Adolescent idiopathic scoliosis (AIS) is the most common form of scoliosis. However, the underlying mechanisms linking spinal curvature in AIS to foot characteristics and walking performance remain unclear.This study aimed to compare walking performance between adolescents with mild, moderate, and severe scoliosis and matched healthy peers with foot posture as covariates.This cross-sectional study of 96 adolescents was conducted between April 2020 to October 2020 in China, with 32 healthy peers in the control group and 64 patients in the AIS group. Foot posture and morphology, plantar pressure distribution, and gait characteristics were analyzed. One-way analysis of variance with Bonferroni correction and a post hoc comparison of the mean differences between the different groups was performed. Multiple analyses of covariance adjusted for age, sex, body mass index, foot posture index (FPI), arch index (AI), and walking speed were performed.Of the 64 adolescents with scoliosis, 18 had mild AIS, 32 had moderate AIS, and 14 had severe AIS. The AI and FPI were much higher in the moderate and severe AIS groups (p = 0.018) and the severe AIS group (p<0.001), respectively, than in the control group. The severe AIS group had advanced and longer midstance (p = 0.014) and delayed propulsion phase (p = 0.013) than the control group. Patients with moderate and severe AIS had asymmetrical gait periods in the left and right limbs (p<0.05). Significant differences in the center-of-pressure excursion index (CPEI) were found between the moderate and severe AIS and control groups (p = 0.003).Moderate and severe AIS significantly influenced walking performance; however, no significant differences were observed between adolescents with mild AIS and healthy controls. Thus, early intervention could target the prevention of specific functional deficits and prevent it from progressing to a severe state. Adolescent idiopathic scoliosis (AIS) is the most common form of scoliosis prevalent in adolescents aged 10–18 years; it is accompanied by an unexplained spinal deformity with a Cobb angle of at least 10° . The maiTo obtain detailed information about the underlying mechanism, a better understanding of the range of physical parameters in patients with AIS is essential, including foot morphology characteristics, plantar pressure distribution, and gait performance. Plantar pressure measurement and gait analysis are widely accepted as vital biomechanical parameters for quantitative assessment of human gait which can provide useful information on foot function and assist with the development of more effective preventive and interventional strategies –9. SeverFoot structure can significantly influence dynamic foot function, and the relationship between foot morphology and function has long been studied ,21. For Based on the above considerations, the purpose of this study was to investigate the differences in plantar pressure distribution and gait in adolescents with mild, moderate, and severe scoliosis compared to controls while considering foot posture as a covariate.This study protocol was registered in the World Health Organization International Clinical Trials Registry Platform of the China Clinical Trial Register under registration No. ChiCTR2000033362.This study was conducted in accordance with the principles of the Declaration of Helsinki and was approved by the ethics committee of the Affiliated Xuzhou Rehabilitation Hospital of Xuzhou Medical University (ethics code No. XKYL2020004). All participants and their legal guardians provided written informed consent before the start of the experiment.Sixty-four patients with AIS and 32 controls volunteered to participate in this study from April 2020 to October 2020 in China. Participants were recruited via offline and online systems concurrently. The participants were recruited from Xuzhou Central Hospital, the Affiliated Xuzhou Rehabilitation Hospital of Xuzhou Medical University, and some community hospitals, and included online platforms , posters, pamphlets, and community advertisements. The main inclusion criteria were as follows: diagnosis of AIS via clinical and radiological methods by professional medical personnel; male and female patients aged 10–18 years who have agreed to undergo longitudinal plain radiography for Cobb angle measurements before the start of the experiment; patients with a left-lumbar and a right-thoracic component of their curves, of varying severity; limb length discrepancy within 1 cm; right limb dominance; no previous specific treatments for scoliosis; no congenital or preexisting musculoskeletal deformities, nervous system deformities, and infectious, traumatic, and psychiatric diseases; no disorder or surgery history of the spine, lower limbs, and feet; able to walk normally without assistance; and able to understand and follow instructions. The exclusion criteria were as follows: diagnosis of non-idiopathic scoliosis; age >18 years; had different deformities affecting normal locomotion; severe deformity of the lower limbs and feet; >1-cm limb length inequality; history of previous treatment for scoliosis; and inability to comprehend and follow task instructions.Demographic information of the participants was obtained via an inquiry and questionnaire. Weight and height were measured using an automatic weight/height measurement system. The Cobb angles on radiographs and ankle ranges of motion were assessed by the same experienced physiotherapist. Patients with AIS were classified as mild (Cobb angle < 20°), moderate (Cobb angle of 20° to 45°), and severe (Cobb angle > 45°) according to the major curve Cobb angles based on the clinical convention. The foot posture index (FPI) was used to measure foot posture, and an experienced investigator examined subjects’ feet manually . The FPIFoot feature parameters were extracted using a laser foot scanner (LSR) system (Vismach Technology Ltd). During scanning, all measurements were performed barefoot in a relaxed standing position with the eyes looking straight forward. Both feet of each subject were scanned using three-dimensional imaging with the LSR 3D Foot Scan V2.5.7.6 software. The scanning system could provide different views of the feet, including the plantar, dorsal, medial, lateral, toe, and heel views. The foot parameters used in the study were foot length, foot width, heel width, arch length, 1–5 metatarsal width, and arch index (AI). The AI, which correlated with foot posture and morphology, and notably, with the navicular height, was measured from the subjects’ standing scans and calculated by dividing the length of the foot into three equal portions (minus the toes) and dividing the footprint in the middle third by the total footprint of all three regions . The refFig 1. 2) The start, end, and peak times of pressure under the medial heel, lateral heel, and 1–5 metatarsals. 3) The stance period during gait was divided into three phases as follows: (i) loading response (0 to 15% of stance), (ii) midstance phase (>15 to 65% of stance), and (iii) propulsion phase  (>65 to 100% of stance); the start time and end time of the gait cycle with the loading response, midstance, and propulsion phases were recorded [The GaitScan pedal pressure system , known to have high reliability and repeatability, has been previously demonstrated to capture plantar pressure and gait characteristics ,29. Gaitrecorded . 4) Footrecorded . 5) The t-test or Wilcoxon nonparametric test was used, where appropriate. The levels of AIS severity compared with healthy status were used as the grouping variable in the univariate analysis of covariance (UANCOVA) and CPEI, indicating that foot function was introduced as a dependent variable. Multiple analyses of covariance adjusted for age, sex, body mass index (BMI), FPI, AI, and walking speed were also performed.Both feet of each participant were selected for the testing and analysis. When comparing the groups, only the left foot was used for the analysis. All statistical analyses were performed using IBM SPSS version 25.0 software , and the significance level was set at p < 0.05. The Shapiro-Wilk test was used to evaluate the normality of distribution, and Levene’s test was used to test the equality of variance. The chi-square test was used for categorical variables, one-way analysis of variance for normally distributed continuous variables, and the nonparametric Kruskal-Wallis test was used for non-normally distributed variables. One-way analysis of variance with Bonferroni correction was performed, and a post hoc comparison of the mean values between the different groups was performed. For within-group comparisons, a paired sample Table 1, when comparing the severe AIS group with the control group, statistically significant differences in weight and BMI were found, revealing that the subjects with severe AIS had lower body weights and BMIs. In the three AIS groups, the proportion of women was much higher than that of men, at 84.37% (54/64) and 15.63% (10/64), respectively. Sex-related differences were observed between the severe AIS and control groups.The study included 96 adolescents and 32 healthy peers in the control group and 64 patients with AIS in the AIS group, consisting of 18 patients with mild AIS , 32 with moderate AIS (30.6° ± 6.5°), and 14 with severe AIS (49.6° ± 3.4°). The mean ± SD age of the participants in the total sample was 13.29 ± 1.75 years, and no significant difference in age was found among the groups. The height of the mild AIS group was higher than that of the control group (p < 0.05); however, no statistically significant difference was found between the other groups. As shown in Table 2. AI was much higher in the moderate and severe AIS groups than in the control group , indicating that patients in the moderate and severe AIS groups tended to have a lower arch than those in the control group. A significant difference was found in the FPI between the severe AIS and control groups , suggesting that individuals with severe AIS tended to have a more pronated foot posture.In contrast to foot length, foot width, heel width, arch length, 1–5 metatarsal width, ankle dorsiflexion, and plantar flexion based on the three-dimensional scanner measurements showed no significant differences among the four groups. The results are presented in Table 3, . Gait parameters such as the loading response, midstance, and propulsion phases were not found to be different between the left and right in the groups . However, the moderate and severe AIS groups had asymmetrical gait periods in both the left and right limbs (p < 0.05), revealing that the patients in the moderate and severe AIS groups might have had abnormal asymmetrical gait patterns. Regarding walking speed, the moderate and severe AIS groups had slower walking speeds than the control group .The severe AIS group had advanced and longer midstance, and a delayed propulsion phase compared with the control group . The medial-to-lateral rearfoot balance (M/L ratio) was significantly different between the moderate and severe AIS and control groups (p < 0.05), being approximately 1.15, 1.32, and 1.48 in the moderate and severe AIS groups, respectively, indicating that impulses were imbalanced across the medial and lateral rearfoot sides. In the forefoot, the percentage of impulses in the first metatarsal was significantly higher in the severe AIS group than in the control group, and the percentages of impulses in the third, fourth, and fifth metatarsals were significantly lower in the severe AIS group than in the control group (p < 0.05). The results demonstrated that the ratio of the impulse from the big toe to the impulse from the first metatarsal (T1/M1 ratio) was significantly higher in the severe AIS group than in the control group (p < 0.05). In terms of the CPEI, significant differences were found between the moderate and severe AIS and control groups (p < 0.05). Univariate analysis of covariance revealed significant differences in CPEI between the groups after adjusting for age, sex, BMI, FPI, AI, and walking speed . No significant plantar pressure difference was found between the mild AIS and control groups. In the intra-group comparison, only the percentage of impulses in the third metatarsal was found to be asymmetrical in the severe AIS group. All of the above-mentioned results are shown in The present study had two purposes. First, this study aimed to compare the plantar pressure distribution, kinematic parameters of gait, foot morphology, and posture between adolescents with mild, moderate, and severe AIS and their matched healthy peers. Second, the foot function of patients with AIS was examined while considering foot posture as a covariate. From the above-mentioned analysis, these following conclusions can be drawn: there were no significant differences between adolescents with AIS (even severe AIS) and their healthy peers in terms of foot length, foot width, heel width, arch length, 1–5 metatarsal width, ankle dorsiflexion, and plantar flexion; however, adolescents with moderate to severe AIS tended to have low foot arches, and those with severe AIS tended to have a more pronated foot posture than the controls. Some abnormal and asymmetrical gait patterns were found in the moderate and severe AIS groups, while these measures were similar between patients with mild AIS and controls. Plantar impulses were significantly imbalanced in adolescents with moderate and severe AIS. In terms of the CPEI, significant differences were found between the AIS particularly moderate to severe cases and control groups after adjusting for age, sex, BMI, FPI, AI, and walking speed.Consistent with the reports of previous studies, female subjects have greater predisposition to AIS ,34. In tIn recent years, researchers have not only focused on the three-dimensional deformity of the spine, but have also focused on different body parts. With the development of medical biomechanics and the overall view of the body structure, studies have shown that biomechanical factors of the pelvis, lower limbs, and feet also have an impact on the formation of scoliosis. The foot structure could significantly influence dynamic foot function, which has been reported previously ,41. In aThis study found that the midstance phase duration was increased and advanced, and the propulsion phase was delayed in patients with severe AIS as compared with their healthy peers. One possible explanation for the prolonged midstance phase was that severe AIS took more time to maintain a steady posture and prepared for the forefoot generating full force to thrust against the ground. The forefoot is known to be the only structure in contact with the ground during the terminal stance . Thus, sThe present study is novel as consideration given to the impact of the different levels of AIS severity since structural deformity might not influence function until the disease has progressed to a severe state . ConsistModerate to severe AIS significantly influenced walking performance; however, no significant differences were observed between adolescents with mild AIS and the controls. These results suggested that early intervention could prevent specific functional deficits such as poor body balance and avert it from progressing to a severe state. Future studies investigating the walking performance of patients with AIS should include foot posture as a concomitant factor not only for scoliosis, as foot posture is likely to influence functional performance. The use of the designated foot orthoses, insoles, and footwear for changing foot loading combined with conventional braces for the conservative treatment of AIS may improve foot function and assist with the development of more effective preventive and interventional strategies.S1 Table(DOCX)Click here for additional data file.


