Electroconvulsive therapy (ECT) is a safe and effective therapy for individuals suffering from major psychiatric disorders, but attitudes towards ECT among patients and caregivers have not been well studied. This study was conducted to elucidate patient and caregiver knowledge and attitudes concerning ECT in South China.n = 92). Participants completed questionnaire measures of knowledge and attitudes related to ECT.The sample comprised 92 patients diagnosed with major psychiatric disorders and their caregivers (p < 0.05). Caregivers reported receiving more adequate information about the therapeutic effects (50.0% versus 44.6%), side effects (67.4% versus 41.3%), and risks (55.4% versus 20.7%) of ECT when compared to patients . However, less than half of patients and caregivers believed that ECT was effective , while more than half of them believed that ECT was beneficial , and approximately half of them believed that ECT was safe . A total of 32.6% of patients and 55.4% of caregivers (p < 0.05) reported that ECT was used only for critically ill patients. A total of 62.0% of patients experienced side effects, with memory impairment being the most commonly reported.Information before ECT was inadequately provided to both caregivers and patients (55.4% versus 37.0%, Clinicians should develop a systematic health education program before ECT treatment and ensure that patients and caregivers have an accurate understanding of ECT, particularly the treatment process, its therapeutic effects and potential side effects prior to administering this treatment. While En = 1,364) focusing on adult patients (≥18 years old) found that 52.1% received ECT, with 53.4% receiving ECT for MDD, 57.8% for BD, 57.0% for schizophrenia, and 32.4% for other diagnoses . Both patients receiving a course of ECT sessions and caregivers who provided informed consent in writing were invited to join the study.The following were inclusion criteria for patients in this study: (1) understanding the questionnaire content focusing on knowledge and attitudes towards ECT; (2) a diagnosis of schizophrenia, BD, or MDD; (3) aged 18 years or older; (4) treated with a course of ECT in the past 6 months; and (5) completion of the last ECT session more than 1 month before this questionnaire. Similarly, caregivers had to fulfill the following inclusion criteria: (1) understanding of the questionnaire content; (2) aged 18 years or older; and (3) a friend or family member  who has provided primary personal care and handled medical-related issues for the patient during the past 6 months.2.2.ECT is available only as an inpatient procedure at the Affiliated Brain Hospital of Guangzhou Medical University. ECT  generally consists of 6–12 sessions per course, performed 2–3 times per week from 8:00 a.m. to noon on weekdays. During each ECT session, the patient’s blood oxygen, heart rate, and blood pressure are monitored with a cardiac monitor.2.3.Face-to-face interviews were performed by one interviewer for all assessments. According to the methodology of previous studies , 19, 21,2.4.p values less than 0.05 (two-tailed) were deemed statistically significant.All statistical analyses in the present study were performed using SPSS version 24.0 . For descriptive analyses, continuous and categorical variables were expressed as the means ± standard deviations (SD) and percentages (%) with regard to the demographic and clinical data of patients. Chi-square tests were utilized to determine the differences in ECT knowledge and attitudes between patients with major psychiatric disorders and their caregivers. 3.3.1.2) of patients were 27.9 ± 10.7 years, 20.9 ± 7.5 years, and 22.3 ± 2.6 kg/m2, respectively.Of the 96 patients and 96 caregivers invited to participate in the study, 92 (95.8%) patients with major psychiatric disorders who received ECT and 92 (95.8%) caregivers fulfilled the inclusion criteria and completed the assessment. Demographic and clinical data of subjects undergoing ECT are summarized in 3.2.p < 0.05). Caregivers (versus patients) reported receiving relatively more adequate information regarding therapeutic effects (50.0% versus 44.6%), side effects (67.4% versus 41.3%), and risks (55.4% versus 20.7%) of ECT . However, patients (versus caregivers) reported receiving more adequate information on the process of ECT (45.7% versus 23.9%) (p < 0.05). A significantly larger percentage of caregivers (71.7%) versus patients (53.3%) believed that ECT was beneficial (p < 0.05). Approximately half of patients (versus caregivers) thought that ECT was more effective (43.5% versus 46.7%) and more rapid than drugs (51.1% versus 45.7%) .As shown in 3.3.p > 0.05). More patients than caregivers disagreed with the statement, “Do you think ECT is dangerous and should not be used?” (66.3% versus 47.8%), and a significantly higher proportion of caregivers than patients endorsed the following queries: “Should ECT be used only for critically ill patients?” (55.4% versus 32.6%), “Should ECT be the last resort?” (51.1% versus 23.9%), and “Would you/your relative like to receive ECT again?” (83.7% versus 55.4%) . Alternatively, a significantly higher proportion of patients (31.5%)versus caregivers (12.0%) responded affirmatively to the question, “Is ECT used for people who do not need it?” (p < 0.05).As depicted in 3.4.Patient experiences of having ECT are summarized in 4.As reported by Tang et al. , China hIn this study, only 37.0% of patients and 55.4% of caregivers reported receiving any information about ECT before ECT was conducted. Similar findings from samples in Hong Kong, China (19.8% in patients and 58.6% in caregivers) , BeijingIn the past, ECT was often misrepresented in the mass media as a punitive measure for aggressive behaviour; such portrayals contributed to negative public attitudes towards ECT , 32. EncA majority of patients 76.1%) reported feeling worried during ECT, which is similar to findings of previous studies 6.1% repo. HoweverApproximately two-thirds of the patients experienced side effects of ECT, including memory impairment (72.8%), short-term confusion (45.7%), and headache (46.7%). Memory impairment was the most commonly reported side effect, while associated rates have varied in the literature from 29% to 79% . More seThe main strength of this study is the first to assess ECT knowledge and attitudes among patients who had undergone the intervention and their caregivers in South China. However, this study has several limitations that should be acknowledged. First, there is no standardized preexisting, interviewer-rated tool to assess ECT knowledge and attitudes. Second, this study with a single-center design was conducted in a psychiatric hospital in southern China. Therefore, the findings of this study cannot be generalized to other countries. Third, the sample size of this study was relatively small, limiting our capacity to detect the differences in attitudes or experiences of ECT among patients with different diagnoses. Fourth, key factors influencing participants’ attitudes towards ECT, such as disease severity and the nature/use of psychiatric medications, were not evaluated. Finally, this cross-sectional study has not been registered.In conclusion, clinicians should develop a systematic health education program before ECT treatment and ensure that patients and their caregivers have an accurate understanding of ECT, particularly the treatment process, its therapeutic effects and potential side effects prior to administering this treatment.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Research Ethics Panel of the Affiliated Brain Hospital of Guangzhou Medical University. The patients/participants provided their written informed consent to participate in this study.WZ was responsible for study design, process supervision, data interpretation, and critical revision of the article. SN and C-JD were responsible for organizing and analyzing the data, data interpretation, and drafting the manuscript. J-XM, XH, and X-BH were responsible for implementing the study, data collection, and data interpretation. All authors contributed to the article and approved the submitted version.This study was funded by Guangzhou Liwan District Science and Technology Plan Project (202201012), Guangzhou Science and Technology Project of Traditional Chinese Medicine and integrated traditional Chinese and Western medicine (20211A011045), China International Medical Exchange Foundation (Z-2018-35-2002), Guangzhou Clinical Characteristic Technology Project (2019TS67), and Science and Technology Program Project of Guangzhou (202102020658). The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the 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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


This work examined the results of different dietary protein levels on indigenous Nigerian guinea fowl growth performance and carcass yield.One hundred and eight unsexed indigenous keets were randomly assigned to three treatments with experimental diets containing 22%, 24%, and 26% crude protein (CP) in a completely randomized design, with 3 replicates with 12 keets in each replicate. The parameters were measured, including proximate composition, feed intake, growth rate, feed conversion ratio (FCR), and carcass yield.p < 0.05) on most carcass traits and prime cuts but not on gastrointestinal tract weight, head weight, dress percentage, or organ yield.The birds fed diets containing 24% CP gained the most body weight at 619.83 gm and had the highest FCR of 3.45. The presence of CP had an impact that was significant (The study concluded that the dietary CP level of 24% was optimal for body weight gain and carcass yield. Numida meleagris) yield is still in its early stages [Even though these birds evolved in Africa and have a ready market on the continent, guinea fowl .The Poultry Unit of Landmark University‘s Teaching and Research Farm in Omu-Aran was used for this study. One hundred and eight unsexed indigenous day old keets hatched in Landmark University‘s Animal Science laboratory were assigned randomly to 3 treatments after brooding, each of which was replicated 3 times in a completely randomized design with 12 keets in each replicate over 13 weeks. Diets were designed to provide the necessary protein for optimal performance, with treatments one (T1), two (T2), and three (T3) having a protein content of 22%, 24%, and 26%, indicating a lower, medium, and upper thresholds, respectively.The Association of Official Analytical Chemists  methodolTwo birds from each replicate of the experimental treatment were chosen at 13 weeks. These birds were slaughtered after being taken off feed for 12 h. The birds chosen had live weights closer to the group‘s average. The birds‘ jugular veins were cut and bled, and then their feathers were softened by soaking them in hot water for around 2 min before being hand-picked. After removing the shanks, head, neck, and viscera, the dressed weight was taken.A one-way analysis of variance was employed in analyzing all the data collected. Internal organs and carcasses were expressed as a percentage of live weight. Duncan’s new multiple range test of GENSTAT Release 10.3DE  softwareThe feed formulated for the experiment is shown in