Despite limited evidence, people with Parkinson's disease (PD) use cannabis for therapeutic purposes. Given barriers to performing randomized trials, exploring real‐world experiences with cannabis in PD is valuable.Investigate the frequency and magnitude of symptomatic effects reported with cannabis use in PD.An anonymous, 15‐question, web‐based survey was deployed on Fox Insight. Cannabis product types were defined (by relative tetrahydrocannabinol [THC] and cannabidiol [CBD] content) and respondents were asked to reference product labels. Questions focused on use patterns and subjective effects on 36 predefined symptoms (rated −2‐markedly worse to +2‐markedly better).1,881 people with PD responded . 73.0% of respondents reported medicinal use, though 30.8% did not inform their doctor. 86.7% knew their type of cannabis product: 54.6% took higher CBD, 30.2% higher THC, and 15.2% took similar amounts of THC and CBD products. Most common use was oral administration, once daily, for less than six months. Frequent improvements were reported for pain, anxiety, agitation, and sleep . Dry mouth, dizziness, and cognitive changes were common adverse effects . Higher THC users reported more frequent improvements in depression, anxiety, and tremor, and more frequent worsening in dry mouth and bradykinesia than other product types.Respondents with PD reported using more CBD products, via oral administration, with mild subjective benefits primarily for sleep, pain, and mood. Higher THC products may be higher risk/higher reward for PD‐related symptoms. Cannabis use increased by approximately 1000% between 2007 and 2016 among adults over the age of 65.There is a wide variety of THC and CBD cannabis products available to consumers in states with legalized cannabis. In addition, hemp products, which are made from a cannabis plant that is high in CBD and have <0.3% THC, are widely available. At present, choosing a specific cannabis product is mostly self‐guided for people with PD given a lack of evidence‐based guidelines. The ratio and individual dosages of cannabinoids, as well as route of administration, may impact symptomatic effects of cannabis products.From a pragmatic standpoint, surveying people with PD who have used cannabis could provide valuable information. Leveraging the reach of the Fox Insight survey platform through the Michael J. Fox Foundation, we aimed to better understand real‐world patterns of cannabis use among people with PD and to determine the frequency and magnitude of self‐reported symptomatic effects, both beneficial and adverse, with various types of cannabis products. With this knowledge, clinicians could more effectively counsel people with PD regarding potential effects of cannabis use and researchers could better design future clinical trials to more objectively study cannabis in PD.Fox Insight, sponsored by the Michael J. Fox Foundation, is an ongoing online clinical study establishing a large cohort of people with and without PD , collecting self‐reported clinical variables. Data collection, management, and validation in Fox Insight are described by Smolensky and colleagues.For this cross‐sectional observational study, all Fox Insight participants with PD received an email invitation to complete our survey . InclusiThe survey introduction provided lay language definitions of cannabis, marijuana, THC, CBD, and hemp to ensure that participants could best identify which type of cannabis they had personally used . Survey Our sampling strategy focused on collecting useful data on the four major types of cannabis products: (1) high CBD/low THC; (2) high THC/low CBD; (3) similar amounts of THC and CBD; and (4) hemp. Based on our clinical experience, we predicted that the proportion of respondents with PD using each cannabis type would vary, estimating about 35% would use hemp, 30% high CBD/low THC, 25% similar amounts of THC and CBD, and 10% high THC/low CBD. A sample size of 200 respondents per cannabis product type group would allow, under the condition of maximum variance, detection of proportion differences of 0.16 with 90% power for categorical variables and differences in means of 0.33 times the within group standard deviation with 90% power for continuous variables. Thus, estimating that 10% of users would be taking high THC/low CBD, at least 2000 respondents were required. Conservatively, we estimated that 20% of Fox Insight participants with PD use cannabis and about 60% of them would complete the survey. Thus, overall response rate would be 12%, requiring a potential respondent pool of at least 16,667 people with PD. At the time of survey deployment, there were approximately 28,000 people with PD registered with Fox Insight.Demographic and clinical characteristics of survey respondents were analyzed using descriptive statistics. Association tests for cannabis use characteristics by the four cannabis product type subgroups were performed using chi‐square, Fisher's exact, or ordinal correlation tests for categorical variables, and with ANOVA type, Kruskal Wallis, or correlation tests for continuous outcomes and scales. For individual survey items, frequencies and means, with 95% confidence intervals, of each response were tabulated for the overall cohort, then also for cannabis