To analyze the level of sex hormone in relation to keratoconus (KC).3), estradiol (E2), progesterone (P), and testosterone (T). Corneal morphological parameters, such as the central corneal thickness (CCT), thinnest corneal thickness (TCT), and maximum simulated keratometry (Kmax), were measured using Pentacam and Sirius.Sixty-two eyes of 62 patients  classified as KC and 120 eyes of 120 patients  with mild to moderate myopia and astigmatism were analyzed. Plasma samples were collected and analyzed using a chemiluminescence immunoassay to determine the concentrations of estriol (EP = 0.044). There were positive correlations between T, CCT  and TCT  in female patients. In male patients, E2 was higher in the KC group than the LVC group , while T was significantly lower . A positive correlation was found between E2 and Kmax  in male patients.The mean age was 23.73 ± 5.16 years for patients with KC and 23.68 ± 6.10 years for patients treated with laser vision correction (LVC). Among the patients with KC, 12 were female (19.35%) and 50 were male (80.65%). The majority of patients with KC were between 20 and 30 years old. In female patients, the concentration of T in the KC group was significantly lower than that in the LVC group (0.86 ± 0.33 vs. 1.18 ± 0.58 nmol/L; 2 was increased in male KC plasma. Different levels of sex hormones are correlated with KC, which, may provide the basis of a new technique for screening and diagnosing KC with or without the assistance of current imaging techniques. Moreover, the correlations between sex hormone alterations and KC provide compelling insight into KC etiopathogenesis.Conclusively, our results showed that T level was reduced in both female and male KC plasma, while E Keratoconus (KC) is a bilateral progressive ectatic disease with central and/or paracentral thinning and steeping of the cornea. It was first described in 1936 by Dr. Benedict Duddell. Since then, extensive research has been conducted to understand the pathophysiology of the disease . The incPatients with KC always develop irregular astigmatism and myopia, leading to mild-to-marked impairment in visual acuity and quality of vision. Central or paracentral stromal thinning, rupture in the Bowman's layer, ring-shaped ferritin deposition at the basal layer of the epithelium (Fleischer's ring), fine vertical lines in the deep stroma of Descemet's membrane (Vogt's striae), and Munson's sign are typical clinical manifestations of KC , 7. NumeThe present study aimed to investigate the association between sex hormones and KC, and analyze their possible mechanisms.This study was conducted at the Peking University Third Hospital Eye Center, Beijing, China. This study was approved by the Medical Science Research Ethics Committee of Peking University Third Hospital, and followed the tenets of the Declaration of Helsinki. Informed written consent was obtained from all subjects.All subjects were enrolled continuously in the Peking University Third Hospital Eye Center in 2020. Patients were divided into two groups: (1) 62 eyes of 62 patients  classified as KC and scheduled to be treated with corneal collagen cross-linking (CXL). (2) Hundred and twenty eyes of 120 patients  with mild to moderate myopia and astigmatism were scheduled to be treated with laser vision correction (LVC). In the KC group, only the operative eye data were included. In the control group, one eye from each patient was randomly selected for comparison. All patients in the control group were examined at least twice before surgery to ensure that the patients' myopia or astigmatism was stable and conformed to the surgical indications. The diagnosis of KC was based on the presence of clinical signs , central or paracentral steepening in the corneal curvature, cornea thickness, and other necessary parameters measured using topographic and tomographic maps. All diagnoses were confirmed by an experienced specialist in CXL and LVC surgery. Patients with a trend of progression in the past 12 months and that met one of the following conditions were treated with CXL: the maximum K reading increased by >1 D, mean corneal refractive power increased by >1 D, astigmatism increased by >1 D, and the spherical equivalent of manifest refraction increased by >1 D with best spectacle-corrected distance visual acuity having lost more than one line. Exclusion criteria included hormone supplementation therapy, previous ocular surgery, trauma, and systemic diseases such as immunodeficiency and desmosis.max) were measured using a Pentacam  and Sirius .All subjects underwent a complete standard ophthalmological examination. The refraction errors were evaluated with a KR-8100 auto kerato-refractometer , visual acuity was measured with the Standard Logarithmic Visual Acuity Chart. The anterior segment and fundus were examined with a slit-lamp microscope , intraocular pressure was measured with non-contact tonometers , and corneal parameters such as the central corneal thickness (CCT), thinnest corneal thickness (TCT), and maximum simulated keratometry (K3), estradiol (E2), progesterone (P), and testosterone (T) using chemiluminescence immunoassay by Immulite 2000 . In consideration of the effect of the menstrual cycle on female sex hormones, blood samples were collected from females during their secretory phase. Once received, the levels of the four hormones are processed within approximately 24 h. For males, normal E2 ranges were 0.00–143.00 pmol/L and normal P ranges were 0.86–2.90 nmol/L. Male androgen had no normal ranges. For females, normal E2 ranges were 111.00–1007.00 pmol/L, normal P ranges were 3.00–68.00 nmol/L, and normal T ranges were 0.00–2.53 nmol/L.Blood samples were obtained at 10:00–13:00 from all subjects. For males, 2–3 ml blood samples were collected from the outpatient venous blood collection area 3 days before surgery, and were sent to the Laboratory of Endocrinology, Reproductive Center, Peking University Third Hospital to test the levels of sex hormones, including estriol . The normality distribution was checked for all parameters using the Kolmogorov-Smirnov test. Age, sex hormone levels, and corneal parameters were analyzed descriptively, and the results were expressed as the mean and standard deviation for the two study groups. Because of the known sex bias for hormones, we thought analysis by sex was essential. The Mann-Whitney U test was used to evaluate differences in parameters between female and male patients. Spearman correlation analysis was used to correlate sex hormone levels, CCT, TCT, KSixty-two patients with KC  and 120 LVC patients  were included in this study. The mean age was 23.73 ± 5.16 years for KC patients and 23.68 ± 6.10 years for LVC patients, with no significant difference between groups. Among the KC cases, there were more males , and the KC patients' ages were concentrated between 20 and 30 years ; Table 1P = 0.044). We performed Pentacam and Sirius examinations, and CCT, TCT, and Kmax were recorded for each patient. The mean CCT, TCT, and Kmax values differed significantly between the groups (P < 001). In the male KC group, E2 was higher than in the LVC group , while T was significantly lower . However, there was no difference between the KC and LVC groups in terms of E3 and P .r = 0.395, P = 0.023) and TCT  in female patients. A positive correlation was found between E2 and Kmax  in male patients. E3 and P were not correlated with any of the KC parameters.in-vitro fertilization always have progression in KC (With the progression of science and technology, the early diagnosis and treatment of KC are more and increasingly accurate and cutting-edge, but the etiopathogenesis of this important kind of corneal ectasia disease has not been completely investigated. Exploring the etiopathogenesis of KCs is difficult, but essential. Hypotheses have proposed that genetic, immunological, metabolic, and endocrinological factors may be involved in the progression of KC. Furthermore, many reports have shown a significant relationship between sex hormones and KC. Some case reports have shown increased tendency for corneal ectasia in pregnancy and hormone replacement therapies and reveal corneal graft rejection in pregnancy. Yuksel et al. found that females treated with on in KC . In Emilon in KC . Spoerl on in KC . Indeed,Plasma concentration screening, clinical observations, and clinical examinations were used to determine the correlation between KC and sex hormones. It was thought that increased levels of sex hormones may be associated with the etiopathogenesis and progression of KC due to some mechanism of action. First, c-fos, an estrogen-target gene, is involved in cell proliferation, differentiation, apoptosis, oncogenesis, and invasion. Pan found that both c-fos and MMP-9 protein levels were higher in females with endometriosis than in those without endometriosis. A positive correlation between c-fos and MMP-9 protein levels was found, which suggests that c-fos may be regulated by sex hormones and promote the gene expression of MMP-9 in the development of endometriosis . Therefo2 and T showed a statistically difference, suggesting that even in the normal range, the sex hormone might have certain influences on the cornea.Although the plasma levels of sex hormones were approximately in the normal range in patients with KC and LVC, the plasma levels of EThis study has several limitations. The present study had a cross-sectional design, and the findings must be confirmed via longitudinal studies that require larger patient populations. Sex hormones were not measured in the tear fluid. In addition, there are changes in many other hormones, such as growth hormone, around the age of 25. Thus, further research is required that examines their role in KC.In conclusion, although the specific mechanism between sex hormones and KC appears not to have been investigated, it is proposed that sex hormones may play an essential role in the development of KC. The results of our study might be helpful as a basis for further studies to elucidate the etiopathogenesis of KC and explore the role of sex hormones in the treatment of KC in the future.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 Medical Science Research Ethics Committee of Peking University Third Hospital. Written informed consent to participate in this study was provided by the participants or their legal guardian/next of kin.XZ, YY, and TS were responsible for the initial plan, study design, data collection, data extraction, data interpretation, manuscript drafting, statistical analysis, and conducting the study. YZ and YC were responsible for data collection, extraction, and critical revisions of the manuscript. YC was the guarantor for this article and has full responsibility for this study. All authors contributed to the article and approved the submitted version.This study was supported by grant of Key Clinical Innovation Program of Peking University Third Hospital, Category A, No. Y65495–05.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.


