Rehabilitation for paraneoplastic cerebellar degeneration (PCD) has been indicated as necessary. However, there are only a few reports on rehabilitation in PCD. We describe the course of physical and cognitive functions and activities of daily living (ADL) in a patient with PCD and examine the effect of rehabilitation, along with a review of relevant literature.A 42-year-old woman experienced rapid deterioration in cerebellar symptoms and was unable to walk. The cerebellar symptoms improved after mastectomy, which was performed 3 months after the onset of symptoms. However, the cerebellar symptoms exacerbated 11 months after the onset of symptoms. She underwent immunotherapy because the level of anti-Yo antibodies was high, and anti-glutamic acid decarboxylase antibodies were detected.She was diagnosed with anti-Yo antibody-positive PCD caused by breast cancer.Rehabilitation was performed preoperatively, postoperatively, and at 1 year after the onset of symptoms.Her physical function and ADL improved after the surgery and at 1 year after the onset of symptoms. Finally, she regained the ability to walk between parallel bars supported with one hand and a walking frame.Given the results of this case and the relevant literature, it appears that rehabilitation improves physical function and ADL after oncotherapy. Particularly, initiating treatment within three months of onset of symptoms may enable patients to walk without assistance. PCD is associated with various cancers, such as lung, ovarian, and breast cancers. Antibody titers  are high, and they react with cerebellar Purkinje cells. The treatment includes oncotherapy and immunotherapy. Although patients are sometimes responsive to oncotherapy, immunotherapy is rarely successful, and the neurological prognosis is generally poor.,2; however, there are only a few reports on rehabilitation in PCD.–8 Furthermore, most reports only describe the changes in ADL after oncotherapy and immunotherapy. This report provides information regarding the course of physical and cognitive functions, and ADL . It also examines the effects of rehabilitation in a patient with PCD along with a review of the relevant literature.Rehabilitation for PCD has been indicated as necessary2This study was approved by the Osaka University Medical Hospital ethical review board , and signed consent was obtained from the patient's family.A 42-year-old woman with an unremarkable medical history, family history, and psychosocial history experienced dizziness, which was a sustained feeling of floating and staggering and consulted a local physician. No neurological findings or abnormal findings on a magnetic resonance imaging scan of the head (head MRI) were observed.She was admitted to the hospital because of a worsening of symptoms 2 months after their onset. She presented with cerebellar symptoms, including dizziness, trunk and limb ataxia, and dysarthria. She could walk between parallel bars supported by 1 hand and could care for self with supervision, but she required assistance while bathing. She had slightly impaired orientation. Miller Fisher syndrome was initially suspected and she received intravenous immunoglobulin (IVIg); however, her cerebellar symptoms continued to deteriorate rapidly. Subsequently, anti-Yo antibody was detected. After that, an abnormal accumulation in the left breast was found on positron emission tomography and it was diagnosed as left breast cancer (T1N0M0). Based on PNS diagnostic criteria in the European Federation of Neurological Societies, it was diagnosed as definite PCD. She underwent left mastectomy 1 month after admission. Histologically, the cancer was microinvasive ductal carcinoma; therefore, she received no postoperative treatment for the cancer, but she did receive IVIg and steroid pulse therapy for immunosuppression. Her cerebellar symptoms improved and she was transferred to a rehabilitation hospital 2 months after the surgery. At transfer, there were no abnormal findings on head MRI.However, her cerebellar symptoms deteriorated again and cerebellar hemisphere atrophy was found on head MRI 4 months after the transfer. Seven months after the transfer, she was discharged, but was readmitted to the hospital for reexamination.At readmission, there was no tumor recurrence or metastasis, but an analysis of her cerebrospinal fluid showed inflammation and high titers of anti-Yo antibodies. In addition, the level of anti-glutamic acid decarboxylase (GAD) antibody was elevated to 35.0 U/mL . The level of anti-glutamic acid decarboxylase (GAD) antibody was elevated to 35.0 U/mL . She received IVIg, 2 courses of steroid pulse therapy, and plasma exchange (PE). The levels of inflammatory markers and anti-GAD antibodies improved, and the progression of cerebellar hemisphere atrophy was halted, but the cerebellar symptoms did not change. She was discharged 3 months after the readmission, and predonine and tacrolimus were prescribed for the suppression of recurrence.Rehabilitation was performed during 3 periods: before and after the surgery, and at 1 year after the onset of symptoms. Before surgery, the rehabilitation program consisted of exercises for strengthening muscles, balance, gait between parallel bars, and ADL. Rehabilitation was performed for approximately 80 min/d, and the rehabilitation period was 2 weeks (every day on weekdays). After the surgery, the same rehabilitation program was followed for 10 weeks. At readmission, the same rehabilitation program was followed for 9 weeks .The assessments of physical and cognitive functions and ADL are described in Table Throughout these rehabilitative interventions, no adverse effects or unanticipated events occurred, and she continued rehabilitation with the goal of re-acquiring gait and ADL. Thus, she hoped to continue rehabilitation after discharge from our hospital, and she decided to continue outpatient rehabilitation at a local hospital.3 as cerebellar plasticity may occur if treatment is started within 3 months after onset. Her surgery was performed 3 months after onset, thus enabling an improvement in cerebellar symptoms after the surgery. However, 8 months after the onset, the cerebellar symptoms deteriorated again, possibly due to cerebellar atrophy, which is sometimes found in PCD a few months after the onset.,2 Alternatively, the possibility of anti-GAD antibody-associated cerebellar ataxia cannot be excluded. However, we thought that it was difficult to judge whether there was a strong association between cerebellar ataxia and anti-GAD antibodies. Thus, the exact cause of the renewed deterioration of cerebellar symptoms remains unclear. However, following immunotherapy the cerebellar atrophy did not advance, and the anti-GAD antibody levels improved as well.In the present case, cerebellar symptoms deteriorated rapidly before the surgery but improved after it. Early oncotherapy is essential for the successful treatment of PCD,; thus, rehabilitation is effective for indolent ataxia. In the present case, the progression of cerebellar symptoms might have outstripped the effects of rehabilitation before surgery and at 1 year after the onset of symptoms.The SARA score improved after the surgery but did not improve before the surgery or at 1 year after the onset. Rehabilitation for ataxia has been reported to improve SARA scores, balance, gait ability, and the Functional Independence Measure (FIM) score in spinocerebellar degeneration Furthermore, at readmission, her muscle strength had deteriorated by 12 points in the MRC sum score and by 13% in knee extension muscle strength compared with that at transfer. Patients with ataxia present with muscle weakness due to reduced activity,12; thus, her muscle weakness might have resulted from disuse because she rarely went out of her home. Therefore, the intensive rehabilitation during hospitalization improved her muscle strength, regardless of the cerebellar symptoms.The Medical Research Council (MRC) sum score and knee extension muscle strength improved for all periods. Although she had participated in skiing competitions a few months before the onset, her knee extension muscle strength at the first admission was approximately half that for a healthy 40-year-old Japanese woman. In addition, balance requires not only coordination, but also a variety of other factors such as muscle strength, sensory function, and cognition, with a particularly profound effect on muscle strength.,14 The BBS score might have deteriorated because of muscle weakness rather than ataxia; therefore, the score improved as muscle strength improved. Gait and FIM score followed the same course as the BBS score; thus, gait and ADL disabilities were mainly due to a balance disorder.The Berg Balance Scale (BBS) score followed the same course as the SARA score did before and after the surgery but improved at 1 year after the onset of symptoms. Compared with the assessments at transfer, the BBS score had deteriorated greatly (10 points) while the SARA score had deteriorated slightly (1 point) at readmission. If the progression of cerebellar symptoms is gradual, balance improves with rehabilitation.,16 In the present case, during the first admission, cognitive functions, as assessed by the Mini-Mental State Examination and Frontal Assessment Battery (FAB) scores, improved even before the surgery. The reason for this improvement is unclear, as there were no significant findings on head MRI. During readmission, the FAB score deteriorated slightly. As the cerebellum transmits output to the prefrontal cortex region, a relationship between the cerebellum and cognitive function, especially prefrontal functions, has been indicated.,18 Head MRI at 8 months after the onset of symptoms indicated atrophy of the cerebellar hemisphere, which may have led to the deterioration of FAB score.There have been reports of cases of PCD with cognitive dysfunction.,4,7,8 rehabilitation was undertaken after oncotherapy. Only the patient in the report by Sliwa et al improved enough to be able to walk independently. That patient received oncotherapy within the 3-month window of cerebellar plasticity, while oncotherapy or PE was started >3 months after the onset of symptoms in the other previous cases where such information was provided. In the current case, surgery was performed 3 months after the onset of symptoms and the patient's balance, gait ability, and ADL improved after the surgery. Subsequently, the patient was able to walk between parallel bars and with a walking frame independently. Fewer than 15% of patients with PCD show improvements in cerebellar symptoms,,20 and >75% with anti-Yo anti-body-positive PCD require wheelchairs to move around. However, rehabilitation appears to improve ADL after standard therapy (mainly oncotherapy), and, in particular, initiating the treatment within 3 months after the onset of symptoms may enable a patient to walk without assistance.Using MEDLINE and Google Scholar, a literature review was conducted to locate reports on the effects of rehabilitation on physical function and ADL in PCD published between 1970 and 2016. A summary of these previous reports is provided in Table This study has some limitations. First, the evidence for the effectiveness of rehabilitation for PCD is limited because this is a case report. Second, the treatment at the time of first admission included mastectomy, IVIg, and steroid pulse. The treatment at the time of readmission included IVIg and steroid pulse; thus, we could not completely exclude the influence of these medical and surgical treatments on the improvement in physical and cognitive functions and ADL. However, muscle strength improved after the first admission and readmission, and BBS and FIM improved despite no changes in SARA during readmission. We believe that those were the results of rehabilitation.In conclusion, physical function and ADL can improve with rehabilitation in patients with PCD when the progression of cerebellar symptoms is halted or improved. In addition, muscle strength can improve independent of the course of cerebellar symptoms. Therefore, in conjunction with early oncotherapy, it is important to continue rehabilitation according to the course of cerebellar symptoms. In the current case, improvement in cognitive function was also observed, which might have resulted due to a synergistic effect with rehabilitation. However, declines in cognitive function have been indicated; thus, awareness of this issue while implementing rehabilitation is also important.www.editage.jp) for English language editing.We would like to thank Editage (