N. meleagris). The effects of varying CP levels were not significantly different (p > 0.05) on the parameters measured. T2 had a higher value (564.67 gm) for total weight gain than the values (530.08 and 516.91 gm) of T1 and T3 (26%) CP, respectively. T2 also had the highest daily weight gain (6.20 gm) value. Furthermore, T3 has the highest value  for total FI, while T2 and T1 had a close range . For daily FI, T3 had the highest value of 21.46 gm. The FCR in all three treatments was insignificant (p > 0.05), with T2 having the best FCR of 3.45.p < 0.05) influenced the carcass trait and prime cut of indigenous guinea fowls, as indicated in p > 0.05) on gastrointestinal weight, head weight, or dress percentage, but T2 had greater values for carcass metrics obtained. The effect of the protein inclusion level in the diet was insignificant (p > 0.05) on the weight of the organs evaluated. T3 had the highest values for gizzard weight (22.46 gm), lung weight (7.51 gm), proventriculus (2.37 gm), and small intestine weight (17.60 gm), whereas T2 had the highest values for heart weight (3.24 gm) and crop weight (4.02 gm).The CP level in the treatments significantly (p < 0.05) in this work [The effect of dietary treatments on weight gain, daily FI, and FCR was shown to be significant , followed by those fed T2  and those fed T1   reportedThe best performance in terms of feed conversion efficiency was in the 24% CP diet, with a ratio of 3.45. In contrast to the results of the present study, the study by Batkowska et al.  reportedThe higher body weight of guinea fowl fed the 24% CP diet may indicate that the birds’ nutritional requirements were met to a greater extent . The impp > 0.05). T2 had the highest live weight value of 654.50 gm across the treatments. This figure is higher than the live weight of 572.54 gm recorded by Amoah et al. [When assessing the impact of incorporating CP in animal feed, live weight is an essential indicator. The impact of CP inclusion on live weight was examined in this study, and it was observed that the mean live weight value was significant  impact on the mean hot carcass weight value, with T2 having the greater value of 511.83 gm and T3 (26%) having the lowest value of 647.83 gm, respectively. This is higher than the findings of Musundire [The CP levels in the feed had a significant (usundire .p > 0.05), with T2 recording the highest value of 56.50 gm and T3 recording the lowest value of 32.33 gm. These figures were lower than those reported by Kgakole et al. [The different CP treatments influenced the mean featherweight value in this study significantly  influenced by dietary protein. T2 had the highest mean value of 591.33 gm, followed by T1, with a value of 573.00 gm, and T3, with the lowest value of 479.50 gm. These disparities could be attributable to dietary variances.The de-feathered weight value was significantly p > 0.05) in this study; hence, the different dietary protein treatments had no effect. However, T2 had the highest value of 57.36 gm, whereas T1 and T3 had 57.08 and 51.84 gm, respectively. The variances could be due to differences in nutrition and feed composition.The gastrointestinal tract weight was not significant (p > 0.05). T2 had the highest mean value of 533.98 gm, while T3 had the lowest value of 427.67 gm. This could be due to the bird’s body conformation.The difference in protein levels in each treatment influences the mean value for eviscerated weight, which is significant (p > 0.05). T2 had the highest value of 119.24 gm and T3 had the lowest value of 98.28 gm. These results are, however, lower than those observed by Chiroque et al. [The mean value was significant . T2 had the greatest value of 62.08 gm, greater than Musundire’s [The dietary protein influenced the mean value of drumsticks, which was significant (undire’s  findingsp < 0.05). T2 had a higher value of 21.31 gm, higher than the findings of Kgakole et al. [The mean shank weight for this study was significant . T2 has the greatest mean value of 79.8 gm, followed by T1 with 76.10 gm and T3 with 63.90 gm, which were higher than 17.19 and 18.60 gm, recorded by Batkowska et al. [The dietary protein levels influenced the mean value of thigh weight, which was significant  effect on the head weight mean value in this study. T2 had a better value of 31.32 gm, followed by T1 and T3, with relative values of 30.84 and 30.65 gm, respectively. These values were higher than those reported by Musundire [Dietary protein levels had no significant  impacted by dietary protein. The heart weight mean values were 3.24 gm in T2, 3.00 gm in T1, and 2.78 gm in T3. The results of Ahaotu et al. [. [The internal organ indices in this study were not significantly  fed a 24% dietary protein inclusion had the best growth performance, with a final mean weight of 619.83 gm; however, birds fed a 22% dietary protein had a similar final mean weight of 586.80 gm, while those fed 26% CP had the least final mean weight of 566.43 gm. Furthermore, birds fed 24% dietary protein had a greater mean weight throughout the treatments regarding carcass features. Even though the energy was isocaloric, the keets appeared to respond to the feed’s protein content. This study showed indigenous guinea fowls could not utilize excessively high CP levels in their diet. Farmers can feed a diet with 24% CP to indigenous guinea fowl, which has been proven to be the best for optimum performance and production in this study. Guinea fowl (N. meleagris) are known to be flighty and wild; hence, chicks should be raised in cages to avoid mortality due to stampedes. When cages are not readily available, the stocking density on the floor should be considered.The dietary protein inclusion levels improved growth performance and carcass attributes. Guinea fowl (


Osteosarcoma is the most common primary malignant bone tumor. The current study was conducted to describe the general condition of patients with primary osteosarcoma in a single cancer center in Tianjin, China and to investigate the associated factors in osteosarcoma patients with lung metastasis.From February 2009 to October 2020, patients from Tianjin Medical University Cancer Institute and Hospital, China were retrospectively analyzed. The Kaplan–Meier method was used to evaluate the overall survival of osteosarcoma patients. The Cox proportional hazard regression analysis was performed to analyze the prognostic factors of all osteosarcoma patients and those patients with lung metastasis, respectively. Furthermore, risk factors for developing lung metastasis were identified in synchronous lung metastasis (SLM) and metachronous lung metastasis (MLM) patients.P = 0.025) and bone metastasis  were risk factors of SLM occurrence. Bone metastasis , good necrosis , elevated Ki-67  and elevated LDH  were proved to be independent risk factors for developing MLM.A total of 203 patients were involved and 150 patients were successfully followed up for survival status. The 5-year survival rate of osteosarcoma was 70.0% and the survival months for patients with SLM and MLM were 33.3 ± 12.6 and 45.8 ± 7.4 months, respectively. The presence of lung metastasis was one of the independent prognostic factors for prognosis of osteosarcoma. In patients with lung metastasis, twenty-one (10.3%) showed lung metastasis at the diagnosis of osteosarcoma and 67 (33%) were diagnosed with lung metastases during the later course. T3 stage (OR = 11.415, 95%CI 1.362–95.677, The overall survival, prognostic factors and risk factors for lung metastasis in this single center provided insight about osteosarcoma management.The online version contains supplementary material available at 10.1186/s12885-023-11024-9. Osteosarcoma is the most common primary malignant bone tumor in young adult, the prevalence was reported to be 8–11 per million people per year , 3.Distant metastasis, especially the lung metastasis, has been a serious issue in osteosarcoma management. To facilitate related studies and improve outcomes of osteosarcoma, four clinical oncology groups in European and American collaborated to construct the EURAMOS (European and American Osteosarcoma Studies) group . Through3 showed a probability of 69% to suffer lung metastasis, compared with 34% in those with the smaller tumor size  months. The 1-, 3-, 5-year survival rate was 94.4%, 77.3% and 70.0%, respectively. For patients without lung metastasis, the average OS was up to 139.2 ± 2.8 (95%CI 133.7–144.6) months. The survival was worse in patients with lung metastasis: 33.3 ± 12.6 (95%CI 8.6–57.9) months for SLM patients and 45.8 ± 7.4 (95%CI 31.3–60.3) months for MLM patients, respectively. The survival curve of different lung metastasis pattern was illustrated in Fig. P = 0.041), the presence of lung metastasis , salvage  and amputation  were four independent prognostic factors.The prognostic factors of all patients with active follow-up were identified and illustrated in the Table The aforementioned results in Fig. P = 0.037] and the presence of bone metastasis . After stratified by multivariate analysis, T3 stage  and bone metastasis  were proved to be two risk factors of SLM occurrence.As shown in Table For MLM, the information of the interval from osteosarcoma diagnosis to lung metastasis was available in 63 patients. The median internal time was 11 2–99) months and the distribution of MLM was illustrated in Fig. –99 monthP = 0.012), good necrosis , elevated Ki-67 , elevated LDH  and elevated ALP more than two times  were associated with the occurrence of MLM. In multivariate analysis, bone metastasis , good necrosis , elevated Ki-67  and elevated LDH  were proved to be independent risk factors for developing MLM. Details information of the Cox regression analysis were summarized in Table In univariate Cox regression analysis, the presence of bone metastasis  patients in the current study showed lung metastasis at the diagnosis of osteosarcoma, which was less than that previously reported , 25. T3 Some limitations should be mentioned. Due to the long internal from osteosarcoma diagnosis, some patients were lost and cannot be reached. Limited size of the included patients and unknown information in some variables caused uncertainty in data statistics. For example, the assessment of HUVOS necrosis rate after neoadjuvant chemotherapy were unavailable in the majority of patients. Furthermore, the limitation of the retrospective study design also leads to weakness to draw confirming conclusions.In summary, the osteosarcoma patients in our institute were effectively treated, with the 5-year overall survival of 70%. The incidences of synchronous and metachronous lung metastasis were 10.3%, and 33%, respectively. The prognostic factors found in the current study can be significant on survival prediction. Risk factors of lung metastasis can be used to identify high-risk patients and guide individualized screening.Additional file 1:Supplementary Table 1. The definitions of T stage and N stage in patients with osteosarcoma.Additional file 2:Supplementary Table 2. Identification of the prognostic factors in osteosarcoma patients with lung metastasis (N=50).Additonal file 3:Supplementary Table 3. The detailed information of osteosarcoma patients who underwent pulmonary metastasectomy.