product type subgroups. For the symptomatic effect question, the frequency of each response was calculated from the Likert scale, only for those respondents who reported presence of that symptom prior to initiating cannabis use. Clinically meaningful frequencies of beneficial and adverse effects were defined as greater than 25% and 5%, respectively. To determine the magnitude of improvement or worsening for each symptom (from mild to marked), means were calculated for those respondents who reported any improvement  or any worsening (−1 or −2). These analyses were performed first for the overall cohort, then also by each of the four cannabis product type subgroups. These analyses were repeated for collapsed cannabis product type subgroups: (1) “higher THC” (high THC/low CBD); (2) “similar THC/CBD” (similar amounts of THC and CBD); and (3) “higher CBD” (included high CBD/low THC and hemp groups). To explore dose‐dependent responses within cannabis types, analyses were also performed by dosage, with high THC dose considered to be >50 mg and high CBD dose >200 mg.P = 0.05. Statistical analyses were performed using the SAS statistical software package .To ensure generalizability of results, comparisons of demographic and clinical characteristics between our respondent group and the broader Fox Insight population with PD who were invited but did not open the survey were performed. Comparisons were also made between the group who completed the entire survey and those who opened the survey but declined to agree to the Certificate of Confidentiality to evaluate for response biases. Statistical significance was set at χ2 = 17.6 [df = 3], P < 0.001) and less likely to identify as Native American for race . In comparison to respondents, the overall PD population of Fox Insight participants who did not complete the survey , was slightly older , less male , in later PD stages , and less likely to be college educated .The survey was deployed on the Fox Insight platform on 1/23/2020 and closed on 6/9/2020, with 1881 people completing the survey. Respondents were an average age of 66.5 ± 9.1 years old, were 58.5% male, and 97.9% white , with broad variability in usage patterns Table . Most re86.7% of respondents could report the specific cannabis product type they were taking . Approximately two‐thirds of respondents were able to report exact THC (65.1%) and CBD (68.8%) dosages from their product labels. For THC, 23% of respondents reported taking <5 mg, 20.1% 6–50 mg, and 3.8% more than 50 mg per day (18.1% endorsed using products with no THC). For CBD, 21.7% were taking <5 mg, 24.7% 6–50 mg, 5.3% 51–200 mg, 2.0% 201–600 mg, and 2.3% >600 mg (12.9% endorsed using products with no CBD).P < .0001). “Higher CBD” and “similar THC/CBD” product users were also less likely to smoke or vape, reporting non‐combustion methods of administration  at higher rates than combustion .Grouping respondents by cannabis product types, “higher CBD” and “similar THC/CBD” product users were more likely to report primary medicinal purpose of cannabis use (89.9%) than “higher THC” users , except for worsening of nausea, which was not significantly different from “higher CBD” product users (P = 0.45).There were significant groupwise differences in reported symptomatic effects Fig.  and 3, bP < 0.05). There were no significant differences in reported symptomatic improvements between the “higher THC” and “similar THC/CBD” groups except for depression , anxiety , and tremor , with “higher THC” more likely to improve than “similar THC/CBD”. Dry mouth  and bradykinesia  were more often reported to worsen with “higher THC” compared to “similar THC/CBD”. Nausea was more likely to worsen with “similar THC/CBD” than “higher THC” . Symptoms were more frequently reported to worsen in “similar THC/CBD” compared to “higher CBD” , except for freezing (P = 0.83), bradykinesia (P = 0.17), and increased appetite (P = 0.17).The “similar THC/CBD” group n = 247; 15.6%) generally reported symptomatic improvement and worsening at frequencies between those reported by the “higher THC” and “higher CBD” groups  and diarrhea (P = 0.002) at significantly higher frequencies than those taking <50 mg of THC . Regarding CBD dose, respondents taking >200 mg of CBD  reported improvements in balance (P = 0.01), dyskinesia (P = 0.007), cognition (P = 0.02), and constipation (P = 0.002) at significantly higher frequencies than those taking <200 mg . There were no significant differences in the frequency of reported worsening of any symptoms between the two THC dose groups (< or >50 mg) or between the two CBD dose groups (<or >200 mg).When comparing effects on symptoms by specific dosages of cannabinoids, respondents taking >50 mg of THC  reported improvements in constipation , anxiety (18.6%), sleep (16.9%), parkinsonism (13.9%), and depression . Over 80% of medicinal users reported using higher CBD products, whereas this pattern is reversed for recreational users, with over 60% reporting use of higher THC products. This finding mirrors that of a prior survey in multiple sclerosis.There could be a synergistic symptomatic effect between THC and CBD,These results should be appreciated in the context of this study's limitations, especially given the bias inherent to