A multivariable-adjusted model showed that patients with high eGFR level (≥ 60 mL/min/1.73 m2) and low albumin concentration (≤ 1.5 g/dL) achieved significantly early remission, whereas those with low eGFR (< 60 mL/min/1.73 m2) and high albumin concentration (> 1.5 g/dL) showed significantly slow remission. In conclusion, lower serum albumin concentration and higher eGFR were associated with earlier remission in MCD, but not with relapse.Previous studies reported conflicting results regarding an association between serum albumin concentration and the cumulative incidence of remission of proteinuria in adult patients with minimal change disease (MCD). The present study aimed to clarify the clinical impact of serum albumin concentration and the cumulative incidence of remission and relapse of proteinuria in 108 adult patients with MCD at 40 hospitals in Japan, who were enrolled in a 5-year prospective cohort study of primary nephrotic syndrome, the Japan Nephrotic Syndrome Cohort Study (JNSCS). The association between serum albumin concentration before initiation of immunosuppressive treatment (IST) and the cumulative incidence of remission and relapse were assessed using multivariable-adjusted Cox proportional hazards models. Remission defined as urinary protein < 0.3 g/day (or g/gCr) was observed in 104 (96.3%) patients. Of 97 patients with remission within 6 month of IST, 42 (43.3%) developed relapse defined as ≥ 1.0 g/day (or g/gCr) or dipstick urinary protein of ≥ 2+. Serum albumin concentration was significantly associated with remission , along with eGFR (per 30 mL/min/1.73 m MCD in adults is highly steroid-sensitive, but steroid resistance is seen in 5–20% of adult patients with MCD4. Epidemiological studies have showed that the incidence of end-stage kidney disease (ESKD) is remarkably lower in patients with MCD than in those with MN and FSGS6, concluding that MCD typically has favorable outcomes. However, compared with the general population, patients with MCD were at significantly higher risk of ESKD and thromboembolism7. Because steroid resistance predicts the incidence of ESKD in adult patients with MCD8, clinical characteristics associated with steroid sensitivity should be clarified to stratify the patients with MCD into several groups with different levels of steroid sensitivity.Minimal change disease (MCD) is one of the major primary nephrotic syndromes10, serum concentrations of creatinine11 and albumin10, selectivity index of proteinuria11, microscopic hematuria11, and acute kidney injury (AKI)12. However, their results may be biased without controlling for potential confounding factors. Recent Japanese retrospective cohort studies, including 14213 or 12514 patients aged ≥ 15 years with MCD, confirmed that young age14, low serum creatinine concentration14, and low urinary protein level14 independently predicted early remission, even after adjusting for clinically relevant factors. Another Japanese retrospective cohort study identified low serum albumin concentration, not urinary protein level, as a significant predictor of early remission, besides young age and no AKI15. The findings of these two studies strongly suggest that high glomerular filtration rate (GFR) level is a predictor of early remission. In contrast, the impacts of urinary protein level and serum albumin concentration on remission were conflicting, which should be examined in a multicenter cohort study with external validity.Early small retrospective cohort studies published between the 1980s and the 2000s including ≤ 62 adult patients with MCD, suggested that several clinical factors were associated with steroid sensitivity in adult patients with MCD without controlling for potential clinical confounders, including age20. The results of the present study provide useful clinical information to identify patients at a high risk of steroid resistance, who might need intensive immunosuppressive therapy (IST).The aim of the present cohort study was to identify the clinical predictors of remission and relapse of proteinuria in adult patients with MCD, with great interest in serum albumin concentration, urinary protein level, and GFR. We used the clinical data collected prospectively in 108 adult patients with MCD in 40 hospitals, who were enrolled in a 5-year prospective cohort study, the Japan Nephrotic Syndrome Cohort Study (JNSCS)2 were 11 (10.2%), 28 (25.9%), 48 (44.4%), and 21 (19.4%), respectively. Approximately, a half of the patients had serum albumin concentration of > 1.50 g/dL (n = 55 [50.9%]). Median level of proteinuria was 7.8 g/day or g/gCr . Within 1 month of IST, most patients (n = 107 [99.1%]) received oral prednisolone (PSL) and intravenous methylprednisolone (mPSL) was administered in an approximately quarter of patients (n = 28 [25.9%]).Clinical characteristics of 108 adult patients with MCD included in the present study were listed in Table 2, respectively. Age, age category, body mass index (BMI), and systolic and diastolic blood pressure, serum creatinine concentration, dipstick hematuria, renin-angiotensin system (RAS) blockade at initiating IST were significantly different among the eGFR groups. Between the patients with eGFR ≥ 60 mL/min/1.73 m2 and those with eGFR < 60 mL/min/1.73 m2, age, age category, systolic blood pressure, serum creatinine concentration, urinary protein level, dipstick hematuria, RAS blockade, and intravenous albumin administration at initiating IST were significantly different  , 28 (100.0%), 46 (95.8%), and 20 (95.2%) patients achieved remission, respectively (Table trend < 0.001)  as significant predictors of remission (Table During the median (interquartile range) observational period of 15 10, 28) days, remission was observed in 12 (100.0%), 40 (97.6%), 28 (100.0%), and 24 (88.9%) patients with serum albumin levels of ≤ 1.00, 1.01–1.50, 1.51–2.00, and > 2.00 g/dL, respectively Table . Patiently Table . Patient8 days, r2, those with eGFR of 60.0–89.9 and ≥ 90.0 mL/min/1.73 m2 were significantly associated with remission at the similar level  (Table 2) and serum albumin concentration (> 1.50 vs. ≤ 1.50 g/dL) and calculated their HRs. Compared with patients with eGFR ≥ 60.0 mL/min/1.73 m2 and serum albumin concentration > 1.50 g/dL, those with eGFR < 60.0 mL/min/1.73 m2 and serum concentration > 1.50 g/dL achieved remission significantly more slowly , whereas those with eGFR ≥ 60.0 mL/min/1.73 m2 and serum concentration ≤ 1.50 g/dL did significantly more rapidly   . Regarding the four eGFR groups, 3 (50.0%), 12 (44.4%), 18 (40.0%), and 9 (47.4%) patients with eGFR of < 30.0, 30.0–59.9, 60.0–89.9, and ≥ 90.0 mL/min/1.73 m2 relapsed after remission, respectively (Table trend = 0.633). Unadjusted and adjusted CPH models showed that no variable was associated with relapse (Table Predictors of relapse of proteinuria were assessed in 97 patients with remission within 6 months of IST. During the median (interquartile range) observational period of 2.2  years, relapse was observed in 3 (25.0%), 21 (53.8%), 11 (45.8%), and 7 (31.8%) patients with serum albumin concentration of ≤ 1.00, 1.01–1.50, 1.51–2.00, and > 2.00 g/dL, respectively Table . No signly Table . The cum2, was associated with slower remission. An advantage of the present study was the detailed assessment of the multivariable-adjusted non-linear association of serum albumin and eGFR with remission, providing clinically useful information to identify the patients who are resistant to IST, namely, those with serum albumin concentration > 1.5 g/dL or eGFR < 60 mL/min/1.73 m2.The present study clarified that serum albumin and eGFR were associated with remission of proteinuria in a non-linear fashion in 108 adult patients with MCD, whereas they were not associated with relapse of proteinuria. Patients with lower serum albumin concentrations, especially ≤ 1.5 g/dL, were likely to achieve remission more rapidly. Lower eGFR, especially < 60 mL/min/1.73 m10, compatible with the results of the present study. A Japanese retrospective single-center cohort study, including 53 adult patients with MCD, verified the inverse association between serum albumin concentration and remission, even after adjusting for potential clinical confounding factors15. In contrast, two cohort studies reported no significant association between serum albumin concentration and remission. A retrospective single-center cohort study in the UK, including 52 adult patients with MCD, showed that serum albumin concentration was not associated with remission in an unadjusted CPH model21. Another Japanese retrospective multicenter cohort study, the STudy of Outcomes and Practice patterns of Minimal Change Disease (STOP-MCD), including 142 adult patients with MCD in five hospitals showed no significant association between serum albumin concentration and remission in a multivariable-adjusted CPH model13. In the STOP-MCD study, high prevalence of intravenous albumin administration (62.0% in the STOP-MCD study vs. 11.1% in the JNSCS) might blunted the association between serum albumin concentration and remission. The present multicenter prospective cohort study with higher external validity than previous studies, including 108 adult patients with MCD in 40 hospitals in Japan, showed that serum albumin concentration was inversely associated with remission.Conflicting associations between serum albumin concentration and remission of proteinuria in patients with MCD have been reported in some retrospective cohort studies. A retrospective single-center cohort study in the UK, including 51 adult patients with MCD at a single hospital, reported that the time to remission was positively correlated with serum albumin concentration21. Inclusion of suspected secondary MCD (11.5%) might potentially dilute the association between eGFR and remission. In contrast, a Japanese single-center retrospective cohort study suggested that higher serum creatinine level was associated with slower remission in 53 adult patients with MCD11. Another Japanese retrospective multicenter cohort study, the STOP-MCD study, including 142 adult patients with MCD in 5 hospitals, confirmed the inverse association between serum creatinine level and remission using a multivariable-adjusted CPH model13. The present study ascertained that patients with lower kidney function, especially eGFR < 60 mL/min/1.73 m2, achieved