To compare the efficacy of two density gradient centrifugation media forretrieving spermatozoa from semen samples by evaluating the total motilesperm count (TMSC) and the percentage recovery.Twenty-two men with different sperm counts participated in the study. Thesamples were divided into two equal aliquots and processed using thecommercial ISolate Sperm Separation Medium  and the GV Gradiente . After separation,samples were counted and evaluated for motile sperm recovery.p>0.05). Thesamples from 6 oligospermic patients (27%) were evaluated separately and theTMSC for ISolate was 4.83±2.92 million/mL, and for the GV Gradiente,it was 4.16±3.12 million/mL (p=0.54). Whenevaluating only normospermic patient samples, the TMSC for ISolate was9.05±7.29 million/mL, and for the GV Gradiente, it was8.47±6.79 million/mL (p=0.83).The mean TMSC in the fresh sample was 19.65±21.08 million/mL. Afterthe ISolate separation the TMSC was 6.71±7.29 million/mL, and for theGV Gradiente it was 6.27±6.82 million/mL. The percentage of motilespermatozoa recovered was 36.47%±21.61 for ISolate and35.22%±21.24 for the GV Gradiente (There was no statistical difference in retrieving motile sperm using the GVGradiente and the ISolate Separation Medium. To perform assisted reproduction techniques (ART), sperm must first be processed byselecting for morphologically normal spermatozoa with high motility in order tomaximize the concentration of good quality sperm. The preparation also removes celldebris, including epithelial cells and leukocytes . However, due to the toxicity of PVP, its use inhumans has been discontinued . The total concentration of sperm and motile formsrecovered were evaluated following semen processing by these DGC methods.This study was approved by the Research Ethics Committee of UNINGA University(Protocol: 4.728.627). Semen samples were obtained from 22 men aged 29 to 47 years,through masturbation into a wide and sterile bottle  after a2 to 7-day period of abstinence. The samples remained at room temperature for 40-60minutes for liquefaction. After complete liquefaction, the samples were counted andthen divided into two equal parts for sperm separation by the DGC method using 1)the ISolate Sperm Separation Medium  and 2) the GV Gradiente.The semen was deposited on a 50/90% ISolate tube and on a 45/90% GV Gradiente tube,and then centrifuged at 300 × g for 20 minutes. The supernatant wasdiscarded, and the pellet consisting of sperm was washed with GV HEPES at 300 × g for 10 minutes. After washing, thesupernatant was again discarded, and the pellet containing the isolated sperm wasresuspended in 0.5 mL of GV HEPES . After the spermseparation methods were applied, the sperm motility parameter in bothsamples(ISolate and GV Gradiente) was evaluated by the manual counting method in aMakler counting chamber under phase contrast microscopy with 400x totalmagnification.p<0.05 was considered statistically significant. GraphPad Prism8.0.1 software was used for the analysis and generating the graphs.Data are presented as mean ± standard deviation. The Kruskal-Wallis testfollowed by Dunn’s multiple comparisons test were used to compare the semenparameters of the fresh samples and after each DGC medium used. The Mann-Whitneytest was used to compare the results from the two different DGC media. The Z testcompared the recovery proportions between the DGC media. The frequency distributionof samples was used to confirm sample equivalence. In all tests,The men participating in the study had a mean age of 37.36 ± 4.35 years(95% CI 35.43-39.30). The fresh total sperm count averaged 36.99 ± 30.95million/mL (95% CI 23.26-50.71). Total sperm count did not correlate with theage of the participants (r=0.20). The total motile sperm count (TMSC) averaged19.65 ± 21.08 million/mL (95% CI 10.30-29.99) and had no correlation withage (r=0.14). The mean percentage of motile sperm in relation to the total was55.18 ± 27.43% (95% CI 43.02-67.34).Of the 22 men, six (27.8%) were characterized with oligospermia (<15 millionsperm per mL of semen) according to the World Health Organization . Among tThe values of motile sperm in the fresh sample and after processing by the twoDGC media were compared and evaluated . The meap=0.93, although there was no significantdifference.The mean rate of motile sperm retrieved was 36.47% ± 21.61 (95% CI26.89-46.05) for ISolate and 35.22 % ± 21.24 (95% CI 25.80-44.64) for GVGradiente . The comAnother evaluation to verify the separation efficiency by the DGC media was theanalysis of samples considered oligospermic and normospermic separately. Spermfrom six patients (27%) with sperm counts below 15 million per mL were comparedafter separation with ISolate and GV Gradiente. The mean TMSC for ISolate was4.83 ± 2.92 million/mL (95% CI 1.76-7.90) and for the GV Gradiente, thevalue was 4.16 ± 3.12 million/mL (95% CI 1.32-19.18). When evaluatingonly patients with normospermia, the mean for ISolate was 9.05 ± 7.29million/mL (95% CI 0.51-12.94)and for the GV Gradiente, the value was 8.47± 6.79 million/mL (95% CI 0.48-12.09).The distribution of samples after separation was evaluated in order to confirmthe equivalence of the DGC media in relation to the retrieval of motile sperm.The frequency histograms show the TMSC distribution after separation with theISolate and GV Gradiente media ,as wellOne of the main challenges of reproductive medicine is to ensure that the highestquality sperm are used in ARTs. The aim of this study was to determine if there wasa difference in sperm recovery after processing with two different DGC media:ISolate and GV Gradiente.Sperm preparation techniques must be operationally simple and cost-effective in orderto suit the routine procedures of human reproduction clinics. It should result in asample that is enriched with high quality sperm in the shortest possible time. Inaddition to removing low-quality sperm, including immotile sperm, sperm preparationtechniques must allow the elimination of other cells, such as leukocytes andbacteria, as well as toxic or bioactive substances like reactive oxygen species(ROS) , Sil-Select® , PureSperm® , or ISolate® . Commercial media are currently used in human clinics to replace Percolldue to its toxicity and side effects on sperm function . The percentage of recovery did not differ either(p=0.93). Furthermore, the recovery percentage for ISolate wassimilar to that demonstrated by The results demonstrated that sperm separation using the two DGC media did not showany significant difference in terms of the recovered motile sperm concentration(p=0.54). When only normospermic samples wereevaluated, there was also no statistical difference between the ISolate and the GVGradiente media (p=0.83). These results demonstrate that the mediahave the same efficiency for preparing sperm from low concentration samples, as wellas normal count or heterogeneous samples.The current study is significant because it assessed sperm quality after twodifferent sperm preparation media used under identical conditions. Furthermore,samples from patients with oligospermia and normosperia were assessed separatelyunder these conditions. It is believed that the effects of gradient media may bemore expressive in samples with a smaller amount of sperm or semen of lower quality. Here itSeveral other methodologies have been developed with the aim of improving the qualityof isolated sperm for use in IVF, such as selection methods depending on spermmotility using microfluidic systems that are based on physical sperm properties suchas chemotaxis, rheotaxis, and thermotaxis. Although promising, these systems havenot yet been shown to be useful for their routine application in clinical practiceand seem to be effective only in some very specific cases of male infertility (The results shown here, in relation to ISolate, GV Gradiente has comparable efficacyin sperm separation, suggesting that this product would be ideal for performing theDGC technique in the country.


Radiographic progression and course of inflammation over 2 years in patients with non-radiographic axial spondyloarthritis (nr-axSpA) from the phase 3, randomized, PREVENT study are reported here.In the PREVENT study, adult patients fulfilling the Assessment of SpondyloArthritis International Society classification criteria for nr-axSpA with elevated CRP and/or MRI inflammation received secukinumab 150 mg or placebo. All patients received open-label secukinumab from week 52 onward. Sacroiliac (SI) joint and spinal radiographs were scored using the modified New York (mNY) grading  and modified Stoke Ankylosing Spondylitis Spine Score , respectively. SI joint bone marrow edema (BME) was assessed using the Berlin Active Inflammatory Lesions Scoring (0–24) and spinal MRI using the Berlin modification of the AS spine MRI (ASspiMRI) scoring (0–69).n = 7) and 2.9% (n = 3) of patients in the secukinumab and placebo-secukinumab groups, respectively, who were mNY-negative at baseline were scored as mNY-positive at week 104. Overall, 1.7% and 3.4% of patients with no syndesmophytes at baseline in the secukinumab and placebo-secukinumab group, respectively, developed ≥ 1 new syndesmophyte over 2 years. Reduction in SI joint BME observed at week 16 with secukinumab  was sustained through week 104 (− 1.73 [3.49]). Spinal inflammation on MRI was low at baseline  and remained low .Overall, 78.9% (438/555) of patients completed week 104 of the study. Over 2 years, minimal changes were observed in total radiographic SI joint scores  and mSASSS scores (0.04 [0.47] and 0.07 [0.36]) in the secukinumab and placebo-secukinumab groups. Most of the patients showed no structural progression  in SI joint score (87.7% and 85.6%) and mSASSS score (97.5% and 97.1%) in the secukinumab and placebo-secukinumab groups. Only 3.3% (Structural damage was low at baseline and most patients showed no radiographic progression in SI joints and spine over 2 years in the secukinumab and placebo-secukinumab groups. Secukinumab reduced SI joint inflammation, which was sustained over 2 years.ClinicalTrials.gov, NCT02696031.The online version contains supplementary material available at 10.1186/s13075-023-03051-5. Axial spondyloarthritis (axSpA) is a chronic inflammatory disease of the axial skeleton predominantly affecting the spine and sacroiliac (SI) joints . Based oSecukinumab, a human monoclonal immunoglobulin (IgG1) antibody that selectively neutralizes human interleukin (IL)-17A, demonstrated rapid and sustained improvement in the signs and symptoms of AS in long-term treatment (up to 5 years) in the phase 3 MEASURE studies –13. PREVStudy design and eligibility criteria have been reported previously in detail . BrieflyThe study was conducted in accordance with the Good Clinical Practice guidelines and the Declaration of Helsinki and was approved by institutional review boards or independent ethics committees at each participating center. Written informed consent was obtained from all enrolled patients.In this exploratory analysis, radiographic progression was assessed by conventional radiography and the course of inflammation by MRI, of SI joints and spine, over 2 years. Radiographs of the spine and SI joints were obtained at baseline and at week 104. MRI images of the spine and SI joints were obtained at baseline and weeks 16, 52, and 104.SI joint radiographs were scored according to the mNY grading method  . The totSpinal radiographs were scored using the modified Stoke AS Spine Score (mSASSS) . The mSASI joint bone marrow edema (BME) was assessed according to the Berlin Active Inflammatory Lesions Scoring (0–24).Spinal MRI images were assessed for signs of inflammation using the Berlin modification of the AS spine MRI (ASspiMRI) scoring (0–69) .At the final reading session (week 104), all baseline and post-baseline images were scored by 2 central independent and experienced readers who were blinded to treatment assignment and image sequence. The cases with the highest between-reader differences for change in score from baseline were identified for adjudication review by a third independent reader. Adjudication review was triggered independently for the spine and SI joint and each imaging modality. The top 5% of radiograph reads with the highest difference in change of scores, the top 5% of discrepant cases of total SI joint edema score, and the top 10% of discrepant cases of total spine edema scores were scored independently by a third reader. All data are reported from the final reading session , which was independent from previous reading sessions  .The change from baseline in the total sacroiliitis score