This study aimed to investigate the effect of a rollator on the prevention of falls in patients with Parkinson’s disease (PD) during outdoor walks.This study examined 30 community-dwelling patients with PD. Factors associated with falls were classified into clinical patient background, physical function, and psychophysiological function factors. The number of falls and subsequent injuries was observed over a period of greater than 6 months, if patients were using rollators while falls happened.Participants who used a rollator had a significantly lower fall rate, number of falls, and injury rate than those who did not use a rollator (p < 0.05).A rollator could protect patients with PD from falls. Additionally, when considering the use of a rollator for patients with PD, it is important to assess the patient’s physical and psychophysiological functions. Patients with PD were defined as those who had been diagnosed by a neurologist per the diagnostic criteria of the International Parkinson and Movement Disorder Society (MDS) This 6-month prospective cohort study was conducted by three medical groups. Approval for this study was obtained from the Ethics Committee of Mukogawa Women’s University . With the help of the research statement, the content and purpose of the study were explained to the participants in writing and orally. The participants provided consent to participate in the study by signing a research participation consent form.3The participants were asked whether they used rollator as a walking aid. Rollator users were asked whether they chose to use it because they felt the need for the same or because others recommended it. They were also asked to freely describe how they felt about using a rollator. Rollator nonusers answered with an open-ended question why they did not use a rollator. Rollator was defined as a walking aid consisting of left and right frames and a central tube connecting them that could be used alone, supported by hands or arms, and equipped with wheels at the lower ends of the left and right frames Clinical background information was obtained based on the following: history of falls, medication status, H&Y scale as an index of PD severity, the Movement Disorder Society-Sponsored Revision of the Unified Parkinson’s Disease Rating Scale (MDS-UPDRS), body mass index (BMI), the Barthel Index (BI), and the Functional Independence Measure (FIM) as an index of ADL. For MDS-UPDRS, the records were measured at the on-time, which was at least 1 h after medication. The subjects were also surveyed for the presence of orthostatic hypotension and dyskinesia using the MDS-UPDRS questionnaire. Regarding medication status, antiparkinsonian drugs were assessed using a conversion table to determine the levodopa equivalent daily dose (LEDD) Physical function was assessed using the following tests and measurements: the Timed Up and Go Test as a measure of balance, the 10-meter walk test as a measure of walking ability, grip strength, the Freezing of Gait Questionnaire (FOGQ), a hand-held dynamometer  to evaluate knee extension muscle strength, and an activity meter  to assess physical activity. Physical activity of the participants was measured by informing them to wear an activity meter on the waist for 1 week, except while bathing and sleeping. Participants who answered “at least once a week” to the third question of the FOGQ—“How much does your foot stick to the ground while walking, changing direction, or when starting to walk?”—were defined as having freezing of gait The psychophysiological functional assessment included five scales. The MMSE, Self-rating Depression Scale (SDS), and Modified Falls Efficacy Scale (MFES) assessed confidence in performing the movement without falling safely. The modified Gait Efficacy Scale (mGES) measured confidence in walking safely, and the Trail Making Test Part A (TMT-A) assessed attention and executive function.Patients with falls were followed up for 6 months from the date of examination and measurement. On the day the participants visited the facilities, they or their caregivers reported the frequency of falls since the date of previous hospital or facility visits. The data were recorded in their communications books and transferred to their medical records to ensure that the number of falls and injuries caused by falls could be assessed. Falls were defined as contact of the knees or hands with the ground or a lower point, not by one’s own will t-test, the Mann–Whitney test, the χ2 test, and the Fisher’s exact test. In addition, risk ratios and confidence intervals for falls and fractures were determined. Statistical analysis was performed using the IBM SPSS Statistics ver. 26.0 (IBM Corp.) with a significance level of 0.05.The participants were classified into two groups based on whether they used a rollator. The rate, frequency, and injury rate after a fall and clinical patient background, physical function, and psychophysiological function were analyzed using a two-sample 4The survey results revealed that 14 individuals used a rollator, whereas 16 did not. Nine (64 %) used a rollator because they felt that its use was necessary, whereas five (36 %) chose to use it because others recommended it. Regarding the use of rollator, eight (50 %) felt that the use was safe, six (38 %) felt that it was stable , and two (13 %) felt that it was functional . Answers to the open-ended question about why they did not use rollator were as follows: nine (56 %) answered that they did not feel the need; five (31 %) answered that they had functional problems ; and four (25 %) answered that they had appearance-related concerns (concern regarding the common use of rollator by older people and regarding others’ perceptions) (duplicate answers were available).In total, 20 (66.7 %) participants experienced falls; 19 (63.3 %) falls were during indoor walks, 13 (43.3 %) were during outdoor walks, and 12 (40 %) were during both indoor and outdoor walks.The clinical background of the participants is shown in A comparison of the fall rate, number of falls, and injury rate after falls between the two groups is summarized in 5In this study, 47 % community-dwelling patients with PD used a rollator. In the living environment in Japan, a rollator is usually used outdoors and rarely indoors. Notably, the participants in this study only used the rollator outdoors.The fall rate of the participants was 67 %, which is similar to that of a previous study of patients with PD, with a fall frequency of 40 %–70 % The patients in this study are enrolled in daycare facilities, day rehabilitation services, and home-visit rehabilitation services. It has been demonstrated that institutionalization is considered a milestone of disease progression in PD, and in some way, falls (another disease milestone) and institutionalization could be two different factors co-contributing to the advanced stage of PD There were no differences in patients’ clinical background in terms of age, gender, medication status, PD severity, ADL, and the presence of dyskinesia and orthostatic hypotension indicating a similar risk of falls in both groups.Orthostatic hypotension is present in 30–60 % of PD Physical function was not significantly different between the two groups. The purpose of rollator use was hypothesized to compensate for the reduction in lower extremity muscle strength and walking ability. However, this study found no significant differences in physical function between the two groups. Therefore, it has been suggested that patients with PD can use rollators regardless of the degree of gait freezing, lower extremity muscle strength, or the severity of PD. Although PD is known to have freezing of gait as a risk factor for falls, our finding that patients with freezing of gait can use a rollator constitutes important evidence.Regarding psychophysiological function, the rollator user group had a greater fear of falling and better attention and executive function than the rollator nonuser group. Previous studies In contrast, there was no significant difference in the rate and number of indoor falls between the two groups, suggesting that the two groups had the same likelihood of falling. However, some patients with PD did not use rollator. Therefore, nonusers of rollator were also examined in this study. In the rollator nonuser group, the fear of falling as well as attention and executive function decreased. In healthy older adults, it has been reported that the fear of falling is felt more strongly by those who fall frequently In older people, attention and executive function has been reported to be associated with falls The present study suggests that attention to falls, rather than physical function, such as the degree of gait freezing, determines rollator use. Patients with PD are more likely to be distracted by the task or environment; therefore, they need better attention and executive function to use a rollator. In contrast, it has been suggested that people with poor psychophysiological functioning have difficulty using a rollator. Therefore, it is important to evaluate physical and psychophysiological functions when considering the use of rollator.The first limitation of this study is that the study was conducted at only three institutions, and the number of cases was limited. It is therefore uncertain whether the results accurately reflect the population. Second, multivariate analysis could not be performed owing to the small number of cases; thus, the influence of other factors could not be eliminated. Third, the impact of the rollator models on the results was not analyzed. So, it is unclear which is more fall-protective than a U-step rollator. Finally, it was found that the severity of PD was high among participants; therefore, it is unclear whether similar results could be obtained in a population with a lower PD severity. Further studies are required to include more cases and various PD severities in many facilities.6This study examined the effect of rollator on preventing falls in patients with PD during outdoor walks. It was found that the fall rate, number of falls, and injury rate after a fall were lower in those using rollator. Therefore, it was suggested that the use of a rollator could prevent falls.The patients’ clinical background and physical function were not significantly different between the two groups. Regarding psychophysiological function, the rollator user group had a greater fear of falling and better attention and executive function.This study suggests that attention to falls, rather than physical function, such as the degree of gait freezing, determines rollator use. Patients with PD are more likely to be distracted by the task or the environment; therefore, they need better attention and executive function to use rollator.In contrast, it has been suggested that people with poor psychophysiological functioning have difficulty using rollator. Therefore, it is important to evaluate physical and psychophysiological functions when considering the use of rollator.In conclusion, this study provides important evidence for the fall-preventive effects of rollators in community-dwelling patients with PD and suggests that better attention and executive functions are required for adaptation to a rollator.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