We introduce a new resource: the SAYCam corpus. Infants aged 6–32 months wore a head-mounted camera for approximately 2 hr per week, over the course of approximately two-and-a-half years. The result is a large, naturalistic, longitudinal dataset of infant- and child-perspective videos. Over 200,000 words of naturalistic speech have already been transcribed. Similarly, the dataset is searchable using a number of criteria . The resulting dataset will be of broad use to psychologists, linguists, and computer scientists. From the roots of children’s language learning to their experiences with objects and faces, naturalistic data about children’s home environment provides an important constraint on theories of development , and are accompanied by monthly parent reports on vocabulary and developmental status. Together, these data present the densest look into the visual experience of individual children currently available.Three families participated in recording head-camera footage see . Two famSam is the child of the family who lived in Australia. He wore the headcam from 6 months to 30 months of age. The family owned two cats and lived in a semirural neighborhood approximately 20 miles from the capital city of Adelaide. Sam was diagnosed with autism spectrum disorder at age 3; as of this writing (at age 7), Sam is fully mainstreamed, has friends, and does not require any special support.Alice is the child of a family who lived in the United States. She wore the headcam from 8 months to 31 months of age. The family also owned two cats throughout the recording period. For the first half of the recordings, the family lived on a rural farm and kept other animals on or near their property, including chickens; the family frequently visited a major city (San Diego). For the second half of the recordings, the family lived in the suburbs in New York State.Asa is the child of a family who lived in the United States. He wore the headcam from 7 months to 24 months of age. Data collection for this child terminated prematurely due to the onset of the COVID-19 pandemic and the birth of a younger sibling. The family owns no pets, and lives in the suburbs in New York State.Parents completed the MacArthur-Bates Communicative Development Inventories (MCDI) inventory each month .On occasions where a child persistently refused to wear the headcam, the camera was used as a normal camera . A spreadsheet detailing which videos are third person accompanies our data on Databrary , and occasional weeks were skipped due to travel or other logistical barriers; additionally, most recordings for the Asa dataset are sampled from between 7 a.m. and 10 a.m. Technical issues, such as memory card or camera failure, although infrequent, resulted in some skipped recording sessions.When recording, we obtained written consent or verbal assent  for participants whose face and voice were featured in the videos. Videos for which we were unable to secure assent/consent for people who were potentially identifiable are omitted from the dataset.Each recording session lasted until the battery on the Veho camera failed, or once 90 min had elapsed. In most cases, this resulted in recording sessions that lasted approximately 60 to 80 min. Each recording session typically resulted in multiple video files, with a maximum single-file duration of 30 min.One goal of our coding efforts was to tag each video with its contents, so that researchers could search for content relevant to their research interests. To do this, videos were skimmed at an effective 10x-speed playback, and visually assessed for locations and location changes. Coding occurred by playing each video in VLC (VideoLAN Client), and coding using custom spreadsheets with macros. Coders were highly trained and deeply familiar with the dataset.https://nyu.databrary.org/volume/564/slot/47832/-?asset=254904). The coder also noted any potential ethical concerns for further screening, and any additional notes.The following pieces of information were coded for each video. First, the coder briefly described the contents of the video . The coder also recorded the location , the activity or activities , the individual(s) present , the body parts visible , and the most salient objects . See Every time there was a significant change in location or in activity, we coded each component of the video anew. For example, if the child moved from the bedroom to the bathroom, we coded all elements of the video  once for the bedroom and once for the bathroom.The intention for coding videos was to allow other researchers to identify videos of interest, not for analysis of the codes themselves. While such an analysis may be both possible and extremely interesting in the future, we encourage caution at this point for several reasons. First, relative frequencies of activities will not be accurately computed until the entire dataset is coded. Second, while our coding scheme remained stable throughout the majority of coding, there are a few items  that were added to our coding scheme after coding began. Their dates of addition are described in the Read Me file. Finally, some objects, activities, or locations may appear briefly in particular videos, but not be tagged . The decision for whether or not to tag a particular entity within the video was subjective, and was guided by the intention of coding the videos in a way that would be helpful for future researchers to identify videos that would be useful for their particular research projects. For example, in the current dataset, the tag “Object: Clothing” labels 114 videos , and researchers who are interested in children’s interactions with clothing will find at least one salient child–clothing interaction in each of those videos. However, the absence of the “Object: Clothing” tag for the remaining videos certainly does not imply that everyone was naked.Researchers interested in contributing to the ongoing coding effort should contact the corresponding author.https://nyu.databrary.org/volume/564/slot/47832/-?asset=254904. Transcribers noted the timestamp for the onset of each utterance, the contents of the utterance itself, the speaker, any notable, significant, or contextually important actions, and any objects that were relevant, visually salient, or interacted with. Transcribers were instructed to record any utterances they heard verbatim. A new entry was created every time there was a change in speaker, there was a meaningful shift in conversation, or there was a meaningful pause between speaking. Each entry begins by recording the time in the video that the utterance in question begins.One goal of this project was to transcribe all of the utterances in each video. Videos were assigned to college students and transcribed. Interested readers can contribute to the ongoing transcription effort by contacting the corresponding author; our original training materials are provided here: There are over 1,800 videos with more than 460 unique recording sessions, yielding well over 500 hr of footage. Currently, over 235,800 words have been transcribed, representing 16.5% of the present dataset; 28.3% of the videos have been coded for location, activity, living things, body parts, and objects and are now searchable.https://skidmoreheadcam.shinyapps.io/SAYcam-TagSearch/. For example, a researcher interested in studying the auditory input surrounding infant nursing could visit our search tool, and search for segments containing nursing. At present, this would yield 53 videos; the output table automatically includes a link to the relevant video, and a link to a Google Sheet containing any available transcriptions. The researcher could then choose to analyze those 53 videos, or to further restrict their search . Or, let’s say a researcher was interested in a child’s exposure to cats prior to acquiring the word “cat.” The researcher would then access the child’s MCDI vocabulary measures on Databrary in order to identify the month at which the child acquired the word “cat.” They could then use our search tool to search for videos from that child from before the child learned the word, and access those videos via the provided Databrary link.Researchers can access our dataset, which contains AVI and MP4 videos, PDFs, and Excel sheets representing our data at the Supplementary Materials online, and can search for relevant videos here: https://nyu.databrary.org/) standard application process. Researchers will need to demonstrate that they are authorized investigators from their institution, that they completed ethics training, and that their institution has an institutional review board.Individuals can apply to access the data using Databrary’s , Award ID: R03 #HD09147.JS, National Institutes of Health (JS: Conceptualization: Supporting; Methodology: Equal; Data Collection: Equal; Funding: Lead; Supervision: Lead; Writing - Original Draft: Equal; Writing - Review & Editing: Lead. MM: Analysis and Coding: Lead; Data Collection: Supporting; Supervision: Supporting; Writing - Original Draft: Equal; Writing - Review & Editing: Equal. AP: Conceptualization: Equal; Data Collection: Equal; Methodology: Equal; Supervision: Supporting; Writing - Original Draft: Equal; Writing - Review & Editing: Supporting. EW: Conceptualization: Supporting; Data Collection: Equal; Methodology: Supporting; Supervision: Supporting; Visualization: Supporting; Writing - Original Draft: Equal; Writing - Review & Editing: Supporting. MCF: Conceptualization: Equal; Data Collection: Supporting; Funding: Supporting; Methodology: Equal; Supervision: Supporting; Writing - Original Draft: Equal; Writing - Review & Editing: Supporting.