and mSASSS score was calculated based on the average of scores assigned by 2 or 3 readers (if available) and presented as a cumulative probability of change from baseline. For binary variables (mNY criteria and syndesmophyte status), the analysis was carried out using a single-reader approach or 2-reader agreement approach. The single-reader approach considered a patient to be mNY criteria-positive when assessed mNY-positive by at least 1 reader. Correspondingly, if at least 1 reader assessed a vertebral unit as 2 or 3, the patient was considered to have a syndesmophyte. The 2-reader agreement approach considered a patient to be mNY criteria-positive or having a syndesmophyte when at least 2 readers assessed the patient as such.Inter-rater reliability was calculated by intraclass correlation coefficient (ICC), based on 2 reading scores made on the same subject by the primary readers. The smallest detectable change (SDC) was calculated using the Bland–Altman analysis at an 80% level of agreement (LoA) based on the data from the 2 primary readers .The full analysis set from week 104 database lock  was used for the analysis. Data from the secukinumab loading and non-loading dose groups were pooled for this imaging post hoc analysis. Data are presented as observed.For radiographs, data are presented according to their original assignment at randomization with patients who switched to standard of care included as originally randomized and placebo patients who switched to secukinumab included in the placebo-secukinumab group. Patients who switched from blinded to open-label secukinumab 150 mg were included and presented as originally randomized (secukinumab group). For MRI, patients originally randomized to placebo who switched to active treatment were not included in the analysis. Patients randomized to secukinumab 150 mg blinded who switched to open-label secukinumab 150 mg were included and presented as originally randomized.A total of 438 of the 555 patients (78.9%) completed week 104 of the study. Overall, 50.8% of the patients in the loading dose group, 47.3% in the non-loading dose group, and 64% in the placebo group switched to either open-label secukinumab or standard of care (only 3 patients switched to standard of care) between weeks 20 and 52. Starting at week 52, all patients (except for 3 patients who had switched to standard of care) received open-label secukinumab 150 mg treatment. Thus, all patients received secukinumab treatment for at least 1 year during the 104-week study period. Demographic and baseline disease characteristics were comparable across treatment groups . Table 1A total of 277 patients (75.1%) in the pooled secukinumab group and 139 (74.7%) in the placebo-secukinumab group had SI joint radiographs evaluable for change in total sacroiliitis score . The mean  baseline total sacroiliitis scores  in the secukinumab and placebo-secukinumab groups were 1.45 (1.53) and 1.47 (1.60), respectively. At week 104, the scores were 1.41 (1.47) and 1.50 (1.61) in the secukinumab and placebo-secukinumab groups, respectively. Thus, only minimal changes in total sacroiliitis scores were observed over 2 years with a mean (SD) change of − 0.04 (0.49) in the secukinumab group and 0.04 (0.36) in the placebo-secukinumab group at week 104.Overall, 87.7% (243 of 277) of patients in the secukinumab group and 85.6% 119 of 139) of patients in the placebo-secukinumab group showed no progression in SI joint scoring by week 104. No progression was defined as an increase in total sacroiliitis score ≤ SDC, which was 0.46 at an 80% LoA. Clinically meaningful progression, defined as a total sacroiliitis score increase of > 1, was observed in only 1.1% (3 of 277) of patients in the secukinumab group and 2.2% (3 of 139) of patients in the placebo-secukinumab group. None of the patients had an increase of > 2 in the total sacroiliitis score  were evaluated as mNY criteria-positive on radiographs obtained at screening, by at least 1 reader. Of these patients, 16.2% (11 of 68) in the secukinumab group and 14.7% (5 of 34) in the placebo-secukinumab group were evaluated as mNY-negative at week 104 . At baseline, the mean (SD) mSASSS scores in the secukinumab and placebo-secukinumab groups were 0.68 (2.35) and 0.81 (2.37), respectively. At week 104, the scores were 0.73 (2.49) and 0.88 (2.60) in the secukinumab and placebo-secukinumab groups, respectively. Thus, there was a minimal change over 2 years with a mean (SD) change of 0.04 (0.47) in the secukinumab group and 0.07 (0.36) in the placebo-secukinumab group at week 104.Overall, 97.5% 273 of 280) of patients in the secukinumab group and 97.1% (132 of 136) in the placebo-secukinumab group showed no structural progression over 2 years, when no progression was defined as a change in total mSASSS score ≤ SDC (which was 0.76 at 80% LoA). An increase of > 2 units in the mSASSS total score was observed in 1.1% (3 of 280) of patients in the secukinumab group and 0.7% (1 of 136) in the placebo-secukinumab group. None of the patients had an increase of > 5  had no syndesmophytes at baseline. Among these, 1.7% (4 of 237) in the secukinumab group and 3.4% (4 of 117) in the placebo-secukinumab group developed at least 1 new syndesmophyte  over 2 years. On the other hand, among patients reported to have at least 1 syndesmophyte at baseline (15.4% of patients [43 of 280] in the secukinumab group and 14.0% of patients [19 of 136] in the placebo-secukinumab group), a higher proportion of patients, i.e., 20.9% (9 of 43) in the secukinumab group and 36.8% (7 of 19) in the placebo-secukinumab group, had developed at least 1 new syndesmophyte by week 104. Also, irrespective of the treatment group, the majority of patients (79% based on an assessment by a single reader) who had at least 1 syndesmophyte at screening were mNY-negative at baseline , and a lower proportion of these patients developed new syndesmophytes (15.4% [2 of 13] in the secukinumab group and 8.3% [1 of 12] in the placebo-secukinumab group) compared with the single-reader assessment. Among patients without syndesmophytes at baseline, only 1 patient (in the secukinumab group) had developed a syndesmophyte at week 104. Importantly, the proportion of patients who had a syndesmophyte at baseline and were considered mNY-negative was even higher when both evaluations required a 2-reader agreement (88% vs 79% based on single reader assessment)  SI joint total edema score decreased from baseline by 1.23 (2.81) in the secukinumab group vs 0.37 (1.90) in the placebo group. This reduction in BME observed at week 16 was sustained through week 104   was low at baseline across the groups. The mean score in the secukinumab group was 0.82, and it was 1.07 in the placebo group. The mean score at week 104 in the secukinumab group was 0.56.This exploratory analysis from the PREVENT study evaluated radiographic changes and spinal inflammation in patients with active nr-axSpA over a period of 2 years. This is one of the largest imaging datasets in patients with nr-axSpA available to date. The overall radiographic changes, along with any changes in spinal inflammation, in a population of patients on an effective biologic therapy were minimal over time, while a reduction in SI joint inflammation was observed as previously reported .Most of the patients in the secukinumab group (87.7%), as well as those originally randomized to the placebo group (85.6%), did not have an increase in total sacroiliitis score above the SDC on SI joint radiographs, indicating no radiographic progression over 2 years. The SDC for the total sacroiliitis score was 0.46. Notably, given that the minimum possible change on mNY grading scale is 1, the smallest change that can be observed in an individual patient when considering the average score by 2 readers is 0.5. None of the patients in either group had an increase in total sacroiliitis score of > 2, and only few had an increase of > 1 (1.1% in the secukinumab group and 2.2% in the placebo group). Furthermore, the proportion of patients who had an increase in the total sacroiliitis score greater than the SDC was similar to the proportion of patients who had a decrease in the score of similar magnitude, suggesting that the observed change may be attributed to reader variability rather than disease progression.As per the eligibility criteria, any patient with mNY-positive status at screening based on the eligibility read by a single reader was excluded from the study. However, when the same images from screening were scored again alongside post-baseline images at week 104, by readers blinded to the sequence of imaging, some of the radiographs were scored as mNY-positive. The poor-to-moderate intra-rater and inter-rater reliability of SI joint damage assessment according to the mNY criteria are well described –21. CaseN = 416) in the DEvenir des Spondyloarthrites Indifférenciées Récentes (DESIR) cohort  [While earlier studies suggested low but evident progression in early axSpA, more recent studies report even lower rates of radiographic SI joint structural progression. In the RAPID-axSpA study, limited changes in SI joint grading were observed after 4 years in patients treated with certolizumab: 4.5% (2 of 44) of patients who were mNY-negative at baseline fulfilled the mNY criteria at week 204 and 4.3% (4 of 93) of patients who were initially mNY-positive were evaluated as mNY-negative at week 204 . There auration) . HoweverLower rates of progression reported in more recent studies may reflect greater awareness of the measurement error in radiograph scoring leading to the implementation of central reading with more than 2 readers and also acknowledging the measurement error in the analysis. Another potential reason for low to negligible progression rates reported here and in other recent studies could be the nr-axSpA population recruited into these clinical trials, who may present with less severe disease and have earlier access to biologic treatment.With regard to the mSASSS score, most patients in both groups showed no progression to week 104. While none of the patients in either group had an mSASSS total score increase of > 5, around 1% had an increase of > 2. The mean change in mSASSS score was 0.04 in the secukinumab group (0.68 at baseline). Previous studies have similarly reported limited spinal progression in patients with nr-axSpA. In the RAPID-axSpA study, which reported 4-year imaging outcomes, the mean mSASSS change was − 0.01  from baseline to week 96 and 0.06  to week 204 in patients with nr-axSpA treated with certolizumab pegol . ProgresIn the present study, approximately 15% of patients had syndesmophytes at baseline identified by at least 1 reader (6% when reader agreement was required). Similarly, 20% of the nr-axSpA patients in the RAPID-axSpA study, 9% of patients in the Swiss Clinical Quality Management cohort (readers’ agreement was required in both studies), and 7% of patients in the DESIR cohort (2 of the 3 readers were required to identify the syndesmophyte) had syndesmophytes at baseline, suggesting that the process of structural damage starts early in the disease at least in some patients , 26, 27.Syndesmophyte formation is thought to be mainly associated with r-axSpA . HoweverThis study had some limitations. Patients were allowed to switch to open-label secukinumab or standard of care leading to a lack of placebo control after 20 weeks. The analyses of images were conducted on patients who continued on the study, thus were likely to respond well to the treatment introducing a potential bias. Also, in this study, inter-rater reliability was modest at best. However, this is not surprising in sacroiliitis assessment where low-to-moderate inter-rater/intra-rater reliability has been reported in previous studies , 38.The value of minor effects of biological disease-modifying anti-rheumatic drugs (DMARDs)  or targeIn summary, in this large study of patients with nr-axSpA, the overall level of spinal inflammation or structural damage at baseline was low. Most patients initially randomized to secukinumab or placebo showed no radiographic progression in SI joints and spine over 2 years. Secukinumab reduced SI joint inflammation (BME) in patients with active nr-axSpA which was sustained over 2 years.Additional file 1: Fig. S1. Study Design. Fig. S2. Mean change in SI joint bone marrow edema score by MRI in the overall population and in patients with baseline score >2 through week 104 . Table S1. Syndesmophyte status at screening by mNY status at screening based on single reader assessment and two reader agreement.