Identifying patients that are less resilient to rehabilitation may promote a centered-patient approach for an individualized and optimized rehabilitation care.Walk speed measured under dual-task conditions (neurocognitive tasks) could reflect patient performance in real-life. Rehabilitation programs are effective in increasing walk speed, but few studies have evaluated the associations between geriatric factors and rehabilitation efficacy under dual-task conditions. Our objective was to investigate the association between geriatric factors and an increase in dual-task walk speed (threshold of 0.1 m/s), after a multidisciplinary rehabilitation program. We performed a retrospective cohort study that included patients aged 75 years and over, who underwent a complete rehabilitation program and who had a neurocognitive assessment at baseline. The primary outcome was the increase in the dual-task  walking speed between pre- and post-rehabilitation assessments. In this study, 145 patients were included, with a mean age of 83.6 years old. After rehabilitation, dual-task walk speed increase in 62 (43%) patients. In multivariate analysis, the following factors were associated with an increase in dual-task walk speed: IADL (OR 2.50, 95% CI ,  Walk speed, often called the sixth vital sign, could be considered as a clinically relevant health biomarker . Indeed,Assessing walk speed under dual-task conditions  could approach locomotion abilities under real-life conditions . In addiSeveral studies have shown that it was possible to create rehabilitation programs to increase dual-task walk speed in older patients. . HoweverConsidering this, the aim of this study was to explore the association between an increase in dual-task walk speed after rehabilitation and the geriatric factors, identified by a geriatric assessment.This retrospective study was conducted in a tertiary, 2600-bed, university-affiliated center, at the Clinical Gerontology Department, Nantes, France. In this department, after a geriatric assessment, a multidisciplinary care program is proposed to older patients who have fallen . The mulFrom October 2010 to March 2019, all patients who were 75 years of age or older were retrospectively included if they met all the following criteria: (i) participation rehabilitation program, (ii) presence of a neurocognitive assessment . For each patient, only the first rehabilitation program participation was considered and there were no other exclusion criteria.All patients were first seen in consultation by a geriatrician who performed a geriatric assessment. Following this consultation, the rehabilitation program was proposed to all patients who agreed to come to the Gerontology Department twice a week . Patient® type mat , which quantifies the spatio-temporal data of the dual-task walk. For dual-fluency walk speed assessment, the patient was asked to walk at normal walk speed, while naming as many animals as possible. The assessment was completed with stabilometry, balance and muscle performance tests. The grip strength was evaluated with a Jamar® dynamometer  and consisted in measuring the dominant hand grip .At the beginning and at the end of the rehabilitation program, walk speed was measured with the GAITRiteThe primary endpoint of the study was the effectiveness of the rehabilitation, defined by a gain of at least 0.1 m/s in the dual-fluency walk speed, between pre and post-rehabilitation. We considered this clinically meaningful change of 0.1 m/s because it was associated with a significant gain of 1 point on the instrumental autonomy scale  at 6 months  and it iThe primary outcome was retrospectively collected from the hospital electronic medical record. The following data were also collected because of their association with walk speed and/or fall risk: age, gender , historyt-test. Univariate and multivariate logistic regression analyzes were performed to determine associations between variation in dual-fluency walk speed and other variables. The variables included in the logistic regression model for multivariate analysis are those variables that were significantly associated with the variable of interest in univariate analysis and/or with p < 0.2, or that are clinically relevant. In addition, strongly correlated variables (Pearson’s correlation coefficient p > 0.7) were not included in the model. Relative risks are expressed as odds ratios with 95% confidence intervals. All p-values < 0.05 are considered statistically significant. Statistical analyzes were performed on SPSS software version 15.0 .Patients’ characteristics were described using means and standard deviations for continuous variables and frequencies and percentages for binary variables. The variable of interest was the change in dual-fluency walk speed between initial and final evaluations, a binary variable defined as follows: gain less than 0.1 m/s on the one hand, variation greater than or equal to 0.1 m/s on the other hand. The comparison between the two groups was made with the Chi-square test for the binary variables. The normality of distribution of continuous variables was determined by the Shapiro-Wilk test. The variables of non-normal distribution were compared by the Mann-Whitney test, the variables of normal distribution by the student’s Because of the retrospective cohort study design, no sampling size calculation was performed and all the patients included were considered in the statistical analysis.The study was conducted in accordance with the ethical principles of the 1983 Declaration of Helsinki and received a positive approval from the Groupe Nantais d’Ethique dans le Domaine de la Santé. Patient identification information was recorded in a secure electronic database.We identified 162 patients  who partn = 62) and those without (n = 83) are described in p = 0.028), the plasma vitamin D level at baseline, which was lower in responders (p = 0.015), the initial dual-fluency walk speed, which was slower in responders (p = 0.001) and the time interval between initial and final assessments , which was shorter in responders (p = 0.001). No significant differences were found for the other parameters.Walk speed increased by at least 0.1 m/s from baseline in 62 (43%) in 145 patients. The characteristics of the patients with an increase in walk speed (p = 0.021), initial dual-fluency walk speed  and the time interval between initial and final assessments .The results of the univariate and multivariate logistic regression analysis are presented in p = 0.009), initial blood vitamin D level , severe sarcopenia , depression , number of drugs , initial dual-fluency walk speed  and time interval between initial and final assessments .In multivariate analysis, the seven following factors were significantly associated with an increase in dual-task  walk speed after a complete rehabilitation program: instrumental autonomy, assessed by the IADL scale  could approach locomotor abilities under real-life conditions . MoreoveIn this study, less than half of the patients had an increase in dual-task walk speed after rehabilitation. This proportion seems lower than that observed in the literature. In particular, Uemura et al. found functional improvement in two thirds of patients receiving identical rehabilitation management in a population of 179 elderly patients . HoweverWe observed a better rehabilitation efficacy in patients with a higher IADL score at inclusion. We assume that this result could be explained by the presence of a greater physiological reserve in patients with a high IADL score at baseline, that could potentiate the impact of rehabilitation. Indeed, the IADL score is a scale strongly associated with the evolution of morbidity and mortality . Thus, pOur study showed a greater effectiveness of rehabilitation in patients who were initially vitamin D deficient. This result is counter-intuitive considering the impact of vitamin D deficiency on the central nervous system and the musculoskeletal system . To explIn this study, patients with a slow initial dual-fluency walk speed benefited more from rehabilitation. We assume that this result can be explained by a combination of several elements. First, we chose a threshold of 0.1 m/s and it is more likely to observe an increase in walk speed in patients whose initial walk speed was slower, as the possibility of improvement is greater. Secondly, in this study, the population analyzed had few comorbidities (low Charlson index) and a relative preserved autonomy . The previous variable are known to be associated with successful ageing, less frailty and better resilience . Thus, wWe found that rehabilitation was less effective in patients with severe sarcopenia. This is not surprising because sarcopenia is a global quantitative and/or qualitative alteration of muscles, associated with a decrease in muscle strength  and the We found a significant association between the presence of depression and the effectiveness of rehabilitation. It is well known that patients with depression have posture, balance and walk impairment, including a reduced walk speed, and studies have shown the effectiveness of physical exercise in improving motor skills in depressed patients ,32,33. TIn our study, patients who have a greater number of drugs respond better to rehabilitation. This is not surprising because Umegaki et al. showed an association between poly-medication and slower walk speed in elderly adults . This asWe found a significant association between a shorter time between pre- and post-rehabilitation assessments and better rehabilitation effectiveness. This result suggests an association between the frequency of rehabilitation and its effectiveness .Our study found no association between executive syndrome, assessed by the FAB, and the effectiveness of rehabilitation. We believe that this result is mainly related to a too great homogeneity of the executive profile of the studied patients. Indeed, it is well described that not all cognitive and executive functions are equally important in walk control . In our We did not find an association between MMSE and magnitude of improvement in walk speed after rehabilitation. This could be explained by the fact that a small proportion of the population included had neurocognitive disorders (high MMSE and high ADL and IADL). Indeed, in a prospective cohort study, Friedman et al.  did showThis study has limitation. First, it lacks power due to the small number of patients and their heterogeneity, which may alter our conclusions by calling into question statistically significant associations. Second, this was a monocentric study, with a retrospective design. Because of this, we were not able to account for the possible existence of potential confounding factors, such as anticholinergic drugs impact, fear of fall, or stabilometry and balance control analysis . In addiRehabilitation may allow an improvement in the dual-fluency walk speed in older adults. Identifying patient profiles that respond to rehabilitation may allow for better referral of patients in the geriatric care system and adaptation to the increasing demand for care. Further studies are called for exploring more precisely the links between functional improvement and different rehabilitation interventions.