Background: Chemotherapy is a cornerstone of treatment in advanced gastric cancer (GC) with a proven impact on overall survival, however, reliable predictive markers are missing. The role of various inflammatory markers has been tested in gastric cancer patients, but there is still no general consensus on their true clinical applicability. High neutrophil-to-lymphocyte (NLR) and low (medium)-platelets-volume-to-platelet ratio (PVPR) are known markers of unspecific immune system activation, correlating significantly with outcomes in advanced GC patients. Methods: Metastatic GC patients (N:155) treated with chemotherapy +/− trastuzumab were enrolled in this retrospective study. Pre-treatment NLR and PVPR, as well as other inflammatory markers were measured in peripheral blood. Univariate Cox regression was conducted to find markers with a significant impact on overall survival (OS) and progression-free survival (PFS). Spearman correlation and Cohen’s kappa was used to analyze multicollinearity. Multiple multivariable Cox regression models were built to study the combined impact of NLR and PVPR, as well as other known prognostic factors on OS. Results: Elevated NLR was significantly associated with increased risk of death , and lower PVPR was significantly associated with improved outcomes . A novel inflammatory marker, based on a combination of NLR and PVPR, allows for the classification of GC patients into three prognostic groups, characterized by median OS of 8.4 months (95% CI 5.8–11.1), 10.5 months (95% CI 8.8–12.1), and 15.9 months (95% CI 13.5–18.3). Conclusion: The NLR and PVPR score (elevated NLR and decreased PVPR) is a marker of detrimental outcome of advanced GC patients treated with chemotherapy. Interactions between the immune system and cancer are attracting considerable interest because of growing evidence of underlying correlations that are sometimes too elusive to be appropriately described. However, many years of intensive evolution of the immuno-oncology field, which finally led to the introduction of checkpoint inhibitors, allowed us to predict, if not fully comprehend, the interplay between the immune system and malignancy in a way that is consistent and beneficial.Despite significant progress in systemic treatment, metastatic gastric cancer (GC) remains incurable. In this palliative setting, systemic chemotherapy still represents the treatment of choice worldwide, with the potential to improve patients’ outcomes  .Platinum-based chemotherapy is a cornerstone of the first-line treatment for stage IV gastric cancer . OxaliplDespite numerous efforts to incorporate immunotherapy into the first-line systemic treatment of GC , the optSystemic inflammation is a well-known phenomenon, although not fully understood, involved in carcinogenesis and progression of cancer, and the modulation of cancer cells’ immunogenicity and sensitivity to systemic treatment . TherefoThe best apprehended and easy-to-measure marker of systemic inflammation is NLR (neutrophil-to-lymphocyte ratio), in which its prognostic potential was demonstrated in multiple malignancies ,16,17. HMany additional markers of systemic inflammation, such as PLR (platelet-to-lymphocyte ratio) , HPR . Our exploratory study aimed to define the optimal inflammatory marker with the highest predictive and prognostic potential.Data on 155 GC patients treated in the Department of Clinical Oncology, University Hospital in Cracow, were retrospectively collected between September 2013 and December 2019. Data were censored on 31 December 2020. Patients were eligible for analysis if at least one cycle of palliative intravenous chemotherapy had been administered. Patients on steroid treatment, defined as a prednisone dose higher than 10 mg per day or equivalent, were excluded. The choice of chemotherapy regimen was left to the discretion of the leading physician; however, the decision was based on the patient’s general condition, HER2 expression, and current therapeutic guidelines at a given time. The allocation of patients to chemotherapy could have been biased by the evolution of guidelines, which occurred over time, promoting the switch from 3-drug to 2-drug regimens in asymptomatic or mildly symptomatic patients or patients with suboptimal performance status. The patient’s treating physician determined the moment of disease progression for each patient, based on clinical or radiological symptoms. The exact date of death was obtained from public databases. Due to the lack of the date of death for some participants, the date of the last visit was used in case of unequivocal progression and health deterioration. Peripheral blood inflammatory markers were measured at the baseline (up to one week before initiation of the first-line chemotherapy). The NLR was calculated by division of absolute neutrophile and lymphocyte counts. The RDW/PLT was defined as the ratio of RDW-SD (red cell distribution-standard deviation) to platelet count. The HPR was calculated by division of Hgb (hemoglobin) and platelet count, and the PLR was calculated by division of platelet and lymphocyte count. The inflammatory marker MPV/PLT, further referred to as PVPR (platelet volume to platelet ratio), was defined as the ratio of MPV (medium platelet volume) to platelet count.p < 0.05.Inflammatory markers were dichotomized based on receiver operator characteristic (ROC) analysis, with a binary state variable split at the median of survival. The best cut-off value was obtained by maximizing the Youden index . In case of nonsignificant results of ROC analysis, the median value of the analyzed marker was used as a tradeoff. Overall survival (OS) was defined as the time from the diagnosis of stage IV disease to death, and progression free survival (PFS) was calculated from the initiation of chemotherapy to either death or disease progression appointed by the leading physician. Otherwise, the subject’s case was censored. Categorical variables were reported with frequencies and percentages, and continuous variables were characterized by medians and interquartile ranges. The Kaplan–Meier method was used to estimate the medians of PFS and OS, and the comparison of survival functions between two or more independent groups was performed by applying a log-rank test. Cox regression analysis was employed to evaluate the impact of selected variables on OS and PFS. Inflammatory markers that were statistically significant in univariable models were designated for further multivariable analysis. Possible redundancy of inflammatory markers was verified based on the pairwise calculated Spearman’s rank correlation coefficients and Cohen’s kappa coefficients under criterion of redundancy detection: rho greater than 0.6 and kappa greater than 0.5, simultaneously. Selected inflammatory markers as well as other plausible clinically meaningful variables were considered as admissible for inclusion in the multivariable Cox regression analysis. The proportional hazard assumption in all multivariable models was tested with visual inspection of Schoenfeld residuals, and was formally complemented with omnibus chi-squared goodness of fit tests, relating failure time to covariate values. Additional evaluation of risk models was performed based on Harrell’s C-index (concordance index) and a value significantly greater than 0.5 was regarded as indicating that the estimated risk scores are good at determining which of the two patients will experience the endpoint first. To get a reliable and robust estimation of the contribution of inflammatory markers to the predictions of OS and PFS, and to verify the consistency of the results, a variety of survival models were fitted covering the wide spectrum of sets of covariates admissible for adjustment. ECOG, the well-known strong risk factor, yet violating the proportional hazard assumption (PH), could not be directly incorporated in multivariable models; instead, the stratification on the ECOG variable was performed. Consequently, it was not possible to obtain a hazard ratio value for the effect of ECOG in multivariable analysis, due to different baseline hazard function for categories 0, 1, and 2. However, roughly parallel log-log survival curves allowed the calculation of the size effect for ECOG in the univariable analysis. The study was exploratory, and no correction was applied for multiple statistical testing. Analyses were done in the SPSS version 27  and R software . All tests were two-sided and statistical significance was defined as Final analysis consisted of one hundred and fifty-five patients. Patient characteristics are reported in p = 0.002). Optimal cut-off ratio calculated with the Youden index for NLR was 3.99  0.678, NPV  0.593). For PVPR, median was used as cutoff . Median OS and PFS for the entire analyzed population were 10.6 months (95% CI 9.4–11.9) and 5.5 months (95% CI 4.3–6.7), respectively. The ROC analysis summarizing the performance of the classifier was significant only for NLR and PDW, for which the best cut-off point was used as a threshold, while the inflammatory markers median value was applied in the remaining cases. The area under curve (AUC) for NLR was 0.651 . The Kaplan-Meier curve with log-rank test  showed sOur study showed that NLR and PVPR score could be used as a novel and unique predictor of survival in stage IV gastric cancer patients receiving standard cytotoxic chemotherapy. Lack of accurate predictive factors hinders optimization of cytotoxic chemotherapy in gastric cancer. Our study defined a promising marker, but the exploratory nature of this analysis requires prospective validation. Chemotherapy might be tailored for individual GC patients when a proven and cheap predictive factor is available. We have shown that both NLR and PVPR had a significant impact on OS and PFS in the first-line treatment of gastric cancer. These markers also did not seem to correlate with each other, so it should be concluded that they reflect two distinct realms of inflammatory response. Accordingly, a combined score of NLR and PVPR seems to enhance each parameter’s prognostic strength. Patients analyzed in the study were treated with cytotoxic chemotherapy appropriate for a given time, with a shift from a 3-drug regimen  to a 2-drug regimen , resulting from changes in gastric cancer treatment guidelines. In our opinion, this is a very interesting setting and further retrospective analysis should be conducted to analyze the difference in predictive factors pre and post shift. The NLR is a well-established marker of inflammation in multiple malignancies and other diseases ,31,32. TRecent insights into the interplay between cancer cells and platelets underline its importance in promoting tumor progression, by decreasing the activity of natural killer lymphocytes , stimulaWe are aware of the limitations of our study. The analysis was retrospective, and all data were gathered manually. There was an initial bias due to only analyzing patients that received at least one cycle of chemotherapy, so the group of patients with exceptionally bad prognosis was omitted. Nevertheless, our study, comprising a real-world population of advanced GC patients, treated with standard chemotherapy, allowed for the definition of a novel, unique, and readily available prognostic factor in clinical practice.In summary, the NLR and PVPR score (high NLR and low PVPR) is a strong predictor of detrimental outcomes (both OS and PFS) in advanced, chemo-naïve GC patients undergoing first-line palliative chemotherapy. Validation of this novel prognostic factor in other populations of cancer patients is warranted.


