urine L-2) diluted 1: 4 with CsLaCl 1 g/L (1 ml control + 3 ml CsLaCl 1 g/L).Once prepared, the samples were analyzed on the Zeenit 700p instrument .A human investigation was conducted according to the Declaration of Helsinki principles after the approval of the study by the Scientific Ethics Committee of AOU Città della Salute e della Scienza di Torino, A.O. Mauriziano, A.S.L. TO1 (Prot. No. 0110644). Each participant released written informed consent before inclusion in the study, and specimens were anonymized after collection.We categorized subjects by risk class based on PSA value, GS, the number of positive biopsies, and TNM staging . We performed univariate and multivariate logistic regression analysis to evaluate the diagnostic performance of urinary zinc in detecting csPCa, alone or in combination with other parameters, such as PSA level, DRE finding, age, and mpMRI. We designed different multivariate logistic models, and the discriminative power was assessed by calculating the area under the receiver operating characteristic (ROC) curves (AUC). We compared the diagnostic performance of four different multivariate logistic regression models. The first model (PSA model) included serum PSA level only. The second model (SOC model) included PSA level, age at diagnosis, and abnormal DRE. The third model (Zinc model) included urinary zinc level. The fourth model (Zinc + SOC model) included urinary zinc level, serum PSA level, age, and abnormal DRE. The fifth model (MRI model) considered PiRADS value. The sixth (SOC + MRI model) included serum PSA level, age, abnormal DRE, and PiRADS. The seventh (Zinc + SOC + MRI model) included urinary zinc, PSA level, age, abnormal DRE, and PiRADS. Comparisons of AUCs of different models were determined using DeLong’s method. Logistic regression coefficients were estimated in Cohort 1 and externally validated in Cohort 2.The Number Needed to Predict (NNP), representing the number of patients who need to be examined in a population to correctly predict the diagnosis of one person, was also calculated .® Statistical Software version 19.8 .; https://www.medcalc.org; 2021Statistical analyses were performed as previously published  with MedRecruited men were divided into training Cohort 1 and validation Cohort 2. In Cohort 1 (n = 394), 187 men had a negative biopsy (100 with no evidence of a tumor or with benign prostatic hyperplasia (BPH), 51 inflammation/prostatitis, 21 high-grade prostatic intraepithelial neoplasia, and 15 atypical small acinar proliferation), while 207 (49.7%) had a positive biopsy outcome.Based on D’Amico’s stratification, 17 patients had low risk, 68 favorable-intermediate (int fav) risk, 67 unfavorable-intermediate (int unfav) risk, and 55 high risk PCa. The percentage of clinically significant (cs) PCa was 48.2% .For Cohort 2, of 201 patients, 74 had a negative biopsy and 127 (63.2%) a positive biopsy outcome, of which 12 were low risk, 38 int fav risk, 38 int unfav risk, and 39 high risk PCa. The prevalence of csPCa was 57.2% .p = 0.02) but a lower prevalence of csPCa (p = 0.01).The average age of subjects was 68 years in both Cohorts. In Cohort 1 and Cohort 2, there was no difference in terms of mean PSA value (7.2 vs. 7.8 ng/)mL. Subjects in Cohort 1 displayed higher DRE abnormality compared to Cohort 2  compared to healthy subjects (1.02 µg/mL) and low-risk PCa (1.93 µg/mL). No differences were observed between low-risk PCa and healthy subjects , Table 2To evaluate the role of prostate massage in extracting prostate contents, zinc was evaluated in the urine collected from men with an indication of biopsy without performing this procedure. In the absence of prostate massage, the average zinc level is lower than that observed in the presence of massage, with no significant differences between healthy individuals and prostate cancer patients .We compared the levels of urinary zinc in subjects without evidence of PCa or low-risk PCa (non-cancer) to that of patients with csPCa. p = 0,0001). The average age . Furthermore, the AUC of zinc + SOC was also higher than the AUC of SOC alone (p = 0.0011).The AUC for blood PSA, the standard of care parameters , urinary zinc alone, or zinc + SOC was 0.551, 0.607, 0.652, and 0.687, respectively. The AUC of both zinc alone and zinc + SOC was significantly higher than the AUC of PSA  and negative predictive value (NPV), hence greater capacity in accurately classifying biopsies results compared to PSA alone and SOC .At 95% sensitivity, the Number Needed to Predict (NNP) was lower for zinc and zinc + SOC than for PSA and SOC .We evaluated if zinc had different diagnostic capabilities in different subgroups of subjects based on PSA value or age. In subjects with a PSA range between 0 and 4 ng/mL, 4.1–10 ng/mL, and 10.1–25 ng/mL, analysis for Zinc levels showed AUCs of 0.589, 0.642, and 0.753, respectively .The AUC of zinc for patients aged below 60, between 60 and 75, and above 75 were 0.629, 0.637, and 0.697, respectively .Four diagnostic models based on PSA, age, DRE, and urinary zinc were developed in Cohort 1 for the capability to assess the probability of csPCa. p = 0.0085, p = 0.0195, and p = 0.0001, respectively. The combination of zinc + SOC showed higher AUC (p = 0.0177) compared to SOC . The AUC for PSA alone, SOC, zinc, and zinc + SOC in predicting the chance of csPCa was 0.558, 0.669, 0.683, and 0.735, respectively . The AUCd to SOC .n = 226), 82 were disease-free, 11 had low-risk PCa, 49 Int-fav PCa, 49 Int-unfav PCa, and 35 high-risk PCa.As mpMRI is becoming the gold standard for prostate biopsy candidate selection, we investigated whether urinary zinc assessment could increase the diagnostic capacity of mpMRI and SOC. In individuals undergoing prostate biopsy for suspected mpMRI .Furthermore, we separately evaluated the diagnostic performance of zinc and SOC in subjects with Prostate Imaging Reporting and Data System (PiRADS) values of 3, 4, and 5. In PiRADS groups 3 and 4, the combination of zinc + SOC displayed an AUC of 0.827 and 0.730, which are statistically different compared to PSA alone (p = 0.1).In PiRADS group 5, the combination of zinc + SOC displayed an AUC of 0.835; that, however, is not statistically different from PSA alone .As shown in The discovery and validation of new biomarkers to avoid unnecessary biopsies and reduce the risk of overdiagnosis and overtreatment for PCa is urgently needed in this field.A multitude of new diagnostic tools have emerged in recent years and have become available on the PCa market, with the intention of providing more accurate information than the standard PSA and DRE test on the actual risk of cancer .Although several tests have shown a potential ability to improve the diagnostic and therapeutic pathway, there is a lack of prospective studies to support their impact on disease outcomes.As biomarkers become available, it is increasingly important to understand how, when, and on which patients they should be used. Integration into routine clinical practice and insurance coverage are still areas where more work needs to be done for the benefit of both doctors and patients.However, the latest update of the European guidelines of the European Association of Urology (EAU-ESTRO-SIOG) recommended none of these tests or tools in support of the standard of care . To date, with the technological advances in MRI, with improvements in the quality and standardization of interpretations with PIRADS-v2 , mpMRI hThe results of this study highlighted that a significant decrease in mean zinc levels is observable in the urine of PCa patients compared to healthy subjects. This evidence is in line with previous studies showing that the level of zinc in prostate tissue and prostatic secretion is substantially lowered in the presence of neoplastic transformation compared to non-malignant pathologies and normal glands ,25,26.Urinary zinc showed an additive value in combination with standard of care and mpMRI. Available evidence suggests that the incorporation of mpMRI in the diagnostic pathways for males should be recommended. However, despite a bought advantage in the use of this approach in the diagnostic path of prostate cancer, the need for instrumentation, specially trained healthcare personnel, and the high costs greatly limit its use as a mass screening method .Furthermore, the high probability that clinically significant prostate cancer will not be detected on MRI leaves many concerns about its use as a first-line test . The latIn this study, we observed that urinary zinc analysis, mpMRI imaging results, and evaluation of standard parameters are complementary. Their simultaneous evaluation could potentially improve accuracy in identifying the correct candidates for prostate biopsy, thus reducing the risk of false positive and false negative results.Several findings provide evidence that strategies combining blood or urine biomarker testing and MRI can improve clinical routine by better detecting prostate cancer early and providing insight for biopsy decision-making .Interestingly, in vivo studies in mice showed that zinc detection by imaging technologies can provide a specific diagnostic method to differentiate non-cancerous prostate tissue from prostate cancer in situations where it may be difficult to detect using current multiparameter MRI protocols .The main limitation of our study is that not all the subjects underwent mpMRI for biopsy decision-making. In further studies, it would be interesting to assess the diagnostic performance of urinary zinc, alone or in combination with other parameters or biomarkers, in a wide range of subjects in different settings, such as symptomatic or general populations undergoing the current diagnostic path, including state-of-the-art magnetic resonance imaging.Future studies will also be performed on men who have not received a prostate biopsy indication, comparing the values of urinary zinc with the results of MRI.The prostatic gland is one of the tissues that particularly accumulate zinc for biological processes. The alteration of zinc absorption and accumulation is one of the key characteristics of PCa. During the progression of the disease, lower zinc levels can be found in comparison to normal tissue. Moreover, PCa appears to be the only prostatic disease in which zinc accumulation is compromised. Other studies highlighted that the decrease in zinc levels is not a characteristic of BPH or prostatitis , suggestUrinary zinc measurement is already in use in some clinical biochemistry laboratories to detect industrial and accidental exposure to zinc or malabsorption. This indicates that the evaluation of urinary zinc is already in use and therefore, there is optimized and dedicated instrumentation and sample preparation methods to carry out this evaluation.PCa is highly heterogeneous, mainly multifocal, and difficult to be fully characterized with a unique signature. It may therefore be necessary to identify and clinically validate a panel of biomarkers that evaluate different aspects of tumor progression to better select suitable patients for prostate biopsy.In conclusion, the loss of zinc accumulation and secretion by the prostate during neoplastic transformation could potentially represent a hallmark of PCa, and its combination with standard diagnostic parameters and mpMRI could represent an interesting approach in the diagnosis of PCa.Although the present study showed a better diagnostic capacity of PCa with the implementation of urinary zinc, results suggest space for improvement, potentially in combination with other urinary biomarkers with similar physiology.