Background: Androgen receptor (AR) expression has emerged as a potential prognostic and predictive marker in patients with triple negative breast cancer (TNBC). We conducted a retrospective analysis to evaluate pathologic complete response (pCR) rates, disease-free survival (DFS) and overall survival (OS) in patients with AR positive and AR negative TNBC treated with neoadjuvant chemotherapy.Methods: 107 patients with TNBC subtype, treated with neoadjuvant chemotherapy between June 2006 and March 2016 were evaluated for AR expression. Androgen receptors were evaluated by immunohistochemical staining  using whole tissue sections from archived paraffin-embedded formalin-fixed (FFPE) blocks. AR positive was defined as ≥10% nuclear stained cells. Correlation of AR expression was examined with age, BMI, race, menopausal status, tumor grade, tumor size, and lymph node involvement, and response and outcomes. Univariate and multivariate analyses were performed to determine an association with AR expression and pathologic response and survival outcomes.Results: Fifty-eight patients with available tumor specimens were stained, with twenty (34.5%) being AR-positive and thirty-eight (65.5%) being AR negative. Median age was 49 years and median follow up was 5.7 years. All patients received anthracycline based neoadjuvant chemotherapy with 13 patients (23%) receiving an additional platinum chemotherapy. BRCA mutation positivity was 7% for the entire group. No differences in age, menopausal status, BMI, race, tumor size and lymph node involvement were observed between the two groups. However, there was a statistically significant difference in tumor grade between the two groups (p=0.008). Overall pCR rate was 28% with no difference between the two groups . There was no statistically significant difference in median DFS (5.9 years vs 5.2 years (p=0.94) and median OS  between the AR positive and AR negative groups.Conclusions: Our study did not find an association of AR status and the pathologic responses or survival outcomes in patients with TNBC treated with neoadjuvant chemotherapy. Further studies exploring the prognostic and predictive role of AR in patients with TNBC are warranted. Triple negative breast cancer (TNBC) is a subtype of breast cancer that accounts for 15-20% of all invasive breast cancers Due to the aggressive biology and fewer options of targeted therapy for TNBC patients, identifying a target aimed treatment that can improve outcomes in these patients is especially important. Androgen receptors (AR) are a class of nuclear steroid receptor that are overexpressed in many major breast cancer phenotypes at an approximate rate of 70-90%. Its' expression in TNBC varies from 10-50% p-value of ≤ 0.05 was considered statistically significant for all of the analyses.This study identified patients with non-metastatic TNBC at the Froedtert Clinical Cancer Center who were treated with neoadjuvant chemotherapy between June 2006 and March 2016. All the TNBC patients included were eligible for neoadjuvant chemotherapy and received the standard anthracycline and taxane containing regimen. The use of platinum was on the discretion of the treating physician. A retrospective chart review was performed, and data was supplemented from the institution's electronic medical record system. Androgen receptors were evaluated by immunohistochemical staining  using whole tissue sections from archived paraffin-embedded formalin-fixed (FFPE) blocks. AR positivity was defined by ≥10% nuclear staining of tumor cells A total of 107 patients with non-metastatic TNBC were identified in this study. Of these patients, 58 patients had available tumor specimens for AR testing, with 20 (34.5%) being AR-positive and 38 (65.5%) being AR negative. Twenty-five patients were premenopausal and 33 were post-menopausal. Patient race comprised: 41 Caucasian, 16 African American, and 1 Hispanic (Table All 58 patients received standard anthracycline and taxane based neoadjuvant chemotherapy with 13 patients (23%), 6 in AR positive group and 7 in AR negative group, receiving an additional platinum chemotherapy. The studies were conducted before the practice of adjuvant capecitabine. Therefore, no patient received capecitabine. BRCA mutation positivity was 7% for the entire group. The average tumor size for the 58 tumor specimens was 3.8 cm ± 1.8 cm, with 14 patients having a tumor larger than 5 cm. Twenty-eight patients (48.2%) had lymph node involvement before surgery with 10 in AR positive group and 18 in AR negative group (Table p=0.56).The overall pCR rate was 28% using standard anthracycline and taxane +/- platinum containing regimens. There was no statistically significant difference between the two groups, with the AR positive group having a pCR rate of 30% and the AR negative group having a pCR rate of 26% (p=0.94).The median DFS was 5.4 years . The AR positive cohort had a median DFS of 5.9 years , whereas the AR negative cohort had a median DFS of 5.2 years , however this difference was not statistically significant (p=0.98).The median OS was 5.7 years . The AR positive cohort had a median OS of 6.2 years , whereas the AR negative cohort had a median OS of 5.4 years . There was also no significant difference between the two groups in terms of overall survival  and overall survival  among AR positive patients. Upon breast cancer subtype breakdown, AR positivity showed similar results of improved prognosis in both ER positive and TNBC patients There has also been growing evidence that AR expression may be a clinical predictor of pCR to chemotherapy. Loibl et al evaluated AR expression and the pCR rate in the phase III GeparTrio trial which showed the pCR rate was 25.4% in patients with AR negative breast cancer but only 12.8% in patients with AR positive breast cancer Our study had several limitations. The first was the AR assay method. Like other clinical studies, we used the immunohistochemical stain method to assay the AR protein expression. However, Lehmann et al reported a study of the use of AR antagonist enzalutamide and PI3K inhibitor taselisib in patients with AR positive metastatic TNBC, where the use of molecular profiling to identify the luminal androgen receptor (LAR) subtype may be a better way to identify patients to benefit from antiandrogen therapy


Hepatitis B virus (HBV) infection is a serious global health issue with more than 250 million chronic carriers. It causes liver diseases such as chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). Persistent suppression of the HBV surface antigen (HBsAg) is necessary for a functional cure of chronic hepatitis B (CHB) virus infection. However, this can hardly be achieved with currently approved drugs. Antibody treatment against HBsAg has shown promise in restoring HBV-specific immune responses and promoting HBV cure. To achieve long-lasting HBsAg suppression, we used an advanced mRNA drug to encode the genes of three anti-HBsAg antibodies, G12-scFv, G12-scFv-Fc, and G12-IgG. Antibody-encoding mRNA-lipid nanoparticles (LNPs), mL (G12-scFv-Fc) and mL (G12-IgG), substantially reduced serum HBsAg levels in treated mice within 30 days after a single dose. In contrast, exogenous antibodies lost effect on reducing HBsAg or HBV DNA levels 9 days postadministration. The high affinity of anti-HBsAg antibodies and the adjuvant activity of mRNA-LNPs resulted in long-term HBsAg seroclearance, which could contribute to the reestablishment of the immune system in HBV carriers. These findings highlight the great potential of antibody-encoding mRNA molecules against CHB infection. Therapeutic antibodies have become the predominant treatments for various pathological states such as cancers and autoimmune and metabolic diseases . CompareIn vitro-transcribed mRNA (IVT-mRNA) synthesis  virus infection . Hepatit25–To deal with such a problem, we used anti-HBsAg antibody-encoding mRNA-lipid nanoparticles (LNPs) in our study. Three anti-HBsAg antibodies  were encoded by mRNA and encapsulated in LNP. The mRNA-LNP of G12 provided persistent passive immunization for at least 30 days. Moreover, mRNA was mainly delivered to the hepatocytes by LNP . These c5 293T cells  was applied for G12-IgG production.Four mRNAs, scFv mRNA, G12-scFv-Fc mRNA, G12 heavy-chain (HC) mRNA, and G12 light-chain (LC) mRNA, were synthesized. Each mRNA comprised a 5′ cap1 structure, 5′ untranslated region (UTR), coding sequence, 3′ UTR, and a poly(A) tail . G12-scF3T cells . To opti3T cells . This rein vivo, we prepared three mRNA-LNPs, mL (G12-scFv), mL (G12-scFv-Fc), and mL (G12-IgG), with particle sizes of 149.6 nm, 154.0 nm, and 186.1 nm and zeta potentials of +13.0 mV, +13.8, and +14.3 mV, respectively (see Cmax]) 15 min after administration. We detected 28.9 ± 2.6 μg/mL G12-scFv, 90.3 ± 32.6 μg/mL G12-scFv-Fc, and 95.3 ± 20.5 μg/mL G12-IgG in the serum, respectively  provided a higher antibody expression level than other LNPs. As shown in To investigate the production of G12 antibody ectively  to C. Thectively  to F. Th10.1128/mbio.01612-22.1FIG S1FIG S1, TIF file, 0.2 MB.Characteristics of G12 IVT-mRNA-LNP. Particle size distribution of mL (G12-scFv) (A), mL (G12-scFv-Fc) (C), and mL (G12-IgG) (E). Zeta potential distribution of mL (G12-scFv) (B), mL (G12-scFv-Fc) (D), and mL (G12-IgG) (F). Download Copyright © 2022 Chen et al.2022Chen et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the in vivo. Antibodies were consecutively synthesized unless the mRNA was degraded. The half-time of endogenous antibodies was positively correlated with that of mRNA -LNP, the adeno-associated virus (AAV)/HBV mouse model with persistent HBsAg expression was established. Mice were intravenously (i.v.) injected with single dose of either 2.5 mg/kg mL G12-scFv or 6.7 mg/kg G12-scFv . Althougt 2 days  and D. Ht 2 days , and aftt 2 days . On the t 2 days  and D, a 15 days .P < 0.05) (P < 0.05) (10) in serum was observed (P < 0.01)  . Compare10.1128/mbio.01612-22.2FIG