remission more slowly. The sample size was comparable to the previous largest Japanese study13 and a large number of participating hospitals provided evidence with high external validity.Previous studies have reported contradictory impacts of kidney function on remission of proteinuria in patients with MCD. A retrospective single-center cohort study, including 52 patients with MCD in UK, reported that eGFR was not associated with remission in an unadjusted CPH model22. A Taiwanese retrospective cohort study of MCD reported that 23 patients with no AKI and creatinine clearance of 88.3 ± 23.6 mL/min had a significantly higher cumulative probability of remission than 20 patients with AKI and creatinine clearance of 31.6 ± 19.2 mL/min12. Another Japanese cohort study, including 53 adult patients with MCD, clarified a dose-dependent association between AKI stage of the Kidney Disease Improving Global Outcomes (KDIGO) criteria and remission, using CPH model adjusting for clinically relevant factors except for eGFR15. In the present study, patients with AKI and, therefore, lower eGFR might achieve remission more slowly than those with no AKI and higher eGFR. The limited number of eGFR measurements available in the present study hindered the identification of the incidence of AKI during the clinical course of each patient. The clinical impact of AKI on the association between eGFR and remission should be assessed in future studies. Second, details of IST, including the time between the onset of symptomatic edema and IST, the initial dose of PSL, and the total duration of prednisolone use, were not available in the present study. Because of the observational nature of the present study, the lack of IST protocol potentially led to biased results. Thus, the associations of use of intravenous mPSL and cyclosporine with remission and relapse in the present study should be interpreted with great caution. The JNSCS is planning to retrieve all laboratory and drug data of each patient during the observational period, which will enable statistical methods for modeling time-updated exposure to IST23 to estimate precise effectiveness of IST in a real-world setting.The present study has several limitations. First, the association between low eGFR and slower remission might be confounded by AKI. Of 716 patients with MCD included in 13 reports, AKI was commonly observed in 235 (33.3%) patientsIn conclusion, this multicenter prospective cohort study clarified that higher serum albumin concentrations and lower eGFR levels were independently associated with a lower cumulative probability remission in adult patients with MCD. The findings of the present study provide a simple risk stratification system for remission in adult patients with MCD, which should be verified in different cohorts.17. Of 455 nephrotic patients who were diagnosed with primary nephrotic syndrome between January 2009 and December 2010 in 56 hospitals and registered in the JNSCS, 81 patients including those with no kidney biopsy (n = 20), kidney biopsy before or the entry period (n = 32), no history of nephrotic syndrome (n = 1), diagnosis of secondary nephrotic syndrome (n = 13), sclerosing glomerulonephritis with unknown etiology (n = 1), incomplete informed consent (n = 7), duplicate registration (n = 3), and unknown reasons (n = 4) were excluded  and the Institutional Review Board of each participating hospital. All procedures performed in the JNSCS involving human participants were in accordance with the ethical standards of the research committee of the institute at which the studies were conducted and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Written informed consent was obtained from all participants and the legal representatives of the participants under 20 years of age in 54 hospitals. A single hospital used an opt-out approach to provide informed consent, according to the Japanese Ethical Guidelines for Medical and Health Research Involving Human Subjects.−0.287 × serum creatinine (mg/dL)−0.094 × 0.739 24. Data pertaining to the use of immunosuppressive drugs within 1 month of IST were also collected, including oral prednisolone, intravenous mPSL, cyclosporine, tacrolimus, cyclophosphamide, mycophenolate mofetil, mizoribine, and rituximab.Baseline characteristics at initiation of IST included age, sex, BMI, systolic and diastolic blood pressure, serum creatinine and albumin concentration, eGFR, 24 h urinary protein (or urinary protein-to-creatinine ratio if 24 h urinary protein was missing), and RAS blockade, including use of angiotensinogen converting enzyme inhibitors and angiotensin II receptor blockers, and intravenous albumin administration. The Japanese equation was used to calculate eGFR: eGFR = 194 × age (year)The outcome of interest was (i) remission of proteinuria defined as 24-h urinary protein < 0.3 g/day or urinary protein-to-creatinine ratio of < 0.3 g/gCr, and (ii) relapse of proteinuria defined as 24-h urinary protein ≥ 1.0 g/day, urinary protein-to-creatinine ratio ≥ 1.0 g/gCr, and/or dipstick urinary protein ≥ 2+ continued two or more times. The observational period to identify the predictors of remission was defined as the period from the initiation of IST to (i) the incidence of remission, (ii) the end of the 5-year study period of the JNSCS, or (iii) loss to follow-up, whichever came first. To identify the predictors of relapse, the observational period was defined as the period from the incidence of remission to (i) the incidence of relapse, (ii) the end of the 5-year study period of the JNSCS, or (iii) loss to follow-up, whichever came first.2; and urinary protein into 4 groups of 3.5–4.9, 5.0–7.4, 7.5–9.9, ≥ 10.0 g/day or g/gCr, baseline characteristics, use of immunosuppressive drugs within 1 month of IST, and the cumulative incidence of remission and relapse were compared among these 4 groups using analysis of variance, the Kruskal–Wallis test, the chi-square test, and the Fisher’s exact test, as appropriate. We also compared these clinical characteristics after categorizing the patients into two categories of serum concentration of ≤ 1.50 and > 1.50 g/dL and eGFR of < 60.0 and ≥ 60.0 mL/min/1.73 m2, using the unpaired t-test, the Wilcoxon rank-sum test, or the chi-square test, as appropriate.After categorizing serum albumin concentration into four groups of ≤ 1.00, 1.01–1.50, 1.51–2.00, and > 2.00 g/day L; eGFR into 4 groups of < 30.0, 30.0–59.9, 60.0–89.9, and ≥ 90.0 mL/min/1.72 m2), systolic blood pressure (mmHg), serum albumin (g/dL), eGFR (mL/min/1.73 m2), urinary protein (log g/day or log g/gCr), dipstick hematuria , intravenous albumin administration, and use of intravenous mPSL and cyclosporine within 1 month of IST as covariates. Because of its skewed distribution, urinary protein was included in CPH models after logarithmic transformation.Cumulative probabilities of remission in the four groups of serum albumin concentration, eGFR level, and urinary protein level were calculated using the Kaplan–Meier method and compared using log-rank test for trend. To identify predictors of remission and relapse, we used unadjusted and multivariable-adjusted CPH models, including age , sex, BMI  and eGFR . The cutoff values between the second and third groups, namely serum albumin of 1.5 g/dL and eGFR of 60 mL/min/1.73 m2, were used as the reference, which were very close to the median values of these variables .To clarify a dose-dependent association of serum albumin and eGFR with remission, we used restricted cubic spline functions using 4 knots placed at 5th, 35th, 65th, and 95th percentileswww.stata.com).Continuous variables were expressed as the mean ± standard deviation or median and interquartile range, as appropriate, and categorical variables were expressed as numbers and proportions. Statistical significance was set at P < 0.05. Statistical analyses were performed using Stata, version 17.0 (Stata Corp, Supplementary Tables.


Atrial fibrillation (AF) is the most common arrhythmia worldwide and is associated with increased morbi-mortality. The prevalence of AF in the Western world is increasing; however, reports on the prevalence of AF in the past decade are scarce, and whether the prevalence of AF increased during the last decade in Switzerland remains uncertain. Therefore, using data from a Swiss population-based sample, we aimed to assess the point prevalence of AF from 2014 to 2017 and to investigate determinants of AF.A cross-sectional analysis of 4616 Caucasian participants aged 45–86 years (55% women) from a population-based sample was designed to explore the point prevalence and determinants of cardiovascular risk factors in the population of Lausanne, Switzerland. AF was assessed using electrocardiography (ECG) between 2014 and 2017.Overall, the point prevalence of AF was 0.9%  and the combined AF + atrial flutter (AFL) point prevalence was 1.1% (95% CI: 8.4–1.5%). The point prevalence of AF was higher among men (81% vs. 19% in women) and increased with age, reaching 3.1% in participants aged ≥ 80. In multivariable analysis, male gender (odds ratio and 95% CI: 4.98 [1.01–24.6]) and increasing age (2.86 [1.40–5.87] per decade) were associated with AF.The point prevalence of AF and of AF + AFL, assessed between 2014 and 2017 in the city of Lausanne (Switzerland), was low but increased with age and in men.The online version of this article (10.1007/s00059-021-05090-7) contains supplementary material, which is available to authorized users. Atrial fibrillation (AF) is the most common arrhythmia worldwide and is associated with increased mortality  and morbSeveral studies performed in Western countries showed variable prevalence of AF, ranging from 0.3% to 7.2%, notably because of heterogeneity in their design and in the age groups studied –8. Indeep = 0.0007; 8.1 to 11.8 per 1000 person-years in women, trend p = 0.009). A population-based survey of individuals aged 35–74 years living in the city of Geneva (Switzerland) at the beginning of 2000 showed an overall prevalence of AF below 1%  for continuous variables. Bivariate analyses were performed using chi-square or Fisher’s exact test for qualitative variables and Student’s Due to its paroxysmal nature, it was possible that some participants already presented with AF or AFL in the past. As it was unknown who had already presented with these conditions, we hypothesized