survey data. There is some evidence of response bias, as respondents were younger, in earlier PD stages, and more likely to be college educated in comparison to the overall Fox Insight population who did not open the survey. However, differences are statistically significant only due to the large number of survey non‐participants within Fox Insight , the effect sizes were small , and responses were qualitatively similar. Also, while we could compare available demographics and clinical features of our respondents to both non‐respondents and non‐finishers of the survey, there are additional characteristics that could have contributed to the decision to respond that are not captured by Fox Insight. For example, it is unclear how the past experiences with cannabis, good or bad, may have influenced their decision to complete the survey, as well as how to respond. However, it is noted that cannabis was rated as having “no effect” on many symptoms overall, tempering the concern that responses were biased in either direction. A further concern is that some responses are based on infrequent cannabis use, with 15.2% of respondents using cannabis less than monthly, and 21.5% using it for less than one month. While limited experience with cannabis may have affected respondents' survey responses, we included all responses to improve generalizability of results at this early stage of research.Despite these limitations, our analyses of the symptomatic impact of more specific cannabis product types  may serve as a guide to future randomized‐controlled trials and product selection based on the PD‐related symptom of interest. This position is bolstered by the fact that almost 87% of respondents reported the THC:CBD ratio in their product and more than two‐thirds knew exact dosages. For example, a clinical trial focused on improving sleep in PD may choose to test a product with similar amounts of THC and CBD, given similar frequency of symptomatic improvement to the higher THC group with lower rates of adverse effects overall, based on our data. However, it may be premature to utilize the results of this study to counsel individual patients with PD regarding specific aspects of cannabis use. Labeling of THC and CBD content in products may be inaccurate,In summary, people with PD report that cannabis subjectively improves some PD‐related symptoms, with higher THC products conferring more frequent benefits than higher CBD products. However, with greater reward there is also greater risk, as higher THC products were associated with more adverse effects. Cannabis product choice among people with PD may be more predicated on avoidance of unwanted side effects, especially for those self‐classifying as medicinal users. These survey results offer a broad overview of real‐world cannabis use patterns and experience among a large group of people living with PD and provide initial results regarding the differential symptomatic effects of higher THC versus higher CBD products. Next steps should include more rigorous, controlled studies, informed by the results herein, to more objectively study the effects of varying types of cannabis on PD symptoms, as well the impact of the different methods of ingestion and specific doses.(1) Research project: A. Conception, B. Organization, C. Execution; (2) Statistical Analysis: A. Design, B. Execution, C. Review and Critique; (3) Manuscript: A. Writing of the first draft, B. Review and Critique.SKH: 1A, 1B, 1C, 2C, 3A, 3B.CD: 1A, 1B, 1C, 2C, 3A, 3B.SS: 1A, 2A, 2B, 2C, 3A, 3B.YL: 1A, 1B, 1C, 2C, 3A, 3B.ML: 1A, 1B, 1C, 2C, 3A, 3B.Ethical Compliance Statement: This study was approved by the New England IRB , in accordance with all applicable national and/or international guidelines/laws.Funding Sources and Conflicts of Interest: This study was funded by the Michael J. Fox Foundation for Parkinson's Research (Grant # 18255). There were no conflicts of interest for any authors.Financial Disclosures for the Previous 12 Months: Dr. Holden has received grant funding from the Michael J. Fox Foundation for Parkinson's Research and a loan repayment award from the National Institute of Neurological Disorders and Stroke (L30 NS103315). Dr. Leehey has received grant funding from the Parkinson's Progression Markers Initiatives, Neuraly Inc., the Michael J. Fox Foundation, the NIH, the Colorado Department of Public Health and Environment, Biogen, US WorldMeds LLC, and Vtesse. All other authors have no disclosures to report.Supplementary Text A: Fox Insight Survey Email Invitations. Three Emails were sent to Fox Insight participants: (1) Initial invitation announcing the survey. (2) Second invitation announcing that the survey can now be completed on a mobile phone or tablet. (3) Third invitation announcing there was only three days left to complete the survey.Click here for additional data file.Supplementary Text B: Fox Insight Survey. (1) Survey Introduction. The Introduction to the survey provided definitions of cannabis, cannabis constituents, and types of cannabis products; and asked respondent to fill out the survey regarding the one type of cannabis product that they took the most. (2) Survey Questions. The 15 questions that were provided in electronic format.Click here for additional data file.