Alterations in nutritional status, in particular sarcopenia, have been extensively associated with a poor prognosis in cirrhotic patients regardless of the etiology of liver disease. Less is known about the predictive value of myosteatosis, defined as pathological fat infiltration into the skeletal muscle. We retrospectively analyzed a cohort of 151 cirrhotic patients with unresectable hepatocellular carcinoma (HCC) who underwent their first trans-arterial embolization (TAE) between 1 March 2011 and 1 July 2019 at our Institution. Clinical and biochemical data were collected. Sarcopenia was assessed using the L3-SMI method while myosteatosis with a dedicated segmentation suite (3D Slicer), using a single slice at an axial plane located at L3 and calculating the IMAC (Intramuscular Adipose Tissue Content Index). The sex-specific cut-off values for defining myosteatosis were IMAC > −0.44 in males and >−0.31 in females. In our cohort, 115 (76%) patients were included in the myosteatosis group; 128 (85%) patients had a coexistent diagnosis of sarcopenia. Patients with myosteatosis were significantly older and showed higher BMI than patients without myosteatosis. In addition, male gender and alcoholic- or metabolic-related cirrhosis were most represented in the myosteatosis group. Myosteatosis was not associated with a different HCC burden, length of hospitalization, complication rate, and readmission in the first 30 days after discharge. Overall survival was not influenced by the presence of myosteatosis. Alterations in body composition have gained clinical relevance in recent years as they have been widely associated with adverse outcomes in a variety of medical conditions, including chronic liver disease (CLD) ,2. The iIn addition to the loss of muscle mass and function, known as sarcopenia, qualitative changes in the structural composition of muscle, such as myosteatosis, have also been associated with worse clinical outcomes in different pathological conditions, especially cancer ,6.Myosteatosis is defined as the fatty infiltration of muscle, both in myocytes (intramyocellular fat) and in muscle fascia (intermuscular fat). It can be diagnosed with invasive techniques, such as muscle biopsy, or non-invasive ones such as computed tomography (CT), magnetic resonance imaging (MRI), and magnetic resonance spectroscopy (MRS) . Most stIn recent years, several studies have focused on the role of myosteatosis in CLD, aiming to assess its impact on prognosis in patients with cirrhosis or in those undergoing orthotopic liver transplantation (OLT). Myosteatosis, similarly to sarcopenia, acts as a negative prognostic factor for morbidity, mortality, and adverse perioperative outcomes ,9,10. WhOn the other hand, data on the impact of myosteatosis on the outcome of patients with hepatocellular carcinoma (HCC) are currently very limited. Kaibori et al. reported a strong association of fat infiltration in skeletal muscle with inferior overall survival in patients suffering from HCC . FurtherBy analyzing a large cohort of patients with HCC who underwent the first transarterial embolization (TAE) in a single center in Milan, and in whom the impact of sarcopenia was already reported, we were able to assess the impact of myosteatosis on overall survival, length of hospitalization, incidence of post-procedural complications, early rehospitalization, and its association with sarcopenia .This is a sub-study of a previously published study where we retrospectively included all consecutive patients with a diagnosis of HCC who underwent the first TAE from 1 March 2011 to 1 July 2019 at our Division of Hepatology . IndicatFor patients meeting the inclusion criteria, a detailed clinical history was obtained, including past medical history, comorbidities, etiology of liver disease, number of HCC nodules, performance status, early complications rate, length of hospitalization, readmission rate within 30 days, and survival time from the first TAE. Biochemical data, such as serum ALT, AST, alpha-fetoprotein, platelet count, INR, albumin, and total and direct bilirubin were recorded, if available. Body mass index (BMI), MELD score, Child-Turcotte-Pugh score (CPT), albumin-bilirubin score (ALBI score), and BCLC stage were calculated for each patient.https://www.slicer.org/ accessed on 15 December 2021) in a semiautomatic fashion. The muscle quality was defined by intramuscular adipose tissue content (IMAC), whose efficacy and feasibility has been widely validated [For the assessment of myosteatosis, a single cross-sectional CT image at the level of the third lumbar vertebra (L3) was used. The segmentation of skeletal muscle and adipose tissue was performed by the same researcher using the 3D Slicer software platform version 4.1 and BC module . Continuous variables were expressed as mean ± standard deviation, while categorical variables were expressed as percentages. Univariate analysis was performed using Student T-test and Chi-square test for continuous and categorical variables, respectively. A value of 2. Liver disease was caused by a viral etiology in nearly half of the enrolled patients, with HCV being represented in 45.3% of them and HBV in 5.3%. In 34 (22.7%) and 23 (15.3%) patients, the etiology was alcoholic or metabolic, respectively, while 11.4% had a rare cause of liver cirrhosis including hemochromatosis, primary biliary cholangitis, and autoimmune hepatitis. The majority of patients had well-compensated liver disease with preserved liver function tests: 82% of patients were in CPT class A, while 17.2% were in CPT class B, and only 0.8% had a decompensated liver disease in class C. The median MELD score was 10 ± 3, and 85.3% of patients had a coexistent diagnosis of sarcopenia. The ALBI score was available in 124 patients: 28 patients (22.6%) presented with Grade 1, 86 (69.3%) had Grade 2, while only 10 patients (8.1%) had Grade 3. BCLC stage was intermediate in the majority of patients , while 43 (28.5%) were considered early stage and 12 (8%) were in the very early stage. The main characteristics of the cohort have already been reported and are showed in HCC was multifocal in 50% of patients, while 28% and 22% of patients presented with a monofocal or bifocal HCC, respectively. The mean diameter of the major lesion was 33 ± 22 mm. In total, 41.1% of patients had already received at least one treatment for HCC other than TAE. The mean length of hospitalization after TAE was 2 ± 1.7 days, with only 11 patients (7.3%) developing a complication after procedure and nine patients (6%) requiring rehospitalization in the first 30 days after discharge. We found that 27.1% of patients had a complete response after the first TAE, and the median overall survival of the cohort was 808 ± 673 days.p = 0.0001) and showed a higher BMI than patients without myosteatosis . Male gender was also associated with myosteatosis as the proportion of male patients was significantly higher in the myosteatosis group . The two groups showed no significant differences in terms of transaminases levels, albumin, bilirubin, INR, CPT class, and MELD score. Interestingly, the etiology of the underlying liver diseases was associated with the presence of myosteatosis, as there was a significantly higher proportion of patients with alcoholic or metabolic-related cirrhosis in the myosteatosis group . Myosteatosis was not associated with sarcopenia defined by SMI at CT scan, as sarcopenia was equally represented in both groups .Of the total cohort, 115 patients had myosteatosis (76%) at CT examination and were included in the myosteatosis group, while the remaining 36 patients did not present with myosteatosis. Patients with myosteatosis were significantly older , as were readmission rates in the first 30 days after discharge . Complication rates were numerically, albeit not statistically, higher in the myosteatosis group  (p = 0.64).Myosteatosis was not associated with different HCC burden since we did not observe any significant differences between the two groups regarding number of HCC nodules and diameter of the largest lesion. Length of hospitalization was similar in the two groups  in patients with HCC who received loco-regional treatment. The prevalence in our study was higher than what previously reported in other cohorts; indeed, in a large cohort of 678 patients with cirrhosis, myosteatosis was present in 52% of patients and was associated with significantly worse median survival, representing an independent predictor of mortality at Cox-regression analysis . The medInterestingly, we could not find any impact of myosteatosis on overall survival following TAE, nor did we observe longer hospitalization, higher incidence of complications, or higher rate of readmissions in the first 30 days after discharge in patients with mysosteatosis.The relationship between body composition parameters and cancer outcomes has been extensively investigated. In a large metanalysis from Aleixo et al., the presence of myosteatosis was associated with significant worse prognosis in patients with different types of neoplasia, including HCC . RegardiAmong the relevant patients’ characteristics that could confound the data is the underlying liver disease etiology as the etiology of cirrhosis has been shown to be a driver in the development of myosteatosis. In our study, alcoholic and metabolic liver diseases were more frequent in the myosteatosis group, while viral liver disease were more represented in patients without muscular abnormalities. Similar findings have been reported by Meister et al. , while nSarcopenia can be ruled out as a confounder in our study, as the prevalence of sarcopenia measured by CT scan was similar in both groups (85.1% in the myosteatosis group vs. 86.1% in non-myosteatosis group). Given that sarcopenia emerged as an independent predictor of outcome, this highlights that the assessment of muscle mass or muscle composition should not be considered equivalent in assessing frailty in patients with HCC.We do acknowledge our study has several limitations mainly due to its retrospective nature and the relatively small number of patients included. Also, our cohort of patients reflects the epidemiology of liver diseases in Italy, where viral hepatitis is the main cause of cirrhosis and liver cancer. Whether our findings are true also in areas with high prevalence of NAFLD/ALD needs validation. Lastly, we included only patients of Caucasian ethnicity, thus, our results are not transferrable to other populations.Despite these limitations, to the best of our knowledge, this is the first study evaluating the role of myosteatosis in the prognosis of cirrhotic patients with intermediate HCC undergoing locoregional treatment. The lack of a significant impact of this variable on safety and outcome of TAE suggests against routine assessment of myosteatosis. On other hand, the observation that in the same cohort sarcopenia plays a major role on survival, calls for individualized treatment/nutritional interventions to improve muscle mass in patients with HCC.