Background: This study explored the influences of electroacupuncture combined with dietary intervention on the intestinal flora in perimenopausal patients with abdominal obesity by using the 16s rRNA sequencing technology.Methods: Perimenopausal patients with abdominal obesity were divided into the Electroacupuncture group and the Control group. Patients in the Control group received healthy lifestyle education, while those in the Electroacupuncture group received electroacupuncture combined with dietary intervention. Before and after treatment, the weight, height, waist circumference, hip circumference, waist-height ratio (WHtR), waist to hip ratio (WHR), and body mass index (BMI) of the patients were recorded; the levels of serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein cholesterol (HDL-C), fasting insulin (FINS), and fasting blood glucose (FGB) were evaluated; and the abundance, diversity, and species differences of intestinal flora were analyzed using 16s rRNA sequencing technology.Results: The body weight, waist circumference, hip circumference, BMI, WHR, and WHtR of patients in the Electroacupuncture group after treatment were lower than those before treatment. Compared with the Control group, patients in the Electroacupuncture group after treatment displayed lower waist circumference, WHtR, WHR, TG, and LDL levels as well as species abundance, higher species diversity, and lager species difference in the intestinal flora. Besides, the proportions of Klebsiella and Kosakonia in the intestinal flora of patients in the Electroacupuncture group after treatment were larger than those before treatment.Conclusion: Electroacupuncture combined with diet treatment generated a therapeutic effect on abdominal obesity in perimenopausal patients by improving the community structure of intestinal flora. With the improvement of people’s living standards and the changes in dietary structure and environment, the number of obese people has maintained an upward trend . ObesityThe routine treatments for obesity normally are dietary intervention and hypoglycemic drugs, which also provide certain references for perimenopausal women with abdominal obesity . In the The prevalence of abdominal obesity has raised people’s curiosity of finding the balance factors that affect the occurrence and development of this disease . The relConsidering that the 16s rRNA high-throughput sequencing technology is the main and cutting-edge method for microorganism analysis , we, in The study has been approved by the Ethics Committee of Zhejiang Hospital [No. 2018 (9K)] and all patients signed the written informed consent.A total of 73 perimenopausal patients with abdominal obesity who received treatment in the Acupuncture Department of Zhejiang Hospital from December 2017 to January 2019 were recruited in this study. The diagnostic criteria for perimenopausal period were formulated according to the chapter of “Menopausal Syndrome” in the 8th edition of “Obstetrics and Gynecology,” combined with “Chinese Obstetrics and Gynecology.” The details are as follows: (1) 40–60 years old; (2) Clinical manifestations of menstrual disorders for more than 3 months, or amenorrhea within a year, etc.; (3) One or more of the following possible related symptoms, such as typical symptoms of unstable vasomotor function, psychoneurological symptoms , physical symptoms  and symptoms of urogenital atrophy ; (4) Imaging examination to exclude organic diseases such as uterus and ovaries. The standards for abdominal obesity were formulated with reference to the “Guidelines for the Prevention and Control of Overweight and Obesity in Chinese Adults”: waist circumference ≥ 80 cm, and (or) WHtR ≥ 0.50, and (or) WHR ≥ 0.78. The general information of the patients including age and height was recorded .The inclusion criteria of patients in this study were as follows: (1) patients were diagnosed with perimenopausal and abdominal obesity; (2) patients had not received weight-loss treatment in the past 3 months. The exclusion criteria of patients in this study were as follows: (1) patients with secondary obesity caused by hypothalamic syndrome, pituitary tumor, Cushing’s syndrome, hypothyroidism, insulinoma, or polycystic ovary syndrome; (2) patients who were diagnosed with uterine or ovarian malformations, have undergone uterine or ovarian resection, or in pregnancy or breastfeeding; (3) patients with severe diseases such as hyperthyroidism, diabetes, psychosis, malignant tumors, heart, liver, and kidney diseases, and hematopoietic system diseases.n = 36) and the Electroacupuncture group (n = 37). The patients in the Control group received healthy lifestyle education, while those in the Electroacupuncture group received electroacupuncture treatment combined with dietary intervention for 8 weeks. Before and after treatment, the stool and blood of the patients were collected for subsequent analyses, and the weight, height, waist circumference and hip circumference of the patients were recorded to calculate waist-height ratio (WHtR), waist to hip ratio (WHR), and body mass index (BMI). Here, the method of measuring waist circumference recommended by the World Health Organization is adopted: the subject takes off his shoes, hat and coat, fasts, and stands upright, with feet being separated by 25–30 cm. The measurement position is at the midpoint of the line between the horizontal anterior superior iliac crest and the inferior edge of the twelfth rib.The patients were randomly divided into two groups: the Control group  and 0.30 mm × 50 mm  disposable acupuncture needles  as well as an SDZ-V electronic acupuncture instrument  were employed. 1.5 inch acupuncture needles were used at the Backshu acupoints and four-limb acupoints. For the Backshu acupoints, the needle was inserted obliquely at 45° into the spine at 1-inch depth, while for the four-limb acupoints, the needle was inserted straight at 0.8–1.0-inch depth . 2.0 inch acupuncture needles were inserted at the acupoints on the abdomen at 2-inch depth. Electroacupuncture was applied to Tianshu, Abdomen Jie, Daheng, and Meridian, and the waveform was a dense wave with a frequency of 2/100Hz. The needle was retained for 30 min. The acupuncture acupoints without electroacupuncture treatment were slightly lifted and twisted3 times every 10 min. Patients in the Electroacupuncture group received electroacupuncture treatment every 2 days (3 times a week) for 8 weeks.Patients in the Electroacupuncture group were given a strict low-sugar, low-fat, low-salt, and high-fiber diet. The diet contained staple foods, meat, and vegetables , fruits, and vegetables (no less than 250 g), and mineral supplements. Besides, the patients drunk 2,000–2,500 ml of water and no more than 20 g of liqueur, ingested less than 6 g of salt per day, and only ate fish products twice a week. Here, dietary intervention was implemented on the basis of menopausal health management strategies .2.Before and after treatment, the weight (kg), height (cm), waist circumference (cm), and hip circumference (cm) of the patients were recorded. Then WHtR, WHR, and BMI were calculated using the following formulae: WHtR = waist/height; WHR = waist/hip; BMI = weight/heightBefore and after treatment, the serum of all the patients was analyzed using an Automatic biochemical analyzer  to evaluate the levels of serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein cholesterol (HDL-C), fasting insulin (FINS), and fasting blood glucose (FBG). Besides, Homeostasis Model Assessment-insulin resistance (HOMA-IR) was calculated according to the formula: HOMA-IR = FBG × FINS/22.5.The intestinal flora in the patients was detected by 16s rRNA sequencing technology according to the previous publication . In brieIn line with the clustering results of OTU, the representative sequence of each OTU was annotated to obtain the corresponding species annotation information and species abundance distribution. Then, the species abundance in the sample and the species differences between samples were obtained through the visual statistical analysis of OTUs, and the Venn and Partial Least Squares Discrimination Analysis (PLS-DA) diagrams were plotted. Next, the information was subjected to Alpha diversity analysis to acquire the community structure information with differences in the sample. The Linear discriminant analysis (LDA) effect size (LEfSe) statistical analysis was applied to further investigate the differences in community structure among samples, and to identify different species via testing the species composition and community results of the grouped samples.P < 0.05. These analyses were performed under SPSS 20.0 software.All data involved in this study except for those from intestinal flora analysis were tested the normality of the variable distribution and analyzed by student’s t-test. Statistical data were presented as mean ± standard deviation. The data were recognized as statistically significant when P < 0.01), waist circumference (P < 0.01), hip circumference (P < 0.01), BMI (P < 0.01), WHtR (P < 0.01), and WHR (P < 0.01) of the patients were decreased after treatment when compared with those before treatment, while the patients in the Control group after treatment showed a larger waist circumference (P < 0.01) and a higher WHR (P < 0.01) than before treatment. Additionally, the waist circumference (P < 0.01), WHtR (P < 0.01), and WHR (P < 0.01) of patients in the Electroacupuncture group were notably lower than those in the Control group, which indicated that electroacupuncture combined with diet treatment might ameliorate abdominal obesity in perimenopausal patients.Given that there was no statistical difference in the general information   of the tP < 0.05) as compared with that before treatment. Moreover, the TG (P < 0.01) and LDL levels (P < 0.01) of patients in the Electroacupuncture group were markedly lower than those in the Control group, which proved that electroacupuncture combined with diet treatment ameliorated the TG and LDL levels of perimenopausal patients with abdominal obesity.Then the levels of TG, TC, LDL, HDL-C, FINS, FBG, and HOMA-IR of the patients in the two groups before and after treatment were evaluated. As illustrated in During the analysis of intestinal flora, the information in the Electroacupuncture group before and after treatment was named Z1 team and Z2 team, respectively, and the information in the Control group before and after treatment was called D1 team and D2 team, respectively. As depicted in In addition, according to the PLS-DA analysis based on OTU determination , the samP < 0.05) and the Enterobacter level was increased in Z2 group, when compared with the Z1 group (P < 0.05), while the Blautia level was lessened (P < 0.05) and the Enterobacter level was augmented in D2 group, when compared with the D1 group (P < 0.05).The phylum-level and genus-level changes of the intestinal flora in patients before and after treatment were exhibited in The Alpha diversity of intestinal flora was evaluated. As shown in The species difference of intestinal flora in the Electroacupuncture group was analyzed. As profiled in More and more researches have proved that acupuncture can safely and effectively treat abdominal obesity, mainly by stimulating the body’s meridians and acupoints, improving body condition as a whole, regulating intake, and enhancing body metabolism . AcupuncIn addition to the physical indexes, there are also a series of serum indexes which are closely related to the glycolipid metabolism of obesity patients. TG and TC are two important factors in the glycolipid metabolism, which both display higher levels in obesity patients than in healthy people . FurtherAs one of the complex ecosystems in the gastrointestinal tract, the intestinal flora is related to metabolism-related diseases and multi-system diseases . AcupuncThe high-throughput sequencing technology has been widely used in microbiome analysis, because it allows scholars to sequence millions of DNA molecules simultaneously and provides a data pool to cover the entire microbiome in the gut . On the Taken together, electroacupuncture combined with diet treatment exerted a therapeutic effect on perimenopausal patients with abdominal obesity through improving the community structure of intestinal flora. Nevertheless, what failed to be underscored in our study is the design of separate dietary intervention group, that is, the same diet restriction was performed without electroacupuncture, which may be further explored in future studies. Consumption patterns may affect obesity, which may be another aspect that we need to pay attention to in future research.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The study has been approved by the Ethics Committee of Zhejiang Hospital [No. 2018 (9K)] and all patients signed the written informed consent.JS and GY: substantial contributions to conception and design and drafting the article or critically revising it for important intellectual content. JZ, CS, YH, ZL, TL, and XJ: data acquisition, data analysis and interpretation. JS, GY, JZ, CS, Y’aH, ZL, TL, and XJ: final approval of the version to be published and agreement to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of the work are appropriately investigated and resolved. All authors contributed to the article and approved the submitted version.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.