Frontal fibrosing alopecia is a distinctive form of scarring alopecia consideredto be a clinical variant of lichen planopilaris. It predominantly occurs inpostmenopausal women and has a slowly progressive course. It was first describedby Kossard in 1994. Since then the number of reported cases has increasedsignificantly. Coexistence of frontal fibrosing alopecia and autoimmunedisorders - such as discoid erythematosus lupus and Sjögren's syndrome -may suggest a common pathogenic background among the diseases. The condition is clinically characterized by a symmetricalfrontotemporal hairline recession .1 It rarely occurs in males, with only one case ofbeard involvement reported.2Familiar involvement has also been suggested.1 The affected skin is pale and atrophic. Perifollicularpapules can be observed in approximately 60% of cases and follicular hyperkeratosis,female androgenetic alopecia, pruritus, and lichen planus in 30% of cases.1 There is a contrast in color betweenthe uniformly pale alopecia area and the sun-damaged skin of the forehead.1 Bilateral loss of eyebrow is acommon finding, which can occur before or after the frontal hair loss.1 Some patients present with hair losson the forearms, armpits, and upper and lower limbs.1Frontal fibrosing alopecia (FFA) is a scarring alopecia considered a clinical variantof lichen planopilaris (LPP).3Mulinari-Brenner et al. (2007) reported 6 cases of FFA, which were the first casesreported in Brazil. The authors observed that all the patients were postmenopausalwomen, with hair rarefaction evolution ranging from 1½-10 years and a meanage of 65.8 years (54-82).1 Serum levelsof androgens, thyroid hormones, and hematological indices are normal. Anti-DNA andFAN can be positive at low titers.1Histological examination of the affected scalp reveals scarring alopecia withmononuclear lichenoid infiltrate involving the infundibulum and isthmus of thefolicle.1 Directimmunofluorescence is negative. Possible diagnoses to be ruled out include alopeciaareata, cutaneous and systemic lupus, traction alopecia, androgenetic alopecia,pseudopelade of Brocq, and familiar high frontal hairline.3Clinical examination reveals hyperkeratosis and erythematous-violaceous coloration insome follicular orifices, which is an early sign that can be observed withmagnifying glasses.6 DLE sharesthe interface dermatitis pattern with inflammatory T-cell infiltrate, which maysuggest a common autoimmune etiology for both diseases.5 We present the second report of association betweenAFF and Sjögren's syndrome (SS).7There is a growing number of FFA reports in association with autoimmune diseases, inparticular with discoid lupus erythematosus (DLE).We report a 50-year-old female menopausal black patient presented with progressivehair loss in the frontotemporal and eyebrow regions for one year. She reports oralmucosa and eye dryness with 10 years of evolution. Clinical examination revealedbilateral patches of scarring alopecia in the frontotemporal region and absence ofeyebrows -3. AlopeTreatment was performed with intradermal injections of triamcinolone  in the frontotemporal region with monthly finasteride 2mg/day PO,chloroquine 250mg/day PO, and minoxidil 5% topical daily. After 4 years of regularfollow up, we observed stabilization with no regrowth of in scarred areas.1 Specific histopathological findings reveal greaterdegree of apoptosis and reduced inflammation in FFA in relation to LPP.6 Clinically, FFA occurs more oftenafter menopause and in the frontotemporal region. These features contrast withclassical multifocal areas of scarring alopecia in LPP.6 Studies also describe the association of the HLA-DR1(DR1 subtype of human leukocyte antigen) system with LPP. HLA is part of the majorhistocompatibility complex class II and is a ligand for T lymphocytes.8 However, cutaneous lesions andmucosal lichen planus have been described in patients with FFA.6The etiology of FFA remains unknown. FFA and LPP share these microscopic features:lymphocytic inflammatory infiltrate around the isthmus and infundibulum of the hairfollicles; presence of apoptotic cells in the outer root sheath; and perifollicularconcentric fibrosis, which result in scarring alopecia.6 Although the disease seems to be self-limiting in mostcases, the degree of progression before stabilization is unpredictable.1 However, the reversal of thefibrosis has never been described. Suggested treatments include: topical,intralesional, and systemic corticosteroids; topical retinoids; oral isotretinoin ;topical minoxidil ; hydroxychloroquine; and finasteride. Oral finasteride (2.5mgdaily) combined with minoxidil 2% halted the progression of alopecia in somepatients after 12-18 months of treatment.9 Some authors suggest that the combination of oraldutasteride with topical calcineurin inhibitor can represent a safe and effectivealterna tive therapy for AFF.9Without therapeutic interference, the progression of the condition is common. Thefrontal recession of the hairline can progress up to half of the scalp or more, butthe progression of the lesion is variable.4 Khan et al. reported the case of a 46-year-oldpatient with clinical and histopathological diagnosis of FFA that later showedphotosensitivity at hair loss sites. A biopsy confirmed the diagnosis of AFF, butdirect immunofluorescence was positive for IgM, IgG, IgA, and fibrin along thebasement membrane zone, consistent with the diagnosis of lupuserythematosus.10 Shapiroet al. reviewed 62 cases and identified 18% of patients with autoimmune disorders ofconnective tissue in association with FFA, which are: systemic lupus erythematosus,DLE, rheumatoid arthritis, and polymyositis.6 Tosti et al. have also recently reported 7 cases of clinicalcoexistence of FFA and DLE on the scalp.5 In 2008 Takahashi et al. described the firstand only case of association between AFF and SS in a postmenopausal Asianpatient.7Gaffney et al. reported a case of discoid lupus erythematosus complicated by FFA in a69-year-old patient. Alopecia developed after two years of evolution of the DLE withmanifestations on the scalp, face, neck and thorax.5 Clearly, FFA isstill a little known condition with few effective treatment options available. Thepresent case report complements the dermatologic literature that highlights theimportance of research of other autoimmune diseases associated with FFA given thetherapeutic and prognostic implications of these associations.Reports of DLE and FFA association, as well as association with other autoimmuneconditions, suggest the evidence of an autoimmune etiology for thiscondition.


AR) is involved in the differentiation and growth of many cancers. We hypothesized that two microsatellite polymorphic variants, AR (CAG)n and (GGN)n repeats, were also associated with the development of Papillary thyroid cancer (PTC) and Osteosarcoma. In current study, we conducted two case-control studies in a Chinese population to investigate the possible relationship between these two AR repeat polymorphisms and the risk of PTC and Osteosarcoma. The AR CAG repeat length was significantly associated with both risk of PTC and Osteosarcoma. Subjects with shorter AR CAG repeats had a higher risk of developing PTC  and Osteosarcoma . Specifically, shorter GGN repeats also contribute a significant increased risk of Osteosarcoma . Our results contribute to a better understanding of the complex hormone related mechanisms underlying PTC and Osteosarcoma.The androgen receptor ( AR) is involved in the differentiation and growth of many endocrine cancers, including breast cancer, prostate cancer, ovarian cancer, endometrial cancer, bladder cancer, thyroid cancer [AR is a nuclear transcription factor that mediates the actions of testosterone and dihydrotestosterone [AR gene, was identified to be associated with the expression level of AR, further the development of many endocrine related cancers [AR CAG and GGN repeat length variation and tumorigenesis of PTC and Osteosarcoma.The androgen receptor n and (GGN)n repeats were also associated with the development of PTC and Osteosarcoma. Thus we conducted this case-control study in a Chinese population to first investigate the possible relationships between these two AR repeat polymorphisms and the risk of PTC and Osteosarcoma.Since GGN and CAG repeats could influence the AR protein yield and its transcriptional activity and then paly roles in the tumorigenesis of PTC and Osteosarcoma, thus, in current study, we hypothesized that Totally included in the current study were 500 PTC cases and 500 matched healthy controls, as well as 500 Osteosarcoma cases and 500 matched healthy controls Table . CompariAR repeats length as continuous variables. Subjects with shorter AR CAG repeats had a higher risk of developing PTC . However, shorter AR GGN repeats didn't show a significant association with the risk of PTC . Then, the repeat length was analyzed as categorical variables, and the median value 22, 23 were selected as cut-points for the CAG repeat and the GGN repeat, respectively. As shown in Table P = 0.001). No significant association was detected for GGN repeat.We first analyzed the P = 0.014) and the GGN repeat  contribute to higher risk of Osteosarcoma Table P = 9.2 × 10–4). While compared to those with the longer (> 23) GGN repeat length, subjects in the category of shorter (≤ 23) GGN repeats had a significant increased risk of Osteosarcoma . Sensitivity analyses were also conducted to eliminate the subjects with extreme age, however, the significant associations were not affected.When analyzed as continuous variables, both shorter CAG repeat  and Osteosarcoma . While compared to those with the longer (> 23) GGN repeat length, subjects in the category of shorter (≤ 23) GGN repeats had a significant increased risk of Osteosarcoma . These findings were consistent with previous studies about epithelial ovarian cancer [In current study, we found that subjects with shorter longer >  GGN repen cancer , TMPRSS2n cancer , breast n cancer , 27, pron cancer , and so AR CAG repeat length as a susceptible factor for PTC and Osteosarcoma risk in Chinese population. Our results contribute to a better understanding of the complex hormone related mechanisms underlying tumorigenesis and add to the current state of knowledge regarding the susceptibility of AR to PTC and Osteosarcoma. However, further prospective studies with larger sample size involving different ethnicities, as well as further functional studies, are needed to confirm our findings.In summary, our study supported short In current study, the patients were recruited from May, 2010, and diagnosed by surgical operation. Controls were recruited from the general population living in the same areas and matched with cases by age at cancer diagnosis and gender. The medical records of healthy controls were also reviewed to ensure that they have no previous or current diagnosis of cancers or related diseases. All participants were interviewed face-to-face by trained professionals  using a structured questionnaire. After the interview, 5 milliliters of peripheral blood was collected from each subject in our study. Both approval from the appropriate institutional review board and written informed consents from patients who were included in this study were obtained.AR repeat region was amplified by PCR from 50 ng template DNA using primers flanking the AR repeat in exon 1 . Then the fragments were tested on denaturing polyacrylamide gels, and PCR products were purified and sequenced using the Applied Biosystems Prism 3700XL and analyzed by Applied Biosystems Prism Genescan automated fluorescence detection . Positive and negative control was used for quality control, and the concordance rate was 100%.Genomic DNA was extracted from peripheral blood samples using the Qiagen Blood Kit  according the instructions of manufacturer. The two t-test. For the AR repeat lengths comparison, the Shapiro-Wilk test was used to verify the normality of distribution, and Levene's test for equality of variances. When the assumptions were met, t-test was used to test the differences for the AR repeat lengths. Or, the Mann-Whitney U test was used. Unconditional logistic regression was used to calculate odds ratios (ORs) as well as confidence intervals (CIs) for PTC risk associated with repeat genotypes. We identified the cut-points  on the basis of repeat number distribution in the control group. All statistical analyses were performed using SAS 9.2 . Statistical significance was determined according to the conventional significance-level of α = 5%.Difference of clinical characteristics  between PTC cases and controls were tested by chi-square test or