S2FIG S2, TIF file, 0.5 MB.mL (G12-scFv) decreased HBV DNA in an AAV/HBV mouse model. (A) AAV/HBV mice were injected with 2.5 mg/kg mL (G12-scFv) or 6.7 mg/kg G12-scFv. (B) Kinetics of HBV DNA levels in 30 days posttreatment. HBV DNA levels were quantified on day 1 (C), day 2 (D), day 5 (E), day 9 (F), day 12 (G), day 15 (H), day 20 (I), day 26 (J), and day 30 (K) posttreatment. Download Copyright © 2022 Chen et al.2022Chen et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the 50) of G12-scFv than that of G12-scFv-Fc and G12-IgG (Neither mL (G12-scFv) nor G12-scFv had a significant effect on HBsAg clearance, despite the statistical difference from the untreated group. This result was due to the low expression of G12-scFv after administration of mL (G12-scFv) and the rapid clearance and lower 50% effective concentration (EC G12-IgG .10.1128/mbio.01612-22.5FIG S550 determinations of G12-scFv (A), G12-scFv-Fc (B), and G12-IgG (C). Download FIG S5, TIF file, 0.1 MB.Affinity of G12 antibodies to HBsAg. ECCopyright © 2022 Chen et al.2022Chen et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the P < 0.0001) (P < 0.001) (P < 0.05) (P < 0.05) and day 5 (P < 0.01) (P < 0.01)  or 6.7 mg/kg G12-scFv-Fc was administered . During  0.0001)  and D, f< 0.001) . From da< 0.001) . In cont< 0.001) . G12-scF < 0.05) . mL (G12 < 0.01) . Also, i < 0.01) .10.1128/mbio.01612-22.3FIG S3FIG S3, TIF file, 0.5 MB.mL (G12-scFv-Fc) decreased HBV DNA in AAV/HBV mouse model. (A) AAV/HBV mice were injected with 2.5 mg/kg mL (G12-scFv-Fc) or 6.7 mg/kg G12-scFv-Fc. (B) Kinetics of HBV DNA levels in 30 days posttreatment. HBV DNA levels were quantified on day 1 (C), day 2 (D), day 5 (E), day 9 (F), day 12 (G), day 15 (H), day 20 (I), day 26 (J), and day 30 (K) posttreatment. Download Copyright © 2022 Chen et al.2022Chen et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Unlike the short effect caused by G12-scFv-Fc, mL (G12-scFv-Fc) induced a persistent suppression of serum HBsAg and HBV DNA levels. Although the serum concentrations of G12-scFv-Fc in the G12-scFv-Fc group are higher than that in the mL (G12-scFv-Fc) group  and E, mP < 0.0001), day 5 (P < 0.001), day 9 (P < 0.01), day 12 (P < 0.05), and day 15 (P = not significant)  (10 on day 15 (mL G12-IgG) or G12-IgG was given to C57 mice at 2.5 mg/kg and 6.7 mg/kg, respectively . During ificant) . With pr12-IgG orn day 15 . On day n day 15 . Noticean day 15 .P < 0.01) (P < 0.05) and day 5 (P < 0.01) (P < 0.05) (G12-IgG reduced HBV DNA levels immediately after treatment ( < 0.01) , followe < 0.01) . mL (G12 < 0.01) . It is n < 0.05) . Further10.1128/mbio.01612-22.4FIG S4FIG S4, TIF file, 0.4 MB.mL (G12-IgG) decreased HBV DNA in AAV/HBV mouse model. (A) AAV/HBV mice were injected with 2.5 mg/kg mL (G12-IgG) or 6.7 mg/kg G12-IgG. (B) Kinetics of HBV DNA levels in 30 days posttreatment. HBV DNA levels were quantified on day 1 (C), day 2 (D), day 5 (E), day 9 (F), day 12 (G), day 15 (H), day 20 (I), day 26 (J), and day 30 (K) posttreatment. Download Copyright © 2022 Chen et al.2022Chen et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Although mL (G12-IgG) took a while to block HBsAg, it was impressive that it could exert an increasingly consistent effect on reducing the HBsAg level. mL (G12-IgG) exhibited a more persistent effect on HBV seromarkers despite the higher serum concentration provided by G12-IgG.HBsAg clearance has been recognized as a long-term goal of anti-HBV therapy. Due to the low efficiency of HBsAg downregulation and the high frequency of viral rebound , current31–in vivo. The mRNA-LNPs and antibodies for G12-scFv, G12-scFv-Fc, and G12-IgG were prepared in vitro. In terms of HBsAg serum clearance, G12-IgG had the best efficacy, followed by G12-scFv-Fc and G12-scFv. This result was consistent with their serum concentration, terminal elimination half-life (t1/2), and EC50 against HBsAg. The efficacy of mRNA (G12)-LNPs on HBsAg serum clearance was highly correlated with serum concentration level of synthesized G12 antibodies on the first 2 days, with efficacy ranking as mL (G12-scFv-Fc), mL (G12-IgG), and mL (G12-scFv). In addition, mL (G12-IgG) and mL (G12-scFv-Fc) provided comparable efficacy in HBsAg reduction from day 9 to day 30, although the serum concentration level of synthesized G12-scFv-Fc was higher in the mL (G12-scFv-Fc) group than that in the mL (G12-IgG) group, consistent with the highest affinity of IgG antibodies to HBsAg.Our study used lipid nanoparticles (LNPs) to deliver mRNA encoding G12 antibodies in vivo. Consequently, mRNA drugs were more effective against HBV than G12 antibodies in a mouse model. The result indicated that mRNA-LNP provided a potent adjuvant activity for passive immunization, which may benefit HBsAg clearance. mRNA molecules could be detected by Toll-like receptor 7 (TLR7) and TLR8 -LNPs exerted more durable HBsAg and HBV DNA suppression and TLR8 . Also, tand TLR8 . Many stTo achieve a functional cure of CHB, G12 antibody-encoding mRNA-LNPs were insufficient. The “sandwich” strategy combines an anti-HBsAg neutralizing antibody with an antiviral drug followed by vaccines for specific active immunization . Shi et The human IgG1 monoclonal antibody G12 against HBsAg was produced as reported previously .4S)3 linker, and the G12-IgG VH domain. Then, the gene was cloned into the pComb3x vector with the N-terminal signal peptide (MKKTAIAIAVALAGFATVAQA) and two C-terminal tags (His6 and FLAG). After the G12-scFv gene sequence was validated, a single fresh colony transformed by HB2151 cells was added to 2 mL of 2× yeast extract tryptone [2YT]) medium (100 μg/mL ampicillin and 2% [wt/vol] glucose), shaken at 250 rpm at 37°C for 4 h, and then transformed with 500 mL of superbroth (SB) medium (100 μg/mL ampicillin). When the optical density at 600 nm (OD600) of culture media reached 0.6 to 0.8, the culture media were incubated overnight at 30°C at 250 rpm with 1 mM isopropyl-1-thio-β-d-galactopyranoside (IPTG). Bacteria were collected at 8,000 rpm for 10 min and lysed by polymyxin B (Sigma-Aldrich) at 30°C for 0.5 h. The supernatant was generated by centrifuging at 8,000 rpm for 10 min and then loaded over nickel-nitrilotriacetic acid resin according to the manufacturer's instructions .The G12-scFv gene was constructed with the G12-IgG VL domain, a (GThe G12-scFv-Fc gene (synthesized by Azenta Life Sciences), composed of G12-scFv and human IgG1 Fc, was subcloned into the pSecTag 2B vector. Once the sequence was confirmed, the G12-scFv-Fc gene was transfected to Expi293 by polyethyleneimine . After 6 days, supernatants were harvested at 2,500 rpm for 15 min and filtered with a 0.22-μm vacuum filter. The recombinant protein was purified using protein G resin . Protein purity was estimated by SDS-PAGE, and protein concentration was measured by the NanoDrop 2000 spectrophotometer (Thermo Fisher).in vitro transcription (IVT) by T7 RNA polymerase. In brief, the coding sequence of the G12 antibody and the flanking 5′ and 3′ UTRs were subcloned into the pUC57 vector. The DNA templates for transcription were prepared by PCR. The poly(A) coding sequence was introduced during PCR. The PCR product was used for the subsequent in vitro transcription. The cap1 structure was introduced to the 5′ ends of mRNA during transcription using the Cap1 analog . Pseudouridine was used to substitute uridine. Reaction mixtures were incubated at 37°C for 2.5 h, followed by DNase I  treatment for 30 min. Finally, mRNAs were purified by oligo(dT) affinity chromatography.The mRNAs were synthesized using sn-glycero-3-phosphocholine [DSPC]/cholesterol/DMG-PEG2000. mRNA was diluted in 50 mM citrate buffer (pH 4.0). For G12-IgG production, LC mRNA and HC mRNA were mixed in the same mRNA solution at the mass ratio of 1:2 (LC mRNA to HC mRNA). The lipid mixture was mixed with mRNA solution at the volume ratio of 1:3 using a microfluidic mixer (Precision NanoSystems). The N/P ratio was about 6:1. LNPs were immediately diluted with an equal volume of 50 mM citrate buffer (pH 4.0) and then dialyzed for about 16 h. Using Amicon ultracentrifugal filters (Millipore), formulations were concentrated and passed through a 0.22-μm filter. All LNP formulations were characterized for particle size, RNA encapsulation, and endotoxin. The size of the particle was characterized using Zetasizer Nano ZS ZEN3600 . mRNA encapsulation efficiency was determined by the Quant-it RiboGreen RNA assay (Thermo Scientific).LNP formulation was prepared as previously described . The lip2 at 37°C. Cells were seeded at a density of 2 × 105 cells per well on a 12-well plate. mRNAs were transfected by Lipofectamine 2000 (Thermo) at a ratio of 1:1.5 (mRNA/Lipofectamine 2000) for 6 h and kept incubated for another 18 h until detection.293T cells were cultured in high-glucose Dulbecco’s modified Eagle medium  supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (P/S) (Gibco) in 5% COC57 mice were divided into groups (5 mice per group). Mice were injected i.v. with mRNA-LNPs at a dose of 2.5 mg/kg or antibodies at 6.7 mg/kg. Blood samples were then taken from the periorbital venous sinus at indicated time points  and centrifuged at 4,000 rpm for 10 min for the plasma collection.10 AAV/HBV1.3 . After 2 weeks, an AAV/HBV mouse model  were coated with 100 ng/well HBsAg  in phosphate-buffered saline (PBS) at 4°C overnight and blocked with 3% milk (wt/vol) diluted in PBS at 37°C for 1 h. Cell supernatant or plasma was serially diluted in 1% bovine serum albumin (BSA) buffer. Plates were washed 3 times in PBS-T (0.05% Tween 20 in PBS) and then incubated with diluted samples at 37°C for 1.5 h. Then, plates were incubated with secondary antibody, horseradish peroxidase (HRP)-conjugated anti-human IgG Fc antibody (Thermo Fisher) for IgG antibody and Fc recombinant protein, and mouse anti-His antibody (GenScript) at 37°C for scFv antibody at 37°C for 45 min after washing for 5 times in PBS-T. Plates were washed 5 times and then incubated with ABTS  at 37°C for 15 min. Analysis was quickly conducted at wavelength of 405 nm by a microplate reader (BioTek).HBsAg levels were rationed by the Roche cobas 6000 . The HBV DNA levels were quantified by the HBV DNA quantitative fluorescent diagnostic kit .t test and one-way analysis of variance (ANOVA). The results were shown as mean ± standard deviation (SD) (n ≥ 3). Statistically, significant differences were defined as *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001.All data were analyzed using GraphPad Prism 8 . Statistical analysis was performed by unpaired All the procedures related to animal handling, care, and treatment were performed and approved by the Ethics Committee of the School of Basic Medical Sciences at Fudan University according to the recommendations in the Guide for the Care and Use of Laboratory Animals, Fudan University.