Flow cytometry analysis revealed that metformin combined with FuOx induced late apoptosis (p < 0.05) by mediating mitochondria-related Mcl-1 and Bim protein expression. Furthermore, in vivo, metformin combined with FuOx more notably reduced tumor volume than FuOx or metformin alone did in BALB/c mice (p < 0.05). These findings demonstrate that metformin may act as an adjunctive agent to enhance the chemosensitivity of CRC cells to FuOx. However, further clinical trials are warranted to validate the clinical implications of the findings.Studies have demonstrated that metformin has antitumor effects in addition to therapeutic effects on hyperglycemia; however, few studies have explored the effects of metformin in chemotherapy. Therefore, we hypothesized that the administration of metformin would enhance the therapeutic effects of 5-fluorouracil and oxaliplatin (FuOx) to inhibit the growth of colorectal cancer (CRC) cells in vitro and in vivo. The results of our in vitro experiments demonstrated that metformin significantly increased the effects of FuOx with respect to cell proliferation ( Colorectal cancer (CRC) is one of the most prevalent malignancies worldwide. According to data from a 2020 statistical analysis, globally, CRC accounts for 10% of all incident cancers (1.9 million cases) in both sexes. Furthermore, CRC has the second highest mortality rate among incident cancers, comprising 9.4% of total cancer deaths (0.93 million cases) /2. The mice were sacrificed under anesthesia with isoflurane and cardiac puncture 1 week after the last chemotherapy treatment. The protocols of this in vivo study were approved by the Institutional Animal Care and Use Committee of Kaohsiung Medical University , and this study was conducted in accordance with the guidelines of the Care and Use of Laboratory Animals.Seven-week-old male BALB/c nude mice were purchased from BioLasco Taiwan . The mice were maintained in a pathogen-free environment and were randomly divided into four treatment groups: (I) control, (II) metformin-only, (III) FuOx-only, and (IV) combination of metformin and FuOx. After 1 week of acclimation, the mice were subcutaneously injected with HT29 cells . Data were analyzed using one-way analysis of variance using Prism 9.0 software , with To assess the antiproliferation effect of metformin, 5-FU, and oxaliplatin on colon cancer cells, we treated HT29, SW480, and SW620 cells with various concentrations of metformin (0–100 mM), 5-FU (0–100 µM), and oxaliplatin (0–100 µM) for 24, 48, and 72 h. The results verified that all of the drugs could decrease the proliferation of the HT29, SW480, and SW620 cells in a dose-dependent and time-dependent manner .p < 0.001) and was inhibited by 35%, 55%, and 65% when treated with 5 mM metformin plus 1, 2, and 5 µM FuOx, respectively . Additionally, compared with the control cells, the SW480 cell proliferation markedly decreased by 45%, 57%, and 63% after treatment with 1, 2, and 5 µM FuOx, respectively , and, respectively, decreased by 57%, 68%, and 71% after treatment with 5 mM metformin plus 1, 2, and 5 µM FuOx . Similarly, the SW620 cell proliferation significantly reduced by 38%, 50%, and 54% after, respectively, treatment with 1, 2, and 5 µM FuOx . After treatment with 5 mM metformin combined with 1, 2, and 5 µM FuOx, the SW620 cell proliferation, respectively, decreased by 46%, 57%, and 59% compared with the control cells . Therefore, the combination of metformin and FuOx was more effective in reducing the cell proliferation of the HT29, SW480, and SW620 CRC cell lines relative to the control cells and the cells treated with FuOx.We further investigated cell proliferation under the combination of metformin and FuOx at 72 h. As presented in p < 0.05, p < 0.05). These results demonstrate that metformin can enhance the ability of FuOx to inhibit clonogenic growth.To investigate the colony formation ability of the CRC cell lines treated with the combination of metformin and FuOx, the CRC cells were incubated with metformin, FuOx, and the combination of metformin and FuOx for 1 week. The clonogenic capacity of the HT29, SW480, and SW620 cells treated with metformin or FuOx alone was lower than that of the control cells . Furthermore, the combination of metformin and FuOx inhibited the migration rate to a greater extent than metformin or FuOx alone did . In summary, metformin combined with FuOx significantly increased the inhibition of cell proliferation, colony formation, and migration, and the inhibition was notably more favorable than that of metformin or FuOx alone.As presented in p < 0.001, p < 0.05, To study the effect of metformin combined with FuOx on cell cycle distribution, cell cycles were analyzed using flow cytometry. In the HT29 cells, metformin-induced cell cycle arrest in the G0/G1 and S phases, whereas FuOx induced cell cycle arrest in the G0/G1 phase . Although treatment with FuOx increased the percentage of cells in the S phase (p < 0.001), compared with FuOx treatment, metformin combined with FuOx did not significantly increase the proportion of cells in the S phase (p = 0.7009). In the SW620 cells, the increase in the cell proportion in the S phase in the combination group was prominent compared with the individual treatment groups . These results demonstrate that combined metformin and FuOx promotes more apoptosis of CRC cells.To evaluate whether metformin and FuOx influence apoptosis, we analyzed the number of apoptotic cells after different treatments by using the Annexin V/7-AAD staining assay through flow cytometry. The results revealed that metformin combined with FuOx increased the percentage of SW480 and SW620 early apoptotic cells compared with the control treatment . The results demonstrated that treatment with the combination of metformin and FuOx downregulated the expression of Mcl-1 and notably upregulated the expression of Bim in the CRC cells.To further extend our findings, we determined the levels of the proapoptotic protein Bim and the antiapoptotic protein Mcl-1 through Western blotting. Bim has multiple isoforms; all of these isoforms are involved in proapoptosis. The expression of Mcl-1 in the HT29, SW480, and SW620 cells in the combination group was significantly reduced relative to that in the control, metformin alone, and FuOx alone groups . Furthermore, compared with the metformin and FuOx alone groups, the metformin and FuOx combination group demonstrated more prominent inhibition of the growth of tumors (p < 0.05). The tumor weight reduction in the combination group was notable relative to those of the untreated control and metformin alone groups . We observed that the combined treatment of metformin and FuOx obviously suppressed HT29 tumor cell growth in BALB/c nude mice.To validate in vivo that metformin could increase the antitumor effects of FuOx, we subcutaneously injected HT29 cells into the right flanks of nude mice. After 7 days, therapy was commenced. As presented in According to the Taiwan Cancer Registry annual report of 2019, among all malignancies in Taiwan, CRC is ranked first in terms of the incidence rate  and third in terms of the mortality rate . From 2009 to 2019, the incidence of CRC cases increased from 12,488 to 17,302, and mortality increased from 4531 to 6436. Furthermore, the age at which patients develop CRC has gradually decreased, with the number of CRC cases involving patients aged <50 years increasing from 1503 to 2136. A growing body of research has reported that hyperglycemia is a key risk factor for CRC development, with hyperglycemia being closely correlated with the progression, incidence, and prognosis of CRC ,28. Our Metformin is a Food and Drug Administration–approved first-line drug for patients with T2DM . In addiEvidence has indicated that metformin has a synergistic effect when used in combination with other chemotherapeutic agents ,35,36. Hp53 mutant cell lines [p53 mutations were used in the present study. As discussed, metformin has the potential to increase the sensitivity of cancer cells to chemotherapy drugs. Evidence has revealed that metformin can enhance the treatment effects of 5-FU on various cancers, and it can significantly inhibit the growth of cancer cells in vitro and in vivo [Studies have reported that metformin has a significant apoptosis-inducing effect on ll lines ,41. The  in vivo ,42,43,44 in vivo ,46. TherZhang et al., demonstrated that the combination of cisplatin and metformin affected mitochondrial function in CRC cells by lowering mitochondrial membrane potential (MMP) and inducing reactive oxygen species (ROS) production, thereby promoting apoptosis through the PI3K/Akt pathway . Chen etIn clinical practice, the recommended daily dosage of metformin for adults is 500–2000 mg. The dosage we used for our BALB/c nude mice was calculated using a previously published dosage conversion formula . AccordiThis study has a few limitations. First, the Mcl-1/Bim signaling pathway was not completely elucidated in our study, although Mcl-1 and Bim are involved in initiating tumor apoptosis. Therefore, the mechanism underlying the combined effect of metformin and FuOx requires further exploration. Second, other studies have reported that metformin can cause mitochondrial dysfunction to preclude the progression of cancer ,52. HencThe aim of the present study was to elucidate the combined effects of metformin and FuOx in CRC cell lines and an animal model. Through a series of in vitro experiments, we obtained results that demonstrated that metformin could enhance the efficacy of FuOx in inhibiting cell proliferation, cell survival, and migration; inducing cell cycle arrest; and promoting apoptosis. Western blot analysis further verified the molecular apoptotic mechanism, which resulted from the underexpression of Mcl-1 proteins and overexpression of Bim proteins.Through in vivo studies, we further verified the effectiveness of the combination therapy. The data revealed that, after 40 days of treatment, the combination of metformin and FuOx had a more pronounced effect on the inhibition of tumor growth and tumor weight than FuOx or metformin alone. We suggest that when combined with FuOx, metformin potentially acts as an adjunctive agent to eliminate CRC in vitro and in vivo.