Pre-treatment biomarkers to estimate overall survival (OS) for malignant pleural effusion (MPE) are unidentified, especially those in pleural fluid. We evaluated the relationship between OS and total protein–chloride ratio in malignant pleural effusion (PE TPClR).A retrospective study was undertaken to identify patients from 2006 to 2018 who had pathologically or cytologically confirmed MPE and received no tumor-targeted therapy. We recorded the pre-treatment clinicopathologic characteristics and follow-up status. OS was estimated by the Kaplan–Meier method, and the association between variables and OS was evaluated by Cox proportional hazards models.P < 0.001). The multivariate analysis between OS and the variables selected from the univariate analysis showed that the levels of neutrophil, alkaline phosphatase, neuron-specific enolase, platelets, albumin in peripheral blood, and white blood cells in pleural effusion were also independent predictors of OS.We screened 214 patients who met the eligibility criteria. The optimal cutoff value for the PE TPClR was set at 0.53. The univariate analysis showed that there was a significant correlation between PE TPClR and OS (In patients with MPE, pre-treatment PE TPClR independently predicts OS. Although further research is necessary to generalize our results, this information will help clinicians and patients to determine the most appropriate treatment for MPE patients. Malignant pleural effusion (MPE) is the second leading cause (only after parapneumonic effusion) of exudative pleural effusion, with more than 125,000 patients hospitalized annually in the United States and with an estimated hospitalization cost of over $5 billion each year , and theSystemic and local inflammatory states have been considered to be associated with outcomes of cancer, and the pattern of inflammatory cell infiltration seems to affect survival , 7. ReceIt is well known that exudate is associated with inflammation or malignant processes resulting in increased capillary permeability. It is formed by active secretions or leakage and contains a high level of protein. As a result, the level of protein in pleural effusion has been studied as a simple marker for local inflammatory response , 14. ChlIn our study, we comprehensively evaluated the prognostic value of clinical and laboratory characteristics in MPE patients before the tumor targeted therapy was taken, especially the markers in pleural effusion, determined the median survival time, and evaluated the prognostic variables associated with OS in MPE patients.A retrospective analysis of a prospectively maintained database was carried out on identifying all patients who were diagnosed according to cancer cells in pleural effusion or pleural biopsy between June 2006 and April 2018 at Beijing Chao-Yang Hospital. In order to avoid the influence of immune-related issues and treatments on various indicators, patients with significant infection, autoimmune disease, or underlying hematological diseases, without pre-treatment complete blood or pleural count values, and who had received anticoagulant or tumor-targeted therapy were excluded. A total of 214 patients met the criteria for admission and were incorporated into our study. The research protocol was approved by the Institutional Review Board of Beijing Chao-Yang Hospital.The demographic data of the patients and their clinicopathological characteristics, including laboratory variables of peripheral blood and pleural effusion, histologic type, and extrapleural metastasis, were retrospectively collected from the database collection of our institution. The patients were routinely followed up every 6 months during the first year after diagnosis. Thereafter, the follow-up should be performed annually and kept for at least 1 year. The follow-up was maintained through personal contact with patients by phone, including asking for information about tumor recurrence and survival status. OS, defined as the interval between the diagnostic date and the date of death or last follow-up, was recorded.t-test or one-way analysis, and categorical data were analyzed by using χ2 test. Survival curves were analyzed by the Kaplan–Meier method, and differences were compared using the log-rank test. The relationship between the PE TPClR and survival was assessed by univariable and multivariable Cox regression models. All statistically significant univariates were included in the multivariable model. Univariate and multivariate Cox regression models provided HR and 95% CI, respectively. A Cox proportional hazards model was used to fit all individual prognostic variables to determine their independent factors. SPSS 23.0 software was used for statistical analysis. The sample power of the study was calculated by PASS 11.0.The PE TPClR values were obtained by dividing the level of total protein by the level of chloride in pleural effusion obtained during diagnostic thoracentesis or thoracoscopy. The optimal cutoff value for age was determined by the median, and those of the other variables were determined by maximally selected rank statistics , which wThe study cohort was comprised of 214 patients, and the sample power of our study reached up to 86.3%, in accordance with the two-sided log-rank test. The significance level was 0.05. The baseline characteristics of the population in this study are shown in P < 0.001), the levels of albumin , calcium (P = 0.001), PE adenosine deaminase (P = 0.002), and PE LDH (P = 0.024) exhibited significant differences between the two groups.According to the best cutoff value of PE TPClR, all patients were classified into low group or high group. The clinical and laboratory characteristics related to the two groups are shown in - in pleural effusion, and LMR were significantly related to worse outcomes .The univariate associations between clinicopathologic factors and OS are shown in P < 0.001). The median survival time was significantly higher for patients with PE TPClR ≤0.53 than for those with TPClR >0.53 , as shown in P < 0.001), platelet (P = 0.004), ALB (P = 0.024), ALP (P = 0.005), NSE (P < 0.001), and PE WBC (P = 0.001) were also independently correlated with OS. Analyses including pre-treatment WBC, NLR, LMR, and CAR in the multivariate model are shown above, but they were not significantly related to OS .Finally, we conducted a multivariate Cox analysis to determine whether PE TPClR independently predicted OS or not Table 3.In our study, we have extensively screened out commonly used clinical laboratory indexes, which included MPE from a variety of causes that are not limited to lung cancer, and found that the PE TPClR is an important prognostic factor for OS in MPE patients. The PE TPClR is effective, safety, easy to calculate, inexpensive, and generally applicable in clinical settings from the different indicators in MPE . TherefoInflammation, which is reported to play a significant role in different stages of oncogenesis, is now considered as a marker of cancer . As we hIn our study, a higher level of total protein in pleural effusion was related to a shorter OS in the univariate analysis. It is different from some previous studies on the relationship between the total protein in pleural fluid and survival time. Abrao et al. showed that, when the pleural fluid total protein value was <3.6 g/dl, the median survival was 74 days, which was statistically significant in the multivariate analysis . They be- level in pleural fluid was an independent indicator for prognosis in MPE patients. Few studies have found that chloride intracellular channels 3 and 4, controlling the intracellular distribution of Cl- to provide ionic counterbalance, are over-expressed in human mesothelioma .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.