There are three main types of cancer in the female reproductive system, specifically ovarian cancer (OVCA), endometrial cancer (EC), and cervical cancer (CC). They are common malignant tumors in women worldwide, with high morbidity and mortality. In recent years, androgen receptors (ARs) have been found to be closely related to the occurrence, progression, prognosis, and drug resistance of these three types of tumors. This paper summarizes current views on the role of AR in female reproductive system cancer, the associations between female reproductive system cancers and AR expression and polymorphisms. AR regulates the downstream target genes transcriptional activity and the expression via interacting with coactivators/corepressors and upstream/downstream regulators and through the gene transcription mechanism of “classical A/AR signaling” or “non-classical AR signaling”, involving a large number of regulatory factors and signaling pathways. ARs take part in the processes of cancer cell proliferation, migration/invasion, cancer cell stemness, and chemotherapeutic drug resistance. These findings suggest that the AR and related regulators could target the treatment of female reproductive system cancer. Cancers of the female reproductive system mainly include ovarian cancer (OVCA), endometrial cancer (EC), and cervical cancer (CC). The 2020 Global Cancer Report shows that these three cancers are the most common malignant tumors in women worldwide . Tumors The AR is a member of the nuclear receptor superfamily and is a well-defined ligand for A. Target gene expression is activated between A and AR through the gene transcription mechanism of “classical A/AR signaling”  or “non-In females, A is mainly produced by the ovaries and adrenal gland. Biologically active A in the body include testosterone (T) and dihydrotestosterone . T can convert to DHT by 5α-reductase. DHT is the most biologically active form of endogenous androgen, and its binding affinity to AR is 2–5 times higher than that of T. A is closely related to female reproductive physiology and pathology, and the main function of A is realized through combination with AR. Furthermore, the level of other androgens links to gynecological cancers. The previous clinical studies found the DHEA, androsterone, and etiocholanolone concentrations were lower in the OVCA patients compared with the age-matched group . AnotherAR has 87 kDA-N terminals truncations (absence of the first 187 amino acids), produced by in vitro proteolysis. AR gene is located on X chromosome (loci: Xq11-q12) and consists of eight exons encoding a 110 kDa protein composed of 919 amino acids . AR consUnliganded bound AR binds to chaperone complex including Hsp90 and is presented in the cytoplasm . After tAR is involved in a variety of functional regulations, such as cell proliferation, differentiation, apoptosis, metabolism, and secretion. It is a key factor in maintaining tissue homeostasis. Meanwhile, A plays an important role in the development of various hormone-dependent cancers ,22, suchThe non-classical AR signaling pathway is not involved in the regulation of A-responsive gene transcription and translation. The non-canonical AR signaling pathway involves multiple pathways. It mainly includes the regulation of signal transduction pathways, such as MAPK/ERK (Mitogen-activated protein kinase/extracellular regulated protein kinases) activation, and mTOR  pathway activation through PI3K/Akt (Phosphatidylinositide 3-kinases/Serine/threonine kinases of the AGC family) pathway ,28. For Src belongs to the tyrosine kinase family and expresses commonly in malignant tumors . Src can6) [Moreover, there are several AR bypass pathways. In prostate cancer, glucocorticoid receptor (GR) can activate part of AR signaling pathway, and inhibition of GR may be a potential target for the treatment of female reproductive system cancer . Previou6) . AR antaFemale reproductive system cancer is a hormone-dependent cancer. However, the molecular regulation of AR in each type of cancer remains unclear. According to GLOBOCAN 2020 database for 36 cancers in 185 countries, there were 19.3 million new cancer cases and nearly 10 million cancer deaths globally in 2020 (excluding non-melanoma skin cancers). Female reproductive system cancer deaths/new incidences both accounted for 13% of the global total: OVCA (207252/313959), EC (97370/417367), and CC (341831/604127) [These three cancers are found in women’s uterus and ovaries which are hormone-dependent organs. Although they differ in organization, they are adjacent to each other and associated with reproductive function. These cancers are speculated to be related. A report shows that young women with EC had an increasing risk of synchronous OVCA, although the prognosis of women with synchronous endometrial and ovarian cancer was good . EstrogeOVCA is a malignant tumor with the highest mortality rate in female. In 2020, the mortality rate of OVCA patients reached 66% 207252/313959), accounting for 4% of the global cancer deaths 3959, acc. ImmunohThe ovary, which produces A, is an important sex hormone synthesis and secretion organ. Recently, the relationship between the A/AR and the OVCA has been explored. Previous studies found that OVCA tissues and cells showed approximately 69~90% AR positive ,25,48. AInterestingly, a large number of studies have found that there was a CAG trinucleotide polymorphism which is about 8–31 repeated segments in exon one of the AR. Genotyping of OVCA cells from 215 Polish–Caucasian patients , 1,800 CMore and more research has focused on the molecular regulation relationship between AR and OVCA. These studies focus on how AR regulates cancer cell proliferation and migration by interacting with a variety of factors in OVCA. These co-regulators including coactivators and corepressors modulate AR activity and activate ARE-mediated transcription in ovarian cancer cells, although most of them also interact with other receptors . OVCA isAR signaling stimulates proliferation, migration, and invasion of OVCA cells . AR is aGenerally, cancer stem/progenitor cells (CSPCs) are considered to be responsible for cancer phenotypes, including stemness/pluripotency, metastasis, drug resistance, and high recurrence rates to stimulate tumor growth and disease progression . NumerouAssociation of AR and Chemoresistance of OVCACurrently, chemoresistance is a major challenge to cancer chemotherapy. The first-line therapy for OVCA is surgery, and the second-line therapy is chemotherapy (primarily platinum or paclitaxel (Taxol)). In the process of chemotherapy, OVCA patients are prone to develop drug resistance, and the overall survival of patients is shortened. Therefore, paclitaxel resistance  has raised great interest. Analysis of genes involved in txr revealed that ERAR is a key gene for resistance . AR exprAssociation of OVCA Cell Migration and PrognosisNumerous studies have demonstrated that high migration rates of cancer cells were often associated with poor prognosis. Clinical studies have shown that matrix metalloproteinase-2 (MMP-2) was associated with AR and reduced overall survival in 88 patients with EOC in Mexico. It was speculated that in the presence of AR, MMP-2 in EOC cells involved in basal layer degradation and promoted ovarian cell adhesion to the peritoneum and momentum, promoting tumor cell migration . In a st6). The patients experienced multiple recurrences with recurrent AR-positive (AR+) ovarian cancer who received daily enzalutamide 160 mg until progression of disease or treatment discontinuation. Assessing long-term survivors of epithelial ovarian cancer, the PFS6 was observed in at least 13 of 59 patients (22%). Including 38.5% of patients with LGS met the PFS6 endpoint. In addition, the patients showed toleration with enzalutamide, while a phase II study about the efficacy of anti-androgen and gonadotropin-releasing hormone with bicalutamide and goserelin in patients suffering from epithelial ovarian cancer found no survival benefit [Recently, there are encouraging clinical results on AR antagonist. A Phase II study has been conducted to evaluate the efficacy of enzalutamide in women with AR positive ovarian cancer . Enzalut benefit . It shou benefit . TherefoHowever, so far, clinical trials on antiandrogen receptor for OVCA have been relatively non-randomized and undersized to draw reliable conclusions. Further clinical trials with larger sample sizes and randomized designs are needed.EC is one of the most common malignancies in women, with low mortality due to its obvious early symptoms . EC origIn 1978, AR was found in EC, and 65–80% of EC was positive for AR , while tSimilar to OVCA, the polymorphism of CAG repeats in AR exon 1 is of interest. Some studies suggested that CAG repeat length had nothing to do with EC , while oHypermethylation of the CpG region of the AR gene spanning the transcription start site is associated with stage III and IV endometrial malignancies . PresumaAR-dependent signaling inhibits the proliferation of endometrial tumor cells . AR can p = 0.041) [The number of AR appears to correlate with the degree of tumor differentiation . By immu= 0.041) . Enhance= 0.041) ,122. Par= 0.041) . MFE-296= 0.041) . This me= 0.041) .It has been reported that A and AR might be involved in the process of endometrial cell proliferation and inhibition of apoptosis by regulating the expression of insulin growth factor I (IGF-I) which was an A-responsive target gene in utero ,127. TheThe proto-oncogenic transcription factor c-Myc has been found to promote tumor growth in various AR-related cancers through AR signaling ,132. AbeIndole-3-methanol (I3C) inhibits the activation of multiple transcription factors including AR, the synthesis of DNA carcinogens and free radical production. It induces cell apoptosis, stimulates 2-hydroxylation of estradiol, and inhibits cancer cell invasion and angiogenesis. Therefore, I3C can inhibit the proliferation of EC, breast cancer, and other tumor cells in vitro .6 in 35 patients with endometrial cancer. Findings demonstrated the safety and promising efficacy of enzalutamide in this setting, with a 71% ORR (95% CI: 54–85%) and 83% PFS6 rate (95% CI: 66–92%).Estrogen induced AR expression . An anasThe evaluation of the therapeutic effected of tumor cells in vitro showed that increased AR expression promoted the migration and epithelial–mesenchymal transition process of EC cells and CSPCs. It also reduced the cytotoxic effect of cisplatin on EC cells .Further research on the occurrence and development of AR and its regulatory factors will help to deepen the understanding of EC, discover new biomarkers and drug targets, and provide new strategies for cancer treatment.CC is the fourth most frequently diagnosed cancer and the fourth leading cause of cancer death in women. According to the 2020 global female cancer statistics, the incidence of CC was 6.5% and the mortality rate was 7.7% . The twon = 30). However, AR expression was observed only in 63% of high-grade cervical intraepithelial neoplasia (HSIL-C 2/3) (n = 30) and 23% of infiltrating squamous cell carcinoma (ISCC) (n = 13). Loss of AR expression was common in HSIL and ISCC. It is prevalent in high-grade squamous intraepithelial lesions and ISCC due to the complex interaction between high-risk HPV and AR [AR-dependent signaling affected uterine growth and ovarian function, and AR was positive varies in different CC subtypes . ImmunohV and AR . HoweverA next-generation sequencing (NGS) analysis of the cancer genome maps in 182 CC patients was conducted and results showed that AR amplifications, mutations, and deletions in 7% of patients , indicatThe ASXL family are epigenetic scaffold proteins that assemble epigenetic regulators and transcription factors into specific genomic loci with histone modifications. They are thought to be tumor suppressor or oncogenic, and play different roles in different types of tumors. ASXL1 assembles various proteins based on protein–protein interactions. It has been found that ASXL1 interacted directly with AR, and promoted AR-dependent transcriptional activation. In CC, ASXL1 expression was increased . It was Cancers of the female reproductive system are a serious threat to women’s health due to their high morbidity and mortality. AR plays an important role on steroid-dependent cancers, especially in pre- and post-menopausal. AR-based combination therapies for the treatment of female reproductive system cancers have gradually become a hot research topic. For example, AR antagonists, such as enzalutamide and bicalutamide, have been used in clinical trials. Enzalutamide has good tolerability and appropriate safety in patients with AR+ recurrent ovarian cancer, especially in the AR+ LGS subpopulation. So, AR antagonists may be a potential treatment option for the specific subtypes of female reproductive system cancers. Although efforts have been made to explore the effects and mechanisms of AR on female reproductive system cancer, further research needs to be conducted in preclinical trials.This review provides a comprehensive overview of AR expression and polymorphisms, AR coregulators, and related mechanisms in the female reproductive system cancers. Findings from clinical, pharmacological, and genetic studies have now converged to demonstrate an important role of AR in these cancers. The mechanisms of AR in these cancers are complex and dynamic, and require the cooperation of many coregulators which are not all identified. In addition, AR is involved in the proliferation and migration of cancer cells and chemotherapeutic drug resistance. Thus, more attention should be paid to AR target genes, activation mechanisms and functions. Further work is required to develop appropriate therapeutic strategies for female reproductive system cancers. Particularly, novel biomarkers and drugs, which degrade or inactivate AR protein through AR co-regulators or antagonists, should be explored.