In this study, atypical choroid plexus papilloma was treated with high-dose rapamycin for 17 days preoperatively in an infant. Rapamycin significantly reduced the blood supply to the tumor while reducing the tumor volume, and most of the tumor was resected successfully. However, the infant developed hyperglycemia related to the rapamycin dose, which was effectively controlled by adjusting the dose and applying insulin. Atypical choroid plexus papilloma (aCPP), a subtype of a choroid plexus tumor (CPT) –3, is a Rapamycin has been widely applied to prevent acute rejection in organ transplantation and successfully treats complicated vascular anomalies and some brain tumors , 5.Here, we report an 11-month-old infant with aCPP treated with high-dose rapamycin for 17 days preoperatively. Rapamycin is effective in reducing the blood supply to the tumor, controlling tumor volume and reducing risk of surgical hemorrhage. However, the infant developed hyperglycemia related to the rapamycin dose, which was effectively controlled by adjusting the dose and applying insulin.An 11-month-old boy (weight 10 kg) was admitted to the pediatric intensive care unit (PICU) on May 4, 2021 because of increasing head circumference for 1 week and a cerebral hernia for 1 day. He presented with irritability, dysphoria, and vomiting, an enlarged head circumference of 50 cm, and an approximate 3*3 cm anterior fonticulus with increasing tension. He was somnolent with obvious nuchal rigidity but had normal pupils, isometric muscle tension, and normal spinal reflexes at admission. He had a negative family history of neurologic or metabolic diseases.Routine blood examination, blood biochemistry assays, functional coagulation assays, endocrine function, and diabetes mellitus–associated laboratory tests were normal, except for a slight increase in cortisol levels at 577 nmol/L  , which returned to normal 1 week later and might be related to the infant’s state of increased stress. Computed tomography (CT) and magnetic resonance (MR) images revealed a primary malignant tumor (60.00*84.01*94.72 mm) with a rich blood supply in the right lateral ventricle and metastatic tumors in the left lateral ventricle, the third ventricle, and cerebellopontine angle (CPA) Figure 22/day rapamycin orally  . The patient became conscious with stable vital signs, and vomiting and neck resistance disappeared on May 13 after 7 days of rapamycin treatment (the 9th day after admission). Therefore, he underwent stereotactic biopsy to confirm tumor pathological diagnosis and Ommaya insertion to alleviate and prevent hydrocephalus  on May 19. The MR images showed that the blood supply to the tumor had notably decreased on May 21 after 14 days of rapamycin treatment (the 17th day after admission), while the volume of the tumor in the right lateral ventricle decreased to 56.82*80.06*92.51 mm  rapamycin orally on June 10 , together with continuous subcutaneous insulin infusion (0.5 IU/h) to prevent hyperglycemia. The patient started receiving CPT-SIOP-2000 protocol chemotherapy   on May 6l doses) , so we icephalus . The pat92.51 mm Figure 2otherapy , 12 on JCPTs with a rich blood supply have a fatal risk of hemorrhaging in the perioperative period and surgery –17, and Rapamycin has been applied in brain tumors , 21, 22 2/day   at the initial recommended dose of complicated vascular anomalies, but the trough concentration of plasma rapamycin was under 10–15 ng/ml after 7 days of rapamycin treatment. According to some reports, the recommended dose for children with brain tumors is 3–5 mg/m2/day (2/day (once a week) without serious rapamycin-induced adverse effects , and the trough concentration of plasma rapamycin reached 14.8 ng/ml. Of note, the trough concentration of plasma rapamycin reached 10–15 ng/ml when the dose of rapamycin was 2–3 times higher than the recommended dose, which was related to individual differences between patients.The doses and courses of rapamycin vary based on the tumor type, location, and size. The recommended dose of rapamycin based on large-scale randomized scale trials of complicated vascular anomalies is 2.6 mg/ml doses)  to achiel doses) . Howeverg/m2/day –27, and  effects . Further effects  and poor effects –32 [the  effects ], the paThe trough concentration of plasma rapamycin finally reached 10–15 ng/ml by adjusting the rapamycin dose in this case, but the patient developed hyperglycemia. After excluding hyperglycemia caused by pancreatic dysfunction, we suspected that the patient’s hyperglycemia was related to rapamycin treatment and that the concentration of plasma glucose was correlated with the rapamycin dose. Rapamycin-induced hyperglycemia could be controlled by insulin or dosage adjustment and returned to normal after rapamycin withdrawal, in this case, demonstrating that rapamycin-induced hyperglycemia was related to high-dose rapamycin. When the patient developed hyperglycemia, there were no other common adverse effects related to rapamycin treatment, such as oral ulcers, hyperlipidemia, liver dysfunction, and bone marrow suppression , 35, becIn conclusion, rapamycin can effectively reduce the blood supply to the tumor and tumor volume but induces reversible hyperglycemia when this aCPP patient received high-dose rapamycin orally to reach the target trough plasma concentration in this case. Therefore, the concentrations of plasma rapamycin and glucose should be monitored during high-dose rapamycin treatment, and dosage adjustment or insulin may be needed if patients develop hyperglycemia.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.Written informed consent was obtained from the minor(s)’ legal guardian/next of kin for the publication of any potentially identifiable images or data included in this article.JL and ML drafted the manuscript. SL, ST, ZW, LYu, DL, LH, LW XLi JZ, XLa and YY collected materials and prepared figures. WZ and LYa critically revised the final manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted without 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.


ROC (receiver operating characteristic curves) analysis revealed that PLR had a significant predictive ability for having PYMS > 1 . The optimal cut-off was 151 with sensitivity of 51.6% (95% CI: 38.6–64.5%) and specificity of 67.3% (95% CI: 60.5–73.6%). Furthermore, 37% of the children (n = 101) yielded a PLR over 151. Our results support a promising value of PLR as a predictive marker for moderate to severe malnutrition in hospitalized children.Malnutrition in hospitalized children represents a significant burden with occasionally detrimental consequences. In this retrospective analysis of pediatric patients aged one to 16 years old, who were hospitalized in the children’s cantonal hospital of Aarau, Switzerland, we investigated the utilization of PYMS  as a routine screening tool for malnutrition in pediatric inpatients. Additionally, we explored the correlation between PYMS and NLR (neutrophil–lymphocyte ratio) and PLR (platelet–lymphocyte ratio), which are two novel biomarkers. Various parameters were analyzed from the medical records of the patients. Most of the sample  was characterized by a low PYMS of 0–1 point. Greater NLR and PLR values were significantly associated with greater PYMS ( Pediautcomes“ ,4. MalnuPediatric inpatients suffering from acute illness or trauma are often exposed to metabolic stress, and they mostly lack a previous setting of malnutrition. The existence of the inflammatory response within the acute phase of trauma or critical illness and also periods of interrupted feeding contribute to the occurrence of malnutrition . It has In this respect, the consensus statement between ASPEN and the Academy of Nutrition and Dietetics recommends the routine malnutrition screening of children upon presentation as well as later throughout their hospitalization .A plethora of screening tools have been developed to identify malnutrition in hospitalized children, with the most prevalent and well-established among them being the Pediatric Yorkhill Malnutrition Score (PYMS), the Screening Tool for the Assessment of Malnutrition in Pediatrics (STAMP), and the Screening Tool for Risk of Impaired Nutritional Status and Growth (STRONG KIDS) .According to the recommendations of the European Society of Clinical Nutrition and Metabolism for nutritional screening, PYMS is a screening tool  that hasA potential disadvantage of PYMS in real-world clinical practice might be the missing data for the calculation of scores upon a child’s presentation, owing, for instance, to overcrowded emergency departments, loss of awareness of possible underlying malnutrition or disease , or patiIn this regard, the neutrophil–lymphocyte ratio (NLR), as well as the platelet–lymphocyte ratio (PLR), represent simple and inexpensive methods for evaluation of a number of pathologies such as acute inflammatory processes, immunosuppression on the ground of transplantation or even psychiatric disorders ,19,20,21Within this retrospective study, we aimed primarily to investigate in real-world clinical practice the demographics of hospitalized children, the utilization of PYMS as a routine screening tool, and its role in the follow-up of pediatric inpatients. Additionally, NLR and PLR constitute novel biomarkers utilized as inflammation biomarkers with clinical impact  and an e® Excel for Mac 2019, Microsoft Corporation, Redmond, WA, USA) and was further managed with the usage of available Excel filters. The eligibility of the retrieved patients was evaluated by two investigators (S.G. and M.D.), following the inclusion criteria . Finally, eligible cases were validated by H.K.This retrospective study included children aged one to 16 years old, who were hospitalized in the children’s cantonal hospital of Aarau for medical or surgical reasons during the year 2019. For the extraction of medical records, the clinical information system “KISIM” has been utilized . The following parameters have been retrieved from the KISIM medical database; hospitalization length, date of entry, date of birth, gender, diagnosis, body