The accuracy of ultrasound in distinguishing benign from malignant adnexal masses is highly correlated with the experience of ultrasound physicians. In China, most of ultrasound differentiation is done by junior physicians.To compare the diagnostic performance of the International Ovarian Tumour Analysis (IOTA) Simple Rules Risk (SRR) and IOTA Logistic Regression Model 2 (LR2) scoring systems in Chinese patients with adnexal masses.Retrospective analysis of ovarian cancer tumor patients who underwent surgery at a hospital in China from January 2016 to December 2021. Screening patients with at least one adnexal mass on inclusion and exclusion criteria. Two trained junior physicians evaluated each mass using the two scoring systems. A receiver operating characteristic curve was used to test the diagnostic performance of each system.P > 0.05 ).A total of 144 adnexal masses were retrospectively collected. Forty masses were histologically diagnosed as malignant. Compared with premenopausal women, postmenopausal women had a much higher rate of malignant masses. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) of the SRR was 97.5% (95% CI: 86.8 -99.9%), 82.7% (95% CI: 74.0 -89.4%), 68.4% (95% CI: 58.7 -76.8%) and 98.9% (95% CI: 92.5 -99.8%). The sensitivity, specificity, PPV, NPV of the LR2 were 90.0% (95% CI: 76.5 -97.2%), 89.4% (95% CI: 81.9 -94.6%), 76.6% (95% CI: 65.0 -85.2%), and 95.9% (95% CI: 90.2 -98.3%). There was good agreement between two scoring systems, with 84.03% total agreement and a kappa value of 0.783 (95% CI: 0.70-0.864). The areas under the curve for predicting malignant tumours using SRR and LR2 were similar for all patients (The two scoring systems can effectively distinguish benign from malignant adnexal masses. Both scoring systems have high diagnostic efficacy, and diagnostic efficacy is stable, which can provide an important reference for clinical decision making. Ovarian cancer (OC), which occurs in ovarian tissue deep within the pelvis, is the third most prevalent type of gynaecological cancer in the world . AccordiEarly diagnosis is important for reducing mortality from OC; however, there is currently no standardised strategy for the screening or early detection of OC .Imaging techniques, such as ultrasound (US) and magnetic resonance imaging (MRI), have been used in the preoperative assessment of adnexal tumours. Because of its low cost and accessibility, US has proved to be the most useful diagnostic tool for adnexal masses . HoweverThe SR model describes five typical features of benign (B) tumours and five typical features of malignant (M) tumours. The model is easy to use and can compensate for junior physicians’ lack of experience . ExternaThe above prediction models were mainly constructed and validated based on data from European populations. To the best of our knowledge, no studies have been published comparing the accuracy of IOTA SRR and IOTA LR2 in discriminating between benign and malignant adnexal masses in Chinese patients. In China, due to the insufficient number of experts in ultrasound examination, most of the judgments of benign and malignant tumors are completed by inexperienced junior physicians and junior physicians also have difficulty in differential diagnosis of tumors . HoweverThe current study retrospectively analyzed the basic information, hospitalization information, and ultrasound images of ovarian cancer tumor patients who underwent surgery in a hospital in China from January 2016 to December 2021 and were diagnosed through postoperative pathological examination. The current study belongs to convenient sampling.According to the Helsinki Declaration, this retrospective observational study was approved by the Ethics Committee of our hospital. All participants have informed written consent to publish the data. Therefore, there is no sensitive data in the current study.The inclusion criteria were as follows: (1) patients diagnosed by US with at least one adnexal mass; (2) patients over the age of 18; (3) patients whose lesions had been removed surgically and evaluated pathologically; and (4) preoperative ultrasound examination was performed according to the requirement and the information of ultrasound image was preserved completely. The exclusion criteria were as follows: (1) pregnant women with adnexal masses; and (2) patients with a history of a malignant gynaecological tumour prior to the diagnosis of an adnexal mass. The study ultimately included a total of 128 patients with 144 adnexal masses.In all cases, transvaginal US examination was the primary scanning modality. A transabdominal US was performed only when the mass was too large to be observed by transvaginal US. A Voluson E8  US machine was used. The intracavitary transducer frequency was set to 5.0–9.0 MHz, and the abdominal probe frequency was 3.5–5.0 MHz. According to previous similar studies by Chinese scholars , the US https://homes.esat.kuleuven.be/~sistawww/biomed/ssrisk/). Following previous studies, the current study used two classifications of SRR risk, i.e. low (≤ 20%) and high (> 20%) [The IOTA SRR scoring system provided a numeric risk estimate of the malignancy of pelvic lesions based on the B- and M-features of the SR model . The B-f (> 20%) .y = 1 / (1 + exp (˗ z)), where z = ˗5.3718 + 0.0354 (a) + 1.6159 (b) + 1.1768 (c) + 0.0697 (d) + 0.9586 (e) ˗ 2.9486 (f). A probability greater than 10% was considered to represent a high risk of malignancy [Next, the IOTA LR2 scores were calculated based on the following six features: (a) patient age; (b) presence of ascites; (c) solid papillary blood flow; (d) maximal solid component diameter; (e) irregularity of internal cyst walls; and (f) the presence of acoustic shadows . The LR2lignancy .Two junior physicians with US diagnostic experience < 2 years analysed the images and evaluated each mass using the SRR and LR2 systems independently. Where results differed, the two physicians worked together with a senior doctor with > 10 years of US diagnostic experience to analyse the images. Prior to the start of the study, two junior physicians were trained in the theoretical and practical aspects of the SRR and LR2 models. The specific content included standardized training for ovarian benign tumors, standardized training for ovarian malignant tumors, and IOTA diagnostic model training. This training was conducted by senior professors in the form of a seminar, where the professor first imparts knowledge and then discusses with the physician. Each training lasts for 2.5 to 3 h and was conducted three times. The entire training was completed within half a month. After the training is completed, two physicians used SRR and LR2 models to diagnose 40 randomly selected adnexal masses, and performed inter group consistency analysis on their diagnostic results. The results showed that the kappa(k) value of the SRR and LR2 models were 0.92  and 0.87 , respectively, with good consistency results. The results of good consistency can increase the reliability and reliability of the study, and provide support for the validity of the study.Both physicians did not participate in patient information and image collection, and both physicians followed the double-blind principle in the image evaluation process.Researchers retrospectively collected basic and clinical information of each patient including age, menopausal status and cancer antigen 125 (CA125) levels using the patient’s medical record homepage and hospital information systems. Experienced physicians used clear histological diagnosis as the gold standard for diagnosing each resected mass.(1) Strictly screen research subjects based on inclusion and exclusion criteria to avoid selection bias; (2) Provide a clear data collection process to ensure the accuracy and completeness of research data; (3) Two junior physicians were trained to ensure that they are familiar with and correctly use the SRR and LR2 scoring systems; (4) After the training, a portion of the samples were randomly selected for simulation, and consistency evaluation was conducted on two junior physicians to ensure the reliability of the study.2) or Fisher’s exact tests. Quantitative data were described as mean ± standard deviation and compared using a t-test. Receiver operating characteristic (ROC) curves were used to evaluate the diagnostic accuracy of the two scoring systems in terms of discriminating between benign and malignant adnexal masses. The ROC curves reflect the relationship between the sensitivity and specificity of the curve [p < 0.05.Data were analysed using the SPSS v.22.0 and MedCalc v.19.0.5 statistical software. Categorical data were described as numbers and percentages and compared using chi-squared  were premenopausal, with a mean age of 28.95 ± 9.51 years, and 54 patients (42.19%) were postmenopausal, with a mean age of 57.96 ± 8.36 years. Sixteen patients (12.50%) had bilateral lesions and 45 (35.16%) patients had increased CA125 levels, including 31 (57.41%) postmenopausal women and 14 (18.92%) premenopausal women. The clinical characteristics of the enrolled patients are summarised in Table p < 0.001). Details of the pathology results are shown in Table As 16 patients had bilateral adnexal lesions, the current study ultimately included 144 adnexal masses, 4027.78%) of which were malignant and 104(72.22%)were benign. These were confirmed by postoperative pathology analysis. The most frequent benign tumour was teratoma , while the most common malignant tumour was serous adenocarcinoma . Compared with premenopausal women, postmenopausal women enrolled in the study had a much higher rate of malignant masses .The ROC curve of malignant adnexal masses diagnosed by physicians using the two scoring systems is shown in Fig. Physicians had good consistency in the preoperative diagnosis of malignant adnexal masses using the SRR and LR2 scoring systems, with a total consistency of 84.03% and a k-value of 0.783 (95% CI: 0.70-0.864). Representative US images with final histopathology diagnoses are shown in Fig. Using the thresholds defined in the current study, 94 (65.28%) of all adnexal masses were classified as low risk  and 50 34.72%) were classified as high risk  by the SRR scoring system. The malignancy rates for each group, according to the SRR score, were 5.32% (5/94) and 70.00% (35/50), respectively. The sensitivity was calculated as being 97.5% (95% CI: 86.8–99.9%), specificity was 82.7% (95% CI: 74.0–89.4%), positive predictive value (PPV)was 68.4% (95% CI: 58.7–76.8%), negative predictive value (NPV) was 98.9% (95% CI: 92.5–99.8%) and youden index was 0.802. See Table 4.72% werUsing the LR2 scoring system to classify adnexal lesions, 97 67.36%) masses were classified as low risk  and 47 (32.64%) were classified as high risk . The malignancy rates for each group, according to the LR2 model, were 4.12% (4/97) and 76.60% (36/47), respectively. The sensitivity was calculated as being 90.0% (95% CI: 76.5–97.2%), specificity was 89.4% (95% CI: 81.9–94.6%), PPV was 76.6% (95% CI: 65.0–85.2%), NPV was 95.9% (95% CI: 90.2–98.3%) and youden index was 0.794. See Table 6% massesThe current study aimed to compare the diagnostic accuracy of two US-based predictive scoring systems, SRR and LR2 (both proposed by the IOTA group), for discriminating between malignant and benign adnexal masses in Chinese patients. The current study’s results showed that both scoring systems exhibited high diagnostic accuracy when performed by junior doctors. The two scoring systems had good agreement in the diagnosis of malignant adnexal tumours, and the diagnostic performance of the two systems was similar.The IOTA LR2 scoring system is one of the best-performing US-based adnexal lesion diagnostic systems currently in use in clinical practice . SensitiThe IOTA SRR scoring system was developed to estimate the risk of malignancy based on the B- and M-features of the SR model and centre type . The IOTThe comparability of the diagnostic performance of the IOTA LR2 and SR systems has previously been established. Niemi et al.  evaluateThe present study has several limitations, the most noteworthy of which was its low sample size. Previous studies with a similar aim showed that a sample size of 100 was sufficient for validating the study objective , 29. AltIn conclusion, the IOTA SRR and IOTA LR2 scoring systems could effectively distinguish benign from malignant adnexal masses. Both scoring systems had high sensitivity and specificity, with SRR system having higher sensitivity and lower specificity than LR2 system. The two systems have high diagnostic efficiency and can serve as important reference for clinical decision-making. They can also serve as a powerful tool to help junior physicians improve their diagnostic ability of malignant adnexal masses and provide patients with earlier intervention and treatment. This will have an important impact on improving the diagnostic level of adnexal tumours and the prognosis of patients in clinical practice. However, it should be noted that the current study is a single center study with a small sample size, and the results may have certain limitations. In future research, we will collect more comprehensive data, conduct prospective studies in multiple centers, and conduct sensitivity analysis to further investigate the diagnostic efficacy of the two systems in clinical practice, and explore the mechanisms and influencing factors of the two systems in assisting junior physicians in diagnosis.