Infantile hemangioma (IH) is a benign vascular tumor, characterized by a unique sequence of non-linear growth and spontaneous involution. Some hemangiomas require intensive treatment to avoid functional and aesthetic insufficiency. Although β-adrenergic receptor (β-AR) antagonists have been increasingly used as the first-line treatment since 2008, the IH rebound still exists with uncertain mechanism. Here, we report three cases of abrupt IH deteriorations that are mainly related to β2-AR agonist administration. Potential IH proliferation induced by β2-AR agonists, especially from oral or nebulized approaches, should be recognized more widely by healthcare providers. Additionally, it is necessary to carry out large sample studies to analyze the influence of β2-AR agonist administration on the deterioration of IH. Infantile hemangioma (IH) occurs in approximately 5%–10% of infants . The potIn this study, we report three cases of increased size and redness in regressed and almost regressed IHs, which were mainly related to oral or nebulized administration of β2-adrenergic receptor (β2-AR) agonists. To the best of our knowledge, it is rarely reported and not widely recognized by healthcare providers . ClinicaA 5-month-old boy was diagnosed with IH and admitted to the Department of Dermatology at Children’ Hospital of Chongqing Medical University for primary OPT. He was a premature infant at 30 weeks gestation with a birth weight of 1.35 kg. The IH had been rapidly growing on his left chest wall since birth, measuring 3.0 × 1.5cm at maximum and 0.6 × 0.4cm at minimum with bright red color (®) therapy after discharge , during which OPT was discontinued to avoid bronchospasm. OPT was restarted as respiratory symptoms improved. Unfortunately, because of worsening bronchitis, OPT only lasted for 6 days and was replaced by topical timolol. Additionally, prednisone acetate tablets (7.5 mg twice a day) and the procaterol hydrochloride oral solution (2.5 ml twice a day) were added to relieve bronchospasm symptoms. After nearly 1 day of comprehensive treatment, the color of the IH progressed into deep red and showed a trend of proliferation  and admitted for OPT . Meanwhile, OPT was replaced with topical timolol to avoid its bronchospasmodic effects. However, after 7 days of respiratory management, the IH showed a trend of proliferation (A 4-month-old boy was diagnosed with IH, which was located on the right periauricular area (OPT is currently the first-line treatment for IH because of its high response rate and safety –20. It uvia the coincident intracellular β2-AR–driven proangiogenic pathway, such as promoting norepinephrine-induced NO release and inhibiting the α-smooth muscle actin expression in pericytes. Their combined effects enhance the vasodilation of IHs, which results in increased size and redness in clinical observations. Furthermore, a recent study showed no significant difference in β2-AR expression among proliferative, involutional, and propranolol-responsive hemangiomas, which may explain the role of β2-AR agonists in the involutional phase (According to published literature, rebound growth was recorded in 6%–25% of IH cases after discontinuation of propranolol –14. In tal phase –14, 27. Even though it is not the first report on IH deterioration associated with β2-AR agonist, it is still not taken seriously by most clinicians. Nicole et al.  reportedThe original contributions presented in the study are included in the article/Supplementary Material. Further inquiries can be directed to the corresponding author.The studies involving human participants were reviewed and approved by Ethics Committee of Children’s Hospital of Chongqing Medica University. Written informed consent from the participants’ legal guardian/next of kin was not required to participate in this study in accordance with the national legislation and the institutional requirements.Conception and design: QC, YZ, and SN. Administrative support: SN and HW. Provision of study materials or patients: QC, LL, XL, and HW. Collection and assembly of data: YZ and CS. Data analysis and interpretation: YZ, QC, and CS. Native English polish and provided critical opinion: CS. Manuscript writing: All authors. Final approval of manuscript: All authors.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Regarding caries lesions, a statistically significant difference (p < 0.05) showed between sound (ICDAS 0) and decayed (ICDAS 3) molars at 24-month follow-up. Our study results supported the resin-based sealant effectiveness in arresting incipient carious lesions, which according to the ICDAS II classification have received codes 1 and 2 but did not support sealant effectiveness in arresting caries lesions classified according to the same classification with code 3.Dental sealants are excellent means to prevent pits and fissure caries. Nowadays, the application of sealants is extended to therapeutic use in arresting non-cavitated carious lesions. This relatively new concept still lacks evidence to support its routine use. The aim of this study was to evaluate the effectiveness of a resin-based sealant applied on first permanent molars with carious lesions (ICDAS 1–3), in comparison with its effectiveness when applied on sound surfaces (ICDAS 0). Included in the study were 114 children aged between six and eight years old, with a high caries risk (according to the CAMBRA system), with all four permanent molars erupted and with deep pits and fissures. A total number of 407 molars were sealed and assessed. A total of 49 were excluded . Out of these 407 molars, 213 were sound (code 0) and 194 had caries lesions as follows: 56 teeth classified as code 1, 79 teeth classified as code 2, and 59 teeth classified as code 3 according to the ICDAS II classification. The retention of the sealant and carious lesions were assessed clinically at 6, 12, 18, and 24 months. Regarding sealant retention, a statistically significant difference ( Dental caries is the most common dental disease affecting people of all ages. Even though its aetiology is well-known, it remains a significant worldwide disease and public health issue. The World Health Organization (WHO) estimated that it affects nearly 3.5 billion people . In deveDue to the high prevalence and early onset of occlusal caries, its prevention has been one of the major goals in dentistry . From alRecently, advances in adhesive materials have facilitated the use of sealants to be extended to therapeutic uses such as managing caries lesions with no cavitation. This is a relatively new concept and still lacks evidence to support its routine use . The hypThe aim of this study was to evaluate the effectiveness of the therapeutic use of a resin-based sealant applied on recently erupted first permanent molars with carious lesions (ICDAS 1–3), in comparison with the effectiveness of the preventive application on sound surfaces (ICDAS 0), over a period of 24 months.This study was conducted from July 2019 to July 2021 (24-month period), in a private medical centre, Denta Aur in Targu-Mures, Romania, with the clinical trial registration number 022/27.06.2019. The written informed consent of the parents of all children involved in the study was obtained before the beginning of the examinations.All the examinations and procedures were performed by two experienced dentists, helped by a trained chair-side clinical assistant, following the 4-handed sitting dentistry model. Before the beginning of the examinations, the dentists completed an ICDAS II calibration course, assessing the condition of tooth surfaces and the presence of caries according to the International Caries Detection and Assessment System (ICDAS II) in a very similar way as previous researchers .In order to recruit for our study, we examined 156 children (624 teeth) with ages between 6 and 8 years. The inclusion criteria were as follows: presence of four erupted permanent first molars without dental abnormalities, with deep pits and fissures, that were at caries risk. For assessing children caries risk, we used the Caries Management by Risk Assessment (CAMBRA) system . A totalAll teeth were cleaned with a prophylaxis cup and a toothpaste without fluoride. The examination was made on clean and wet/dry teeth. We use a ball-ended explorer to remove any remaining plaque and debris and to check for surface contour, cavitation, fillings, or sealants.Each occlusal surface was examined and received classification ranging from 0 up to 6 according to ICDAS II criteria  Table 1Table 1:A total of 64 teeth classified as code 1;A total of 79 teeth classified as code 2;A total of 59 teeth classified as code 3 according to the ICDAS II classification.Out of the total of 476 teeth, 427 molars were sealed by the operators; 49 teeth were excluded because they had caries, which according to the ICDAS II classification were classified with codes 4–6, or had older sealants or fillings. Of these 427 molars that we sealed, 225 were sound (code 0) and 202 had caries lesions as follows:The sealing material used was a light-curing resin-based sealant, Helioseal F™, Ivoclar Vivadent Schaan, Liechtenstein. According to the manufacturer’s instructions, the application steps were: professional tooth cleaning, rinsing with water then air-drying, isolation with cotton rolls and air-drying, application of the enamel etching with phosphoric acid gel (37%) for 30 s, rinsing with water and air-drying, control of the acid-etched dental surface, bonding application, light curing of the bonding, sealant application, light curing of the sealant, control of marginal adaptation, and occlusion control.The follow-up interval was every six months, over a period of 24 months. We evaluate sealant retention and the incidence of new caries lesions or the progression of the lesions that were already present. The integrity and marginal adaptation of the sealant were assessed through visual and tactile examination.I: completely retained;II: partially retained;III: missing sealant.For assessing sealant retention, we used Simonsen’s criteria :I: complThe required sample size was determined to be 407 teeth using G-power software™, Heinrich Heine University, Dusseldorf, Germany, for Windows, for a power of 95%  .p was considered significant when p ≤ 0.05. All data were recorded using GraphPad Prism™ V6.01 software for Windows™ 2017. For evaluation of the categorical data, we used Fisher’s exact test and chi-squared test. The chosen significance level was set at 0.05, and For 5 children, we could not perform all the periodic check-ups, so we had to exclude them throughout the study period. The final sample of the study included 114 children and 456 first permanent molars which were sealed and assessed. Of these molars, 213 were sound (ICDAS 0) and 194 had caries lesions (ICDAS 1–3). The follow-up intervals were at 6, 12, 18, and 24 months. All the obtained data were systematised in tables  and one p > 0.05) regarding sealant retention between the two types of sealed teeth, sound (ICDAS 0) and decayed (ICDAS 1–3) (p = 0.02) between sound (ICDAS 0) and decayed (ICDAS 3) sealed teeth (p = 0.0086) regarding sealant retention, between sound (ICDAS 0) and decayed (ICDAS 3) molars  . The 18-ed teeth . The 24-) molars .p > 0.5) between the two types of sealed teeth concerning carious lesion development  concerning carious lesion development only between two types of sealed teeth—sound (ICDAS 0) and with caries lesion (ICDAS 3)  .Some of the carious lesions initially classified with codes 0, 1, or 2 according to ICDAS II classification required the reapplication of a new sealant on the 6, 12, 18, and 24-month follow-up assessments. Three of these lesions progressed to code 3. None of these lesions progressed to code 4–6, which would indicate the application of fillings.Some of the carious lesions initially classified with code 3 according to ICDAS II criteria required the reapplication of sealants at 6- and 12-month assessments. At 18- and 24-month follow-up assessments, 4 and 9 lesions, respectively, required the application of fillings, due to their transformation into lesions classified with codes 4 or 5.The idea of sealing and arresting a carious lesion in the pits and fissures is not new, as many researchers have pointed this out since the 1970s. Handelman was the first to support the possibility of arresting dental caries with the help of sealants and without the need for an invasive intervention to remove the caries lesion . Since tAnalysing these studies’ results, we found out that the information provided is very general, and it is difficult to draw exact conclusions based on which practitioners have clear recommendations about which caries lesions could be sealed and arrested .Out of the desire to clarify certain aspects regarding the situations in which sealants have the highest efficiency, related to the depth of carious lesions that can be arrested with dental materials, in our study, we used precise criteria. The children participating in the research were divided according to the CAMBRA classification  in the cIt is very important to notice that not every tooth would necessarily become affected, so the selection of children and teeth which must receive a sealant is one of the most important steps. For the sealants to be effective, they have to be placed in only those pits and fissure that are at caries risk. Regarding caries risk, one of the best predictors of caries in permanent dentition is the presence of carious lesions in temporary dentition. This is due to the accumulation of factors such as the high level of cariogenic bacteria found in the mouth of these children, poor oral hygiene, and carbohydrate consumption ,40. MoreSealants placed only on children (teeth) that are at caries risk have shown a positive outcome (cost-effective and less time-consuming) in clinical practice, especially in public health programs ,47.The results of our study are similar to those of studies from the literature, supporting the effectiveness of sealants in the prevention of caries in the pits and fissures of sound teeth, classified according to the International Caries Detection and Assessment System (ICDAS II) with code 0 ,10,11,12Regarding the lesions classified according to ICDAS II with code 3, our study results showed that sealants are not so effective in arresting this stage of carious lesion. This may be due to the lower retention of the sealant when applied on these lesions, which leads to a higher rate of reseals and the progression of carious lesions that have lost the covering material. It is generally accepted that the efficacy of resin-based sealants in preventing caries is based on good retention ,49,50.The poor retention of sealants in the case of demineralised enamel and dentin which surround caries fissures has been also reported by other researchers. These demineralised tissues have reduced adhesive properties ,52 and cBecause of the small sample size and short period of time, the results of our study must be interpreted with caution.The results of this study supported the effectiveness of dental sealant materials in the prevention of caries in the pits and fissures of sound teeth, classified according to ICDAS II with code 0, and also supported their effectiveness in arresting incipient carious lesions classified according to the same classification with codes 1 and 2.Our study results did not support sealant effectiveness in arresting caries lesions classified according to ICDAS II with code 3.