weight, height, BMI, head circumference (percentile und z-score), pediatric Yorkhill malnutrition score (PYMS), co-morbidities, nutritional intervention, platelets, leukocytes and their subsets , gastrointestinal endoscopies, imaging studies, psychological interventions, request for weight control and other routine laboratory tests recorded in the chart. Curves for z-BMI have been calculated automatically in the clinical information system “KISIM,” which is based on the BMI-WHO  Growth Charts for boys and girls. The dataset was then extracted to an empty Excel sheet (MicrosoftQuantitative variables were expressed as mean ± SD (standard deviation) or as median (interquartile range). Qualitative variables were expressed as absolute and relative frequencies. A Mann–Whitney test was used for the comparison of NLR and PLR between children with PYMS 0–1 and children with PYMS > 1.p values are tailed by two.To investigate the association of two continuous variables, the Spearman correlation coefficient (rho) has been used. The ROC curve (a receiver operating characteristic) was applied to evaluate the predictability of NLR and PLR. To determine the sensitivity and specificity for optimal cut-offs, the area under the curve (AUC) was computed. All reported p 0.05, and analyses were conducted using SPSS Statistics for Windows, version 22.0 .Logistic regression analysis was used to investigate whether levels of PLR were associated with having PYMS > 1. Gender, age, and infection were also included in the model. Adjusted odds ratios (OR) with 95% confidence intervals (95% CI) were computed from the results of the logistic regression analyses. Statistical significance was set at A total of 273 children were eligible and considered for our data analysis.2 (12.5 kg/m2), whereas the mean p BMI percentile was 50.4% (32%), as well as 13.2% of children were found to have a BMI less than the 10th percentile. Furthermore, the mean duration of hospitalization amounted to 4.2 days (2.5 days). Regarding the disease spectrum, infectious diseases were diagnosed in 151 children (55.1%) of the sample, whereas gastrointestinal diseases amounted to 18 (6.6%). One-third of them  had to undergo a gastrointestinal endoscopy. The second most common diagnosis was traumatic head injury, which was mostly mild. No patient with head trauma injury had received anticonvulsant therapy or was in a coma. Cumulative patients’ basic characteristics are illustrated in detail in Of this pediatric population, 138 patients were boys (50.5%). The mean age for the whole study population was 7.2 years (4.2 years). The mean BMI was 18 kg/mThe median NLR was 2.4 (25th–75th percentile: 1.3–5), and the median PLR was 127.4 (25th–75th percentile: 88.4–180). For further information regarding participants’ biochemical indices and biomarkers, refer to Most of the sample  was characterized by a low PYMS, namely 0 or 1, whereas 57 children (20.9%) were graded as PYMS 2–3, and the rest of the sample  was found to have a PYMS of 4 or greater. Overall, 62 (22.7%) of the children had a PYMS of more than one, indicating mild or moderate undernutrition. PYMS values are depicted in p = 0.004) among all three PYMS levels, with the percentage being lower in cases with a PYMS of 0–1. Similarly, the percentage of a nutritionist being present was significantly lower in children with a PYMS of 0–1. Patients with gastrointestinal disease or having undergone a digestive endoscopy were not significantly associated with PYMS . Moreover, the percentages of children being referred to a pediatric psychiatrist or having asked to have their weight controlled were significantly greater for children with PYMS values greater than 3 .Information about participants’ hospitalization is presented in p = 0.030 and p = 0.004, respectively, Furthermore, greater NLR and PLR values were significantly associated with greater PYMS values  (PLR was significantly higher in children with a PYMS of more than one (= 0.024) .p = 0.580).The optimal cut-off was 151, and it yielded an estimated sensitivity of 51.6% (95% CI: 38.6–64.5%) and specificity of 67.3% (95% CI: 60.5–73.6%). Furthermore, 37% of the children (n = 101) yielded a PLR of over 151. PYMS > 1 was not significantly predicted by NLP   .Within this retrospective study, we screened initially a total of 1043 children being hospitalized in the cantonal hospital of Aarau for the calendar year 2019, and 450 were found to have an available PYMS (43.15%). In this respect, Gerasimidis et al. [2) is slightly higher than that of other published studies [Moreover, in our study, half of the population (50.5%) was boys, which is comparable with yielded percentages of other pediatric malnutrition studies with an inclusion rate of boys 56.9–58% . Further studies ,26. Rega studies ,26,27.Regarding the diagnoses’ spectrum, we found that infectious diseases were the most predominant ones (cumulatively 55.1%), which was followed by traumatic head injury (8.4%) and appendicitis (6.3%). Likewise, Malekiantaghi et al.  includedThe deviations in findings between our results and the ones of the above-mentioned studies might be explained through possible differences in methodology and characteristics of the included population, such as different genetic or cultural backgrounds.Considering the fundamental tool of PYMS, most of the sample (77.3%) was characterized by a low PYMS (0–1), whereas 57 children (20.9%) were graded as PYMS 2–3. Only a minority (1.8%) of children had a PYMS greater than 3. Cumulatively, 22.7% of the children had a PYMS greater than one, indicating mild, moderate, or severe undernutrition. These findings contrast with the publication of Gerasimidis et al. , where ap = 0.030 and p = 0.004, respectively). Additionally, using ROC analysis, we exhibited that PLR had a significant predictive ability for having PYMS > 1 with an optimal cut-off of 151, which is a finding that remained statistically significant (p = 0.009) even after adjusting for potential confounders .A further substantial novel finding of our study is the calculation of NLR and PLR, which were found to be statistically significantly associated with greater PYMS .According to the literature we have access to, this is the first study to link these inflammatory biomarkers with validated screening equipment (PYMS) for pediatric inpatient malnutrition. In a neonatology study, Can et al.  investigFurther pediatric studies outside the spectrum of malnutrition, albeit under the prism of PLR and NLR, have been previously published, particularly regarding appendicitis, transplanted children, or even psychiatric disorders ,19,20,21Concerning adult scientific evidence of PLR, NLR, and malnutrition, the scarce existing literature confines itself to the following studies: Kaya et al.  screenedNLR and PLR are considered acknowledged inflammatory markers . InflammParticularly regarding PLR, it has been established as a useful marker indicating inflammatory shifts of lymphocyte and platelet counts. PLR has been thoroughly investigated in oncologic, transplanted as well as hematologic and rheumatologic patients. The significance of PLR as an inflammatory marker is exceptional, once its variations are considered together with other complementary indices, especially NLR, which yields further useful information regarding the activity of disease or inflammation. PLR is believed to better predict the clinical outcomes of individuals with systemic inflammation compared to other counts . The extent of stress-induced hypercortisolemia with a secondary release of platelets into the bloodstream and transient lymphopenia affect the degree of PLR rise across a plethora of proinflammatory and prothrombotic disease states. Such a non-specific pathogenetic cascade of PLR increase might be counteracted by intensified platelet destruction or consumption at the sites of immune inflammation and thrombosis, requiring cross-checks of all blood cell counts and various inflammatory and immune markers .Despite the previously mentioned novelties and findings, our study is characterized by several limitations: namely, the retrospective nature of the study design as well as the relatively small sample have to be acknowledged. In addition, an association does not imply causality. Furthermore, confounding effects of glucocorticoids might affect CBC, and a single “snapshot” of CBC does not necessarily reflect their dynamics. Additionally, the specificity of the shifts in PLR is rarely neglected in individuals with chronic comorbidities, such as metabolic, vascular, autoimmune, and neoplastic ones. It has also to be emphasized that those molecular pathways of activation of platelets and the rest of the blood cells were not estimated to unravel the associations to their full extent with cellular markers of inflammation. In addition, there are no up-to-date valid age-, ethnic-, and gender official recommendations for the usage of CBC . Lastly,Pediatric malnutrition, especially in inpatient children, represents a serious and often underestimated medical problem, which may conceal long-term detrimental consequences if not early diagnosed and correctly treated. PYMS remains one of the most validated and reliable screening tools for children with or jeopardizing to the development of undernutrition , which is suggested to be performed routinely on all patients upon admission. Nevertheless, the occasional lack of awareness or the insufficient training of health care providers, the crowded emergency departments, or further potential difficulties such as linguistic and cultural particularities of referred children, may limit its utilization. PLR and NLR are regarded as practical and easy-to-perform indices which have been previously well-established for adult and pediatric clinical outcomes, including inflammatory and immunomodulatory ones. Our results support a promising value of PLR as a predictive marker for moderate to severe malnutrition in children. Further large-scale relevant pediatric studies are highly encouraged in order for the usefulness and validity of this simple marker to be further clarified.