The increase in clinical trials with androgen receptor (AR)-targeting drugs emphasizes the need of clarifying the role of AR expression in different breast cancer subtypes. AR confers good prognosis in estrogen receptor positive (ER+) breast cancer, but its role in ER-negative (ER−) breast cancer is unclear. The aim of this study was to elaborate on previous findings of a differential prognostic role for AR depending on ER status, using breast cancer mortality (BCM) as endpoint, in a population-based cohort from the Malmö Diet and Cancer Study.Immunohistochemical AR expression was assessed in 910 women with invasive breast cancer diagnosed 1991–2010, supplemented with clinicopathological information, vital status, and cause of death, with the last follow-up in December 2014 (median 10 years). Survival analyses according to AR status and AR/ER combinations were performed.n = 573, 85%) was associated with favorable established tumor markers and lower BCM in univariable analysis, especially during the first 5 years following diagnosis . Multivariable analysis for short-term follow-up indicated higher BCM among patients with AR+ER− tumors  than other AR and ER combinations.AR expression was available for 671 tumors. AR+ (AR expression added prognostic information to ER expression with respect to short-term prognosis. The worst prognosis was seen for patients with AR+/ER− tumors in short-term follow-up, supporting the pre-specified hypothesis. However, larger cohorts are needed for further characterization of the role of AR expression in ER− breast cancer. The heterogeneous nature of breast cancer highlights the importance of identifying prognostic and predictive markers for clinical implementation of tailored treatment. Recently, there has been increasing interest in the androgen receptor (AR) as a potential prognostic biomarker and treatment target , 2. The The most recent meta-analysis reported AR to be prognostically favorable irrespective of ER expression, but only included studies published until early 2015 . In lateThe aim of this study was to validate and elaborate the previous findings of a differential role for AR on the outcome depending on ER status, using breast cancer-related death as endpoint, in an independent cohort with longer follow-up, the population-based Malmö Diet and Cancer Study (MDCS). We hypothesized that AR positivity would be associated with overall favorable tumor characteristics and prognosis and that analyses stratified by tumor ER status would reveal a differential role of AR depending on ER. The primary endpoint was breast cancer mortality (BCM), and the secondary endpoint was all-cause mortality. Another aim was to investigate whether AR could add long-term prognostic information.The population-based prospective MDCS was initiated to examine associations between diet and cancer  and inclThe female MDCS cohort consists of 17,035 women (born 1923–1950). Information on incident breast cancer and vital status were annually retrieved from the Swedish Cancer Registry, the South Swedish Regional Tumor Registry, and the Swedish Cause of Death Registry. Ethical permission was obtained from the Ethical Committee at Lund University (Dnr 472/2007). All participants signed a written informed consent form.n = 576) were therefore excluded  and patients who received neo-adjuvant treatments (n = 4) were excluded. Patients were also excluded if they had distant metastasis at diagnosis (n = 14) or died from breast cancer-related causes ≤0.3 years from diagnosis (n = 2). Patients with bilateral cancers (n = 17) were excluded due to difficulties in evaluating the relation between tumor characteristics and prognosis. Finally, one patient who declined treatment for four years before accepting surgery was excluded. Thus, the final study population consisted of 910 patients. Information on cause of death and vital status was retrieved from the Swedish Causes of Death Registry, with last follow-up December 31st, 2014.This is a case-only analysis of incident primary breast cancer within the MDCS, and patients with a previous breast cancer diagnosis at enrollment  was retrieved from pathology reports for tumors from 2005 and onwards. From 2008 onwards, information was also obtained for the immunohistochemistry (IHC) based markers ER, progesterone receptor (PR), proliferation index (Ki67), and human epidermal growth factor receptor-2 (HER2) status. From 1991 to 2007, ER, PR, Ki67 and HER2 were assessed using tissue microarrays (TMAs) . A pathohttp://www.pathxl.com). Following assessment of tumor invasiveness using hematoxylin and eosin stained slides, a semi-quantitative scoring of AR fractions  of positively stained nuclei, irrespective of nuclear staining intensity, was performed by one observer (KE), Fig. A TMA was constructed with two 1 mm cores from each tumor . Among the 910 patients included in the study population, tumor tissue was available from 718 patients . Distributional differences were assessed by The association between AR and prognosis was examined with BCM as the primary endpoint. BCM was defined as the incidence of breast cancer-related death (when breast cancer was the cause of death or the contributing cause of death). The secondary endpoint was all-cause mortality, i.e., death from any cause. Follow-up was calculated from the date of breast cancer diagnosis to the date of breast cancer-related death, date of death from another cause, date of emigration or the end of follow-up as of December 31, 2014.Firstly, AR expression in relation to BCM was assessed by comparison of cumulative BCM (CBCM) for patients with AR+ and AR− tumors in the overall population and stratified by ER status. Rather than analyzing cause-specific survival, this method  takes coP = 0.06). The follow-up time was subsequently divided into categories of 0–5 years and >5 years, where the proportional hazards assumption was better met, and survival analyses by AR status were repeated for 0–5 years and >5 years of follow-up.Adjustments were made in two multivariable models including the following covariates. Model 1 included age at diagnosis (continuous), tumor size >20 mm (yes/no), ALNI (≥1 yes/no), histological grade (III vs. I–II), and ER status (±). Model 2 included the covariates from Model 1 with the addition of planned adjuvant treatments, chemotherapy (yes/no), radiotherapy (yes/no), and endocrine treatment (yes/no). An interaction term between AR and ER was added to the Cox models to evaluate the strength of evidence against the null hypothesis of no interaction between these factors on outcome. Proportional hazards assumptions were evaluated visually by inspecting the log minus log survival curves and formally using Schoenfeld’s test. There was weak evidence for a non-proportional hazards effect on AR ± for the overall follow-up  was used to estimate cause-specific cumulative mortality. Stata was also used to draw the cumulative mortality graphs and to test proportional hazards assumptions. All other statistical analyses were performed in SPSS 22.0 (IBM).All tests were two-sided, and Tumor AR expression was assessable in 671 of 718 cases (93%) where tumor tissue was available . When stratified by ER status, the CBCM observed among all patients with AR+ tumors was attributable to patients with AR+ER+ rather than AR+ER− tumors. However, the evidence for an association between AR+ER+ and lower CBCM was weak (LogRank P = 0.20) compared to AR−ER+. During the first 5 years of follow-up, no difference in CBCM by AR+ was observed for patients with ER− tumors. AR+ was weakly associated with higher all-cause mortality (LogRank P = 0.08), but this association was not seen in CM analyses stratified by ER status  regarding BCM.For the overall follow-up period, the incidence of BCM for patients with AR+ tumors was half that of patients with AR− tumors in univariable Cox analyses . The corresponding figures for the interval from five years and onwards pointed in a similar direction, although non-significant . In the adjusted models for ER+ versus ER− strata during the short-term follow-up, the estimated hazard ratios for BCM by AR diverged to a higher degree compared to the corresponding estimate in the overall follow-up period. However, no interactions between AR and ER status were observed (Pinteraction ≥ 0.35). After adjustments for age and tumor characteristics , there was moderate evidence that patients with AR+ER− tumors had higher BCM during the short-term follow-up compared to patients with AR+ER+ tumors. In this model, BCM for patients with AR+ER− tumors was more than threefold that of patients with AR+ER+ tumors. Furthermore, patients with AR+ER− tumors had almost double the BCM compared with patients with AR−ER− tumors. When further adjustments for adjuvant treatment were incorporated into the model (model 2), the patients with AR+ER− tumors still had worse prognosis compared to all other AR/ER combinations.The lower BCM in the AR+ group observed in the overall period seemed driven by a lower BCM relative to AR− during the first 5 years . This interpretation was supported by the unadjusted analysis of combined AR/ER. However, in line with our hypothesis, AR added short-term prognostic information after adjustment for confounders. The poorest prognosis was seen among patients with AR+ER− tumors. This was also true after adjustment for treatment, but the adjusted HRs were closer to 1.0, and the evidence for higher BCM compared to the other subgroups was weaker after adjustment. If AR+ER− breast cancer is confirmed to have inferior prognosis, this may impact the choice of AR-targeted treatments and reveal a need for closer surveillance of this patient group.There are several suggested reasons as to why we could not fully confirm our hypothesis. Our previous results on AR expression considered DFS in a cohort of median follow-up of 5 years. The time from diagnosis to breast cancer-related death may not be comparable with the event of local or regional recurrence, contralateral cancer, or distant metastasis, as evaluated previously . There mAR+ was confirmed to be associated with favorable tumor characteristics, as consistently shown previously , 29, 30.The strengths in this study were the long-term follow-up and a study population from a population-based, well-characterized cohort , 31. SinA lack of standardization of AR antibodies and cut-offs used has been common and may explain some discrepancies across studies , 11. In The tumor marker data were derived from TMAs for patients diagnosed up until 2007. Thereafter, routine clinical pathology reports were used. For the PR variable, this introduced a systematic bias due to higher positive frequency in clinical data as compared to TMA data. Thus, PR was only addressed in the descriptive analyses and should be interpreted with caution. In contrast, ER status did not depend on the diagnostic period. The missing ER was addressed by presenting the associated HRs separately.This study indicates that cohorts with larger sample sizes are needed to further elucidate the prognostic role of AR in ER− breast cancer. Indeed, there is a need to incorporate several hormonal receptors in future studies of prognosis and treatment prediction . AR may The number of ongoing clinical studies involving AR targeting is steadily expanding and has included patients with both early and advanced breast cancer and of different subtypes –39 (ClinThere is yet no consensus on whether AR antagonists or agonists are preferable as targeted treatments in any breast cancer subtype. The forthcoming reports of ongoing clinical trials—combined with observational studies and mechanistic studies on treatment response/resistance –44—will In conclusion, AR+ was associated with lower BCM in the overall study population and for the overall and short-term follow-up intervals, but not for long-term follow-up. The poorest prognosis was seen among patients with AR+ER− tumors after adjustment for confounders in the short-term follow-up. Future studies on the role of AR in breast cancer require larger cohorts, especially in the ER− subset, and the inclusion of gene expression analyses may add valuable information.