that participants taking anticoagulants might have AF or AFL. Therefore, we decided to exclude them from the sensitivity analysis.Commission cantonale d’éthique de la recherche sur l’être humain (www.cer-vd.ch) approved the baseline CoLaus|PsyCoLaus study (reference 16/03) and the approval was renewed for the first (reference 33/09) and second (reference 26/14) follow-ups. The study was performed in agreement with the Helsinki Declaration and all participants gave their signed informed consent before joining the study.The institutional Ethics Committee of the University of Lausanne, which later became the On the initial 4881 participants, 4616 (94.6%) were retained for analysis. The exclusion criteria are indicated in Fig. Overall, 42 cases of AF were detected, resulting in an AF point prevalence of 0.9% (95% CI: 0.7–1.2). The point prevalence of AF by sex and age is presented in Fig. Furthermore, 10 cases of typical AFL were recorded, but no atypical AFL was detected. Therefore, the combined point prevalence of AF + AFL is 1.1% (95% CI: 0.4–1.5%). The point prevalence of combined AF + AFL by sex and age is presented in Fig. The bivariate comparisons of the clinical and sociodemographic characteristics between AF and non-AF cases are summarized in Table Similar findings were obtained when comparing participants with and without AF + AFL .Our study reported an AF point prevalence of 0.9% (95% CI: 0.7–1.2%). This finding is similar to the population-based survey conducted in Geneva, Switzerland, using data from 2000, where the prevalence of AF among subjects aged ≥ 50 years was 0.9% . Later, The point prevalence of AF was higher among men, which is in agreement with previous studies , 12. In Increasing age is one of the leading risk factors of AF . In partn = 2421 participants) and therefore could explain the reduced statistical power. The trends of the OR still confirm the results previously published [Hypertension, higher BMI, personal history of CVD, diabetes, dyslipidemia, and physical activity were associated with AF in the bivariate analysis, which was expected as they are known risk factors for AF , 32, 33.ublished , 32, 33.ublished . Thus, eNo association was found between alcohol drinking and AF. An explanation is the relatively low level of alcohol consumption in our study (median of 7.5 units/week among AF cases), as AF is mostly associated with high levels of alcohol consumption .No association was found between OSA and AF. For a sizable number of participants there were no data for the Berlin Questionnaire and the statistical power was therefore reduced. The link between OSA and AF should be examined better, using a larger number of patients with polysomnography data.For AF + AFL, the point prevalence was 1.1% (95% CI: 0.4–1.5%) in our study, which is in line with previously published reports , 12. TheThis study has some limitations. First, our study potentially excludes participants with more comorbidities, since participation in population-based studies are selective and healthy people participate more frequently . Thus, oThe prevalence of atrial fibrillation and of atrial fibrillation + atrial flutter, assessed between 2014 and 2017 in the city of Lausanne (Switzerland), was low but increased with age and in male participants.Supplementary Table 1: characteristics of included and excluded (without ECG) participants at 2nd follow-up, CoLaus|PsyCoLaus study, Lausanne, SwitzerlandSupplementary Table 2: characteristics of included and excluded (without ECG or missing covariate(s) ≥ 1) participants at 2nd follow-up, CoLaus|PsyCoLaus study, Lausanne, SwitzerlandSupplementary Table 3: characteristics of participants with and without AF, CoLaus|PsyCoLaus study, Lausanne, Switzerland. Analysis restricted to participants with all covariatesSupplementary table 4: multivariable analysis of the factors associated with AF or with AF + AFL at 2nd follow-up, CoLaus|PsyCoLaus study, Lausanne, Switzerland. Analysis including risk of sleep apnea


To the Editor,IL‐4 and IL‐13 are Type‐2 (T2) inflammatory cytokines and key drivers in T2 immune response, which is considered to play a central role in the pathogenesis of several atopic diseases such as atopic dermatitis (AD) and asthma. Dupilumab, a fully human monoclonal antibody, binds to the α‐subunit of the interleukin (IL)‐4 receptor and blocks the signaling pathway of IL‐4 and IL‐13.This study consecutively included adult AD patients with comorbid asthma and at least one measurement of the Asthma Control Questionnaire (ACQ)‐5 , and/or FEV1, who started dupilumab treatment for AD and participated in the BioDay registry from October 2017 to June 2022. The ACQ‐5 was used as patient‐reported outcome, consisting five questions on symptom control of asthma. Following The Global Initiative for Asthma (GINA)‐guidelines controlled asthma in a real‐life setting is defined as ACQ‐5 < 0.5.n = 236) at baseline, and significantly improved over time (p < 0.00), with −0.24  at week 16 and −0.26  at week 52. Mean FEV1 at start of treatment was 2.96 L  and significantly improved over time (p < 0.00) with 0.10 L (95% CI 0.03–0.16) and 0.12 L (95% CI 0.05–0.19) at week 16 (n = 81) and 52 (n = 64), respectively. No significant change for ACQ‐5 and FEV1 was found between week 16 and 52. Secondary effectiveness endpoints are presented in Figure n = 22) was 23.43 ppb (95% CI 16.37–33.53) and significantly decreased over time (p < 0.00), to 13.13 ppb  at 16‐weeks and to 15.24 ppb  at 52‐weeks of treatment  investigating the efficacy and safety of dupilumab in patients with uncontrolled asthma showed a more impressive effect on the primary endpoints.As shown in the Appendix, no profound effect of T2‐indicator, blood eosinophilia, at the start of treatment was found regarding the effectiveness of dupilumab on FEV1. On the contrary, in the asthma studies,A limitation of the study is the missing data due to the daily practice setting and COVID‐pandemic. Additionally, spirometry measurements were only conducted in patients using inhaled corticosteroids thereby excluding patients with mild asthma.In conclusion, 1 year of dupilumab treatment primarily indicated for AD resulted in a significant improvement of comorbid asthma with the largest effect in the first 16 weeks. Dupilumab treatment in AD patients provides an additional advantage for patients with comorbid asthma.All authors have made substantial contributions to conception and design of this study and have been involved in drafting or revising the manuscript. All authors have given final approval of the version to be published an agreed to be accountable for all aspects of the work. Lotte S. Spekhorst had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.Lotte S. Spekhorst is a speaker for Abbvie. Marlies de Graaf is an advisor, consultant, speaker or investigator for Sanofi‐Genzyme, Regeneron Pharmaceuticals, LEO Pharma and Eli Lilly. Lisa P. van der Rijst has nothing to disclose. Nicolaas P. A. Zuithoff has nothing to disclose. René C. Schweizer has nothing to disclose. Marijke Kamsteeg has nothing to disclose. Inge Haeck is an advisor, consultant, speaker or investigator for Sanofi‐Genzyme, LEO pharma, AbbVie and Eli Lilly. Anneke M. T. van Lynden‐van Nes has nothing to disclose. Paula van Lumig has nothing to disclose. Geertruida L. E. Romeijn has nothing to disclose. Marie‐Louise Schuttelaar is an advisor, consultant, speaker and/or investigator for AbbVie, Pfizer, LEO Pharma, Regeneron, Sanofi Genzyme, Eli Lilly and Galderma. She has received grants from Regeneron, Sanofi Genzyme, Novartis and Pfizer. Marjolein S. de Bruin‐Weller has been a consultant, advisory board member, and/or speaker for AbbVie, Almirall, Arena, Aslan, Eli Lilly, Galderma, Janssen, Leo Pharma, Pfizer, Regeneron, and Sanofi‐Genzyme.Regeneron Pharmaceuticals; AbbVie; Sanofi; LEO Pharma; Eli Lilly and Company; Pfizer.Supplementary Material S1Click here for additional data file.


Establishing reference ranges of the complete blood count (CBC), reticulocyte hemoglobin content (Ret-He), immature reticulocyte fraction (IRF), and reticulocyte production index (RPI) helps diagnose a disease related to the changes in erythrocyte indices, white blood count, platelets, and reticulocytes, especially in babies. Therefore, the study aims to establish a reference range for CBC and reticulocyte parameters in healthy babies aged 1–4 months. The study design was a cross-sectional study with descriptive analysis of CBC and reticulocyte in babies aged 1–4 months. Three hundred forty-eight babies met the inclusion criteria. This study recruited 89 babies aged 1 month, 87 babies aged 2 months, 86 babies aged 3 months, and 86 babies aged 4 months. The P5-P95 reference range of healthy babies for hemoglobin (Hb) aged 1 month, 2 months, 3 months, and 4 months was 9.95 to 15.45 g/dL, 9.74 to 13.42 g/dL, 9.51 to 12.40 g/dL, and 10.04 to 13.10 g/dL respectively. The P3-P97 reference range of healthy babies for Hb aged 1 month, 2 months, 3 months, and 4 months was 9.60 to 15.90 g/dL, 9.46 to 13.97 g/dL, 9.26 to 12.82 g/dL, and 10.00 to 13.33 g/dL respectively. This study also defined reference ranges for CBC, Ret-He, IRF, and RPI. The reference range of CBC, Ret-He, IRF, and RPI for healthy babies aged 1–4 months in this study can be used as a benchmark. Anemia is still a major global health problem. Thereby, the early detection of anemia is crucial considering its long-term effects that can interfere with a baby’s growth and development. Several studies have shown the impact of iron deficiency anemia (IDA) on cognitive and behavioral deteriorations4. Then, early diagnosis and treatment of impending brain dysfunction in the pre-anemic stage are necessary to prevent neurological deficits. Iron deficiency and IDA are caused by not giving oral iron supplementation and low iron stores at birth; the baby’s growth rate is swift, and inadequate iron intake.All ages anemia prevalence was 22.8% globally in 2019. The prevalence was highest among children under five years, 39.7%, with the most contributing cause being dietary iron deficiency (ID)Reference range data for complete blood count (CBC), reticulocyte hemoglobin content (Ret-He), immature reticulocyte fraction (IRF), and reticulocyte production index (RPI) for babies 1–4 months old are limited. Therefore, determination of the reference range