Methods: A retrospective analysis of patients treated for scalp region reconstruction was conducted with 16 patients. All patients underwent the same procedure: scalp full-thickness tumor excision with simultaneous reconstruction with Matriderm® and the application of a split-thickness skin graft in the same surgical time. During follow-ups, the surgical outcome was evaluated by accurate clinical examination of the wound, adopting the Vancouver Scar Scale (VSS). Results: The outcomes obtained were satisfying: wound healing at the end of the procedures was optimal, grafted skin resulted similar to surrounding tissue, and pigmentation and vascularity showed a decrease in the period between 6 months and 1 follow-up. Conclusions: The use of Matriderm® and split-thickness skin grafting for scalp full-thickness defects reconstruction resulted in an optimal, stable, and safe procedure, suitable for elderly patients.Background: The scalp region represent a common area affected by benignant and malignant skin tumor, and it represents a surgical challenge when it is needed to be reconstructed. The aim of this study is to present our experience with full-thickness scalp skin defects, reconstructed using Matriderm The sca® dermal substitute, 1 year of follow-up. Patient data were collected from the department database, including age, sex, comorbidities, surgical procedures, histopathological examination, and follow-up records.A retrospective analysis of patients treated for scalp region reconstruction in the Oral and Maxillo-Facial Department of the “Federico II” University of Naples was conducted during the period between 2017 and 2021. Inclusion criteria for the selected study were tumor excision in scalp region with full-thickness defect, implantation of Matriderm®, and the application of a split-thickness skin graft in the same surgical time , considering: pigmentation (from 0 to 2 points), vascularity (0 to 3 points), pliability (0 to 5 points), height (0 to 3 points). The lower the score, the better the scar status.® use, split-thickness defects, full-thickness defects closed by local flaps, reconstruction not related to tumor excision but locoregional flap harvesting. Sixteen patients were conclusively selected and enrolled in the study. All enrolled patients filled in written informed consent for the study. The mean age of patients was 75 years old, with a range from 68 to 94. The ratio between males and females was 3:1. A total of 62.5% of patients (10/16) were treated with local anesthesia, while six patients (37.5%) underwent general anesthesia. Only two patients were treated with a two steps technique, and both margins were clear. The shape of the full-thickness defect was approximately round, with an average maximum diameter of about 5.86 cm and with a range from 4.5 cm to 8.3 cm. The most frequent histopathologic finding was basal cell carcinoma (10/16) with 62.5% of cases, followed by squamous cell carcinoma . Among the SCC cases, two of them were reported as actinic keratosis with SCC in situ. Follow-ups were recorded at 1 month, 6 months, and 1 year. Clinical follow-ups showed good recovery in the majority of patients , and Integra®  [® substitute is bilayered and it acts differently; the outer layer is made by silicone that simulates the epidermis and must be removed 3 weeks after the application, while the inner layer is the dermal matrix.A diffused technique is to adopt a dermal substitute in the first layer of the reconstruction, followed by a second layer where an STSG is positioned above the granulating wound ,7. In thNJ, USA) ,9. The I®, a highly porous membrane based on freeze-dried acellular collagen with a thickness of 1 or 2 mm as a dermal prosthesis. It is composed of type I, III, and V bovine dermal collagen and hydroxylated elastin [® resorbs in 6 weeks, after collagen production via fibroblasts [In our case, the choice was Matriderm elastin . Dermal  elastin . The his elastin . MatrideThe adoption of dermal substitutes in the treatment of large wounds, burns, and multilayer reconstructions is widely diffused in the literature. Considering, in particular, the reconstruction of scalp defects, it is well established that the combination of dermal substitutes and STSGs offers a more durable and thicker coverage than STSGs only procedure, giving better aesthetical and functional outcomes ,5. In ouBased on our experience and what is reported in the literature, the only disadvantages of this procedure are represented by the cost of the membrane and its thickness sometimes not satisfactorily filling the gap height . In fact® and split-thickness skin grafting for scalp full-thickness defects reconstruction resulted in an optimal, stable, and safe procedure, suitable for elderly patients. The availability of dermal matrices has created the possibility of overcoming important surgical limitations, such as bone exposure in large defects, suboptimal aesthetic results, and residual scarring. The possibility of obtaining promising results with a simple, effective, and safe intervention makes dermal substitute implantation the first choice for scalp full-thickness defects reconstruction.Reconstructive surgery must consider benefits and disadvantages to offer a reasoned solution adapted to the patient. The use of Matriderm


Population-based cancer screening has raised many controversies in recent years, not only regarding the costs but also regarding the ethical nature and issues related to variant interpretation. Nowadays, genetic cancer screening standards are different in every country and usually encompass only individuals with a personal or family history of relevant cancer.Here we performed a broad genetic screening for cancer-related rare germline variants on population data from the Thousand Polish Genomes database based on 1076 Polish unrelated individuals that underwent whole genome sequencing (WGS). We identified 19 551 rare variants in 806 genes related to oncological diseases, among them 89% have been located in non-coding regions. The combined BRCA1/BRCA2 pathogenic/likely pathogenic according to ClinVar allele frequency in the unselected population of 1076 Poles was 0.42%, corresponding to nine carriers.Altogether, on the population level, we found especially problematic the assessment of the pathogenicity of variants and the relation of ACMG guidelines to the population frequency. Some of the variants may be overinterpreted as disease-causing due to their rarity or lack of annotation in the databases. On the other hand, some relevant variants may have been overseen given that there is little pooled population whole genome data on oncology. Before population WGS screening will become a standard, further studies are needed to assess the frequency of the variants suspected to be pathogenic on the population level and with reporting of likely benign variants. It has been over 20 years since the very first version of the entire human genome was released. Not a long time afterward, the scientific community agreed that cancer is a disease of the genome. Today, although advanced sequencing methods are available at a reasonable price and the role of significant genetic variants localized along the whole genome is quite well defined, the clinical implementation of whole-genome sequencing (WGS) in diagnosis and treatment remains in its infancy. Tumor sequencing (somatic variants) can provide valuable information regarding treatment response and outcomes, however, cannot explain all cancer cases. Around 5-14% of all cancers are caused by germline (inherited) variants , BRCA1 5382insC (c.5266dupC), or BRCA2 6174delT (c.5946delT)  (BRCA1 5382insC and C61G variants in Poland was 0.25% (1/400)  variants carriers  . The folcarriers . In the carriers . Given tThe aim of this study is to investigate germline cancer-related gene variants on the population level in Poland, to assess the type of variation , to compare the frequencies with the frequency of corresponding variants estimated for the non-Finnish European population (NFE), and to interpret their relevance in the context of the family history-based testing. Our study is an extension of the study presented in Kaja et al. , howeverhttps://naszegenomy.pl/ after accepting the policy and filling in a short form.The cohort was described in detail in Kaja et al., 2022 . BrieflyThe core cohort consisted of 1076 unrelated individuals of Polish origin. The median age of participants was 45.4 (2–99) years and there was a slight predominance of males (697 vs. 525). This is comparable to the median age in Poland. The core cohort characteristics are presented in DNA was isolated from the patient’s peripheral blood. The library has been prepared using a TruSeq DNA PCR-free kit  and 550 bp inserts. Sequencing was performed on the Illumina NovaSeq 6000 platform with the following parameters: 150 bp paired-end reads, an average read depth of 35.26×. The quality has been assessed with FastQC v0.11.7 and mapped to the GRCh38 human reference genome. Single-nucleotide variants and short indels have been identified with DeepVariant v0.8.0 and genotyped all together with GL nexus v1.2.6-0-g4d057dc. Multiallelic variant calls were decomposed into monoallelic and normalized groups using BCFtools v1.9 . The varPLAF is alternative allele frequency in polish cohort and NFEAF is alternative allele frequency in gnomAD NFE. Statistical significance of odds ratios for each variant was estimated with Fisher’s exact test and corrected for Bonferroni adjustments. The prevalence of carriers was counted as a ratio of positive samples to the total number of samples tested.The variants associated with cancer were chosen based on the COSMIC database  and lithttps://www.internationalgenome.org/, accessed on 2023-01-13). The African (AFR), Admixed American (AMR), East Asian (EAS), South Asian (SAS) and European, gnomADe and gnomADg NFE and EUR for 1KGP, ancestries were considered. Variants were filtered according to Combined Annotation-Dependent Depletion CADD , which is a deleteriousness measure.To analyze genetic diversity AF from our database have been compared to exomes from gnomAD (gnomADe), genomes from gnomAD (gnomADg) and 1000 Genomes Project (1KGP) database  in the Polish population. Moderate impact variants with the highest OR (p<0.05) in the Polish population were: NUTM2B, MUC4, NOTCH1, NUTM2B, USP6  and other ancestries presented as follows: NFE (n=5/7), AMR (n=3/7), AFR (n=2/7), EAS (n=1/7), SAS (n=1/7).To present variants related to oncology identified in our database in regard to global genetic diversity and other genetic and genomic databases, we compared the number of variants identified in the Polish population to different ancestries and from three databases. In the first step the analysis was performed for the In the second step, we analyzed the variants related to oncology in Poland in the broader perspective. Not to exclude common variants, but at the same time to use computational resources effectively, we analyzed the variants with CADD > = 20 from the variants related o oncological diseases list. These included 8491 variants, among them 840 rare (MAF<0.01) and 7651 common (MAF>0.01). Among 8491 variants, 5647 were unique for Polish population (monomorphic) according