Strategies adopted by individuals in a social network significantly impact the network, and they strongly affect relationships between individuals in the network. Links between individuals also heavily influence their levels of cooperation. Taking into account the evolution of each individual’s connection, we explore how sensitivity and visibility affect the prisoner’s dilemma game. The so-called ‘sensitivity’ and ‘visibility’ respectively present one’s self-protection consciousness and the ability of gaining information. We find that at moderate levels of player sensitivity cooperative behavior increases, but that at high levels it is inhibited. We also find that the heterogeneity of the weight of individuals at the end of the game is higher when sensitivity and visibility are increased, but that the successful-defection-payoff has less impact on the weight of individuals and on the relationship between the heterogeneity of the weight of individuals and the density of cooperators. This framework can be used to clarify the interaction mechanism between the micro-level of individual behavior and the macro-level of individual co-evolutionary processes. Robert May has said that the most important unanswered question in evolutionary biology and in the social sciences in general is how cooperative behavior evolved and how it is maintained in human societies and in animal groupings2356891011121415161719202122Although cooperation is ubiquitous in social life and an important topic in a wide variety of economic interactions, field data rarely enable us to discriminate among competing theories. Although our understanding of cooperative behavior among humans is thus limited, carefully designed laboratory experiments may enrich our knowledge of human cooperation27In an adaptive dynamical environment, choosing an advantageous strategy is the key factor in winning. A chosen strategy can significantly affect relationships with other individuals, but the closeness among individuals also heavily influences their chosen level of cooperation. To understand the interaction mechanism between the micro-level of individual behavior and macro-level of individual co-evolutionary processes, we construct a model that takes into consideration the evolution of the links of individuals and explore how sensitivity and visibility affect the prisoner’s dilemma game. Although we find the sensitivity and visibility of individuals can persuade more players to adopt a cooperative strategy and increase individual weight heterogeneity, we also find that a higher level of sensitivity has a negative impact on cooperative behavior. We also find that the successful-defection-payoff has less impact on individual weight heterogeneity and on the relationship between individual weight heterogeneity and cooperator density.C ↔ C, C ↔ D, and D ↔ D, respectively. Here we adopt edge weight wij to measure the closeness of two players. A large edge weight in general indicates a better relationship between the two players, and a small one the opposite. The edge weight wij evolves with player strategy. Note that a win-win strategy promotes the relationship between two individuals. The edge weight increases when both ends of an edge choose a cooperation strategy, but when a cooperator connects with a defector, the cooperator loses heavily and the closeness is significantly weakened. The evolution of closeness isWe distinguish the edge types between any two players as cooperation-cooperation, cooperation-defection, or defection-defection, and we mark them w is the variable used to reflect the individual’s sensitivity awareness. When the value of Δw is large, the players are extremely sensitive to changes in their payoff. The result is that when a cooperator connects with a defector, the closeness decreases heavily in the next game round. In particular, the link between players i and j breaks when wij ≤ 0, and the player who receives the smaller payoff will choose to connect to another player. Note that self-connections and multi-edges are avoided. i and j.where ΔZ = 1000. Initially the topology is a Erdos-Renyi random network with an average degree <k> = 6. Because the relationship between any two players differs, we set a weight U stands for a uniform distribution). We then use the Monte Carlo method and perform a prisoner’s dilemma game29i is initially designated with equal probability either a cooperator (C) or defector (D) (see Methods). To alleviate randomness, we obtain the equilibrium by averaging over 100 independent runs.We consider the evolution of social dilemmas on a network of size w, ρc evolves as the game progresses. The unique Nash equilibrium in the prisoner’s dilemma game is defection-defection. The optimal Pareto configuration is cooperation-cooperation. Because cooperation brings an overall increase in benefit, the equilibrium of the optimal behavior of a single individual does not coincide with the Pareto optimum. We can find this through comparison. Although players often defect in hopes of obtaining a larger income, this weakens relationships and raises the probability that opportunistic defectors (free-riders) will be quickly abandoned, causing a significant decrease in their income. Under a different sensitivity awareness ΔT produces a higher return, the higher the sensitivity awareness Δw of the players, the more quickly they change their relationships with each other. Thus when a player adopts a betrayal strategy, his links with cooperative players disconnect. Defectors with more neighbors may get a larger income, but in general defectors have fewer neighbors, most of which are also defectors. This eventually reduces the total income of the defectors. Although the successful-defection-payoff strategy i by adding up all the edge weights that link to its partners. Thus the weight of node i is Ni denotes the neighbor node sets. Variance informally measures how far a set of numbers is spread out from their mean, we here use w2〉 is the mean of the square of the weight, 〈w2〉 is the square of the mean of the weight. w increases. When the sensitivity of a player increases to a certain level, the edges connecting them to betrayers break easily. Although the betrayal strategy can bring greater benefits to an individual, changes in the network topology and node connectivity accelerate as player sensitivity increases. Eventually most of the cooperative players are linked with other cooperative players, and the closeness between the win-win players is enhanced. At the same time, players who choose defection have a low probability of connecting with other players. Thus network heterogeneity increases.The importance of the node within a system is its weight, and node weight is strongly associated with the weight of its edge. We calculate the weight of node p, the final cooperator density ρc(∞) is significantly higher than when there is a low initial visibility level. p increases, network heterogeneity slowly increases. At a higher visibility level p, players can obtain greater amounts of information from their neighbors. When a player can see that their income is lower than that of their neighbors, they change their current connection mode and cause an imbalance in weight distribution.The visibility level of individuals in the network reflects the information flow. In a social network, the level of visibility is also an expression of the fairness of information acquisition. The greater the proportion of visible individuals, the greater will be the ability of the individuals to obtain information. When there is a high initial visibility level p exhibiting qualitatively different behavior. The density of cooperators ρc(∞) increases with individual income sensitivity Δw irrespective of the p value. Note that when the sensitivity increases to a certain value, the final cooperator density decreases. The Δw value significantly affects the final heterogeneity level of the system. When individual sensitivity Δw increases to a certain level, small changes in individual payoff strongly impact the relationships among the players. Although it encourages players to connect to optimal “partners,” relationships between the individuals change rapidly. The network becomes “unstable.” It is unstable in terms of network topology and network connectedness. When network topology changes rapidly, players connect to new neighbors more quickly irrespective of the strategy the neighbor has adopted. However, here individuals may link to defectors more frequently, which causes more players to take the betrayal strategy and eventually decreases the proportion of cooperative players.Dw reflects the differentiation of the system. A large Dw value means that the distribution of node influence is more heterogeneous. In order to study the relation between ρc(∞) and Dw, we use the value of Δw from 0 to 1  in our model. We find that the density of cooperators first increases and then decreases with network heterogeneity under differing successful-defection-payoff T values. Dw values facilitate cooperative behavior between selfish individuals. At the same time the sensitivity of players to their payoffs hinders cooperative behavior. The successful-defection-payoff T has less influence on the relation between the final network heterogeneity wD and the density of final cooperators ρc(∞).The final network heterogeneity p and successful-defection-payoff T. T, irrespective of individual sensitivity values. However when individual sensitivity is low Δw, the cooperator density decreases as the visibility level increases for a large T. At the same visibility level and successful-defection-payoff T value, a larger Δw promotes cooperation more effectively. Note that when Δw is large enough, the value of T has little effect on the density of cooperators ρc(∞), but when Δw is small the value of T strongly influences the density of cooperators ρc(∞).We see that an individual cooperative behavior can be controlled by a player’s past experiences and present environment. T has a smaller impact on the heterogeneity of player weights and on the relationship between heterogeneity of player weights and cooperator density. We have run our model using different parameter values, and we have found that the final results indicate a high level of robustness. The framework developed here could be used to clarify the interaction mechanism between the micro-level of individual behavior and macro-level of individual co-evolutionary processes.To investigate some of the consequences of cooperation in the prisoner’s dilemma game, we have examined the effects of sensitivity and visibility. We find that although high player sensitivity promotes cooperative behavior, extremely high levels inhibit it. We also find that the heterogeneity of player weights in the final network is higher at higher levels of sensitivity and visibility, but that the successful-defection-payoff indirectly affect individual behavior, i.e., we focus on how people influence others when there is a connection, irrespective of linking weight. Our model is nevertheless realistic. In general, conflicts of interest exist when there are connection, and vice versa. We here use a simple but practical model to show the evolution of human behavior. Further experimental research is needed to clarify our results, and we hope that our work will motivate work in that direction.To summarize, in a complex social network, choosing a strategy is important. A player’s chosen strategy strongly affects their relationship with other players, and the links between individuals strongly affect their decision of whether to cooperate. This study offers a new perspective on the prisoner’s dilemma game in a dynamic social network: how player sensitivity and visibility affect its evolution. Our analysis provides a framework for studying the co-evolution of games and strategies, and suggests that maintaining cooperation may be more difficult than previously thought. Although in some real-world systems