of CBC, Ret He, IRF, and RPI in babies 1–4 months is essential to determine whether a baby is anemic or not. Combining several parameters, Hb (hemoglobin), MCV (mean corpuscular volume), MCH (mean corpuscular hemoglobin), RDW (red cell distribution with), reticulocyte count, Ret He, IRF, and RPI will undoubtedly make it easier to determine if a baby has ID before IDA occurs. In addition, to monitor oral iron therapy, these combining parameters can be used too. For example, a lower value than the lower limit of the Hb reference range according to age indicates a baby is anemic.On the other hand, other erythrocyte indices reference ranges are needed to diagnose anemia, especially suspected IDA. The main problem is determining if a baby has IDA with a simple, cheap, and fast laboratory test; a small amount of blood is drawn and can be done in almost any health facility. It is even better if the ID can be detected before IDA occurs.7. Therefore, a lower value than the lower limit of the Ret-He reference range according to age indicates that an infant is iron deficient.Conventionally, ID diagnosis is based on determining biochemical parameters, such as ferritin, transferrin saturation, and soluble transferrin receptor. Nevertheless, these biomarkers are strongly influenced by the diurnal phase, inflammation, infection, and malignancy, and of course, they are expensive, and not every health facility can check them. In addition, many studies have shown that Ret-He can show iron deficiency without anemia8. In addition, RPI can be used as a diagnostic guide to the etiology of anemia9. Uniquely, all erythrocyte indices and reticulocyte parameters can be seen automatically on the Sysmex XN-450 Hematology Analyzer .The clinical utility of IRF applied in various circumstances, such as monitoring anemia treatment and neonatal transfusion needs, can be a prognostic factor in prematurity anemia and diagnosing and monitoring aplastic anemias. Furthermore, RPI monitors bone marrow response to iron-treated anemiaThe purpose of this study was to obtain the lower limit of the baby's hemoglobin value related to the age so that it can be used to declare an anemic baby or not. Moreover, establishing reference ranges of the CBC, Ret-He, IRF, and RPI helps diagnose a disease related to the changes in erythrocyte indices, white blood count, platelets, and reticulocytes, especially in healthy babies aged 1–4 months.Three hundred forty-eight healthy babies  met the inclusion criteria. Therefore, this study recruited 89 babies aged 1 month , 87 babies aged 2 months , 86 babies aged 3 months , and 86 babies aged 4 months . P5-P95 and P3-P97 of the complete blood count, reticulocyte count, Ret-He, immature reticulocyte fraction, and reticulocyte production index for healthy babies aged 1–4 months can be seen in Tables The P5-P95 reference range of healthy babies for hemoglobin (Hb) aged 1 month, 2 months, 3 months, and 4 months was 9.95 to 15.45 g/dL, 9.74 to 13.42 g/dL, 9.51 to 12.40 g/dL, and 10.04 to 13.10 g/dL respectively. The P3-P97 reference range of healthy babies for Hb aged 1 month, 2 months, 3 months, and 4 months was 9.60 to 15.90 g/dL, 9.46 to 13.97 g/dL, 9.26 to 12.82 g/dL, and 10.00 to 13.33 g/dL respectively.10. Determination of the lower limit of Hb with P3-P97 and P5-P95, of course, by considering race, ethnicity, age, gender, and altitude above sea level11. This study accommodates all reference range limits to be used flexibly.Establishing the lower limit of P3-P97 and P5-P95 on examining specific parameters is commonly used statistically. For example, the WHO  and CDC (Centers Of Disease Control and Prevention) growth charts set the lower limit of Weight-for-age, Length/height-for-age, Head circumference-for-age, and Arm circumference-for-age at P3The results of this study showed the reference range of CBC, reticulocyte count, Ret-He, IRF, and RPI at P3-P97 and P5-P95. Values below the lower limit of the reference range of Hb according to age can be used as a limit to determine if a baby has anemia. In addition, the value below the lower limit of the reference range of Ret-He according to age can be used as a limit to determine whether a baby has ID. The baby is suffered from IDA; if the value of Hb and Ret-He is below the lower limit of the reference range. Thus, for example, this study indicates that the lower limit of Hb for healthy babies aged 4 months  and increased absolute reticulocyte count (ARC) generally indicated an adequate erythroid response to anemia. Meanwhile, an IRF of 0.23 or less in patients with anemia reflects bone marrow that is nonresponsive or under-responsive to the anemia21. Immature reticulocyte fraction may be the first sign of hematologic recovery. It is a powerful indicator of post-chemotherapy aplasia in children with cancer, serving as an additional parameter of impaired bone marrow function22. A low total count with a relatively high IRF indicates a regenerating marrow, whereas reticulocytopenia with low IRF is typical of severe aplastic anemia or renal failure. A high total count with high IRF occurs in acute hemolysis and blood loss. In contrast, a low to average total count with a high IRF occurs in dyserythropoietic and during the early response to haematinics. It may also help decide whether macrocytic anemia is megaloblastic or nonmegaloblastic23. Mullier et al. showed that IRF, combined with reticulocyte count, may be helpful in hereditary spherocytosis (HS) diagnosis as a high reticulocyte count characterizes HS without an equally elevated IRF. Therefore, a reticulocytes/IRF ratio higher than 7.7 is a precondition for screening HS cases24. The IRF increases earlier than the reticulocyte number. It helps monitor the efficacy of therapy in nutritional anemias such as megaloblastic or IDA. Therefore, the reference value of the IRF range in this study can be considered to assess the responsiveness of the anemia therapy given.Immature reticulocyte fraction indicates the younger fraction of reticulocytes, reflecting erythropoietic activity. Several studies have demonstrated the clinical utility of IRF. For example, an increase in IRF within a few days indicates recovery of bone marrow following bone marrow transplantation, erythropoiesis-stimulating agent therapy, or chemotherapy25.The reticulocyte count reflects the erythropoietic activity of bone marrow. In addition, it is valuable in diagnosing anemias and monitoring bone marrow response to therapy. RPI is currently used in pediatrics to assess bone marrow responsiveness to anemia therapy given. However, considering that “conventional” RPI was developed from a study conducted in adults, it is necessary to have an RPI based on standard values adjusted for different pediatric ages, especially in babies. Therefore, the reference value of the RPI range in this study can be considered to assess the responsiveness of the anemia therapy given. However, according to Bracho et al., the RPI is an inadequate tool for evaluating the bone marrow response in the presence of anemia due to differences in hematologic values between children and adults. In addition, the absence of information on the maturation time of reticulocytes in childrenThe power of this study is that it parades all reference ranges of CBC, Ret He, IRF, and RPI at P3-P97 and P5-P95 for healthy babies aged 1–4 months. The weakness of this study is that the altitude above sea level, race, genetics, ethnicity, and population of this study is different from other areas. Hence, it certainly affects the reference range of CBC and reticulocyte parameters. In this study, 96.5% reside at altitudes less than 100 m. Again, however, race, genetics, and ethnicity differed from other countries. Therefore, this reference range for CBC and reticulocyte parameters in healthy babies aged 9–11 months can be used in countries with demographic, socioeconomic, and population structures, such as Southeast Asia, especially Indonesia.26. All of the baby’s parents had signed the informed consent. This study obtained ethical clearance from the Research Ethics Commission of the Medical Faculty of the University of Lambung Mangkurat No. 272/KEPK-FK ULM/EC/VIII/2020. All methods have been performed under the relevant guidelines and regulations of the Declaration of Helsinki.The study design was a cross-sectional study with descriptive analysis of CBC and reticulocyte in babies aged 1–4 months. The study was conducted at 10 Community Health Centers in Banjarbaru, South Kalimantan, from August 2020 to August 2021. Inclusion criteria are babies born at term  with birth weight ≥ 2500 g, babies are not twins, not taking a hematinic drug, and normal nutritional status. Exclusion criteria were hematological diseases and congenital anomalies. In addition, gestational age and birth weight had obtained from the mother’s medical record . The doctor in charge declared the baby healthy. At the time of recruitment, the baby’s weight, length, and head circumference were measured by health personnel. Nutritional status is assessed based on body weight and length, divided into good nutrition and undernutrition. Good nutrition if the z-score is − 2 SD to + 3 SD in this study. In contrast, undernutrition if the z-score is < − 2 SDEvery baby who meets the inclusion and exclusion criteria will be taken a blood sample of 1 ml from the median cubital vein. First, the blood sample was put in a tube with EDTA anticoagulant, homogenized by turning it over, and stored in a storage box. Then the blood sample was sent to the Banjarbaru Idaman Hospital Laboratory. The Sysmex XN-450 Hematology Analyzer  was doing complete blood count and reticulocyte examinations.All baby anthropometry measurements were analyzed by SPSS ver.25 for P3-P97 and P5-P95. Moreover, the CBC and reticulocyte laboratory findings were analyzed by SPSS ver.25 for P3-P97 and P5-P95. All data are presented in narrative and table. Reference range P3-P97 means that 94% of normal individuals have normal laboratory results, while the other 6% may not be sick outside the normal limits. Reference range P5-P95 means that 90% of normal individuals have normal laboratory results, while 10% may not be sick outside the normal limits.Supplementary Table 1.Supplementary Table 2.Supplementary Table 3.Supplementary Table 4.Supplementary Table 5.Supplementary Table 6.Supplementary Table 7.