to 1KGP database, 4392 according to gnomADe and 2315 according to gnomADg. The number of variants related to oncology shared between Polish cohort and particular ancestries is shown in BRCA1 gene and 72 in the BRCA2. Among them in BRCA1 two were of a high impact, 30 of a moderate impact and 13 of a low impact. Among BRCA2 five were of high impact, 46 of moderate impact and 21 of low impact. According to ClinVar three variants in BRCA1 were classified as P/LP ) and four variants in BRCA2  were classified as P/LP. The BRCA1/BRCA2 variants and their AF are shown in BRCA1/BRCA2 P/LP variants according to ClinVar  associated with lung adenocarcinoma, myeloproliferative neoplasm, breast invasive ductal carcinoma, polycythemia vera, and colon adenocarcinoma  have been identified in 32 individuals with tumor from the extended cohort and classified as pathogenic, likely pathogenic, and VUS  were carried by several individuals with cancer in extended cohort. Ribophorin 1 (RPN1) is a major part of oligosaccharyltransferase complex, which takes part in the glycosylation process. RPN1 is upregulated in breast cancer and has also been associated with several other cancers, such as high-grade ovarian serous adenocarcinoma, colon adenocarcinoma, oesophageal adenocarcinoma, and bladder urothelial carcinoma. As much as 7 out of 12 individuals from our cohort with cancer had a likely pathogenic variant on both alleles. Another recurrent variant was localized in HRAS gene. HRAS is a known oncogene related to several different cancers, among them Costello syndrome, a rare condition predisposing to tumours in different parts of the body, epidermal tumours, head and neck cancer, bladder carcinoma, and with ovarian carcinoma patients, but was also identified in the healthy controls The variants frequency is not annotated in ClinVar so that it also requires further characterization on the populational level. The variant p.Ile1035Val in PALB2, associated with breast and ovarian cancers, was identified in two out of eight individuals with breast cancers and in one ovarian cancer person from the extended cohort. All these variants will be further followed up in the affected cohorts in Poland in further studies.Variants in 5 genes  may contribute to the individual high or moderate risks of developing cancer or alternatively may imply a lower risk of developing the malignant disease. Polygenic Risk Score (PRS) aggregates the effects of many genetic variants across the human genome into a single score, being an estimate of an individual’s genetic liability to a trait or disease . PRS canBRCA1/BRCA2 analysis, the pathogenic variants identified in this study for the Polish population were mostly shared with the NFE ancestry and just one common variant within the EAS and SAS ancestry was identified. As Polish BRCA1/BRCA2 P/LP variants have a strong founder effect that is why as expected there are shared mainly with the European NFE population, which is the closest genetically. Most of these variants are not represented in the other ancestries. In the context of variants with a highest CADD (mostly common variants) located in the genes associated with cancer, when compared to the gomADg database, encompassing most of the variants, as many as 27% were monomorphic for Poles, but this was strongly dependent on the database. The result highlights the importance of creating a local database due to the fact that a large part of the variants related to oncological diseases is population specific and not shared not only between the various ancestries, but also between the same continent (PL vs. NFE). Moreover, the size of the database has a considerable meaning, while a low number of individuals can lead to bias. For example, 1KGP database encompasses around 3202 WGS individuals from different ancestries ) gene reported as having conflicting interpretations of pathogenicity in ClinVar and suggested of being pathogenic in the literature ) variant in the Polish core cohort in a very high AF (12.3%).The Thousand Polish Genomes database can be used as a reference database for germline cancer variants. To the best of our knowledge, this is the first study on the Polish population using a whole-genome sequencing approach for cancer predisposition screening. Previous studies focussed on selected genes and known founder mutation, although they have encompassed large cohorts, such as 16 229 healthy persons. The Thousand Polish Genomes database encompass also genetic modifiers and genetic biomarkers related to treatment outcomes. In Poland several projects targeted the discovery of new genes related to cancer and cancer susceptibility genes , 44. Theterature  was presBRCA1/BRCA2 variants only in the unselected Polish population. However, especially on the individual level we are not able to interpret data reliably, for example in terms of PRS, and that is why we cannot make any prediction based on them. As a scientific and clinical community, we should further try to provide results of the negative association in databases like ClinVar, not only the positive ones. We should be aware of differences between populations, even within the same ethnicity group, and to be able to identify risk alleles we advocate for large cohorts. For now, cancer screening with WGS on a research basis in larger populations will show us how feasible the method could be in clinics.Are we ready for population cancer screening using WGS? Given the sinking costs of sequencing technologies and the rising availability of prevention management alternatives, we are heading towards it. We identified nine P/LP variants in the https://1000polishgenomes.com.The datasets presented in this study can be found in online repositories. The names of the repository can be found below: The studies involving human participants were reviewed and approved by the Ethics Committee of the Central Clinical Hospital of the Ministry of Interior and Administration in Warsaw (decision nr: 41/2020 from 3 April 2020 and 125/2020 from 1 July 2020). The patients/participants provided their written informed consent to participate in this study.Conceptualization, MM, PD, BI, JL. Formal analysis, MM, TS, JL. Data curation, TP, PD, PT, TS, JSt. Data generation, PD, PT, PZ, ZK, EK, MD, MW. Methodology, MM, MatS, TS, JSt, BP-S. Visualisation and writing—original draft preparation, MM, JL, BI, JSz, BP-S, MarS. Writing—reviewing and editing: JSz, PD, JSt, MD, BP-S, TP. Funding acquisition, ZK. Supervision: PD, JSz. All authors contributed to the article and approved the submitted version.


Mechanical forces are an inherent element in the world around us. The effects of their action can be observed both on the macro and molecular levels. They can also play a prominent role in the tissues and cells of animals due to the presence of mechanosensitive ion channels (MIChs) such as the Piezo and TRP families. They are essential in many physiological processes in the human body. However, their role in pathology has also been observed. Recent discoveries have highlighted the relationship between these channels and the development of malignant tumors. Multiple studies have shown that MIChs mediate the proliferation, migration, and invasion of various cancer cells via various mechanisms. This could show MIChs as new potential biomarkers in cancer detection and prognosis and interesting therapeutic targets in modern oncology. Our paper is a review of the latest literature on the role of the Piezo1 and TRP families in the molecular mechanisms of carcinogenesis in different types of cancer. Cancers are undoubtedly one of the most urgent problems in present and future medicine ,2. AccorFor the purposes of this narrative review, we conducted comprehensive English language literature research for original and review articles using the PubMed and Scopus databases through September 2022. We searched for the combination of the following terms: Piezo channels (Piezo); Piezo in cancers; transient receptor potential channels (TRP); TRPM (melastin); TRPV (vanilloid); TRPC . We found 2608 related articles, and the final number of papers that were selected for this manuscript was 159. Studies with the highest level of evidence and relevance to the discussed topics (32) were selected by the authors’ consensus.Mechano-gated ion channels are a unique class of channels that respond to mechanical forces such as pressure, stretch, or touch. These channels play a crucial role in various physiological processes, including sensation, muscle contraction, and blood pressure regulation. In the neurosensory field, mechano-gated ion channels are particularly important for the detection of touch and pressure, hearing, and the function of other mechanical senses .There are several families of mechanosensitive ion channels that have been identified, including Piezo or TRP  among others. These channels have different properties and functions and are activated by different mechanical stimuli. For example, Piezo channels are activated by mechanical stretching of the cell membrane, while TRP channels are activated by changes in temperature, pressure, or chemical stimuli .Mechanically activated ion channels, such as MscL, MscS, TREK-1, Piezo1, OSCA1.2, TRP, and NOMPC , share a similar structural feature—the presence of amphipathic helices, which are amino acid segments with both hydrophobic and hydrophilic regions. These amphipathic helices are connected directly or indirectly to the pore-lining regions of the channels. The structure of MscL, for example, has been determined using X-ray crystallography and is composed of a pore-lining transmembrane segment (TM1) connected to an amphipathic S1 helix ,14. The The structure of TREK-1 is composed of pore domains that are gated by a C-type mechanism, with an amphipathic C-tail extending below the M4 helix . Piezo1’Another type of mechanosensitive channel is Tmc, also known as transmembrane channel-like 1 (TMC1) and TMC2, as a key component of the auditory system, responsible for converting mechanical vibrations into electrical signals that can be interpreted by the brain.Recent research has focused on understanding the systematic properties of Tmc, including its structure, function, and regulation. TMC-1 is composed of two subunits, TMC-1A and TMC-1B. The structure of TMC-1 was determined using cryo-electron microscopy (cryo-EM) and X-ray crystallography. The TMC-1A subunit is composed of two transmembrane helices, TM1 and TM2, and an extracellular domain that forms a pore through which ions can flow. The TMC-1B subunit is composed of a single transmembrane helix, TM3, and an intracellular domain that is involved in signaling . AdditioWhile much of the research on Tmc has focused on its role in the auditory system, recent studies have also investigated its expression and potential role in cancer cells. Some studies have suggested that Tmc may be overexpressed in certain types of cancer, such as lung and breast cancer, and may play a role in cell proliferation and migration . HoweverAccording to activation, two mechanisms have been established for mechanosensitive ion channels: the force-from-lipids model and the force-from-filaments model . The firAnother important aspect of mechanosensitive ion channels is their