We found an ever-increasing relationship between LTL quartiles and survival. Hazard ratio analysis showed that for each unit increase in LTL and Short Physical Performance Battery (SPPB) scores, the mortality risk was reduced by 22.41% and 8.78%, respectively. Conversely, male gender, Charlson Comorbidity Index, and age threatened survival, with mortality risk growing by 74.99%, 16.57% and 8.5%, respectively. Determinants of LTL elongation were SPPB scores  and years of education , while male gender  and increased Disease Count Index  were determinants of LTL attrition. Longer LTL predicts a significant survival advantage in elderly people. By identifying determinants of LTL elongation, we provided additional knowledge that could offer a potential translation into prevention strategies.Leukocyte telomere length (LTL) represents a key integrating component of the cumulative effects of environmental, lifestyle, and genetic factors. A question, however, remains on whether LTL can be considered predictive for a longer and healthier life. Within the elderly prospective TRELONG cohort ( The aging of the population, also named the “gray” tsunami, is a growing social and public health issue. The impact of the phenomenon is forecasted by worrying data in Italy, one of the most elderly nations in the world . In partAging is a peculiar and manifold process characterized by a gradual deterioration of the body’s capability to react to internal and/or external stress factors, resulting in an increased risk of illness and death . This deAccumulating evidence has revealed that leukocyte telomere length (LTL) reduction can be regarded as the early pillar of biological aging and may be the origin of cellular dysfunction . TelomerHowever, one of the most discussed questions in gerontology research is whether LTL is prognostic for a long and healthy life. The problem in interpreting the results of several studies arises from the heterogeneity of the factors that affect LTL, including genetics and environment, and in particular the availability of follow-up data both of LTL and survival of enrolled subjects.The Treviso Longeva (TRELONG) study is a longitudinal cohort study initiated in 2003, which encompassed 668 seniors, women and men, over the age of 70 years, living in Treviso, the town with the highest longevity in North-East Italy ,18,19,20In view of the above, we investigated LTL within the prospective TRELONG cohort with two main aims: to ascertain whether this biomarker is predictive of longevity and to identify the main determinants in LTL among the personal characteristics , physiological and lifestyle features, physical performance and frailty measures, chronic diseases, biochemical measurements and apolipoprotein E (APOE) genotyping.A complete description of the TRELONG study has been previously reported . EligiblGender, age at the time of baseline in 2003, marital status, education in years, institutionalization, Body Mass Index (BMI), hypovisus, hearing loss, smoking status and alcohol intake were detected through home interview. To assess physical activity, participants were asked whether they took walks and/or did gardening every day. Data on eating habits were collected in 2003 (baseline) by questioning the participants about the regular intake (yes/no) of a list of foods. The statistical analyses showed that answers were substantially the same for the whole sample, suggesting all participants had the same eating habits .The Short Physical Performance Battery (SPPB) was used to measure frailty via standing balance, walking and chair stand tests . StandinPhysical strength was also measured via handgrip using a Collins dynamometer , lifting a 2 kg weight over the head and taking the average of three assessments with the stronger hand . PhysicaIn-person interviews and examination of medical records provided information on the history of chronic diseases such as cerebrovascular diseases, chronic renal insufficiency, tumors and others . Using tThe presence of cognitive decline was measured globally using the Mini-Mental State Examination (MMSE), range 0/30 . Raw scoA fasted morning blood was collected at home from each participant and transported to the Clinical Chemistry Laboratory of Treviso Hospital, within 30 min before 9.00 a.m. (12h fasting) as previously described . BrieflySerum biochemical markers, such as albumin, total and high-density lipoprotein (HDL)-Cholesterol, creatinine, etc., were determined by commercial methods  on an Aeroseth Analyzer. Hematologic indices were assessed with the ADVIA 120 Hematologic System . Coagulation tests were performed using a plasma-citrate CA-7000  and erythrocyte sedimentation rate (ESR) using an automatic analyzer Test 1 TH . Immunoglobulins and high sensitivity C-reactive protein (hsCRP) in serum were measured with Behring Nephelometer Analyzer II (BNA) . Plasma interleukin-6 (IL-6)  was quanFasting peripheral blood samples (30 mL) were collected by venipuncture; one aliquot was used to separate mononuclear cells (PBMC) by a standard Ficoll centrifugation procedure as previously described ,19. PBMCAPOE genotype was determined by PCR using the following primers: for 5_-tcggccgcagggcgctgatgg-3_; rev 5_-ctcgcgggcccccggccccggcctggta-3_; the resulting 332-bp PCR product was digested with CfoI  as previously described . BrieflyLTL in genomic DNA was appraised by the real-time quantitative polymerase chain reaction (PCR) as previously described ,27 accorThe distribution of LTL at the first wave (2003) according to characteristics of the study population was appraised with Mann–Whitney U Test and Kruskal–Wallis test.n = 162 subjects with available three telomere measurements , we used the Friedman test to compare the longitudinal distribution of the LTL at the three waves.Considering the subgroup of Since LTL could be measured more than once on the same person (repeated measures taken over time) to account for both within-person and across-person variability, we used the linear mixed model to appraise the influence of chronological age and gender (independent variables) on the logarithmically-transformed LTL (dependent variable), since the telomere distribution was non-Gaussian. The addition of the random effects for each subject was useful to take into account the individual heterogeneity and the dependence between the measurements made on the same subject.Survival analysis was performed using the non-parametric Kaplan–Meier curves analysis on all 613 subjects. The survival time was calculated for each person starting from the date of the interview of the year corresponding to the first LTL measurement available until the date of death or end of follow-up (25 June 2019). The survival curves were estimated stratifying the subjects in two groups based on the median LTL, or in four groups based on the quartiles . The difference between survival curves was estimated by the log-rank test and the Peto–Peto log-rank test, which is preferable when there are intersections among the survival curves.p = 0.92) for each covariate and for the model as a whole; therefore, the hypothesis of proportionality of the risks was accepted. Based on the relevant literature, gender, age, CCI and SPPB were used as covariates besides LTL in the Cox regression model. For the interpretation of the estimates, the hazard ratios of each variable were reported in the tables. Since LTL varied over time and there was more than one measurement for some subjects over the three waves, we used an extension of the Cox model that allows for the inclusion of time-dependent variables by the technique of episode splitting.To evaluate the effect of LTL on survival adjusting for other factors, a semi-parametric Cox proportional hazards model was constructed using the same sample of 613 subjects. Before, the adequacy of the proportionality assumption of risks was assessed by analyzing the Schoenfeld residues. The analysis of the Schoenfeld residuals gave a non-significant result  and the second (2010) waves or between the second (2010) and the third (2013) waves and 0 in absence of elongation. This was the outcome variable of a logistic regression model where the predictors were the variables reported in p-Value < 0.05 was obtained. The analyses were performed using the statistical software R version 3.6.3 [Results were considered significant when a on 3.6.3 .p < 0.05) with age, institutionalization, hearing loss, moderate and severe physical frailty and presence of chronic diseases (DCI and CCI). LTL increased (p < 0.05) with increasing physical activity, better cognitive profile (MMSE score > 26), greater functional autonomy (ADL score = 6 and IADL score = 8), better physical performance (SPPB score ≥ 6 and effective placement of one’s hands behind the shoulders) and physical strength (hand gripscore ≥ 12.2 kg and full lifting of the weight of 2 kg above head). Furthermore, LTL increased with blood chemistry variables such as albumin, hemoglobin and insulin-like growth factor-1 (IGF-1).The distribution of LTL (2003) according to the characteristics of the study population is reported in p < 0.001).p = 0.594). The estimated model indicates that for each additional year of age, LTL dropped by 7.96% (p < 0.001). The average trend estimated by the model on the original scale is shown in p = 0.064).p < 0.001) and the Peto–Peto log-rank test (p < 0.001), which was better because a slight intersection between the two survival curves was observed in the first period (see riod see .p <0.001). The median survival time was over 4 years longer in the group with the longest telomeres (in purple in p < 0.001). The coefficient associated with LTL and SPPB had a negative sign, indicating a protective effect of these variables with respect to mortality risk, while the sign was positive for other variables , CCI), indicating an increase in risk. The interpretation of the hazard ratios for each variable can be the following: (1) age, for each additional year of the subject, there is an increase of 7.75% in the risk of mortality, all other factors being equal; (2) LTL, for each unit increase in the LTL, the risk of mortality decreases by 16.99% with the other explanatory variables being equal, age included; (3) Charlson index score, for each unit increase in the index, the risk of mortality increases by 9.65%, all other factors being equal; (4) SPPB index score, for each unit increase of the index, the mortality risk, all other factors being equal, is reduced by 10.01%; (5) gender, males have a 73.16% higher risk of mortality, all other factors being equal, compared to females.Out of 613 subjects, overall 73 subjects (11.9%) presented an increase in LTL: they were 38 in the first period (2003–2010) and 35 in the second period (2010–2013). No subject registered an increase during the whole observation period (2003–2013).p = 0.0143) along with increase in the DCI values . By contrast, SPPB scores  and years of education  were determinant of telomere elongation.A key finding from our study is the steadily increasing relationship between LTL and survival: the longer the LTL, the greater the survival and, therefore, a lower mortality risk is estimated. In particular, as shown in LTL has often been related to life expectancy, but contradictory results have been reported in older adults. Whereas some studies have reported that LTL attrition is a marker of mortality ,31,32,33Key survival determinants were not only LTL but also SPPB: for each unit increase in the index, the mortality risk was reduced by 8.78%, considering all other factors being equal, age included . It is kOn the other hand, the male gender, CCI and age were factors that threatened the survival, with a risk of mortality growing by 74.99% for males, by 16.57% per unit increase in CCI and by 8.5% for each additional year of age . These dLTL distribution at the baseline in 2003 showed that LTL attrition was related to age . AnotherAnother noteworthy key finding of our study was the paradoxical increase in LTL in almost 12% of the study population. This finding, with almost exactly the same percentage, has been previously reported . Loss ofAmong determinants eliciting LTL elongation were SPPB and years of education, while being male and DCI were factors that contributed to reduced LTL. Until now, few studies have explored the association between LTL and SPPB, with conflicting results. Pereira et al. , in a stThe positive relationship between LTL and years of education agrees with previous findings on a positive association between educational attainment and LTL ,58. The Instead, the role of the male sex as a negative factor for telomere elongation is consistent with the majority of previous cross-sectional and longitudinal studies establishing that women have longer telomeres than men  and thatConvincing evidence in the literature suggests the association between shorter LTL and several age-related diseases  ,64,65. TWhether LTL can be considered a predictor for longevity and healthy life is one of the most questioned issues in the field of gerontological research. Problems in results’ interpretation stem from the heterogeneity of factors influencing LTL, including genetics and environment, and in particular from the available follow-up data for both LTL and survival for recruited samples.Our study was conducted in Treviso, in Veneto, a region of North-East Italy, which is characterized by the highest longevity, in particular among women . PrevaleIn our study, we determined LTL during ten-year follow up in the prospective cohort “Treviso Longeva (TRELONG)”, a very well-characterized study population. In particular, this is a systematic sample of subjects aged 70–106 years, drawn from the list of residents of the municipality of Treviso. Therefore, the study sample is highly representative of the community and of the real world . FurtherWe used the q-RT PCR technique to assess LTL in triplicate. This fast and highly sensitive method requires relatively small quantities of DNA and allows high throughput. Since an almost totally automated workflow from DNA extraction to LTL analysis was used, pre-analytical factors and analytical factors did not vary amongst analyses and the same pool DNA was used as a reference standard in all q-RT PCR reactions. In this way, the interbach variation was under control, minimizing measurement error and warranting a CV of less than 9%.Moreover, in this study, the start date of the survey was the day of the interview rather than the date of birth so that the measurements relating to the lengths of the telomeres of the interviewees who had very different ages were comparable. In this case, then, the duration of the episode indicates the remaining survival time for the individual with that LTL.Another strong point is that the Cox model estimate for time-dependent variables was further improved considering that for all 613 subjects a statistical weight was computed so that the TRELONG study population was representative per gender and age of the Italian population over seventy. Therefore, the results could be more easily extended to the entire population over seventy at the national level. It should be noted, however, that while the sample of the present study was found in a single city, a sample detected at the national level would be affected by the different geographical distribution of the subjects recruited.Our results demonstrate that LTL could be a powerful tool for predicting survival in our elderly population. By identifying SPPB as a key survival determinant, our study suggests that the regular application of the SPPB in clinical practice may offer helpful prognostic information regarding the risk of all-cause mortality. This study also confirms that male gender, CCI, and, therefore chronological age, are threats for survival.Furthermore, by identifying SPPB scores and years of education as positive and DCI as a negative determinant of LTL elongation, we recognized factors on which to converge to improve prevention strategies.