Finally, cytotoxicity against murine fibroblasts L929 was tested. All of the analyzed compounds permeated pig skin and accumulated in it. LPr[FA] and PPr[FA] were characterized by much better permeability compared to the parent ferulic acid. Additionally, it was shown that all the analyzed derivatives are characterized by high antioxidant activity and lack of cytotoxicity. Therefore, they can be considered as an interesting alternative to be applied in dermatologic and cosmetic preparations.Ferulic acid (FA) has been widely used in the pharmaceutical and cosmetics industry due to its, inter alia, antioxidant, antiaging and anti-inflammatory effects This compound added to cosmetic preparations can protect skin because of its photoprotective activity. However, the usefulness of FA as a therapeutic agent is limited due to its low solubility and bioavailability. The paper presents the synthesis, identification, and physicochemical properties of new FA derivatives with propyl esters of three amino acids, glycine (GPr[FA]), L-leucine (LPr[FA]), and L-proline (PPr[FA]). The NMR and FTIR spectroscopy, DSC, and TG analysis were used as analytical methods. Moreover, water solubility of the new conjugates was compared with the parent acid. Both ferulic acid and its conjugates were introduced into hydrogel and emulsion, and the resulting formulations were evaluated for stability. Additionally, in vitro penetration of all studied compounds from both formulations and for comparative purposes using Franz diffusion cells was evaluated from the solution in 70% ( One of the main factors responsible for the harmful effect of ROS on the skin is long-term skin exposure to ultraviolet (UV) radiation —glycine propyl ester ferulate.1H NMR  δ in ppm: 7.44 , 7.25 , 7.05 , 6.78 , 6.36 , 4.00 , 3.80 , 3.32 , 1.58 , 0.88 . 13C NMR  δ in ppm: 173.61 (C12), 168.37 (C10), 148.98 (C3), 147.92 (C4), 143.74 (C8), 125.94 (C1), 122.65 (C6), 116.66 (C9), 115.52 (C5), 111.02 (C2), 65.54 (C13), 55.67 (C7), 43.06 (C11), 21.59 (C14), 10.28 (C15). FT-IR (ATR) ν cm−1: 517, 572, 699, 731, 815, 868, 909, 931, 981, 992, 1036, 1059, 1118, 1171, 1228, 1275, 1378, 1427, 1452, 1508, 1558, 1587, 1602, 1637, 1745, 2880, 2968, 3215.Yield: 97%; LPr[FA]—L-leucine propyl ester ferulate.1H NMR  δ in ppm: 7.46 , 7.26 , 7.06 , 6.78 , 6.36 , 3.99 , 3.81 , 3.33 , 1.71 , 1.58 , 1.44–1.29 , 0.84–0.90 . 13C NMR  δ in ppm: 175.98 (C11), 168.22 (C10), 149.02 (C3), 147.91 (C4), 144.02 (C8), 125.87 (C1), 122.72 (C6), 116.27 (C9), 115.51 (C5), 111.05 (C2), 65.45 (C17), 55.66 (C7), 52.34 (C12), 43.66 (C14), 24.20(C16), 22.78 (C15), 21.95 (C13), 21.58 (C18), 10.30 (C19). FT-IR (ATR) ν cm−1: 453, 508, 525, 571, 588, 604, 713, 744, 815, 863, 926, 958, 978, 1036, 1060, 1089, 1126, 1160, 1195, 1209, 1267, 1304, 1368, 1404, 1454, 1467, 1517, 1591, 1645, 1736, 2834, 2865, 2934, 2959, 3203.Yield: 98%; PPr[FA]—L-proline propyl ester ferulate.1H NMR  δ in ppm: 7.46 , 7.26 , 7.06 , 6.78 , 6.36 , 3.99 , 3.81 , 3.68 , 2.90 , 2.77 , 1.98 , 1.74–1.54 , 0.88 . 13C NMR  δ in ppm: 174.60 (C11), 168.23 (C10), 149.01 (C3), 147.91 (C4), 144.06 (C8), 125.86 (C1), 122.73 (C6), 116.22 (C9), 115.51 (C5), 111.05 (C2), 65.61 (C16), 59.07 (C12), 55.67 (C7), 46.34 (C15), 29.64 (C13), 25.12 (C14), 21.57 (C17), 10.26 (C18). FT-IR (ATR) ν cm−1: 455, 494, 529, 570, 581, 604, 697, 744, 778, 806, 825, 858, 885, 897, 931, 965, 986, 1022, 1055, 1121, 1159, 1177, 1215, 1275, 1373, 1422, 1453, 1518, 1585, 1602, 1635, 1739, 2966.Yield: 94%; 3 or in DMSO-d6 on a BRUKER DPX-400 spectrometer  operating at 400.13 MHz (1H) and 100.62 MHz (13C). CDCl3 as solvent and TMSCl as chemical shift standard were used for amino acid propyl ester hydrochlorides and DMSO-d6 for ferulic acid and its derivatives. The NMR spectra for ferulic acid and its derivatives were recorded in CDCl−1. The resolution was 4 cm−1.The FTIR spectra were recorded using a spectrometer model ‘Nicolet 380′  with Diamond ATR in transmission mode, in the range of 4000–400 cm® from NETZSCH . Samples of 2–5 mg, loaded in aluminium crucibles, were heated from 30 °C to 600 °C at the heating rate of 10 °C·min−1, in the air atmosphere (25 mL·min−1) with nitrogen flow (10 mL·min−1) as the purge gas. Thermogravimetric (TG) analysis was carried out using thermomicrobalance TG 209 F1 Libra−1. Indium and mercury were used as standards to calibrate the temperature. Differential scanning calorimetry (DSC) was performed using TA Instruments, model DSC 250 . The sample was loaded on an aluminum pan with a lid. The analysis was carried out in nitrogen atmosphere. The sample was first heated from 20 °C to specified temperature , then cooled to 20 °C and again heated to the specified temperature. The rate of heating/cooling/heating was 10 °C·minD20 was measured in ethanol solution using AUTOPOL IV Polarimeter  for the concentration of compound of 0.01 g·cm−3. A polarimeter precision was 0.001° of angular rotation, and accuracy of temperature determination was 0.1 °C.The specific rotation [α]®F5 column  was used in solubility experiments. Analyses were performed at 30 °C under isocratic conditions, with mobile phase consisting of water/acetonitrile mixture  and a flow rate of 1 mL min−1. Detection wavelength was 215 nm. Instrument control and data acquisition and processing were performed using an Agilent ChemStation software. Injections were repeated at least three times for each sample and the results were averaged. Concentration of ferulic acid and its salts were calculated based on peak area measurements using a calibration curve method.The Agilent 1200 Series HPLC system of Agilent Technologies Inc. , consisting of Quaternary pump, Vacuum degasser, Manual Injector, Diode Array Detector and Thermostatted Column Compartment equipped with KinetexSolubility in deionized water was determined for ferulic acid and its salts with amino acid propyl esters and defined as saturation concentration—a maximum quantity of a substance that may be dissolved at a given temperature per volume unit of solvent. An excess of the compound was added to 2 mL of water in screwed vials and was stirred vigorously at 25.00 ± 0.05 °C for 24 h. Then mixture was centrifuged and liquid above the solid was taken, diluted, and analyzed by the HPLC method to determine the concentration of the compound. Three independent solubility test replicates were performed for each compound.n-octanol/water partition coefficient (log PO/W) was determined for ferulic acid and its derivatives by shake flask method. n-Octanol saturated with water and water saturated with n-octanol were prepared at 25 °C, 24 h prior to their use in the experiments. Then, a sample of 10 mg of compound was weighed with the accuracy of 0.01 mg in a 16 cm3 screwed vial and dissolved in 5 cm3 of n-octanol. Next, 5 cm3 of water was added to the vial. The resulting two-phase mixture was kept in water bath shaker at 25 °C for 2 h and was strongly shaken to reach the partition equilibrium of the compound between the phases. Then the mixture was left unshaken for 30 min and 1.0 cm3 of water phase was carefully taken out to centrifuge tube. The sample was next centrifuged for 10 min at 7500 rpm at 25 °C to ensure the most precise phase separation. The concentration of compound in water phase was then determined using HPLC method. The partition coefficient, log PO/W was calculated according to the formula:0—initial concentration of the compound in n-octanol, calculated based on the weighed mass of compound, mg·cm−3; V0—volume of n-octanol, cm3; CW—concentration of the compound in water after partition equilibrium, mg·cm−3; VW—volume of water, cm3.The v/v) ethanol were also prepared at the same concentration of 1 g FA per 100 cm3.For the ethanol solution, the following abbreviations were used: EtOH-[FA], EtOH-GPr[FA], EtOH-LPr[FA], and EtOH-PPr[FA].The hydrogel was prepared according to a modified procedure by Zagórska-Dziok et al. and Nowak et al. ,27. The g at 25 °C for 10 min. Moreover, the stability of all preparations was also evaluated using the heating/cooling test: incubation at 45 °C  for 48 h, followed by incubation at 4 °C (in the refrigerator) for 48 h. The test was performed for six cycles. Stability of all preparations kept at heating/cooling condition were confirmed by visual appearance  ethanolic solutions of ferulic acid and its derivatives—GPr[FA], LPr[FA], PPr[FA]—were prepared. The antioxidant activity of the prepared solutions was measured as follows: 2.85 mL 0.3 mmol dm−3 ethanolic solution of the DPPH radical of absorbance about 1.000 ± 0.020 at λ = 517 nm was placed in the tube, and an appropriate amount of the ethanolic solution of ferulic acid or its derivative was added to obtain the molar ratio of the compound to DPPH 0.1, 1.0 and 2.0. Blank samples without antioxidant were prepared in the same way. The tubes were wrapped in aluminum foil and were sealed with a stopper and then incubated for 10 min at room temperature. After this time, spectrophotometric measurements were carried out in triplicate. For determination of antioxidant activity of the acceptor fluid after 24 h of permeation and solutions obtained by extraction of the skin obtained after penetration study, 0.15 mL samples of each solutions were separately added to 2.85 mL of DPPH solution and spectrophotometric measurements were performed. The antioxidant activity was expressed as mmol Trolox·dm−3.To determine the antioxidant capacity of ferulic acid and its derivatives the DPPH method was used ,32. The −3.Total polyphenols content in acceptor fluid taken after 24 h of permeation and in solutions after skin extraction was determined by the Folin–Ciocalteu method, as described previously . The antIn the experiment, abdominal porcine skin coming from the local slaughterhouse was used. Pig skin was chosen because of its similar permeability to human skin ,35. The v/v/v) and was pumped at flow rate of 1 mL min−1. The column temperature was set at 25 °C, and the injection volume was 20 µL.The concentration of FA and its derivatives in the acceptor fluid and in the fluid obtained by skin extraction after permeation experiment was assessed using liquid chromatography system  consisting of a model 2600 UV detector, Smartline model 1050 pump and Smartline model 3950 autosampler. HPLC was controlled by ClarityChrom 2009 software. The detector was operated at 280 nm. The separations were performed on 125 mm × 4 mm ID chromatographic column filled with Hyperisil ODS (C18), particle size 5 µm, in isocratic mode. The mobile phase consisted of acetonitrile, 1% acetic acid, and MeOH (47/47/6 2. The volume of the acceptor chamber was 8 mL. This chamber was filled with PBS solution (pH 7.4). During experiment, a constant temperature of 37.0 ± 0.5 °C was maintained in each diffusion unit  (because of limited solubility). The cell biocompatibility was evaluated using a PrestoBlue HS Cell Viability Reagent  based on the reduction of the resazurin to highly fluorescent resorufin, of which the amount is directly correlated to the number of living cells. The L929 cells were seeded in 96-well black microplates  at a density of 5 × 103 cells/well and allowed to adhere for 24 h. Afterwards, cell culture medium was removed and replaced with 90 µL of the fresh medium containing 0.03125, 0.0625, 0.125, 0.25, 0.5, 1.0, and 2.0 mM . The final concentrations of ethanol did not exceed 1.0% and its effect on cell viability was also evaluated. The cells without tested compounds were used as control. The interaction of tested compounds without cells and Prestoblue Reagent (blank wells) was excluded. After 24 h of treatment, optical microscopy imaging of L929 cells was performed using Smart Fluorescent Cell Analyzer Microscope JuLi . Next, the PrestoBlue reagent (10 μL) was added to each well and incubated for 30 min. The fluorescence was measured using a spectrophotometric microplate reader  at ex/em: 560/594 nm. The readings were acquired from at least three independent experiments (each conducted in triplicate) and normalized to the control (100% viability).Murine fibroblasts (L929) were cultured in a humidified incubator (5% COt-test (p < 0.05). Statistical calculations were performed using Statistica 13 PL software .Results are presented as the mean ± standard deviation (SD). A one-way analysis of variance (ANOVA) was