Background: The p16/Ki67 technique has been poorly studied in postmenopausal women with ASC-US cytology. The objective of this study was to compare the accuracy of p16/Ki67 staining, HPV testing and HPV 16 genotyping for the identification of CIN2 + lesions in postmenopausal women with ASC-US cytology. Method: A total of 324 postmenopausal women with positive ASC-US were included. The women underwent HPV test, colposcopy, and biopsy. The slides were discolored and then stained with the CINtec Plus Kit for p16/Ki67. The HPV test results were classified as HPV16 +, hrHPV+ (other hrHPV genotypes), or HPV negative. Results: The p16/Ki67 sensitivity for CIN2+ was 94.5%, the specificity 86.6%, PPV of 59% and NPV of 95.9%. The HPV test showed a sensitivity of 96.4% for CIN2+, a specificity of 62.8%, a PPV of 35% and a NPV of 98.8%. In postmenopausal women, the prevalence of genotype 16 decreases in favor of the other high-risk genotypes. Conclusion: Given the low sensitivity of cytology and the low percentage of HPV16-positive cancers among elderly women, triage via cytology and genotyping is not the best strategy; double staining cytology shows high profiles of sensibility and specificity for CIN2+ in ASCUS postmenopausal women. The age-related incidence of cervical carcinoma reported in Germany  in 2013 The management of ASC-US was a problem until the publication of the ASC-US/Low-grade Squamous Intraepithelial Lesion Triage Study (ALTS Study), in which the HPV test was incorporated into the interpretation and management process . The AmeIn the literature, the vast majority of ASC-US hrHPV-positive cases did not have underlying high-grade abnormalities .Short-term estrogen treatment, followed by a repeat Pap smear test, can correct false positive interpretations of cytology ; howeverThe co-expression of p16 and Ki67 within the same cell occurs only under abnormal conditions . Many stThe objective of this study was to compare the accuracy of p16/Ki67 staining in the HPV DNA test and in HPV 16 genotyping for the identification of CIN2+ lesions in postmenopausal women with ASC-US cytology.Between 2017 and 2020, 856 women were diagnosed with ASC-US on cytology. From the database of our HPV Unit center, we enrolled 493 post-menopausal women (age: 54.9 ± 5.0 years) for at least two years, with a cytological diagnosis of ASCUS and subjected to HPV test triage. The results of the HPV test were classified into HPV16 positive, hrHPV positive (other hrHPV genotypes) or HPV negative. Patients with a history of cancer, those with chronic or autoimmune diseases, those who had received hormone replacement therapy, and those who had not completed the diagnostic pathway were excluded.The p16/Ki67 test was performed retrospectively on all conventional pap smears diagnosed with ASC-US and where discoloration could be performed. All slides were reviewed by two gynecological pathologists.The results were compared with the histological results of the patients’ biopsy or cone biopsy with LEEP. We performed 176 targeted biopsies in Transformation Zone 1–2 and 149 diagnostic LEEPs in Transformation Zone 3.The histological diagnosis of CIN was established according to the criteria of the World Health Organization on the basis of morphological criteria using H&E staining.The primary outcome was Sensitivity (SE), Specificity (SPE), NPV and PPV of p16/Ki67 for CIN2+ compared with those of the HPV test in post-menopausal ASC-US women. The secondary outcome was the significance of HPV16 genotyping in the triage of post-menopausal women with ASC-US.The study protocol was notified, according to the current legislation on observational studies provided by AIFA, to the Catania1 Ethics Committee of the Catania University Hospital, which did not request additions or changes to the protocol. Furthermore, the Catania1 Ethics Committee of the Catania University Hospital found the consent of the study participants unnecessary as the study concerned only the retrospective review of the medical databaseAfter cytological sampling for HPV DNA, samples were sent to the laboratory for DNA extraction and viral DNA genotyping via genetic amplification followed by hybridization with genotype-specific probes capable of identifying most of the HPV genotypes of the genital region . The commercial method used was the MAG NucliSenseasy system . The technique was previously described .Colposcopy was performed using a Zeiss OPM1F colposcope  and applying acetic acid and a Lugol iodine solution. Any colposcopic anomaly was classified according to the nomenclature proposed by the International Federation for Colposcopy and Cervical Pathologies (IFCPC) into three degrees of increasing abnormality according to severity: (i) Abnormal Transformation Zone (ATZ) grade 1 (ATZ1); (ii) grade 2 (ATZ2); or (iii) cancer. We evaluated the visibility of the squamous-columnar junction (SCJ) and specific biopsies were taken from the portion.Double p16/Ki67 immunostaining was performed retrospectively on all Pap smears interpreted as ASC-US with the CINtec Plus Kit  according to the manufacturer’s instructions. One of the salient features of the test is that it does not require additional samples and can be used on the same slide set up for the Pap smear. We performed the discoloration of the Pap smears, and then, we colored the entire slide.2O for 5 min and begin the first passage in 70% alcohol (5–10 min), then in 80% alcohol and then in H2O.Disassemble the slide with xylene for about 12 h and perform four passages in alcohol of 5 min at decreasing concentrations ; then, perform the decolorization: 5 min in 70% alcoholic solution, keep in HA positive reaction was defined if a red-colored nucleus and brown-colored cytoplasm were present in the same cell, no matter how abnormal the cell morphology was.Cells with cytoplasmic brown staining only and/or nuclear red staining only were defined as negative.t-test for the difference between averages or proportions, the association between p16/Ki67 and the presence of CIN2+ using the Chi-square test and Fisher Exact test for tables with dimensions <5. Using a histological diagnosis of CIN2+ as an end-point, we examined the sensitivity and specificity with widths of 95% confidence intervals (95% CI) of the HPV Test and p16/Ki67 immunocytology in post-menopausal women with ASC-US. We also analyzed the significance of HPV16 genotyping in post-menopausal women. The value of p < 0.05 was considered as statistical significance for all analyses.The analyses were conducted with the STATA software version 13 . The sample was analyzed using descriptive statistics, using the median, mean and standard deviation for quantitative variables, absolute frequencies and percentages for categorical ones. The results are summarized in figures and tables. The homogeneity of the groups was evaluated using the Forty-two of the 493 enrolled women were excluded due to inadequate slides. In addition, the ASCUS slides, before double staining, were reevaluated with the following result: 49 cases of atrophy, 39 cases of reactive alterations and 3 cases of HSIL were excluded. Therefore, a total of 324 patients were considered .2 = 64.188; p < 0.001) OR = 44.79 (95% CI 10.68–187.77)  were HPV positive and 171 (52.7%) were HPV negative . Conside–187.77) . In conc2 = 2.482; p > 0.05) OR: 0.58 (95% CI 0.30–1.14)  cases were HPV16+ and 79 (51.6%) cases were other high-risk genotypes (hrHPV+) positive (χ30–1.14) .Among women positive for genotype 16 only, 21 cases (28.4%) were positive for a CIN2+ lesion.Among other hrHPV-positive women, we found 32 cases (40.5%) of CIN2+ lesions. The most frequent genotype was 31 (8 cases), followed by HPV33 (7 cases), HPV52 (5 cases), HPV51 (5 cases), HPV35 (4 cases), HPV45 (2 cases) and HPV18 (1 case).The prevalence of genotype 16 in postmenopausal women with CIN2+ histological lesions was 38.2% (21/55), and that for hrHPV+ genotypes was 58.2% (32/55), while 3.6% of CIN2+ lesions were negative to HPV .2 = 153.913; p < 0.001) OR: 114.34 (95% CI 33.93–385.37)  with histologically confirmed CIN2+ had positive immunostaining for p16/Ki67. A total of 15.1% of patients  with histologically proven CIN1 and 11.5% of patients (15/130) with negative histology had positive staining for p16/Ki67 (χ–385.37) .The overall sensitivity of p16/Ki67 immunostaining for the detection of CIN2+ was 94.5% (95% CI: 91.3–96.7), the specificity (%) was 86.6 (95% CI: 82.3–90.0), the PPV (%) was 59% (95% CI: 53.5–64.5) and the NPV (%) was 98.7 (95% CI: 96.6–99.6) .The HPV test showed a sensitivity (%) of 96.4 (95% CI: 93.5–98.0%) for CIN2+, a specificity (%) of 62.8 (95% CI: 57.3–68.1), a PPV (%) of 35 (95% CI: 29.5–40.1) and a NPV (%) of 98.9 (95% CI: 96.7–99.6).The diagnosis of CIN2+ in post-menopausal age is also made more difficult by the decline in the role of colposcopy in elderly women due to the low disease rate, the predominance of non-HPV16 genotypes and the lack of visualization of SCJ . The useOther authors have demonstrated that cytology is not an appropriate method for the diagnosis of CIN2+ in this age group, confirming the poor correlation between cytology and histology . In HermMost CIN2+ histological lesions in post-menopausal women are not detected using the Pap smear test.An additional marker is needed as a triage for post-menopausal ASC-US women such as p16/Ki67 double-stained cytology and HPV16 genotyping.In the literature, numerous authors have studied the double staining cytology, and reported that it has a sensitivity comparable to the HPV test but with a higher specificity ,22,23. Tp < 0.001) for p16/Ki67 positivity and increasing histological severity of the lesion: this was observed in 11.5% of histology-negative cases, in 18.7% of CIN1 cases, and in 94.6% of CIN2+ cases detected on histology.In our study population, 55 cases (16.9%) showed CIN2+ histological lesions. The high detection rate of CIN2+ can be attributed to several factors. One reason is that our study population comes from a second-tier center where only positive cases are referred. Secondly, the sample under study consists of postmenopausal women in whom cytology loses its effectiveness for diagnosis. There was a significant trend (2 = 153.913 p < 0.001). In particular the use of p16/Ki67 provides a 94.5% sensitivity level (95% CI 91.3–96.7) to detect CIN2+, very close to the 96.4% sensitivity of the HPV test (95% CI 93.5–98) but with greater specificity (86.6%) (95% CI 82.3–90.0), leading to a higher PPV (59%) for CIN2+.Our study found that the specificity of p16/Ki67 immunocytochemistry for CIN2+ detection was higher than the HPV DNA test  .In our study of post-menopausal women, 39.6% (21/53) of CIN2+ lesions were HPV16 positive, while 60.4% (32/53) had hrHPV genotypes; the proportion of cervical lesions that were HPV16 positive seems to decrease substantially with age (the two cases of invasive cancer were not HPV 16 positive) . Anotherp = 0.139 does not allow us to discriminate between CIN2+ women and CIN2-  women; therefore, the genotyping may not be the best strategy in older women [The presence of the HPV16 genotype in our previous study (women of childbearing age) was associated with a five times greater risk of developing a high-grade lesion (CIN2+) OR = 4.62 (95% CI: 3.13–6.82); the current study  makes us deduce that HPV16 genotyping with an OR = 0.60 (95% CI 0.30–1.18), er women .Based on our data, we can conclude that given the low sensitivity of cytology and the low percentage of HPV16-positive cancers among elderly women, triage by cytology and genotyping is not the best strategy in elderly women; the double staining cytology shows high profiles of sensitivity and specificity for CIN2+ in ASC-US triage both in young and post-menopausal women.The strength of this study is that it is the first retrospective analysis to study the utility of dual staining in ASCUS triage in postmenopausal women.We also considered a sample of post-menopausal women for at least two years with restrictive inclusion criteria, with cervical and conical biopsy samples instead of curettage samples; this allowed the evaluation of both cytology and the architectural relationship with the basement membrane.We investigated the genotype and correlation between HPV16 and CIN2+.The limitations of our research are mainly related to the small sample size and its retrospective nature. Another potential bias could be that the study population selected was from a second-tier center; this could lead to slightly different results than a general population.Further prospective studies with multiple post-menopausal women are needed to confirm the current findings.


Milk contaminated with mycotoxins is a significant issue affecting human health, especially in infants. The current study aimed to investigate the presence of mycotoxins in milk collected from women farmers' vendors (WFV), and to evaluate certain herbal plant fibers as green mycotoxin binders. Moreover, explore the binding efficiency ratios of mycotoxins using shaking or soaking process incorporated with herbal extracts. Furthermore, compare the taste evaluations of tested milk are enriched with herbal extracts. Results indicated that the fumonisins were not detected in the collected cow milk samples but realized a 25% occurrence ratio in buffalo's milk samples. A high occurrence ratio of aflatoxin M1 (aflaM1) was observed in buffalo and cow milk samples. The soaking process of plant fibers in contaminated milk overnight significantly degrades and adsorbs mycotoxins particles. The shacking process incorporated with plant fibers exhibited more effectiveness in mycotoxins degradation than soaking or shacking processes alone. The speed of shacking process played an important role in the mycotoxin's binding process. All the tested plant fibers effectively reduced all mycotoxin presence in contaminated milk, especially green tea, during the soaking or shacking process. Moreover, the shacking process incorporated with plant fibers promoted and supported the mycotoxins degradation process. Milk contaminated with mycotoxins is a highly significant issue affecting human health. The study investigated the role of the shacking process on plant fibers binding process. The shacking process with plant fibers promoted and supported the mycotoxins degradation process. It is also a rich source of protective agents such as antibodies, enzymes and growth factors. Animal species, genetics, physiological status, the environment, and managerial considerations or handling procedures all affect the composition of milk in different ways. For instance, there are significant differences between the composition and characteristics of fresh cow's milk and buffalo milk. Buffalo's milk has a higher level of fat (7.64%), total protein (4.36%), total solids (19.3%), lactose (5.0%) and ash (0.76%) than cow's milk   by toxigenic molds, the milk can become contaminated with mycotoxins  epicarp fruit), which comprises a substantial amount of saponins, coumarins, hydroxycinnamates, essential oils and flavonoids  is a perennial herbaceous plant used as a medicinal sweet drink by ancient Egyptian Pharaohs. It contains flavonoids , saponin triterpenes   agents that bind the mycotoxin particles. Also, this study tested the effect of shacking process and its speed on the efficiency of the adsorption and binding process of mycotoxins and the new taste of the treated milk.22.1Sixty‐four samples of unheated raw milk (2 kg each), where buffalo's milk (16 samples) and cow milk (48 samples) were obtained once per week from farmer's vendors at four open‐air rural big markets in west Alexandria . The samples were collected from November 2021 to January 2022. Each sample was packed in polyethylene bags and immediately kept under freezing conditions (−10°C) until analyzed  (NYSE: TMO) coupled with a Mass spectrometer (HPLC–MS/MS Pacific Biolab)  were brought from Ebn Sina medicinal plant market after estimation of any mycotoxin's contamination occurrence using multi‐screening TLC method according to  coupled with a mass selective detector  equipped with Agilent HP‐5 ms capillary column. Apparatus (GC/MS/MS) made in (Germany) in Mass spectrophotometer Lab, at RCFF Cairo, Egypt, according to  then the filtrated samples were taken for mycotoxins estimation. The control samples (without any addition) were used for mycotoxins analysis using HPLC coupled with mass spectrometry according to  were used. Samples (100 mL each) were taken in a conical flask of 250 mL. The ground plant parts were added individually in three replicates at three concentrations of 0.5%, 1% and 2%. All samples were gently homogenized and kept overnight at 10°C. At the end of the incubation time, all samples were filtered by Watman filter paper (2.6.2n = 1) filter paper. The supernatants were taken for multi‐mycotoxins estimation using HPLC coupled with mass spectrometry according to , respectively. At the end of the shacking process, the samples were filtered again by Watman . The milk experiments were performed in triplicate in a completely randomized design with four variables Duncan, .33.11 (aflaM1) was the highest contaminant in both buffalo and cow milk samples, with 62.5% and 83.34% contamination ratios, followed by Zear 56% and 79.17% for buffalo and cow milk samples, respectively. The frequency of CPA and Don was similar (43.75%) in buffalo milk samples, while they were 20.83% and 10.42% in cow milk samples, respectively. Conversely, the total fumonisins were not found in the tested cow milk samples, but it was 25% in buffalo milk samples. Animals that have consumed feeds contaminated with aflaM1, or other types such as Zear and CPA will be formed due to the metabolic process in the liver of ruminants and excreted in milk   and CPA (6348%) with statistical differences, followed by the green tea leaves (GTL) treatment in binding or eliminating Zear (90.63%) from milk samples.At a dose of 0.5%, DOU fibers showed the highest ratio in binding and degrading aflaMIn this sense, GTL exhibited the best bio‐binders in degrading Zear (100%). Regarding the Dias, co‐treating with DOU presented the most effective bio‐binder and degrading agent for Dias (97.20%), followed by licorice (LIC) (91.03%). All the plant fibers used as bio‐binders in this trial exhibited significantly moderate efficacy in binding and degrading CPA at a dose of 0.5% (63.48% and 61.89%) for DOU and LIC, respectively.1 with a ratio of 96.75%, followed by GTL (95.74%), but LIC was the best treatment in binding and degrading for CPA (93.43%) followed by DOU treatment (78.71%). All the tested bio‐binders and degrading treatments were exhibited significantly higher efficient in getting rid and removing Dias from milk. In this sense, medicinal plants' efficiency in removing the Dias in milk samples was 100%, 95.59% and 82.35% for LIC, DOU, and GTL, respectively. On the other hand, GTL was the best treatment for completely binding and degrading Zear (100%), followed by DOU (93.71%) then LIC (82.24%), respectively.At dose, 1%: DOU produced the best bio‐binder and degrading treatment in reducing aflaMAt the dose of 2%: the detoxification of aflaM1 rations in milk samples were 97.89%, 92.49% and 91.70% for GTL > DOU > LIC, successively. For the removal of CPA in milk samples, results indicate that LIC > GTL > DOU efficiencies were 96.12%, 80.30% and 72.83%, respectively. A similar trend was detected for Zear, whereas the best results were produced by LIC (95.52%), followed by GTL (94.68%) and then DOU (72.17%). For the Dias biodegradation, the efficacies of LIC > DOU > GTL were 91.03%, 85.59% and 72.20% respectively. Our data exhibited that the detoxification process using bio‐binding and biodegradation treatments was affected by the treatment's concentration, the bio binder's variety and its phenolic components content, and the type or structure of mycotoxin (Table 3.5The shacking and filtration processes of milk samples have shown certain moderate effects in degrading mycotoxins (without any plant fiber addition) except in the Dias case Figures , 3, 4, 51 (13.90%), CPA (25.27%), Zear (6.85%) and Dias (70%). After that, increasing the shacking speed by more than 200 rpm might reduce the efficiency of biodegradation of mycotoxins. This variation in mycotoxin behavior as a response to different shaking speeds could be associated with the differences in their molecule's chemical