restrictive assumptions may not be germane, our work sheds some light on the framework of evolutionary games in networks. Although here we focus on how links directly affect individual behavior, we do not consider how links A composed of Z players in which A = (aij)z×z is the adjacency matrix. When two players i and j interact, aij = 1, otherwise aij = 0. In the classical prisoner’s dilemma game, each player decides to be either a cooperator or a defector. When a cooperator links with a defector, the cooperator receives payoff S and the defector receives payoff T (we designate this scenario successful-defection-payoff). When both are cooperators, both receive payoff R, and when both are defectors, both receive payoff P. The prisoner’s dilemma game thus satisfies the conditions: T > R > P > S and 2R > T + S31R > P mutual cooperation produces a higher payoff than mutual defection. When T > R and P > S the player with a defection strategy will obtain a higher payoff, regardless of what choice the opponent makes in a single round of the game. An addition, 2R > T + S is required to ensure that cooperation exists. We consider the prisoner’s dilemma game in a network i can be described using the payoff matrix of the prisoner’s dilemma game,Because an individual’s gain or loss depends on the actions of other players, the payoff to player i. If player i adopts a cooperation strategy, then R = 1, S = 0 and P = 0. We can also calculate the total payoff fi of player i using Here p and that they know both their own payoffs and the payoffs of each of their directly connected neighbors. Blind players only have knowledge of their own payoffs. A seeing player i decides whether to adopt the strategy of their neighbors. Player i with strategy Si chooses neighbor j who has the maximum payoff and then imitates the strategy of j. We apply the Fermi rule that individual i adopts the strategy of selected neighbor j with a probabilityBecause real-world individuals differ in their ability to acquire information, we divide players into two groups, those who are seeing and those who are blind. We assume that the initial proportion of seeing players is β ≥ 0, which in physics corresponds to the inverse temperature, here measures the strength of natural selection. When β = 0, we get β = 10, which means that it is highly likely that the strategy of the better performing players will be adopted by their neighbors, and less likely (but not impossible) that the strategy of the less successful players will be adopted by their neighbors.The quantity t−1 player i strongly benefited using their chosen strategy, they will continue with that same strategy, but if in the previous round t−1 player i weakly benefited using their chosen strategy, they will adopt the opposite strategy. Typically player i randomly chooses a strategy that is either the same or the opposite of their previous strategy if there is no change in payoff. The adopted strategy of player i at game round t isIf players are blind, they will choose a strategy for the next round of the game based solely on their previous experience. If in the previous round si indicates a strategy opposite to si, and fi(t−1) and fi(t−2) are the individual’s payoffs at game round t − 1 and t − 2, respectively.Here −We use nonparametric regression procedures to obtain a smooth set of points from each set of scattered data Kh(k − ki) is a Gaussian kernel of the form,Here h suggested by Bowman and Azzalini isThe optimal bandwidth Where How to cite this article: Li, D. et al. The co-evolution of networks and prisoner’s dilemma game by considering sensitivity and visibility. Sci. Rep.7, 45237; doi: 10.1038/srep45237 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


Seventy-four per cent of YLDs are the result of health conditions for which rehabilitation may be beneficial.,–Factors contributing to the unmet need for rehabilitation services include poor accessibility, transport barriers, high out-of-pocket expenses and long waiting times.The role that rehabilitation plays in maximising the impact of other health services – such as surgical interventions, trauma care and management of noncommunicable diseases – and its potential for significant cost savings are also frequently misunderstood and underestimated.Rehabilitation 2030: a call for action, a commitment to key actions to strengthen rehabilitation services in Member States.Rehabilitation is part of universal health coverage and should be incorporated into the package of essential services, along with prevention, promotion, treatment and palliation. To this end, on 7 February 2017, WHO, Member States, international and professional organizations, nongovernmental organizations and rehabilitation experts issued


We design an axiomatization for reversible computation called reversible ACP (RACP). It has four extendible modules: basic reversible processes algebra, algebra of reversible communicating processes, recursion and abstraction. Just like process algebra ACP in classical computing, RACP can be treated as an axiomatization foundation for reversible computation. Reversible computation , algebra of reversible communicating processes (ARCP), recursion and abstraction. While in CCSK, recursion and abstraction are not concerned.one atomic action. Also auto-concurrency is out of scope for our work here.In comparison to ACP, it is almost a brand new algebra for reversible computation which has the same advantages of ACP, such as modularity, axiomatization, etc. Firstly, in RACP, the alternative composition is replaced by choice composition, since in reversible computing, all choice branches should be retained. Secondly, the parallel operator cannot be captured by an interleaving semantics. Thirdly, more importantly to establish a full axiomatization, all the atomic actions are distinct, the same atomic action in different branches  will be deemed as the same RACP is an axiomatic refinement to CCSK:The paper is organized as follows. In section “For convenience of the reader, we introduce some basic concepts about equational logic, structural operational semantics and process algebra ACP  A signature f has an arity ar(f), being its number of arguments. A function symbol a, b, c, …of arity zero is called a constant, a function symbol of arity one is called unary, and a function symbol of arity two is called binary.(Term) Let s, t, u, …over (Substitution) Let x in t by x.(Axiomatization) An axiomatization over a signature  An axiomatization over a signature f is a function symbol with (Model) Assume an axiomatization (Term rewriting system) Assume a signature s is not a single variable; (2) all variables that occur at the right-hand side t also occur in the left-hand side s. A term rewriting system (TRS) is a finite set of rewrite rules.(Rewrite relation) A TRS over a signature f is a function symbol with  A term is called a normal form for a TRS if it cannot be reduced by any of the rewrite rules.(Termination) A TRS is terminating if it does not induce infinite reductions (Weak confluence) A TRS is weakly confluent if for each pair of one-step reductions u such that Commutativity and associativity) Assume an axiomatization f is commutative if (Convergence) A pair of terms s and t is said to be convergent if there exists a term u such that , transition system specification (TSS), transition rule and its source, source-dependent, conservative extension, fresh operator, panth format, congruence, bisimulation, etc. These concepts are coming from Fokkink , and areS of states, a finite, non-empty set of transition labels A and a finite set of predicate symbols.We assume a non-empty set Labeled transition system) A transition is a triple s, P) with P a predicate, where (Transition system specification) A transition rule H a set of expressions tP with tP with (Proof) A proof from a TSS T of a closed transition rule l, and K is the set of labels of nodes directly above this node, then (a) either K is the empty set and T.(Generated LTS) We define that the LTS generated by a TSS T consists of the transitions T. hold for a set A set Three-valued stable model) A pair T if it satisfies the following two requirements: (1) a transition T proves a closed transition rule N contains only negative premises and T proves a closed transition rule N contains only negative premises and  The ordinal numbers are defined inductively by: (1) 0 is the smallest ordinal number; (2) each ordinal number (Positive after reduction) A TSS is positive after reduction if its least three-valued stable model does not contain unknown transitions.(Stratification) A stratification for a TSS is a weight function tP of  A process (graph) p is an LTS in which one state s is elected to be the root. If the LTS contains a transition sP, then pP. (1) A process (Reverse transition) There are two processes p and a[m] is a kind of special action constant a, and (Bisimulation) A bisimulation relation pP, then qP; (4) if qP, then pP. Two processes p and q are bisimilar, denoted by (Forward–reverse bisimulation) A forward–reverse (FR) bisimulation relation pP, then qP; (6) if qP, then pP. Two processes p and q are FR bisimilar, denoted by (Congruence) Let (Panth format) A transition rule  A branching bisimulation relation pP, then there is a sequence of (zero or more) qP, then there is a sequence of (zero or more) p and q are branching bisimilar, denoted by (Branching forward–reverse bisimulation) A branching forward–reverse (FR) bisimulation relation pP, then there is a sequence of (zero or more) qP, then there is a sequence of (zero or more) pP, then there is a sequence of (zero or more) qP, then there is a sequence of (zero or more) p and q are branching FR bisimilar, denoted by (Rooted branching bisimulation) A rooted branching bisimulation relation pP, then qP; (4) if qP, then pP. Two processes p and q are rooted branching bisimilar, denoted by (Rooted branching forward–reverse bisimulation) A rooted branching forward–reverse (FR) bisimulation relation pP, then qP; (6) if qP, then pP. Two processes p and q are rooted branching FR bisimilar, denoted by (Lookahead) A transition rule contains lookahead if a variable occurs at the left-hand side of a premise and at the right-hand side of a premise of this rule.