used. In the case of the cumulative mass after 24 h permeation and the cumulative mass in the skin the significance of differences between individual groups was evaluated with Tukey test (α < 0.05). A cluster analysis was carried out to determine similarities between all compounds tested and all vehicles considering all time points. In the case of the viability of cells, the significance of differences between treated and control cells was evaluated with Student’s The derivatives of ferulic acid and amino acid propyl esters were prepared by a three-step method . Three a1H and 13C NMR and FT-IR.In the first synthesis step , the hyd1H NMR, 13C NMR, and 1H-13C HMQC spectra for each compound are included in the 13C NMR spectra of ferulic acid and its derivatives are gathered in 13C NMR spectra of the derivatives, we can see signals characteristic of the ferulic acid part and amino acid ester moiety. The differences between values of chemical shifts from comparison of 13C NMR spectra show that amino acid propyl esters affect the chemical shift of carbon atoms of FA. For almost all carbons, except for the C9 carbon atom, the signals are a little upfield-shifted, which means higher electron density. Only the signal of the C9 carbon, directly bound with a carbonyl carbon, is downfield-shifted and its chemical shift changes from 115.91 ppm for ferulic acid to the range of 116.22–116.66 ppm for FA derivatives. This change indicates a decrease in the shielding of the nucleus of the C9 carbon and a change in the nature of the vicinity carboxyl group to the more ionic. The most distinct changes concern the electronic charge distribution in the region of the double bond between the carbonyl atom and the aromatic ring. The differences between chemical shifts of the C8 and C9 signals are 29.18 ppm for FA and in the range of 27.08–27.84 ppm for FA derivatives. Moreover, in the downfield region in 13C NMR spectra of FA derivatives, two carbonyl carbon signals are detected at chemical shifts between 173.61 ppm and 175.98 ppm for the ester moiety of the amino acids and between 168.22 and 168.37 ppm for the carboxyl groups of FA in its derivatives. The latter is a little upfield-shifted compared to this signal for unmodified FA (168.61 ppm).Individual −1 for the derivatives of ferulic acid and propyl esters of glycine, L-leucine, and L-proline were collated with the spectrum of the parent acid in −1 as well as ν(C–O) at 1323 cm−1 and ν(O–H)COOH at 3430 cm−1, which are strong in the spectrum of FA, disappear in the spectra of FA derivatives. The absence of these bands in the spectra of derivatives proves that new structures were formed. In addition, the formation of carboxylate anions can be evidenced by the appearance of low intensity bands at 1452–1454 cm−1 (νs (COO−)) and 1602 cm−1 (νas(COO−)), characteristic for salts of carboxylic acids. Moreover, olefin C=C stretching vibrations band occurs in the spectrum of FA at 1618 cm−1 is shifted toward higher wavenumbers 1635–1645 cm−1, in the spectra of FA derivatives. This suggests an increase in electronic charge density around the olefinic double bond for the vicinity of the carboxylate anions. Such a type of shift was also found in the interaction of FA with metals  is 60 times more soluble (42.1828 ± 3.3793) than ferulic acid (0.6729 ± 0.0828). The solubilities of GPr[FA] and LPr[FA] are close to each other and are seven times lower than PPr[FA] but still much better compared to the unmodified acid. The lipid matrix in the outer layer of the skin is crucial for the skin barrier function. However, an effective action of bioactive ingredients involves their dissolution, distribution between vehicle and skin, and diffusion into skin. Therefore, besides solubility in water, the partition coefficient between n-octanol and water is the important physical parameter for prediction of transdermal permeation in topical applications. Moreover, both the solubility in water and the partition coefficient can also be crucial for the selection of an appropriate formulation for active ingredients. In O/W for ferulic acid is 1.643 ± 0.003, which means that almost 98% of ferulic acid is partitioned to the n-octanol. The prepared salts of ferulic acid with amino acid propyl esters are characterized by partition coefficient value between 0.447 for PPr[FA] and 0.582 for GPr[FA], which is 3–4 times lower than for unmodified acid. For these values of the partition coefficient, above 70% of the compound is still located in octanol.Wide application of ferulic acid in skin care and cosmetics products is limited by its poor solubility in water and also in oils. In −3) to 86.5 ± 0.001 %RSA (0.763 ± 0.001 mmol Trolox dm−3 in the case of FA), from 17.6 ± 0.001 %RSA (0.099 ± 0.001 mmol Trolox dm−3) to 86.6 ± 0.001 %RSA (0.764 ± 0.001 mmol Trolox dm−3 in the case of GPr[FA]), from 17.6 ± 0.001 %RSA (0.098 ± 0.001 mmol Trolox dm−3) to 87.8 ± 0.001 %RSA (0.776 ± 0.001 mmol Trolox dm−3 in the case of LPr[FA]), and from 19.3 ± 0.001 %RSA (0.115 ± 0.001 mmol Trolox dm−3) to 87.7 ± 0.001 %RSA (0.775 ± 0.001 mmol Trolox dm−3 in the case of PPr[FA]); see The penetration tests were carried out using hydrogel and emulsion as vehicles for the FA and its derivatives. After preparing the emulsions and hydrogels with and without active compounds, their stability and separation was tested. Both centrifugation tests and temperature shocking tests demonstrated that control samples and preparations with FA and its derivatives are uniform in color and do not separate . v/v) ethanol and as hydrogel and emulsion preparations in the amount equal to 10 mg expressed as ferulic acid. The cumulative mass of the tested compounds in acceptor fluid, considering all time points, is presented in −2 and 415.12 ± 8.71 µg FA·cm−2, respectively. A similar result was observed in the case of the hydrogel, where the highest penetration was found for PPr[FA], 396.86 ± 42.13 µg FA·cm−2. Other derivatives as well as pure FA penetrated from this vehicle in a significantly smaller amount. Percutaneous permeation of all analyzed compounds from the emulsion was the least effective. The cumulative mass determined in case of this vehicle after 24 h of penetration was also the highest for PPr[FA]—128.80 ± 19.86 µg FA·cm−2 , while in the case of hydrogels and emulsion this value was between 0 and 3 h of the study.The permeation rate determined at each time interval is presented in Images from transverse sections of pig skin after application of all vehicles are shown in −3 for FA to 0.098 ± 0.011 mmol Trolox dm−3 for LPr[FA]. For hydrogels, the values were from 0.063 ± 0.009 mmol Trolox dm−3 for LPr[FA] to 0.075 ± 0.012 mmol Trolox dm−3 for PPr[FA], whereas in the case of emulsions, the values were from 0.053 ± 0.005 mmol Trolox dm−3 for LPr[FA] to 0.061 ± 0.007 mmol trolox dm−3 for [PPr[FA]. The content of total polyphenols in the acceptor fluid collected after permeation from the ethanolic solution was the highest for solution of the derivative PPr[FA] and LPr[FA] amounting to 0.122 ± 0.002 and 0.125 ± 0.002 mmol GA dm−3, respectively. For the acceptor fluid taken after the hydrogel and emulsion application, the content of total polyphenols was the highest after penetration PPr[FA], i.e., 0.082 ± 0.001 and 0.061 ± 0.002 mmol GA dm−3, respectively—Antioxidant activity and total polyphenol content were evaluated in the samples obtained during the in vitro skin penetration study. The determinations were performed in acceptor fluid collected after 24 h penetration  and in t−3 for FA to 0.255 ± 0.038 mmol Trolox dm−3 for [Leu[FA]. For hydrogels, activities were from 0.381 ± 0.011 mmol Trolox dm−3 for FA to 0.298 ± 0.009 mmol Trolox dm−3 for LPr[FA], whereas in the case of the emulsions, the values were from 0.300 ± 0.024 mmol Trolox dm−3 for [FA] to 0.235 ± 0.037 mmol Trolox dm−3 for GPr[FA]. The total content of polyphenols in the fluid skin extraction collected after penetration from the ethanolic solution was significantly the highest for fluids containing FA and derivative GPr[FA], with values of 0.275 ± 0.015 and 0.234 ± 0.004 mmol GA dm−3, respectively. A similar tendency was observed when the hydrogel formulations were applied., while in these cases of emulsion, the total content of polyphenols was the highest after penetration of derivative GPr[FA], namely 0.264 ± 0.000 mmol GA dm−3; see The fluid obtained after skin extraction showed high antioxidant activity after each vehicle used. In the case of ethanol solution, the fluid collected after the skin extraction exhibited antioxidant activity from 0.394 ± 0.004 mmol Trolox dmThe biocompatibiliy of FA and its derivatives were evaluated using resazurin-based cell viability assays. The L929 cells were treated with increasing doses of FA, GPr[FA], LPr[FA], and PPr[FA] for 24 h. No statistically significant differences in viability between control and FA-treated cells (at any dosage) were observed . SimilarFerulic acid is a well-known bioactive substance with multiple functions. It shows its main action as antioxidant and anti-inflammatory factors but also has a protective effect on the kidney as well as the circulatory system . In receIn our study, we attempted the synthesis and evaluation of new ferulic acid derivatives, such as glycine propyl ester ferulate (GPr[FA]), L-leucine propyl ester ferulate (LPr[FA]), and L-proline propyl ester ferulate (PPr[FA]). In the first stage of our study, compounds were identified by NMR, FTIR, and solubility studies. O/W for ferulic acid is 1.643 ± 0.0033, which means that almost 98% of ferulic acid is partitioned to n-octanol. Similar values of partition coefficient for ferulic acid were determined by other authors, i.e., 1.69  to 79% for GPr[FA]. Much higher solubility in water than ferulic acid, with a high proportion still partitioning into octanol, may suggest that these compounds easily overcome the lipid organization of the intercellular space of the skin and simultaneously more easily penetrate by intracellular pathway into deeper layers of the skin.The log Pe., 1.69  using ree., 1.69  and 1.32e., 1.69  using thTheir antioxidant activity by the DPPH method was also determined. All derivatives were characterized by high antioxidant activity, which was confirmed by other works on other FA derivatives. For example, it was found that sodium ferulate could protect human corneal endothelial cells from the oxidative damage induced by lidocaine, which causes corneal thickening, opacification, and corneal endothelial cell loss . In anotIn our study, we estimated the permeation of pure FA and its derivatives from two popular and frequently used pharmaceutical/cosmetics vehicles and, for comparison, from an ethanol solution. The FA belongs to a large group of polyphenols that are attractive as ingredients in cosmetics formulations as result of their ability to delay the aging process. In addition, it can be used as an antioxidant to prevent damage from ultraviolet (UV) radiation and skin photodamage . Howeverv/v) ethanol is evident because such solvent is an optimal vehicle for transepidermal application of the drug , exhibited the highest permeation among all compounds. The derivative PPr[FA] permeated more effectively from the ethanol solution and from the hydrogel compared to the unmodified FA in these vehicles. The vehicle used may be of key importance in the penetration of active substances. In the case of topical penetration, it is very important to release the given substances from the preparation form and to reach all layers of the skin as well as the underlying layers . We obsethe drug ,61. Highthe drug . Ethanolthe drug . Comparithe drug ,65. Due the drug ,66,67. The greater permeation of the derivatives was probably also due to their greater solubility. Modification of the substance to increase its solubility is probably one of the reasons to increase its permeation through the skin, which has been shown in previous studies on anti-inflammatory drugs ,68. Our study also assessed the accumulation of FA and its derivatives in the skin after 24 h penetration. It was observed that all the analyzed compounds accumulate in the skin. No significant differences were found between FA and the derivatives, with the exception of PPr[FA] which, when released from the ethanol solution, accumulated