To evaluate the clinical effectiveness of a novel CellDetect staining technique, compared with fluorescent in situ hybridization (FISH), and urine cytology, in the diagnosis of urothelial carcinoma (UC).A total of 264 patients with suspicious UC were enrolled in this study. All tissue specimens were collected by biopsy or surgery. Urine specimen was obtained for examinations prior to the surgical procedure. CellDetect staining was carried out with CellDetect kit, and FISH was performed with UroVysion detection kit, according to the manufacturer’s instructions. For urine cytology, all specimens were centrifuged using the cytospin method, and the slides were stained by standard Papanicolaou stain.In this study, there were 128 cases of UC and 136 cases of non-UC, with no significant difference in gender and age between the two groups. Results for sensitivity of CellDetect, FISH, and urine cytology were 82.8%, 83.6%, and 39.8%, respectively. The specificity of the three techniques were 88.2%, 90.4%, and 86.0%, respectively. The sensitivity of CellDetect and FISH are significantly superior compared to the conventional urine cytology; however, there was no significant difference in specificity among three staining techniques. In addition, the sensitivity of CellDetect in lower urinary tract UC, upper urinary tract UC, non-muscle-invasive bladder cancer (NMIBC), and muscle-invasive bladder cancer (MIBC) were 83.3%, 81.8%, 83.5%, and 72.0%, respectively. The screening ability of CellDetect has no correlation with tumor location and the tumor stage. The sensitivity of CellDetect in low-grade UC and high-grade UC were 51.6 and 92.8%. Thus, screening ability of CellDetect in high-grade UC is significantly superior compared to that in low-grade UC.CellDetect and FISH show equal value in diagnosing UC, both are superior to conventional urine cytology. Compared to FISH, CellDetect is cost effective, easy to operate, with extensive clinical application value to monitor recurrence of UC, and to screen indetectable UC. Urothelial carcinoma (UC) is the second most common urologic malignancy after prostate cancer, which accounts for approximately 90% of all bladder cancer . Upper uUrine cytology is the most common noninvasive examination for UC detection, however, a large scale study provides further evidence that cytology has low sensitivity for UC detection . In receCellDetect is a novel cell staining technique by Zetiq Technologies Ltd (Israel) for cancer screening and diagnosis . CellDetIn this study, we compared the clinical effectiveness of CellDetect, FISH, and urine cytology in the diagnosis of UC. The results showed that CellDetect provided a unique and useful tinctorial clue for the detection of UC. CellDetect used in urine exfoliated cell screen provides an effective technique for early diagnosis of human UC.A total of 264 patients with suspicious UC were enrolled in the Department of Urology of Beijing Friendship Hospital from January 2020 to March 2021. All subjects had hematuresis, irritative bladder symptom, abdominal pain, and hydronephrosis on the affected side. No patient received neoadjuvant chemotherapy. All tissue specimens were diagnosed through biopsy or surgery. Histological cell type of the resected UC samples was determined by two experienced pathologists, tumor stage was evaluated according to the Union for International Cancer Control (UICC) 2017 TNM classification system, and histological grade was also assessed according to the World Health Organization (WHO) 2004 grading system for UC. Prior to the surgical procedure, urine was obtained for CellDetect, FISH, and urine cytology. On the onset of the study, study protocol approved and provided by the Research Ethics Committee of Beijing Friendship Hospital and informed consent were provided by all patients.Cell staining was carried out with CellDetect kit , according to the manufacturer’s instruction. Urine sample (minimum of 50 ml) was collected from each patient and smeared onto the slide. After mixing with 10% of trichloroacetic acid, nuclease was stained first with hematoxylin, followed by a differentiation with hydrogen chloride/ethanol. Followed by staining with the red and the green dye in the kit. In between staining, conditioning was performed using the plant extract, and further differentiation was carried out as mentioned above. Normal, inflammatory, and malignant cells can easily be differentiated. Malignant cells show a red nucleus and pink cytoplasm; normal urothelial or squamous epithelial cells show a dark purple or green nucleus and green cytoplasm; inflammatory cells show a purple nucleus and red cytoplasm, and can be distinguished morphologically. Each slide was accessed under a microscope by two cytologists (a third cytologist was on standby for any discrepancies), and assigned two categories according to the best practice of pathology department: negative, and positive .®5000 processor , one slide was used for FISH, and the other was used for urine cytology stained using Papanicolaou staining. FISH was performed with UroVysion detection kit  according to the manufacturer’s instructions. Positivity of the FISH test was determined by the criteria as listed in the package after all cells were evaluated. Abnormal cells of ≥ 4 had chromosomal gain of at least two of the chromosomes 3, 7, or 17; with ≥ 10 tetraploid cells with normal morphology; or ≥ 12 cells with homozygous loss of 9p21 locus were considered positive. All cytology specimens were centrifuged and obtained by cytospin method, and slides were stained by standard Papanicolaou method. For diagnosis of negative or positive UC, refer to CellDetect staining.A total of 200 ml urine was collected for FISH and cytology examination. After centrifugation for five minutes at 2000 rpm, two slides were prepared with the ThinPrepp value of ≤ 0.05 was considered significant.Data were collected and compiled using SPSS 16.0. All results were expressed as mean ± standard deviation. Statistical significance was determined by Student’s t-test and Chi-squared test for comparisons among CellDetect, FISH, and urine cytology examination. A A total of 264 patients, 190 male (72%) and 74 female (28%) with suspicious UC; received CellDetect, FISH, and urine cytology examination. All participants received cystoscopy or ureteroscopy, and pathological results were derived from biopsy, TUR, radical cystectomy, and nephroureterectomy. Baseline characteristics of the study population are listed in Table Compared to the conventional urine cytology, CellDetect showed superior features: 1) nuclear/cytoplasmic ratio is maintained; (2) nuclear irregularity is clearly seen; (3) hyperchromic nucleus is evident. The unique plant extract adds color and valuable feature that can easily and accurately detect the suspicious areas. The urine smears stained by CellDetect were shown in Fig.  nuclear/We evaluated the diagnostic value of CellDetect, FISH and cytology in UC Table . The totp = 0.975). Tumor staging detection on NMIBC, and MIBC by CellDetect were 86/103 (83.5%), and 18/25 (72.0%), respectively (p = 0.301). It demonstrated that the screening ability of CellDetect has no correlation with the tumor location and clinical stage. However, total sensitivity of CellDetect in low-grade UC and high-grade UC were 16/31 (51.6%), and 90/97 (92.8%), respectively (p < 0.001). Thus, we can conclude that the screening ability of CellDetect in high-grade UC is significantly superior than in low-grade UC.We further evaluated the diagnostic value of CellDetect in UC Table . The totCurrently, an estimate of 429,000 new cases of UC were diagnosed, with 165,000 deaths per year in the world . China aAlthough some noninvasive examinations have been applied to UC detection and screening, the recurrence of UC such as urine cytology and biomarkers, still show low sensitivity to UC diagnosis , 22. TheCellDetect staining is a unique platform for cancer diagnosis,the proprietary plant extract and dyes enable the color distinction between benign and malignant cells based on staining and morphology. CellDetect was able to spot carcinoma in situ (CIS) cases, even missed by cystoscopy. And for some cases of high-grade UC, the nucleus of the tumor cells shrinked while inflammatory cells enlarged, usually confuses pathologist. However, we found that the nuclei of high-grade UC tend to be smaller, lose their round shape and smooth nuclear membrane, which can be easily observed with CellDetect. A previous study indicated that 94% sensitivity and 89% specificity to detect UC using CellDetect, which had overall superior sensitivity compared to urine cytology . AnotherIn this study, we compared diagnostic value among CellDetect, FISH, and urine cytology in UC. Our results indicated that CellDetect and FISH have equal-level sensitivity in the diagnosis of UC, and both are significantly superior to conventional urine cytology. However, there was no significant difference in specificity between the three staining techniques. In addition, the sensitivity of CellDetect has no correlation with tumor location and clinical stage. However, the sensitivity of CellDetect in low-grade UC and high-grade UC were 51.6 and 92.8%, respectively, which suggests that the screening ability of CellDetect in high-grade UC is significantly superior to that in low-grade UC. In a previous study on the diagnostic value of FISH and cytology in UC, the sensitivity of FISH in low-grade UC and high-grade UC was 25 and 73%, respectively. The sensitivity of cytology were 36 and 75% , 27. TheDue to the high recurrence of bladder cancer after TUR, bladder infusion chemotherapy combined with regular cystoscopy is the conventional strategy in the management of lower urinary tract UC. However, cystoscopy has some recognized limitations, such as small or occult tumor lesions that are not easy to visualize and diagnose . The morIn conclusion, CellDetect and FISH show equal value in diagnosing UC, both are superior to that of the conventional urine cytology. CellDetect can obviously improve the ability in monitoring recurrence of UC, and it can be applied in the screening of indetectable UC of the upper urinary tract. CellDetect technique is also cost effective and easier to operate. Thus, it provides an extensive clinical application value in the diagnosis of UC and postoperative follow-up in the future.