high sensitivity to changes in matrix stiffness. The cellular microenvironment is crucial in tumorogenesis and metastasis. In tumor development, mechanical cues, acting via mechanosensitive ion channels, affect both the cancer cells directly and the microenvironment as well. These signals are transduced into a cellular response  or to a response affecting the matrix microenvironment  ,40. A ch2+-permeable channels that transduce diverse mechanical forces into biochemical signals [Piezo are mechanically activated (MA) non-selective Ca signals . These c signals . Both Pi signals ,50,51,52 signals ,44,45,46 signals . However signals . Piezo1  signals ,54,55. Recently, the roles of Piezo channels in human physiology have been deepened by registering the consequences of mutations in Piezo genes. Abnormalities due to Piezo1 gene mutations are reported in generalized lymphatic dysplasia, dehydrated stomatocytosis, hereditary xerocytosis, heart failure, and diabetes mellitus ,59,60,61In breast cancer cells, functional mechanosensitive ion channels of Piezo1 type) were discovered for the first time in an MCF cell line by Li C et al. . However type werPiezo1 induces both collective and single-cell migration by modulating cells’ adhesion, stiffness, and contractility . The dyn2+ influx which consequently activates ERK1/2 and p38 MAPK that are involved in cell proliferation. Moreover, the presence of Ki-67 and PCNA (proliferation markers) in the cell is in direct proportion with Piezo1 expression. However, Piezo1 expression induction through YAP signaling is not relevant for the clinical T (tumor size) stage, histological grade, or lymph node metastasis [Piezo1 plays an important role in the proliferation of oral squamous cell carcinoma (OSCC). When Piezo1 expression is upregulated, the proliferative feature of the tumor is promoted . This coNot Piezo1, but Piezo2 and the hypermethylation of its promoter support the development of laryngeal squamous cell carcinoma (LSCC). In tumor tissues, Piezo2 promoter hypermethylation is significantly more frequent than in normal tissues and correlates with gender, differentiation, tumor (T) stage, lymph node metastasis, and clinical stage. Consequently, Piezo2 promoter hypermethylation is not only an LSCC risk factor but also a marker for tumor progression and the presence of metastases, a prediction marker of a patient’s poorer survival, and a possible therapeutic target .2+ influx. This signaling pathway remains unknown. Piezo1 knockdown cells also presented a decreased expression of integrin β1. This type of integrin has been reported to be highly expressed in malignant phenotypes of gastric cancer and supports cell migration [2+ influx, upregulates the expression of hypoxia-inducible factor 1α (HIF-1α) which in turn induces the production of vascular endothelial growth factor (VEGF) and accelerates epithelial–mesenchymal transition (EMT). Consequently, angiogenesis, tumor growth, and, finally, invasion with metastasis are endorsed by Piezo1. The expression of Calpain 1/2 which is responsible for angiogenesis together with the division of focal adhesions, is induced by Piezo1 through activation of HIF-1α [Piezo1 expression is upregulated in the majority of gastric cancer cell lines, especially those metastasizing to the omentum and lymph nodes, causing poor disease-specific survival ,78. Therigration ,81,82. Aigration . By knocigration . Moreoveigration . Furtherigration . Apoptosf HIF-1α ,77,78. If HIF-1α . It is af HIF-1α . 2+ influx and ATP release which lead to muscle contraction [Little is known about the involvement of Piezo1 and 2 in bladder cancer. However, Piezo 1/2 are expressed in bladder cells and even at higher levels in bladder cancer cells ,84. In atraction . If thisThe exception to the rule of Piezo1 supporting tumorigenesis is Piezo1’s role in lung cancer. In fact, there is more evidence of an omnidirectional role of Piezo1 in tumors. Piezo1 is highly expressed in lung tissue . NonetheFunctional Piezo1 channels are reported in leukemia cells (human myeloid leukemia K562 cell line), but their influence on tumor cell behavior remains unknown . Other sEsophageal squamous cell carcinoma (ESCC) is the most common malignant neoplasm of this organ. During the swallowing process, the esophageal tissue is subjected to constant changes in mechanical forces. Interestingly, the expression of Piezo1 in esophageal cells is relatively higher than in the rest of the body’s tissues . Gao L eThe cellular component and the cerebrospinal fluid help to maintain the brain tissue mechanics in homeostasis. Mechanical forces are significantly reorganized under brain tumor conditions. How tumor cells sense and regulate tissue mechanics is largely unknown . For thiProstate cancer (PCa) is one of the most common cancers in men worldwide . It is aColon cancer (ColCa) is believed to arise from two types of precursor polyps via two independent pathways: conventional adenoma associated with a mutation in the APC gene and serrated adenoma with unknown genetic ethology . Interes+ loading [Mechanosensitive ion channel Piezo1 was discovered in mesenchymal tumors such as osteosarcoma (OsS) and synovial sarcoma (SynS) ,112. OsS loading   expansion . MDSCs aTRP is a type of mechanoreceptor widely expressed in particular tissues and displays numerous functions. Eight TRP families can be distinguished: TRPC , TRPV (vanilloid), TRPM (melastatin), TRPML (mucolipin), TRPP (polycystin), TRPA (ankyrin transmembrane protein), TRPN (NomPC-like), and the recently reported TRPY (yeast) . Among tTRPV1Sensory neurons and plasma membranes are the main tissue localization of TRPV1. This family of channels is sensitive to heat, pH, and compounds  and is also involved in thermal nociception ,142,143.TRPV2Oncogenicity in different types of cancers is associated with the expression of the TRPV2 channel . It has TRPV3 2+ channel that is highly expressed in epithelial tissues, especially in keratinocytes of the skin. There, TRPV3 can be triggered by heat and can regulate various phenomena such as skin barrier formation, wound healing, sensing of temperature, itching, and pain [TRPV3 is a non-selective Caand pain . Little and pain ,152.TRPV42+ influx [2+ influx is produced, increased endothelial cell migration for breast tumor-derived endothelial cells is observed [The high expression level of TRPV4 is especially characteristic for epithelial tissues, where TRPV4 reacts to, a.o. heat, osmotic changes, and mechanical stretching by inducing Ca+ influx ,153,154.+ influx ,156 or t+ influx . TRPV4 i+ influx ,159. In + influx . These p+ influx  or the E+ influx . Interes+ influx . After Tobserved .All the evidence suggests that the functionality of TRPV4 in tumor cells is truly complex and it needs to be further studied.TRPV5 and TRPV62+-selective, responsive to 1,25-dihydroxyvitamin D3, and they regulate the homeostasis of Ca2+ in organisms. The role of TRPV5 in cancer is barely known, contrary to TRPV6 [The TRPV5 and TRPV6 families are very similar both in structure and their functions . These cto TRPV6 ,167. Theto TRPV6 ,169,170.TRPM members are divided into four groups: TRPM1/M3, M2/M8, M4/M5, and M6/M7. TRPM channels are involved in plenty of physiological functions. However, they are also linked to diverse pathophysiological human processes, including cardiovascular and neurodegenerative alterations, organ dysfunction, and cancer . When itThe seven mammalian TRPC members, which can be divided into four subgroups  based on their structure and functional similarities, contribute to plenty of cellular functions and physiological roles . TRPC5 iMechanosensitive ion channels play a crucial role in the communication between cells and the surrounding environment. Therefore, they are essential for the adaptation and survival of cells—the cancer ones as well. The most important mechanosensitive ion channels for tumors are the Piezo and TRP families. TRPV, TRPM, and TRPC are the families mostly associated with tumor development. They play a wide role in the development of many types of cancers such as breast, prostate, and colon cancer. They affect tumor growth and spread on multiple levels.Piezo 1 and 2 channels, although nowadays widely studied, influence cells by diverse mechanisms which are yet to be known in depth. In breast cancer cells, there is a high level of Piezo expression. The channels’ activation enhances tumor invasion and confined migration while it suppresses unconfined migration. The influence on proliferation is inconclusive. In oral cancer, Piezo induces tumor proliferation via Hippo and Yap signaling. In gastric cancer, there are numerous pathways connected to Piezo1 and its involvement in tumor proliferation, migration, invasion, and angiogenesis. The most essential is said to be Piezo 1’s correlations with integrin β1, RhoA, Rac1, and HIF-1α. The exact mechanisms of Piezo1 on bladder cancer cells are yet to be discovered; however, this channel has a positive impact on tumor stage, grade, and size. Piezo’s incoherent role in cancer is presented in lung cancer. In this cancer, it is Piezo inhibition, not activation, that supports tumor migration. In the case of esophageal squamous cell carcinoma, the downregulation of Piezo1 reduces the migration and invasion of cancer cells. Moreover, the expression of Piezo1 is associated with the activity of the TP53 pathway. The inhibition of TP53 significantly upregulates Piezo1. The expression of Piezo1 within prostate tumors is significantly higher than in normal tissues. In this cancer, the mechanism is closely associated with higher tissue pressure in the tumor and in bone metastasis. A germinal mutation in Piezo1 can cause the formation of polyps in the large intestine. Furthermore, the expression of Piezo1 in colon cancer is significantly higher and associated with a poorer prognosis. Piezo1 is overexpressed in aggressive human gliomas and it is associated with a poorer prognosis. Moreover, a correlation between Piezo1 upregulation and mutations in genes related to extracellular matrix reorganization, cell adhesion, and angiogenesis was also shown. The association between aggressive sarcomas such as synovial and osteosarcoma and Piezo1 expression was also demonstrated.The family of Piezo mechanosensitive channels is essential in the development of normal as well as abnormal vessels. The role of Piezo in tumor angiogenesis has been so far evidenced in rat squamous cell carcinoma, where endothelin occurs, and gliomas, where β-catenin-dependent signaling is essential. It has also been shown that Piezo1 plays a crucial role in the development of cancer metastases on multiple levels of this process. Furthermore, Piezo1 can be a very important immunological checkpoint. Leukocytes without Piezo1 expression were unable to transduce mechanosensory signals. Moreover, it has been demonstrated that Piezo1 signaling in inflammatory cells promotes cancer progression and myeloid-derived suppressor cell expansion. The connection between mechanosensitive ion channels and cancer development is wide, multi-faceted, complex, and not fully explored. Further studies in this area are necessary to find among them new, interesting, and important therapeutic targets for modern oncology.