to a lesser extent compared to pure FA. The accumulation of this compound in the upper layers of the skin is desirable due to its UV-protective effect. In the case of a cosmetics preparation, the accumulation of active substances in the skin is beneficial. The accumulated active substances on the skin may have an antioxidant effect for a longer time, showing beneficial anti-aging or anti-inflammatory effects, among others, while in the case of ferulic acid and its derivatives, the photoprotective effect is also important. In our study, the acceptor fluid obtained after 24 h penetration showed antioxidant activity, which could also suggest such action in deeper layers. Photodamage of skin is one of the most frequently occurring dermatological problems. A majority of UVB is absorbed in the epidermis of the skin, but UVA reaches deeper skin layers, such as the dermis, and produces reactive oxygen species (ROS). Excessive amounts of ROS could be the cause of the aging, inflammation, and other severe problems such as melanoma. Application of exogenous antioxidants to the skin can be counteracted ,70. The −1 i 1000 μg mL−1 (corresponding to 2.57 mM and 5.15 mM), reduced cell viability to 67% and 28%, respectively , GPr[FA], LPr[FA], and PPr[FA] were tested in vitro using the L929 murine fibroblasts as model cells in a cytotoxicity study. Our data confirmed the biocompatibility of [FA] and PPr[FA] at a concentration ranging from 0.03125 to 2.0 mM, and for GPr[FA] and LPr[FA], the range was from 0.03125 to 1.0 mM. Similar safety of FA was confirmed by Borges et al., who tested the effect of various isothiocyanates and phenolic compounds on the viability of L929 cells. Namely, after 72 h of treatment, FA, at the concentration of 500 μg mLectively . In anot0 mg L−1 , who shoL1 cells . Two indL1 cells  and cervL1 cells  were assL1 cells .The study presents the synthesis, identification, and characterization of three new FA amino acid derivatives, namely glycine propyl ester ferulate (GPr[FA]), L-leucine propyl ester ferulate (LPr[FA]) and L-proline propyl ester ferulate (PPr[FA]). The obtained compounds showed significantly improved solubility in water compared to unmodified ferulic acid. Additionally, the FA derivatives usually provided increased permeability of the active substance through porcine skin. The highest permeability was observed for PPr[FA] from ethanolic solution and hydrogel formulation. Furthermore, the high antioxidant activity of both the acceptor fluid collected after 24 h of permeation and the fluid obtained after skin extraction were demonstrated for all compounds. The total polyphenol content was usually higher in the acceptor fluid for the best permeating PPr[FA], while in the skin extracts it was least permeating for unmodified FA and GPr[FA]. The more effective percutaneous permeation of PPr[FA] may increase the probability of broader antioxidant action in deeper tissues. Interestingly, all the analyzed compounds accumulated in the skin, usually in its upper layer or just below it. This is also very beneficial because the accumulated active substances in the skin may have an antioxidant effect for a longer time, showing beneficial effects such as antiaging, anti-inflammatory, or photoprotective effects, among others. Therefore, the derivatives of FA and amino acid propyl esters after topical application can be an interesting alternative in skin protection and also systemic action. In addition to providing the active substance of ferulic acid, the presented derivatives are also a source of amino acids , which are important for the structure of the skin, for both the construction and strengthening of proteins and myofibril contractile proteins. Thus, they can enhance antiaging effects and attenuate skin wrinkles. It is worth emphasizing that ferulic acid and its amino acid derivatives have a little effect on the viability of L929 murine fibroblasts, which decreased to 80% for all compounds only at the highest concentration.


Several studies have shown that various concentrations of low-concentration atropine can reduce myopia progression and control axial elongation safely and efficiently in children. The aim of this study was to evaluate the effects of 0.02% and 0.01% atropine on ocular biometrics.N = 120) wore only SV spectacles. Ocular and corneal astigmatism were calculated using Thibos vector analysis and split into J0 and J45.Cohort study. 138 and 142 children were randomized to use either 0.02% or 0.01% atropine eye drops, respectively. They wore single-vision (SV) spectacles, with one drop of atropine applied to both eyes nightly. Controls (P < 0.05). Both anterior chamber depth (ACD) and ocular astigmatism (including J0) increased, and lens power decreased in the three groups . However, there were no differences in the changes in ACD, ocular astigmatism, and lens power among the three groups . Intraocular pressure (IOP), corneal curvature, ocular astigmatism J45, and corneal astigmatism (including J0 and J45) remained stable over time in the three groups . The contributions to SER progression from the changes in AL, lens and corneal power of the three groups were similar (P > 0.05). The contribution of AL change alone to the change in SER was 56.3%, 63.4% and 78.2% in the above corresponding three groups.The changes in cycloplegic spherical equivalent refraction (SER) and axial length (AL) were −0.81 ± 0.52D, −0.94 ± 0.59D, and −1.33 ± 0.72D; and 0.62 ± 0.29 mm, 0.72 ± 0.31 mm, and 0.89 ± 0.35 mm in the 0.02% and 0.01% atropine and control groups, respectively (all After 2 years, 0.02% and 0.01% atropine had no clinical effects on corneal and lens power, ocular and corneal astigmatism, ACD or IOP compared to the control group. 0.02% and 0.01% atropine helped to control myopia progression mainly by reducing AL elongation. Myopia, also known as short-sightedness, continues to worsen over time because of changes in lifestyle and behavior, leading to high myopia. Prominently, high myopia, also called pathologic myopia, is relevant to excessive axial length (AL) growth, which can exponentially increase a person's risk of developing sight-threatening eye diseases such as macular hemorrhage, retinal detachment, glaucoma and cataracts –3. ThereSeveral studies have shown that various concentrations of low-concentration atropine can reduce myopia progression and control axial elongation safely and efficiently in children –12. To dCurrently, only one 1-year study  found thThe research approach has been published previously and is briefly described here , 7. FourEligible children had the option of atropine or no atropine, and then the atropine groups were subsequently divided into either 0.01% or 0.02% in a double-blinded and randomized manner. This study conforms to the ethical norms and standards in the Declaration of Helsinki. The possible risks were thoroughly explained before the commencement of the treatment.One clinician examined all subjects, who was unaware of the experimental groups of each participant. The children in the control group wore the full-correction single-vision (SV) spectacles with the least negative power, consistent with the best visual acuity of long-term wearing. Both experimental groups were prescribed SV spectacles according to the same protocol as the control group. The children in the experimental groups were given one drop of atropine eye drops in both eyes every night at bedtime.After instilling one drop of compound tropicamide in both drops (0.5% tropicamide and 0.5% neo-synephrine)  4 times at 10-minute intervals. Cycloplegic autorefraction was performed 10 min after the last administration of the eyedrop. Taking three autorefraction measurements , the mean was obtained. The degree of myopia was expressed as the SER. Using Thibos vector analysis –18 calcuLP,BR-lens power using Bennett–Rabbetts method; L-axial length, Scv-spherical equivalent refraction at the corneal vertex ; K-corneal power, n = 1.336 (the refractive index of aqueous and vitreous humors); ACD-anterior chamber depth; cBR = 2.564 mm (distance between thin lens position and anterior lens surface).Here: R2 value was used to represent the proportion of the variance of the dependent variable predicted by the independent variable in the regression model. Statistical analysis was performed using the Empower software (www.empowerstats.com); X & Y Solutions, Boston, MA, United States) and SPSS 25.0 . Statistical significance was set at P < 0.05.Only data from the right eye was used for analysis. Continuous baseline variables were expressed as the mean ± SD and evaluated using an analysis of variance. The chi-squared test was used to compare the number of male and female. A generalized additive mixed model was used to estimate the longitudinal trend with time  for dependent variables such as SER, corneal curvature, ocular astigmatism (OJ0 and OJ45), and corneal astigmatism (CJ0 and CJ45), and differences in the rate of change among the three groups. The change represents the slope for each treatment group of dependent variables over time, and the change difference represents the difference in the slope of dependent variables over time between the groups. Multivariate linear regression analysis was used to detect the relationship between the dependent variable (change in SER) and independent variables . The adjusted In total, there were 138, 142, and 120 children in the 0.02% atropine, 0.01% atropine, and control groups, respectively . There wP < 0.05) (P > 0.05), and AL showed a tendency similar to that of SER every year.Over the two-year period, the changes in SER were −0.81 ± 0.52 D, −0.94 ± 0.59 D and −1.33 ± 0.72 D and changes in AL were 0.62 ± 0.29 mm, 0.72 ± 0.31 mm, and 0.89 ± 0.35 mm in the 0.02% and 0.01% atropine, and control groups, respectively. There were significant differences in the changes in SER and AL among the three groups  . In the P > 0.05). The ACD increased over time in all three groups, but the degree of change was similar  but no change in J45 .The ocular astigmatism changes were −0.38 ± 0.29 D, −0.47 ± 0.38 D, −0.41 ± 0.35 D and ocular astigmatism J0 changes were 0.19 ± 0.28 D, 0.22 ± 0.36 D, 0.18 ± 0.31 D in the 0.02%, 0.01% atropine and control groups, respectively. There was a small but significant increase in ocular astigmatism (including J0) . Considering the ocular biometric changes , sex, and age, the multiple linear regression model explained 80.6%, 77.1%, 84.7% of the SER change in the three groups (Model 4).The contributions of ocular biometric changes  to SER progression were summarized in This two-year cohort clinical trial showed that 0.02% atropine had a better effect on controlling myopia progression and axial elongation than 0.01% atropine. Additionally, no significant differences in the changes in corneal and lens power, ocular and corneal astigmatism, ACD, and IOP between the 0.02% atropine, 0.01% atropine, and control groups. Contributions to SER progression from AL, corneal and lens power in the three groups were similar, axial elongation contributed to most of the SER progression, and 0.02% and 0.01% atropine helped to control myopia progression mainly by reducing axial elongation.Several studies , 20, 21,Furthermore, 0.02% and 0.01% atropine did not affect corneal and ocular astigmatism (after vector decomposition) and ACD compared to with the control group in the current two-year study. LAMP study  also fouThe strength of this study was to include a control group. The human ethics committee advised that subjects were to be offered either atropine or no atropine, and double-blinded randomization is carried out only for the two active arms of the study. The control and test groups had similar demographic and clinical parameters. The subjects were recruited using identical inclusion criteria contemporaneously and from the same population. A limitation of the current study is that a changing trend in ocular biological parameters after atropine withdrawal was not observed. This topic is currently under further investigation.Use of 0.02% and 0.01% atropine had no clinically significant effect on corneal curvature, corneal and ocular astigmatism, lens power, ACD, or IOP. 0.02% and 0.01% atropine mainly help control myopia by reducing axial elongation over the two-year follow-up period.