and physical properties and molecular weights  were as follows: Dias (366 g/mol) > CPA (336.4 g/mol) > aflaM1 (328.27 g/mol) > Zear (318 g/mol) > Don (296.31 g/mol).At shacking speed of 200 rpm (SH200) for 1 h, the reduced concentrations of each mycotoxin were as the following: aflaM1 (25.67%), CPA (28.95%), Zear (25.03%) and Dias (58.23%). All the mycotoxins studied (except Dias) were decreased after the treating shacking process at the speed 500 rpm (SH500) as follows: aflaMts Omar, . This nots Omar, , who repAt a concentration of 0.5%, our present study showed that GTL was the best treatment in binding and removing all the occurred mycotoxins as shown in the following order aflaM1 (100%), Zear (99.02%), Dias (95.89%) and CPA (92.14%) followed by DOU which realized efficacy ratios in the following order: aflaM1 (97.98%), CPA (97.01%), Zear (95.24%) and Dias (84.86%). Moreover, the application of LIC fibers treatment was not very efficient as a mycotoxin bio‐binder and biodegraded agent for Zear and aflaM1 as follows: CPA (97.21%), Dias (95.44%), aflaM1 (83.41%), and Zear (79.02%). The outcomes of this trial are consistent with other research . Furthermore, these efficacy ratios were augmented with licorice 2% to achieve the following ratios: 100% for both CPA and Dias, which possess a higher molecular weight (https://pubchem.ncbi.nlm.nih.gov/compound/54682463), then (99.44%) for aflaM1. On the other hand, the removal efficacy ratio of Zear dropped at concentrations of 2 percent of both LIC and DOU from 100 percent (at 1 percent concentration) to 98.74 percent and 95.24 percent, respectively, which may be associated to an imbalance enhancement between the constituents of this employed bio‐binder and their related substituents at high concentrations . CPA has reduced with 92.44% and 97.41% efficacies at 1% and 2% concentrations, respectively. At the same time, LIC was the second‐best treatment which realized at a concentration of 1% the following removing efficacy ratios for Zear (100%), CPA (99.20%), Dias (98.90%) and aflaMAt shacking speed 500 rpm: In case of LIC, the efficacy ratio (ER %) of this bio‐binder in adsorbing and degrading mycotoxins was as follows: At concentration 0.5%, aflaM1 (97.87%) > Zear (92.59%) > CPA (90.25%) > Dias (88.53%). At concentration 1%, aflaM1 (97.75%) > CPA (93.34%) > Dias (90.29%) > Zear (83.92%). At concentration 2%, CPA (98.21%) > Dias (95.29%) > aflaM1 (91.14%) > Zear (75.80%). In all cases, our results illustrated that LIC was very effective in removing all the mycotoxins naturally contaminated cow milk samples, specially aflaM1, at all the tested concentrations. Our data followed those of , the ER % of DOU in removing the following toxins were as the following, Dias (100%) > AflaM1 (97.53%) > Zear (94.40%) > CPA (83.78%). Whereas at concentration 1%, AflaM1 > Dias (93.97%) > CPA (89.55%) > Zear (70.49%). But at concentration 2% AflaM1 (93.95%) > CPA (93.63%) > Dias (81.91%) > Zear (49.09%). Similar to our data,  > aflaM1 (97.31%) > Dias (96.32%) > CPA (94.13%). While, at concentration, 1%, the efficacy ratios order is: Dias (100%) > Zear (97.62%) > CPA (96.62%) > aflaM1 (83.41%). On the other hand, at the concentration of 2%, the efficacy ratios were changed to the following order: CPA and Dias (100%) > Zear (88.25%) > aflaM1 (77.13%). Our findings were globally matched with the study of  > Don (92.60%) > CPA (88.75%). Otherwise, the behavior of DOU in removing mycotoxins from milk samples differed depending on their concentrations. The efficacy ratios of DOU in removing mycotoxins were the best at a concentration of 2% for all the mycotoxins contaminants, but 1% was the best for removing CPA. At the same time, the worth efficacy ratios (ERs %) were 57.28% and 83.53% for both Don and aflaM1, respectively, at 0.5% and 88.25% for Zear at 1%. The order of the efficacy ratios was as follows: aflaM1 (98.19%) > Don (98.06%) > CPA (97.72%) > Zear (93.03%). The behavior of GTL in removing mycotoxins was ascendant and proportional to its concentration in all the mycotoxins contaminants except in the Zear case. At a concentration of 5%, the best‐removed mycotoxin ratio was detected for Zear (96.09%), followed by Don (88.11%). At the concentration of 1% of GTL, Don then CPA were the higher removed mycotoxins with ERs % 96.36% and 94.12%, successively. On the other hand, the best concentration in removing mycotoxins was detected at a concentration of 2% as the following order: Zear (99.55%) > Don (97.33%) > CPA (96.47%) > aflaM1 (95.47%). Overall, our results illustrated that GTL showed the best efficacy ratio in removing Zear (99.55%) and better in removing Don, CPA and aflaM1 from cow milk samples. Our results agree with  of LIC in removing mycotoxins are presented in Table 3.8Our results exhibited that shacking process and then filtration moderately induced the mycotoxins degradation process, which was augmented by increasing the shacking speed with Zear and Don or decreased as augmented speed such as aflaM1 and CPA.Shacking speed at 200 rpm (SH200): In the case of LIC, the concentration of 2% realized the highest efficacy ratios in binding CPA (99.70%) and entirely binding both Zear and Don (100%). The efficacy of removing aflaM1 in buffalo milk samples was 96.38% and 95.58% for 0.5% and 1.0%, respectively.In the case of DOU, 0.5% of DOU exhibited degradation of both aflaM1 and Don completely, while Zear was entirely removed by 2% treatment and the CPA was highly removed at a concentration of 1% with an efficacy ratio (92.43%).In the case of GTL, the ERs were 93.32% and 98.30% for aflaM1 and Don at 2% treatment, respectively. Furthermore, the CPA (95.66%) and Zear (98.76%) were highly removed by 1% and 0.5%, respectively. The data obtained revealed that the bio‐binder's efficacy ratio depended on the concentration of the used plant and the type of mycotoxins. Our results are in harmony with Becker‐Algeri et al.  and IsmaShacking speed at 500 rpm (SH500): In the case of LIC, the removal process was ascendant according to the augmented concentrations in the case of aflaM1, CPA and Don but descendant in the case of Zear. The best ER % in the case of aflaM1 and Don was at 2% (87.77%) and (94.90%), respectively. But CPA was entirely removed in 1% and 2% concentrations. The best‐removed concertation of Zear (95.29%) was found at level 0.5%.In the case of DOU, both aflaM1 and Don were highly removed at level 0.5% with ERs % (97.39%) and (97.57%) successively. At the same time, Zear and CPA were removed at 0.5% and 1% concentrations, respectively.In the case of GTL, only aflaM1was highly removed at the level 0.5% with ERs % (98.19%), while at level 2%, GTL exhibited a highly effective tool in removing CPA and Zear with ERs % (98.38%) and (98.22%), respectively. Regarding Don, the ERs was 97.94% at a concentration of 1%.All the present data indicated that both levels of GTL (0.5% and 1.0%) exhibited the best binder in removing aflaM1 and Don in buffaloes' milk samples, respectively. Moreover, CPA was entirely removed by LIC and DOU at level 1.0% and Zear which was completely removed at DOU 0.5%. The differences in mycotoxins responses to phyto‐ treatment levels might be due to the number of active compounds in each studied medicinal plant. Authors suggested that the isolation of several active compounds from those plants could lead more precisely to explain the potential role in removing the capacity of mycotoxins in milk. In this sense, our results were matched with those of , then GTL at 2% (96.36%). At the same time, the plant fibers' binding process efficiency decreased in SH500 groups except in the case of GTL at 2%, which entirely binding and removed the CPA. Our results indicated that the shacking speed process played a very important role combined with the plant fibers binding to eliminate the toxins from contaminated milk. Moreover,  then LIC at level 2% (98.32%). Furthermore, the behavior of plant fibers (types and their different concentrations) in binding mycotoxin was completely changed according to the applied shacking speed, duration time, and the type of the mycotoxin  and GTL (1% or 2%) completely could remove Dias with shacking speed 500 rpm. Plant fiber concentrations in the binding Dias process were still different from the other mycotoxins binding processes. Our findings have coincided with those of  were included in this test. People preferred the milk taste with LIC because it is sweet and almost like milk without any addition Table . Moreove4The current study demonstrated how the soaking procedure significantly aids the mycotoxin adsorption in cow and buffalo milk. The mycotoxin kind, the plant adsorbent agent, the applied concentration, and the milk constituents all impacted the method's effectiveness. At concentrations of 0.5% and 1%, GTLs completely adsorbed Zear particles. Contrarily, at a level of 1% in cow milk, LIC absorbed Dias particles; however, its efficiency was considerably diminished in the case of buffalo milk. Due to the mycotoxin particles' possible aggregation during the shacking process, the milk shacking method followed by filtering has only a modest impact in reducing mycotoxin particles. The incorporated plant fibers with the shacking process in contaminated milk augmented the effectiveness capacity to bind and adsorb the different particles of mycotoxins. In the cow milk case, GTL (1%) and shacking speed 200 rpm were adsorbed aflaM1, Zear and Dias particles. Still, the same plant fibers at level 2% and shacking speed 500 rpm were completely adsorbed CPA and Dias in milk‐tested samples. Whereas LIC (2%) combined with shaking speed 200 rpm were adsorbed CPA and Dias particles, at level 1% entirely adsorbed Zear particles. In the buffalo milk case, LIC and DOU exhibited the best efficacy ratios in removing mycotoxins than GTL under all the tested concentrations combined with shacking speed (200 or 500 rpm), where aflaM1 and Don were highly removed with GLT at levels 0.5% and 1%, respectively. Overall, the bio‐adsorption and biodegradation of mycotoxins using plant fibers have been proven to be a low‐cost and safe way to remove mycotoxins.The authors declare no conflict of interest in this paper.