Escherichia coli significantly increased during rehabilitation, particularly ciprofloxacin and tetracycline resistance, and seven of the rehabilitated wild birds acquired multidrug resistance. The diet and habitat changes experienced by wild migratory birds during rehabilitation may have induced the observed gut microbiome dysbiosis and acquisition of antibiotic resistance. These rehabilitation-induced alterations might affect the adaptability of wild birds to their natural environments and contribute to the spread of antibiotic resistance after their release.Wild migratory birds are essential for sustaining healthy ecosystems, but the effects of a rehabilitation period on their gut microbiomes are still unclear. Here, we performed longitudinal sampling, 16S rRNA sequencing, and antibiotic resistance monitoring of the gut microbiome of six species of wild migratory birds protected as natural monuments in South Korea that are subject to short- or long-term rehabilitation periods. Overall, gut microbiome diversity was significantly decreased in the early stages of rehabilitation, and it did not recover to a level comparable to that of wild birds. Moreover, while the abundance of short-chain fatty acid-producing bacteria decreased, that of zoonotic pathogens increased, indicating rehabilitation-induced dysbiosis. The metabolic pathways involved in the degradation of aromatic pollutants were significantly downregulated, suggesting the depletion of pollutant-degrading microorganisms. Antibiotic resistance of IMPORTANCE Wild migratory birds are key for ecosystem health but highly sensitive to anthropogenic activities. Therefore, wild migratory birds often undergo rehabilitation to prevent species extinction or biodiversity monitoring. However, the impact of rehabilitation on the gut microbiome of wild migratory birds, which is closely associated with host fitness, remains unclear. For the migratory bird species considered natural monuments in South Korea evaluated here, such impacts could include rehabilitation-induced gut microbiome dysbiosis and acquisition of antibiotic resistance, with possible repercussions on the adaptability of wild birds and spread of antibiotic resistance in the environment after their release. Therefore, the dynamics of the gut microbiome and antibiotic resistance should be considered for implementing sustainable rehabilitation strategies. Wild migratory birds play an essential role in sustaining a healthy ecosystem. For instance, wild migratory birds contribute to shaping the distribution of global biodiversity, as they are involved in the long-distance dispersal of various organisms, including seeds and microorganisms . Moreove5–The gut microbiome is a symbiotic community of microorganisms, including bacteria, fungi, and viruses and their genomes –10. It i–11–16–Wildlife rehabilitation is a human-related activity in which a variety of wild animal species are bred or grown and then reintroduced into their natural habitats; this is mostly performed to prevent species extinction and for biodiversity monitoring . In the 21–Falco tinnunculus, Falco subbuteo, Otus bakkamoena, Otus scops, Ninox scutulata, and Accipiter gentilis kept in a rehabilitation center were evaluated from the wild state (immediately after rescue) to the release state (immediately before release). Our study revealed the dynamics of the gut microbiome, including changes in the taxonomic composition, diversity, bacterial network, and potential metabolic pathways, as well as changes in antibiotic resistance, which may affect host fitness after release to the natural habitat. This study provides information for developing sustainable rehabilitation strategies for wild birds.It is essential to improve the understanding of the dynamics of the gut microbiome of wild birds in response to rehabilitation to establish a sustainable conservation strategy for these species. Thus, in the present study, we investigated the effect of rehabilitation on the gut microbiome of wild migratory birds. We performed longitudinal sampling, 16S rRNA sequencing analysis of the gut microbiome, and antibiotic resistance monitoring on six species of wild migratory birds that are protected as natural monuments in South Korea. Individuals of The overall scheme of our sampling and study design is shown Firmicutes, Proteobacteria, Bacteroidota, Actinobacteriota, Fusobacteriota, and Patescibacteria were the six dominant phyla in both the wild and release states . In the wild state, Firmicutes was the most enriched phylum across all samples , followed by Proteobacteria  and Bacteroidetes . In contrast, in the release state, Proteobacteria was the most enriched phylum across all samples , followed by Firmicutes and Bacteroidetes .Ralstonia and Enterococcus were the most enriched genera in the wild state , followed by Staphylococcus and Bacteroides . In the release state, Escherichia-Shigella and Ralstonia were the most enriched genera , followed by Bacteroides and Enterococcus .The dominant genera differed between the wild and release states. For instance, P < 0.05) in release birds than in wild birds (P < 0.05) during long- and short-term rehabilitation (P > 0.05) between the short- and long-term rehabilitation groups (P < 0.05) in the first 2 weeks of rehabilitation, and it did not recover to the wild-state level during long-term rehabilitation  and Shannon’s index. Both indices showed significantly lower values (ld birds  and b. Nlitation  and d. Hn groups  and f. Llitation  and h.P < 0.05). Moreover, samples taken from wild birds immediately before their release were clustered, regardless of the period of rehabilitation  based on the unweighted UniFrac distance, followed by permutational multivariate analysis of variance (PERMANOVA). There was a significant difference in the gut microbiome composition between the wild and release states (litation .Proteobacteria was the only phylum significantly enriched in the release state compared to the wild state (adjusted P < 0.05). Twelve genera . In contrast, only five genera  were significantly enriched in the release state compared to the wild state (adjusted P < 0.05).To explore the specific components of the gut microbiome that contributed to this shift, and thus to the disruption of the microbiome composition and function (dysbiosis), we performed differential abundance analysis between the wild and release states. e genera , namely,P < 0.05, r > 0.7) in the wild and release state networks, respectively  were considered significant (ectively . Moreoveectively . The numP < 0.05) in the wild state. On the other hand, 11 metabolic pathways, including carbohydrate biosynthesis, cofactor, carrier, and vitamin biosynthesis, lipopolysaccharide biosynthesis, glycan biosynthesis, amino acid degradation, fatty acid and lipid biosynthesis, amine and polyamine degradation, and carbohydrate biosynthesis, were significantly enriched (adjusted P < 0.05) in the release state.The impact of rehabilitation on the metabolic pathways of the gut microbiome was analyzed using PICURSt2 software  and b. DEscherichia coli from fecal samples. The 30 E. coli strains isolated from 15 of the 17 wild birds (one isolate each per bird for the wild and release states) were then tested for antibiotic susceptibility. In the wild state, E. coli showed the highest resistance rate to ampicillin (46.67%), followed by tetracycline (33.33%), amoxicillin (13.33%), and ciprofloxacin (6.67%). In the release state, E. coli showed the highest resistance rate to ampicillin and tetracycline (both at 66.66%), followed by ciprofloxacin (60.00%), amoxicillin (26.67%), and cefotaxime (6.67%). E. coli showed no resistance to colistin, imipenem, and cefoxitin in both the wild and release states (P < 0.05) in the release state compared to that in the wild state  producers, such as t health . A decret health . Thus, ot health . For birt health –39. As tCampylobacter and Peptoclostridium, indicating that pathogenic species were able to colonize the guts of wild migratory birds during rehabilitation. If an external infection was the source of this increase in zoonotic pathogens, clinical signs, such as lethargy, diarrhea, or behavioral changes, would have been observed; however, none of the birds in the present study showed obvious signs of infection. Considering the absence of clinical signs and the results of the 16S rRNA sequencing analysis, these pathogens may have colonized the gut of wild migratory birds during rehabilitation as common members of the gut microbiome. Our result is consistent with that of a previous study, which showed that several zoonotic pathogens, such as Campylobacter, increased in the gut microbiome of wild animals during rehabilitation  and captive-bred quail were provided to birds that were alert and had the ability to feed themselves. The health status and behavior of the rescued wild birds were monitored daily by veterinary staff at the Seoul Wildlife Center. Injured and convalescent wild birds, birds showing abnormal clinical signs, and juvenile birds not capable of self-feeding were excluded from the study. Thus, all birds used in the present study were clinically healthy. When birds were physically and behaviorally able to forage and breed in the wild, and therefore considered ready for release by veterinarians, they were released into a native forest or park. Wild birds that were rehabilitated for less than 4 weeks were considered to have undergone short-term rehabilitation, while those that were rehabilitated for 4 weeks or more were considered to have undergone long-term rehabilitation. Information regarding the wild birds used in the present study is provided in Table S1 in the supplemental material.Fecal samples of the wild birds were collected by veterinarians, immediately transferred to the laboratory, and processed for gut microbiome 16S rRNA sequencing analysis and antibiotic resistance monitoring. Sample collection was performed in the wild state, within the first 2 weeks of rehabilitation, and then at 4-week intervals until release. Fifty fecal samples were used for DNA extraction and microbiome 16S rRNA sequencing analysis .16S rRNA V3-V4 hypervariable gene region was performed using the primers 341F (5′-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG-3′) and 805R (5′-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC-3′) from Illumina Inc. . PicoGreen was used for pooling and normalizing the amplified products. All sequencing procedures were conducted using the Illumina MiSeq platform at Macrogen, Inc. .Fecal DNA extraction and sequencing were performed as previously described . Brieflyhttps://mafft.cbrc.jp/alignment/software/). The taxonomy profile of ASVs