Dendrocalamus sinicus, endemic to Yunnan Province of China, is the strongest woody bamboo in the world. In the present study, the MaxEnt model was performed to simulate the distribution of different types of D. sinicus in China and neighboring countries or regions. The results suggested that the suitable distribution range of “straight type”, the main type for cultivation and utilization, was 8°–30°N and 73°–122°E under the current climate conditions, while the potential distribution range of “bending type” was 6°–31°N and 79°–109°E. The two most key climate variables associated with distribution of “straight type” were “Temperature Annual Range” with 36.6% contribution rate and “Temperature Seasonality” (32.4%), while “Isothermality” (47.8%) and “Precipitation of Driest Month” (24.8%) for “bending type”. Under different climate change scenarios  and periods , the potential distribution area of the “straight type” were apparently different, indicating that the distribution area of D. sinicus will be affected significantly by climate changes in the future. Our findings would be not only beneficial to understanding limiting factors for natural distribution of D. sinicus, but also helpful for further germplasm conservation, introduction and cultivation planning of this rare woody bamboo.Climate change will significantly affect the distribution area of species. Through establishing distribution model, we can simulate the current and future potential distribution range and provide reference for the introduction and cultivation planning of rare or economic plants.  In geneDendrocalamus sinicus, endemic to southern and southwestern Yunnan, is the largest woody bamboo species documented in the world. It is a subtropical and tropical sympodium bamboo with a diameter at breast height (DBH) of 30 cm and a height of nearly 30 m. The peak period of bamboo shooting is from July to August, depending on the beginning date of rainy season (Phyllostachys edulis), which is the main economic bamboo species in eastern Asia , with about 80 genera and more than 1,500 species . Under ny season . The aveern Asia . Therefoal value .D. sinicus, two natural types of culm, namely “straight type” and “bending type”, are detected , National Science and Technology Infrastructure (http://www.nsii.org.cn/2017/home.php), Flora Reipublicae Popularis Sinicae (http://www.iplant.cn/frps), Global Biodiversity Information Facility (http://www.gbif.org) as well as our investigation and sample collection in the field, a total of 25 distribution locations of D. sinicus were obtained  (r = 0.8) and bio7 (r = − 0.9), then bio6 was dropped and both bio5 and bio7 were reserved based on the PCA result.In this study, bioclimatic variables from the WorldClim website (ignette)  was usedignette) . The envignette) . In ordehttps://www.worldclim.org/data/cmip6/cmip6_clim2.5m.html). The associated scenarios included the SSP1-2.6 (ssp126), SSP2-4.5 (ssp245), SSP3-7.0 (ssp370), and SSP5-8.5 (ssp585), following 2 in 2100) and SSP5-8.5  as future test scenarios according to Future climate projections were extracted from the Shared Socioeconomic Pathways (SSPs) of the BCC-CSM2-MR global climate  database in the Coupled Model Intercomparison Projects 6 (CMIP6) (https://github.com/marlonecobos/kuenm). The receiver operating characteristic curve (ROC) of all environmental variables was calculated, and the importance of different environmental variables was measured by Jackknife test, following For each type, we created 493 candidate models by combining 17 values of regularization multiplier , and all 29 possible combinations of five feature classes . The performance of the candidate model was evaluated on the basis of significance , omission rates (E = 5%), and model complexity . The best models were selected according to the significant models and the omission rate ≤ 5%. The models of Delta AICc ≤ 2 were selected as final models in this model set . The comD. sinicus.After kuenm R package screening, the feature class was LP and regularization multiplier was 2 in the final model parameter combination of the “straight type”. As for the “bending type”, the feature class was LQTH and regularization multiplier was 3. We rebuild the model using the optimized parameter combination. The AUC values of the “straight type” and “bending type” were 0.978 and 0.996  respectiD. sinicus, four most key climate variables affecting the geographical distribution were screened out using the MaxEnt model. As for the “straight type”, four key climate variables with a cumulative contribution rate of 94.8% were: Temperature Annual Range , Temperature Seasonality , Annual Precipitation  and Precipitation of Wettest Month   . On the 3; 9.9%) , with a 3; 9.9%) . Because3; 9.9%) , S3, cliD. sinicus were analyzed using the Jackknife test in the MaxEnt model. Within the “straight type”, the order of the four key climate factors was Temperature Seasonality (bio4), Temperature Annual Range (bio7), Annual Precipitation (bio12) and Precipitation of Wettest Month (bio13)  , based o (bio13) . As for  (bio13) , and the (bio13) . The resD. sinicus included 12 countries or regions under the current environmental conditions. The potential distribution range of the “straight type” was between 8°–30°N and 73°–122°E , Thailand (12.82%), and Bangladesh (3.94%)  . By and As for the “bending type”, its potential distribution range was between 6°–31°N and 79°–109°E , and theAs an excellent timber bamboo species and main type for industrial utilization and cultivation, the “straight type” is our concerning focus, and in this study, we predicted the potentially suitable distribution area of the “straight type” under SSP1-2.6 and SSP5-8.5 future climate change scenarios . Under tCompared with the current distribution, three types of the predicted suitable habitats, namely poorly, moderately and total highly suitable regions, exhibited different change trends under the climate change scenarios SSP1-2.6 and SSP5-8.5 in 2050. The total highly suitable regions displayed a decreasing trend, namely 27.42% and 8.19% under SSP1-2.6 and SSP5-8.5 scenarios respectively . But theOn the other hand, comparison with the current distribution, almost predicted suitable habitats under two climate scenarios in 2090 exhibited dramatically decreasing trends, except the highly suitable area slightly increased 0.59% under the SSP1-2.6 climate scenario in 2090 . MeanwhiD. sinicus has outstanding performance in producing timber. Meanwhile, because of narrow distribution and provenance scarcity, its germplasm conservation and introduction planning have raised extensive attentions since publication as a new species  for the “straight type” and Isothermality (47.8%) for the “bending type” respectively  and Xinping County (24°04′N), in which clumps could bear new shoots normally and the culms grew up to 20 cm in diameter. Moreover, approximate 10% introduced clumps bloomed and died in 1–2 years after cultivating. The above results suggested the strict conditions for the introduction and cultivation of D. sinicus. On the other hand, based on the MaxEnt model, we predicted the optimal climate conditions of highly suitable areas for “straight type” were: (1) the Temperature Annual Range was <23.9 °C; (2) the Precipitation of Wettest Month was >282 mm and (3) the Annual Precipitation was >1,418 mm, respectively. Compared with the empirical climate variables for highly suitable areas of “straight type”, our climate conditions based on MaxEnt model were more strict. However, our results were also more consistent with the actual outcomes, indicating our results had a higher reliability.On the aspect of introduction and cultivation planning of  habitat . The pre habitat . From 20 habitat . After 1The adaptation to climate and climate change is vital to plant growth, geographical distribution and biodiversity . In the D. sinicus. In addition, we should also realize that the MaxEnt model does not consider the factors such as soil and soil microorganisms at specific sites, so the accuracy of this prediction is limited.In summary, our results confirmed that the “straight type” had poorer heat resistance than “bending type”, which is important to instruct future introduction and cultivation planning of D. sinicus occurrence records and bioclimatic variables, the current and future suitable habitat of D. sinicus in China and adjacent regions was modeled using MaxEnt model for the first time. The bioclimatic variables of temperature annual range and isothermality were revealed to have crucial effect on D. sinicus distribution. In the next 70 years, the habitat suitability of this woody bamboo may be different with climate change. The prediction of this study is of strategic significance for further germplasm conservation, introduction and cultivation planning of this rare and precious bamboo species.Based on 10.7717/peerj.13847/supp-1Supplemental Information 1Click here for additional data file.10.7717/peerj.13847/supp-2Supplemental Information 2Click here for additional data file.10.7717/peerj.13847/supp-3Supplemental Information 3Click here for additional data file.10.7717/peerj.13847/supp-4Supplemental Information 4Click here for additional data file.10.7717/peerj.13847/supp-5Supplemental Information 5Click here for additional data file.10.7717/peerj.13847/supp-6Supplemental Information 6Click here for additional data file.10.7717/peerj.13847/supp-7Supplemental Information 7Click here for additional data file.10.7717/peerj.13847/supp-8Supplemental Information 8Click here for additional data file.10.7717/peerj.13847/supp-9Supplemental Information 9Click here for additional data file.