(Patience rule) A patience rule for the ith argument of a function symbol f is a panth rule of the form(RBB cool format) A TSS T is in RBB cool format if the following requirements are fulfilled. (1) T consists of panth rules that do not contain lookahead. (2) Suppose a function symbol f occurs at the right-hand side the conclusion of some transition rule in T. Let f in T. Let tP with (Source-dependency) The source-dependent variables in a transition rule of t are source-dependent, then all variables in (Freshness) Let is source-dependent. (2) For eacheither the source ofis fresh, orhas a premise of the formortP, whereall variables intoccur in the source ofandorPis fresh.ACP Fokkink  is a kinBisimulation or rooted branching bisimulation based structural operational semantics is used to formally provide each process term used the above operators and constants with a process graph. The axiomatization of ACP  for BRPA as follows.The first transition rule says that each history of an atomic action s or t, but the other branch remains.The next four transition rules say that s or t is reversed successfully, we define The next four transition rules say that t in the first and leads to a successful atomic action, after successful reverse of t, then reverse of s follows. When both s and t are reversed successfully, we define The last four transition rules say that We give the reverse transition rules under transition system specification (TSS) for BRPA as follows.We design an axiomatization mentclass2pt{minimThe following conclusions can be obtained.FR bisimulation equivalence is a congruence with respect to BRPA.The forward and reverse TSSs are all in panth format, so FR bisimulation equivalence that they induce is a congruence. is sound for BRPA modulo FR bisimulation equivalence.s and t to basic reversible process terms, then we need to check that Since FR bisimulation is both an equivalence and a congruence for BRPA, only the soundness of the first clause in the definition of the relation = is needed to be checked. That is, if RA1 (commutativity of +) says that RA2 (idempotency of +) is used to eliminate redundant branches.s, t, u.RA3 (associativity of +) says that RA4  of the + , and this equivalence relation is denoted by s then represents the collection of basic reversible process term t such that s modulo AC of the + can be represented in the form s.We consider basic reversible process terms modulo associativity and commutativity  for the static parallel s and t are forward executed successfully, we define The above eight transition rules are forward transition rules for the static parallel s and t are reversed successfully, we define The above eight transition rules are reverse transition rules for the static parallel The forward transition rules under TSS for communication merge are as follows and say that the communication can be merged. Where a communication function The reverse transition rules under TSS for communication merge are as follows and say that the communication can be merged.RPAP is a conservative extension of BRPA.Since the TSS of BRPA is source-dependent, and the transition rules for the static parallel FR bisimulation equivalence is a congruence with respect to RPAP.The TSSs for RPAP and BRPA are all in panth format, so FR bisimulation equivalence that they induce is a congruence. We design an axiomatization for RPAP illustrated in Table Then, we can obtain the soundness and completeness theorems as follows.is sound for RPAP modulo FR bisimulation equivalence.s and t to reversible process terms, then we need to check that Since FR bisimulation is both an equivalence and a congruence for RPAP, only the soundness of the first clause in the definition of the relation = is needed to be checked. That is, if s and t.RP1 says that s.RP2 says that RP3-RP7 say that the static parallel operator satisfies associativity, left distributivity and right distributivity to + and s and t.RC8-RC15 are the defining axioms for the communication merge, which say that RC16-RC17 say that the communication merge We only provide some intuition for the soundness of the axioms in Table These intuitions can be made rigorous by means of explicit FR bisimulation relations between the left- and right-hand sides of closed instantiations of the axioms in Table is complete for RPAP modulo FR bisimulation equivalence.To prove that (1) We consider the introduction to the static AC) of the + , and this equivalence relation is denoted by s then represents the collection of reversible process term t contains s modulo AC of the + can be represented in the form s.We consider reversible process terms contains +, Then RP2-RP7 are turned into rewrite rules from left to right:AC of the +.Then these rewrite rules are applied to the above reversible process terms modulo We let the weight functionAC of the +.We can see that the TRS is terminating modulo n and n and a of n. Then a.Consider a summand a[m] of n. Then a[m].Consider a summand n. Then Consider a summand n. Then Consider a summand n. Then Consider a summand n. Then Consider a summand The summands Next, we prove that normal forms n is also a summand of n. In other words, Hence, each summand of s and t contains +, AC of the +, so it reduces s and t to normal forms n and AC of the + can be derived from the axioms, Finally, let the reversible process terms (2) We prove the completeness of the axioms involve the parallel operator The axioms RP1 and RC8-RC17 are turned into rewrite rules, by directing them from left to right.AC of the +.Then these rewrite rules are applied to the above reversible process terms modulo We let the weight functionAC of the +.We can see that the TRS is terminating modulo n.n is an atomic action, then it does not contain any parallel operators.If s and t do not contain n does not contain Suppose n cannot be of the form s and t do not contain s and t, which all lead to the conclusion that one of the directed versions of RC8-RC17 can be applied to n. We conclude that n cannot be of the form Suppose We prove that normal forms n do not contain occurrences of the remaining two parallel operators Hence, normal forms do not contain occurrences of parallel operators s and t be FR bisimilar. The TRS is terminating modulo AC of the +, so it reduces s and t to normal forms n and AC of the + can be derived from the axioms, Finally, let the reversible process terms H of atomic communicating actions and their histories, which renames all actions in H into A mismatch in communication of two actions t of which the labels are not in H, which is expressed by the following two forward transition rules.The encapsulation operator The reverse rules are as follows.ARCP is a conservative extension of RPAP.Since the TSS of RPAP is source-dependent, and the transition rules for encapsulation operator FR bisimulation equivalence is a congruence with respect to ARCP.The TSSs for ARCP and RPAP are all in panth format, so FR bisimulation equivalence that they induce is a congruence. The axioms for ARCP are shown in Table The soundness and completeness theorems are following.is sound for ARCP modulo FR bisimulation equivalence.s and t to reversible process terms, then we need to check that Since FR bisimulation is both an equivalence and a congruence for ARCP, only the soundness of the first clause in the definition of the relation = is needed to be checked. That is, if RA6 says that the deadlock RA7-RA8, RP8-RP9, RC18-RC19 say that the deadlock RD1-RD5 are the defining axioms for the encapsulation operator t labelled with atomic actions from H are blocked.RD6-RD8 say that in We only provide some intuition for the soundness of the axioms in Table These intuitions can be made rigorous by means of explicit FR bisimulation relations between the left- and right-hand sides of closed instantiations of the axioms in Table is complete for ARCP modulo FR bisimulation equivalence.To prove that The axioms RA6-RA8, RD1-RD8, RP8-RP9, RC18-RC19 are turned into rewrite rules, by directing them from left to right. The resulting TRS is applied to process terms in RPAP modulo AC of the +.AC of the +.Then these rewrite rules are applied to the above reversible process terms modulo We let the weight functionAC of the +.We can see that the TRS is terminating modulo n.If If If If If We prove that normal forms n do not contain occurrences of Hence, normal forms do not contain occurrences of s and t be FR bisimilar. The TRS is terminating modulo AC of the +, so it reduces s and t to normal forms n and AC of the + can be derived from the axioms, Finally, let the reversible process terms To capture infinite computing, recursion is introduced in this section. In ARCP, because parallel branches cannot be merged, the static parallel E, F, G are guarded linear recursion specifications, X, Y, Z are recursive variables. We first introduce several important concepts, which come from Fokkink  A recursive specification is a finite set of recursive equations(Solution) Processes (Guarded recursive specification) A recursive specification(Linear recursive specification) A recursive specification is linear if its recursive equations are of the form and the RSP (Recursive Specification Principle) are shown in Table RP2–RP9 + RDP + RSP is sound for ARCP-RP with guarded recursion modulo FR bisimulation equivalence.s and t to reversible process terms, then we need to check that Since FR bisimulation is both an equivalence and a congruence for ARCP-RP with guarded recursion, only the soundness of the first clause in the definition of the relation = is needed to be checked. That is, if Soundness of RDP follows immediately from the two transition rules for guarded recursion, which express that Soundness of RSP follows from the fact that guarded recursive specifications have only one solution modulo FR bisimulation equivalence.We only provide some intuition for the soundness of RDP and RSP in Table These intuitions can be made rigorous by means of explicit FR bisimulation relations between the left- and right-hand sides of RDP and closed instantiations of RSP in Table RP2–RP9 + RDP + RSP is complete for ARCP-RP with linear recursion modulo FR bisimulation equivalence.The proof is similar to the proof of “ Fokkink . \documeE a linear recursive specification:E consist of the recursive equationsE, RSP yields Firstly, each process term Then, if A program has internal implementations and external behaviors. Abstraction technology abstracts away from the internal steps to check if the internal implementations really display the desired external behaviors. This makes the introduction of special silent step constant Firstly, we introduce the concept of guarded linear recursive specification, which comes from Fokkink .Guarded linear recursive specification) A recursive specification is linear if its recursive equations are of the form the TSS of ARCP-RP with guarded linear recursion is source-dependent; (2) and the transition rules for the silent step Rooted branching FR bisimulation equivalence is a congruence with respect to ARCP-RP with silent step and guarded linear recursion.The TSSs for ARCP-RP with silent step and guarded linear recursion are all in RBB cool format, by incorporating the successful termination predicate The axioms for silent step are shown in Table RP2–RP9 + RB1–RB4 + RDP + RSP is sound for ARCP-RP with silent step and guarded linear recursion, modulo rooted branching FR bisimulation equivalence.s and t to reversible process terms, then we need to check that Since rooted branching FR bisimulation is both an equivalence and a congruence for ARCP-RP with silent step and guarded recursion, only the soundness of the first clause in the definition of the relation We only provide some intuition for the soundness of axioms in Table The axioms in Table This intuition can be made rigorous by means of explicit rooted branching FR bisimulation relations between the left- and right-hand sides of closed instantiations of RB1–RB4. RP2–RP9 + RB1–RB4 + RDP + RSP is complete for ARCP-RP with silent step and guarded linear recursion, modulo rooted branching FR bisimulation equivalence.The proof is similar to the proof of “ Fokkink .E a guarded linear recursive specification:E consist of the recursive equationsE, RSP yields Firstly, each process term Then, if Abstraction operator t that are in the set I into Abstraction operator with guarded linear recursion is a conservative extension of ARCP-RP with silent step and guarded linear recursion.Since (1) the TSS of ARCP-RP with silent step and guarded linear recursion is source-dependent; (2) and the transition rules for the abstraction operator contain only a fresh Rooted branching FR bisimulation equivalence is a congruence with respect towith guarded linear recursion.The TSSs for The axioms for abstraction operator are shown in Table Before we introduce the cluster fair abstraction rule, the concept of cluster is recaptured from Fokkink .Cluster) Let E be a guarded linear recursive specification, and X and Y in E are in the same cluster for I if and only if there exist sequences of transitions (a or aX is an exit for the cluster C if and only if: (1) a or aX is a summand at the right-hand side of the recursive equation for a recursion variable in C, and (2) in the case of AX, either ∉C Table .Table 7C+ RSP + RDP + CFAR is sound forwith guarded linear recursion, modulo rooted branching FR bisimulation equivalence.s and t to reversible process terms, then we need to check that Since rooted branching FR bisimulation is both an equivalence and a congruence for RTI1–RTI5 are the defining equations for the abstraction operator t labelled with atomic actions from I are renamed into RTI6–RTI7 say that in We only provide some intuition for the soundness of