was generated using the q2-feature-classifier implemented in QIIME2 against the SILVA database   software package . Raw seqef NR99) . Sequencr > 0.7 and P < 0.05 as the cutoff values for significant correlations. Metabolic pathways were analyzed using the phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt2) (https://metacyc.org).Downstream analysis of sequence data was performed using the MicrobiomeAnalyst R package . The alpICRUSt2)  and the E. coli for monitoring antibiotic resistance in wild birds during rehabilitation, fecal swabs from 17 wild birds in the wild state and release state were inoculated into 2 mL of E. coli broth  and enriched overnight at 37°C. After enrichment, 100 μL of the culture broth was spread on MacConkey agar (Oxoid) and incubated at 37°C for 24 h. Cultures were then streaked on eosin methylene blue agar , and colonies exhibiting the culture characteristics of E. coli were pure cultured and confirmed by matrix-assisted laser desorption ionization–time of flight mass spectrometry. As a result, E. coli was isolated from 15 birds and further analyzed for antibiotic susceptibility.To isolate Disk diffusion susceptibility tests (Kirby-Bauer method) were conducted for eight antibiotics: amoxicillin-clavulanic acid (20/10 μg), ampicillin (10 μg), cefotaxime (30 μg), imipenem (10 μg), tetracycline (30 μg), ciprofloxacin (5 μg), colistin (10 μg), and cefoxitin (30 μg). Antibiotic susceptibility results were interpreted following the Clinical and Laboratory Standards Institute guidelines.The rehabilitation program at the Seoul Wildlife Center was conducted with the authorization of the Seoul Metropolitan Government and Ministry of Environment of Korea. All procedures in this study strictly followed the guidelines and ethical principles of the following South Korean legislation: the Wildlife Protection and Management Act, the Guidelines for the Operation of the Wildlife Rescue and Management Center by the Ministry of Environment of Korea, and the Cultural Heritage Protection Act by the Cultural Heritage Administration of Korea.PRJNA814404.Sequence data from this study are deposited in the National Center for Biotechnology Information Short Read Archive database under accession number


Adequate nutrition is of utmost importance for athletes, especially during rehabilitation after injury in order to achieve fast healing and return to sports. The aim of this narrative review is to define the proper nutritional elements for athletes to meet their needs and facilitate their fast return to sports after surgery or injury, as well as determine the effects of specific nutrients intake. Studies on antioxidants, which are substances that protect against free radicals, for the injured athlete are few and unclear, yet poly-phenols and especially flavonoids might improve healing and inflammation following an injury. Benefits of vitamin C or E on muscle damage are disputable in relevant studies, while optimal levels of vitamin D and calcium contribute to bone healing. Minerals are also essential for athletes. Other supplements suggested for muscle damage treatment and protein synthesis include leucine, creatine, and hydroxymethylbutyrate. Diets that include high-quality products, rich in micronutrients  bio-active compounds and other nutritional elements (like creatine) are suggested, while an individualized nutrition program prescribed by a trained dietitian is important. Further studies are needed to clarify the underlying mechanisms of these nutritional elements, especially regarding injury treatment. Sports injuries have a significant economic  and psycProlonged injury leads to sedentary periods that cause a decrease in muscle mass, muscle strength, and function . In addiRehabilitation refers to the techniques and services for individuals with physical disabilities  trying to make them reach and keep a functional level in their everyday life as normally as possible . Sports The use of sports rehabilitation strategies is essential for a fast rehabilitation process in athletes’ health and their return to training and competition. In the early stages of rehabilitation, nutritional intervention is crucial to ensure adequate intake of energy and nutrients, prerequisites for wound healing and management of inflammation and oxidative stress caused by the injury .Adequate and balanced nutrition is essential for rehabilitation progress . The rolβ-carotene being one of the most common [β-carotene supplements. Instead, it is widely suggested that β-carotene should be consumed through carotenoid-rich foods, as part of a diet that includes consisting of the appropriate macro-, micronutrients and other nutritional elements [The role of vitamin and other micronutrients on sport rehabilitation is presented in t common . Some cat common . Recent elements . Findingelements   are expressed in the muscle stem cells depicting muscle regeneration after injury . A recenVitamin D supplementation has different outcomes according to muscle groups and functions . A meta-Vitamins A, E and C have frequently been called “antioxidants” since their deficiency tends to lead to oxidative stress . There iVitamin E supplementation in rats following spinal cord injury improved the hind limb locomotor function, and reduced spinal cord histopathological and morphological damage, while decreasing inflammation . BelisleVitamin A is an essential fat-soluble vitamin that has been used topically in dermatology for several years to control conditions such as photo-damage, psoriasis, and after procedures such as dermabrasion to facilitate healing  Table 1Table 1. The results of creatine on muscle strength and muscle mass gain have been examined regarding muscle atrophy, during immobilization or rehabilitation. Evidence for the impact of creatine on muscle loss during immobility is not very clear . As showMoreover, creatine supplementation at 20 g/day for 7 days has been shown to strengthen exercise capacity in individuals with complete cervical-level spinal cord injury . ConversEarly research data on the prevention and treatment of injuries to tendons and ligaments highlight the role of nutritional support from collagenous proteins and micronutrients such as vitamin C, which have shown potential benefits  Table 1Table 1. Minerals, such as manganese (Mn), copper (Cu), zinc (Zn), iron (Fe), and selenium (Se), act as antioxidants and, specifically as co-factors of important antioxidant enzymes, inactivate ROS and repair oxidative damages ,35 Tabl. AdequatUnsaturated fatty acids have anti-inflammatory and immune-modulatory effects. A prolonged inflammatory reaction, often seen after a serious injury or surgery, can be detrimental to rehabilitation. It has been suggested that n-3 PUFA lead to a decrease of muscle inflammation through the reduction of the level of pro-inflammatory cytokines, such as TNF-α and IL-6, reduced production of arachidonic acid (AA) and reactive oxygen species (ROS), resulting in a reduction in the inflammatory response . Fish oiCurcumin, a constituent of the spice turmeric, is a highly pleiotropic molecule with anti-inflammatory and anti-oxidant properties that have shown a variety of beneficial effects on human health . A systeIn general, a multi-disciplinary team, consisting of a doctor and a nurse, a physiotherapist, and a dietitian are necessary for optimal rehabilitation. The role of the dietitian is crucial, to plan an individualized nutrition program, that takes into consideration the athlete’s needs and preferences, and according to the current guidelines and recommendations, regarding nutrition during rehabilitation. Micronutrients, such as carotenoids, polyphenols, flavonoids, vitamin E or C are greatly supported for their properties against oxidative stress and they might have anti-sarcopenic properties. Although findings on the effects of antioxidants for the injured athlete are few and unclear, it has been shown that polyphenols and especially flavonoids might improve healing and inflammation following an injury. Additionally, optimal levels of vitamin D and calcium contribute to bone healing, thus they are also beneficial during the rehabilitation process. Furthermore, both the antioxidant and the anti-inflammatory properties of ingredients such as omega-3 fatty acids and curcumin could be beneficial during athletic rehabilitation. Other supplements suggested for muscle damage treatment and protein synthesis include leucine, creatine and HMB. Mineral with antioxidant function, such as manganese (Mn), copper (Cu), zinc (Zn), iron (Fe), and selenium (Se), are also essential for athletes. Athletes’ requirements and supplementary doses should be considered carefully, preferably through a diet that includes high-quality products, rich in nutrients.The anti-inflammatory effects of the Omega-3 polyunsaturated fatty acids (n-3 PUFA) have been observed to attenuate during various rehabilitation interventions of muscle injuries. The level of supplementation, in this case, should remain much higher than in the case of a typical diet .Rehabilitation of bone injuries after an athletic injury may benefit by sufficient intake of nutrients like calcium, and vitamins like D and A, along with other macronutrients (proteins) . FurtherClose et al. conducted an extended study regarding the micronutrients that promote the health and rehabilitation of tendons and ligaments. The main micronutrients in that area are vitamin C, copper, glycine, gelatin/hydrolyzed collagen, turmeric/curcumin, taurine, arginine, bromelain, or boswellic acid ,68.During various exercise-based rehabilitation interventions, high-quality proteins, fat, vitamins, antioxidants, minerals, and other supplements can play a major role in supporting athletes’ anabolism. Although there are several studies on the ergogenic effect of nutrients and supplements before and during training and competition, there is a research gap on the effectiveness of these nutrients in the rehabilitation of athletes after injury or surgery, in maintaining muscle mass and in reducing rehabilitation time. For this reason, it is not possible to make definitive recommendations on the use of the nutrients and supplementations. Future research is warranted to clarify the underlying mechanisms of nutrients, especially regarding injury treatment, as their efficacy has not yet been assessed satisfactorily. Monitored evaluation should follow, in order to assess nutrient indicators and to avoid levels above sufficiency. Diets that include high-quality nutrients, rich in macro- and micro- and bioactive compounds are suggested. Biomedical indices and vitamin and mineral levels should be assessed and monitored, to avoid unnecessary supplementation.