axioms in Table This intuition can be made rigorous by means of explicit rooted branching FR bisimulation relations between the left- and right-hand sides of closed instantiations of RTI1–RTI7. + RSP + RDP + CFAR is complete forwith guarded linear recursion, modulo rooted branching FR bisimulation equivalence.The proof is similar to the proof of “ Fokkink .E a guarded linear recursive specification.Firstly, each process term Then, if RACP has many applications, for example, it can be used in verification for business protocols with compensation support. Since a business protocol is usually cross organizational boundaries and survives for a long period of times. The failure of a business protocol can be remedied by a series of compensation operations. A business protocol with compensation support means that each atomic operations in the business protocol is corresponding to an atomic compensation operation, and the computation logic of the business protocol can be reversed.The user plans a travel on UserAgent.He/she submits the travel plan to the travel corporation TravelCorp via UserAgent.TravelCorp receives the travel plan.It books traffic tools and hotels according to the travel plan.It sends the pay order to UserAgent.UserAgent receives the pay order.UserAgent sends the pay information to TravelCorp.TravelAgent receives the pay information.TravelAgent sends the business order to the Bank.The Bank receives the business order and does paying.We take an example of business protocols as Fig. The above business protocol as Fig. We define the following communication functions.After the successful forward execution of the above process term, the following reversible process term can be obtained.After the successful reverse execution (Compensation) the above process term, the original process term can be obtained.RACP can be used in correctness verification under the framework of reversible computation for business protocols with compensation support.UTB and let interactions between UserAgent, TravelCorp and Bank be internal actions. UTB receives external input A by communicating action D by communicating action In Fig. entclass1pt{minimaThen the state transition of UserAgent can be described by RACP as follows.The state transition of TravelAgent can be described by RACP as follows.And the state transition of Bank can be described by RACP as follows.We define the following communication functions.U, T and B in parallel, then the system UTB can be represented by the following process term.Let Then we get the following conclusion.The business protocol as Fig. 2showsexhibits desired external behaviors under the framework of reversible computation.Let Then we apply abstraction operator We get entclass1pt{minimaOne of the most fascinating characteristics is the modularity of RACP, that is, RACP can be extended easily. Through out this paper, we can see that RACP also inherents the modularity characteristics of ACP. By introducing new operators or new constants, RACP can have more properties. It provides RACP an elegant fashion to express a new property.In this section, we take an example of renaming operators which are used to rename the atomic actions.t, and assumes a renaming function Renaming operator with guarded linear recursion and renaming operators is a conservative extension ofwith guarded linear recursion.Since (1) the TSS of Rooted branching FR bisimulation equivalence is a congruence with respect towith guarded linear recursion and renaming operators.The TSSs for The axioms for renaming operator is shown in Table + RSP + RDP + CFAR + RRN1–RRN5 is sound forwith guarded linear recursion and renaming operators, modulo rooted branching FR bisimulation equivalence.s and t to reversible process terms, then we need to check that Since rooted branching FR bisimulation is both an equivalence and a congruence for RRN1–RRN3 are the defining equations for the renaming operator t are renamed by means of the mapping f.RRN4–RRN5 say that in We only provide some intuition for the soundness of axioms in Table This intuition can be made rigorous by means of explicit rooted branching FR bisimulation relations between the left- and right-hand sides of closed instantiations of RRN1-RRN5. + RSP + RDP + CFAR + RRN1-RRN5 is complete forwith guarded linear recursion and renaming operators, modulo rooted branching FR bisimulation equivalence.t in E a guarded linear recursive specification. Namely, then the desired completeness result follows from the fact that if It suffices to prove that each process term t can be applied. The only new case (where RRN1-RRN5 are needed) is E, we prove the case of E consists of guarded linear recursive equationsF consists of guarded linear recursive equationsStructural induction with respect to process term F. So by RSP, Replacing In this paper, we give reversible computation an axiomatic foundation called RACP. RACP can be widely used in verification of applications in reversible computation.For recursion and abstraction, it is reasonable to do extensions based on ARCP-RP (ARCP without static parallel Any computable process can be represented by a process term in ACP (exactly n et al. . That isSame as ACP, RACP has good modularity and can be extended easily. Although the extensions can not improve the expressive power of RACP, it still provides an elegant and convenient way to model other properties in reversible computation.


Androgen deprivation therapy with antiandrogens to reduce androgen biosynthesis or prevent androgens from binding to AR are widely used to suppress AR-mediated PCa growth. However, most of ADT may eventually fail with development of the castration resistance after 12-24 months. Here we found that a natural product baicalein can effectively suppress the PCa progression 50 were determined by MTT assay after 48 hrs treatment. RT-PCR was used to evaluate the mRNA levels of AR target genes including PSA, TMPRSS2, and TMEPA1. Western blot was used to determine AR and PSA protein expression.PCa cells including LNCaP, CWR22Rv1, C4-2, PC-3, and DU145, were treated with baicalein and luciferase assay was used to evaluate their effect on the AR transactivation. Cell growth and ICThe natural product of baicalein can selectively inhibit AR transactivation with little effect on the other nuclear receptors, including ERα, and GR. At a low concentration, 2.5 μM of baicalein effectively suppresses the growth of AR-positive PCa cells, and has little effect on AR-negative PCa cells. Mechanism dissection suggest that baicalein can suppress AR target genes  expression in both androgen responsive LNCaP cells and castration resistant CWR22Rv1 cells, that may involve the inhibiting the AR N/C dimerization and AR-coactivators interaction.Baicalein may be developed as an effective anti-AR therapy via its ability to inhibit AR transactivation and AR-mediated PCa cell growth. Prostate cancer (PCa) is the second leading cause of cancer death among American men. In 2015, approximately 228,800 men were diagnosed with PCa, and 27,540 men were expected to die from this disease in the United States . Since tADT aims to reduce the biosynthesis of androgens and/or prevent from androgen binding to androgen receptor (AR) –6. HowevScutellaria baicalensis Georgi  (MTT) were purchased from Sigma-Aldrich  and stored at −20°C in the dark. All other chemicals and solvents used in this study were of reagent grade or high performance liquid chromatography (HPLC) grade.Commercial compounds and reagents include 5α-dihydrotestosterone (DHT), dexamethasone (DEX), RU486, progesterone, hydroxflutamide (HF), 17β-estradiol (E2), ICI 182,780 (ICI), methanol, dimethyl sulphoxide (DMSO), chloroform  were extracted with 70% ethanol at effluent temperature for three hrs twice. The solvent was evaporated to obtain crude extract (49.8g), which was applied to the silica gel column chromatography, eluted by n-Hexane- Ethyl acetate mixture and petroleum ether- Ethyl acetate as mobile phases to obtain different fractions, based on the TLC pattern. Baicalein (28 mg) was separated and purified under silica gel column chromatography with a solvent system of petroleum- Ethyl acetate mixture, and identified by comparison of NMR and MS spectral data with reference values  was used at room temperature for 1 hr. Immunoblot analysis was performed with horseradish peroxidase- conjugated anti-rabbit or anti-mouse IgG antibodies using enhanced chemiluminescence Western blotting detection reagents (Amersham Biosciences). The quantification of Western blotting results was done by Image Lab statistic software (Bio-red).2.5×104 LNCaP cells, 1.5×104 CWR22RV1 cells, 6.0×103 C4-2 cells, 5.0×103 PC-3 cells, 5.0×103 DU145 cells were plated in triplicate in 24-well culture plates. Cell medium was replenished and cell growth was determined by MTT assay (Sigma) and direct cell count. Serum-free medium containing MTT (0.5 μg/ml) was added into each well. After two hrs incubation at 37°C all crystals had solubilized and the optical density of the solution was determined spectrophotometrically at 570 nm [To determine cell growth, 2.0×10t 570 nm –34.50 value, 1.0×106 LNCaP cells, 5.0×105 CWR22RV1 cells, 6.0×104 C4-2 cells, 5.0×104 DU145 cells, and 5.0×104 PC-3 cells were plated in triplicate in 5% CS-FBS RPMI medium in 24-well culture plates. The cells were incubated with serial concentrations of baicalein for 2 days and cell viability was determined in triplicate by MTT assay. Baicalein + medium only (no cells) were included as controls. Baicalein treated cells were compared to untreated cell control wells. IC50 values were analyzed with the program CompuSyn (Developer).Cytotoxicity assay was performed according to the protocol in our laboratory . To deteData are presented as the means ± SDs for the indicated number of separate experiments. The statistical significance of differences between two groups of data were analyzed by paired t-test and P-values <0.05 were considered significant.Scutellaria baicalensis Georgi extract, is a novel potential AR signaling inhibitor, which can block AR regulated gene expression and cell growth in androgen-sensitive cells and in CRPC cells. The inhibitory effects of baicalein are due to its interference with the AR N-C dimerization and AR-coregulator complex formation. Our data also indicated that baicalein could inhibit the E2 or Adiol-induced AR transactivation in the absence of androgens. These results suggest that baicalein may represent a promising anti-AR compound to better suppress the CRPC.In conclusion, the results of the present study demonstrated that baicalein, a natural compound isolated from


Momordica charantia) is widely cultivated as a vegetable and medicinal herb in many Asian and African countries. After the sequencing of the cucumber (Cucumis sativus), watermelon (Citrullus lanatus), and melon (Cucumis melo) genomes, bitter gourd became the fourth cucurbit species whose whole genome was sequenced. However, a comprehensive analysis of simple sequence repeats (SSRs) in bitter gourd, including a comparison with the three aforementioned cucurbit species has not yet been published. Here, we identified a total of 188,091 and 167,160 SSR motifs in the genomes of the bitter gourd lines ‘Dali-11’ and ‘OHB3-1,’ respectively. Subsequently, the SSR content, motif lengths, and classified motif types were characterized for the bitter gourd genomes and compared among all the cucurbit genomes. Lastly, a large set of 138,727 unique in silico SSR primer pairs were designed for bitter gourd. Among these, 71 primers were selected, all of which successfully amplified SSRs from the two bitter gourd lines ‘Dali-11’ and ‘K44’. To further examine the utilization of unique SSR primers, 21 SSR markers were used to genotype a collection of 211 bitter gourd lines from all over the world. A model-based clustering method and phylogenetic analysis indicated a clear separation among the geographic groups. The genomic SSR markers developed in this study have considerable potential value in advancing bitter gourd research.Bitter gourd ( Over the past half century, the molecular markers utilized in research have shifted from enzyme-based to various DNA-based types. More than twenty kinds of DNA marker systems have been established . Of thesMomordica charantia) belongs to the family Cucurbitaceae and is an important vegetable and medicinal herb in parts of Asia and Africa. The species has gained widespread attention as a result of its nutritional quality and incomparable hypoglycemic action, which has inspired the nickname ‘vegetable insulin’ for the crop . Utilizing the recently released ‘OHB3-1’ bitter gourd genome from Japan (C. sativus), watermelon (C. lanatus), and melon (C. melo). The distribution frequency of SSR motifs among the four cucurbits were comprehensively determined and compared. Furthermore, a total of 138,727 unique SSR primer pairs were identified from the bitter gourd line ‘Dali-11’ genome, and 71 of them were validated by polymerase chain reaction (PCR) amplification. To verify their usefulness, 21 out of 71 unique SSR primer pairs were used to evaluate the genetic diversity of a collection of 212 Momordica samples with various origins. This work provides a valuable set of genome-wide SSR marker resources and will be useful for further molecular genetics applications in bitter gourd.We initiated the bitter gourd whole genome sequencing project in early 2014 and conducted om Japan  as well,Momordica charantia) samples and one Momordica balsamina sample (as an outgroup), originating from 16 countries as well as the World Vegetable Center (AVRDC) and seed companies, were used to validate the usefulness of SSR primer pairs that were identified in the present study (Supplementary Table S1). Apart from the 12 samples from AVRDC and 10 from seed companies, the other 189 bitter gourd samples can be divided into five distinct geographic origins, namely South Asia (47), Southeast Asia (62), Latin America (4), China (69), and Tanzania (7). Plants were grown in an open field at Zengcheng Teaching and Research Base, South China Agricultural University, Zengcheng  in Guangdong Province, China.A collection of 211 bitter gourd (1 was used to identify the SSRs from whole genome data. We constrained the length of SSR motifs to a range from 1 to 6 bp corresponding to mononucleotides (Mono-), dinucleotides (Di-), trinucleotides (Tri-), tetranucleotide (Tetra-), pentanucleotide (Penta-), and hexanucleotide (Hexa-), respectively. The detailed search criteria are described in our previously published study . The general primer picking conditions were as follows: primer size, 18–27 bp with an optimum of 20 bp; primer melting temperature (Tm), 57.0–63.0°C with an optimum of 60°C; product size, 100–500 bp with an optimum of 250 bp; and primer GC content, 40–60% with an optimum of 50%. Then all the designed primer pairs were aligned to the ‘Dali-11’ bitter gourd reference genome. Unique primer pairs were defined only if both the forward and reverse primers were uniquely aligned (with a 100% match rate) to the reference genome.In order to design primers flanking the SSR loci, two Perl scripts2. PCR amplification was conducted under the following conditions: an initial denaturing step was performed at 94°C for 5 mins; followed by 25 cycles at 94°C for 30 s, 60°C for 30 s, and 72°C for 1 min; and finally followed by an extension step at 72°C for 5 min. The amplification products were resolved on a 6% polyacrylamide gel at 300 V for 2 h in 0.5× TBE buffer (pH = 8.3). Bands were visualized by silver staining.To validate the predicted motifs, we selected 27 primer pairs for SSR markers developed in two previous studies  and aligMomordica samples. The model-based clustering program STRUCTURE2.3.4 , was used to infer population groups. The statistic K was determined under the admixture model with correlated alleles, with a K that ranged from 2 to 10. Twenty independent runs of 10,000 Markov Chain Monte Carlo replicas and 100,000 generations of burn-in were used to estimate each K value. The optimal K value was determined by the log likelihood of the data [LnP(D)] in the STRUCTURE output and an ad hoc statistic ΔK, which was based on the rate of change in LnP(D) between successive K values  ranged from 10,150 (MC07) to 23,915 . The number of SSR motifs identified in bitter gourd genome  was moderate in cucurbit species, which was higher than that found in cucumber genome , but lower than that of watermelon genome  and the melon genome . Considering the different sizes of examined genomes, we calculated the density of SSR motifs for all cucurbit species. Bitter gourd genome, ‘Dali-11’ and ‘OHB3-1,’ had SSR densities of 639.73 and 585.27 SSRs/Mb, respectively, displaying comparable values to those of cucumber  and melon (624.78 SSRs/Mb), but lower values than those of watermelon .In this study, a total of ∼2.15 Gb of sequence data from seven genomes of four cucurbit species, including two bitter gourd genomes, two cucumber genomes, two watermelon genomes, and one melon genome, was analyzed (Supplementary Table S2). According to the search criteria, we identified 129,947–331,062 SSR motifs among seven cucurbit genomes (Supplementary Tables S4–S8); 188,091 and 167,160 SSR motifs were identified within the ‘Dali-11’ and ‘OHB3-1’ genomes, respectively. SSR motifs in the ‘Dali-11’ genome are summarized in Supplementary Table S4 and Table 1). As the repeat number increased, the number of SSR motifs for each type was dramatically reduced (Supplementary Table S9). All SSR motifs could be classified into one of 424 kinds of motif types according to their occurrence (Table 2 and Supplementary Table S10). Of these, 335 and 333 kinds were identified in ‘Dali-11’ and ‘OHB3-1’ bitter gourd genomes, respectively; both of which exhibited more kinds of motif types than the other cucurbit genomes. The theoretical number of motif types for Mono-, Di-, and Tri- repeats were 2, 4, and 10, respectively, and all 16 of these kinds of motif types appeared in the seven cucurbit genomes. Whereas, inter-specific differences in numbers of motif types occurred among Tetra-, Penta-, and Hexa- repeats (Table 2). Notably, the classified motif types were not completely uniform between the two genomes even belonging to the same species. For example, the two motif types AGCT/AGCT and CCCCCG/CGGGGG were identified in ‘Dali-11,’ but not detected in ‘OHB3-1.’Our search results revealed that Mono- repeats were dominant in cucurbit genomes, followed by Tri-, Di-, Tetra-, Penta-, and Hexa- repeats . Briefly, with regard to the Mono-, Di-, Tri-, and Tetra- repeats, A/T, AT/AT, AAT/ATT, and AAAT/ATTT were overrepresented, respectively, which totally accounted for 63.86–76.30% of all motifs identified in the seven cucurbit genomes. The AAAAT/ATTTT motif was found to be the most frequent motif of Penta- repeats in bitter gourd and watermelon genomes, whereas AAAAG/CTTTT was the most frequent one in cucumber and melon genomes. The AAAAAG/CTTTTT and AAAAAT/ATTTTT motifs were most representative among Hexa- repeats, and their numbers were roughly uniform across bitter gourd and watermelon genomes, whereas the former motif was several times more numerous than the later in cucumber and melon genomes.Furthermore, the frequency of classified SSR motifs in the four cucurbit species were variable  unique primer pairs (Supplementary Table S11).Table 3). Overall, 23 out of 27 (85.19%) of these motifs were consistent, but markers N9 and N12 mapped to the same position within S18 and S15, respectively. Hence, we designed 21 unique primer pairs (Table 3). In combination with 50 additional randomly selected primers, each of the examined 71 primers produced clear bands in at least one of the two bitter gourd lines ‘Dali-11’ and ‘K44’ DNA samples, showing a 100% amplification rate .We selected 27 pairs of primers for SSR markers from published reports and verified their motifs in the present study (M. charantia) samples and one M. balsamina sample. All primer pairs successfully amplified multiple bands, ranging from 2 (MC01_3973) to 16 (MC06_73242) across the bitter gourd population. In total, 122 alleles were detected with a mean number of 5.30 alleles per locus (Table 3). PIC values of each marker ranged from 0.20 (MC01_3973) to 0.84 (MC08_106502), with a mean value of 0.54.In this study, 21 primer pairs for markers that have motifs consistent with those of previous reports were used for the genetic diversity assessment of 211 bitter gourd . At K = 3, the clustering of samples was the most appropriate because it produced the highest ΔK value . Accordingly, the total panel was divided into three main populations that were labeled P1, P2, and P3, consisting of 23, 85, and 103 samples, respectively . This result was consistent with the NJ tree and geographic origins of the samples .STRUCTURE was used to infer population structure of the bitter gourd samples .From a geographic origin perspective, samples from Southeast Asia had wider distributions that were mainly localized in the P1 and P3 populations. Samples from South Asia and China were largely divided, and mainly grouped with P2 and P3, respectively. All four samples from Latin America were included in P1. One and six samples from Tanzania were assigned to P1and P2, respectively. Additionally, all the samples from AVRDC and seed companies were localized in P2 and P3 populations . Conversely, SSR density is usually negatively correlated with genome sizes . Notably, cucurbit genomes are more compact, which is consistent with our results showing that the SSR density of four cucurbit species was comparable to that of Arabidopsis and rice, but two-fold higher than that of pepper, tomato, and potato (We examined the distribution of SSRs (composed of 1–6 bp) within seven genomes from four cucurbit species. Similar to other plant species , the numme sizes ; howeverd potato . Therefod potato .We first compared the classified types of SSR motifs among cucurbit genomes. Overall, bitter gourd had more motif types than did the other three cucurbits. Whether this difference is related to the complexity or evolution of the genomes of each species is unclear. Variation in selective constraint on sequence repeats might differ among the SSR motifs . In geneThe genome-based SSR development is far more than the efficiency of traditional methods, including FIASCO  and SSR-Because of limited germplasm collection and molecular marker availability, scant attention has been paid to studying the genetic diversity of bitter gourd. Until recently, the best characterization of genetic diversity in bitter gourd was conducted using 114 bitter gourd accessions that originated from South and Southeast Asia by using 50 SSR markers . We collin silico unique SSR primer pairs were designed for bitter gourd based on ‘Dali-11’ reference genome. A selected set of 71 SSR primer pairs were validated by PCR amplification with a successful rate of 100.00%. We then used 21 polymorphic SSR markers to assess the genetic diversity of 211 bitter gourd samples, yielding three distinct populations that displayed clear geographic differentiation. These genome-wide SSR markers and the clustering results for the bitter gourd panel will play an important role in bitter gourd genetic improvement and breeding programs.In this study, we identified 129,947 to 331,062 SSR motifs among four cucurbit genomes, including 188,091 and 167,160 SSR motifs within the bitter gourd ‘Dali-11’ and ‘OHB3-1’ genome, respectively. Of these, the dominant SSR motif type in cucurbits is rich in A/T, AT/AT, AAT/ATT, and AAAT/ATTT. A large set of 138,727 JuC, JiC, and KH conceived and designed the experiments. DN, QZ, and ND contributed to germplasm collection. JuC and JP performed the experiments. YH and WH performed the bioinformatics analyses. JuC and JiC wrote the manuscript, and HK revised the manuscript. All authors read and approved the final manuscript.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.