Physalis angulata is a renowned traditional Chinese medicine for the treatment of various conditions. Physalin is the major type of bioactive constituents conferring medicinal properties of P. angulata. Despite the medicinal importance, the pathways leading to physalin are largely unknown. In this study, we employed a transcriptomic approach to identify a Pa24ISO gene from P. angulata. Through heterologous expression in yeast, Pa24ISO was revealed to catalyze an isomerization reaction in converting 24-methylenecholesterol to 24-methyldesmosterol. Real-time PCR analysis showed that the abundance of Pa24ISO transcripts correlated with the accumulation pattern of physalin B in different tissues of P. angulata. A direct role of Pa24ISO in channeling of 24-methylenecholesterol for physalin B biosynthesis was illustrated by suppressing the gene in P. angulata via the VIGS approach. Down-regulation of Pa24ISO led to reduced levels of 24-methyldesmosterol and physalin B, accompanied with an increase of campesterol content in P. angulata. The results supported that 24ISO is involved in physalin biosynthesis in plants. Physalis angulata L, popularly known as Kuzhi in Chinese, has been prescribed for centuries for treatments of a variety of diseases, such as fever, malaria, liver disorders, and diabetes , catalyzed by sterol side chain reductase 1 (SSR1), or withanolide via 24-methyldesmosterol that is catalyzed by a sterol Δ24-isomerase  system for P. angulata. A direct role of Pa24ISO in physalin biosynthesis was discovered by silencing it in P. angulata through the VIGS approach. The Pa24ISO expression pattern also matched well with the accumulation of physalin B across different tissues.To investigate whether 24ISO is involved in physalin biosynthesis P. angulata were harvested from the Langxi county, Anhui province of China in August 2018. The identity of the plant material was confirmed by Dr. Xiaodong Li at the Wuhan Botanical Garden, Chinese Academy of Science. P. angulata plants were grown in a growth chamber at 22°C under long-day (16 h of light/8 h of darkness) conditions. Transgenic plants were grown at an open field, located at the campus of Shanghai University, China.Seeds of Ws24ISO (GenBank accession: AXG64144.1) as a query, a BLASTP analysis of our recently constructed P. angulata calyx-derived transcriptome (unpublished data) revealed the candidate Pa24ISO. Deduced amino acid sequences of Pa24ISO were aligned with the previously reported sequences of SSR1 and SSR2 (see their accession numbers in Arabidopsis DWF1 was used to root the tree.Using PDS (phytoene desaturase) gene, associated with the photobleaching phenotype, was used as a reporter gene to test the TRV-based VIGS system for P. angulata, The cDNA sequence of PaPDS, as shown in P. angulata calyx transcriptome, and a 382 bp-fragment of PaPDS was amplified from the P. angulata calyx cDNA. For silencing the Pa24ISO gene expression, a 297 bp-region, which is specific to Pa24ISO, but is not conserved in the putative PaSSR1 and PaSSR2, was selected , pTRV1, and pTRV2-PaPDS were separately transformed into the Agrobacterium GV3101. The transgenic Agrobacterium strains were inoculated into LB medium supplemented with kanamycin at 50 mg/ml and rifampicin at 50 mg/ml at 28°C, and used for infection when OD600 value of the Agrobacterium culture reaching 1.5. For infiltration of the P. angulata leaf, the TRV1-contained Agrobacterium culture was mixed at a 1:1 ratio with the cultures containing pTRV2-PaPDS, as well as the empty vector pTRV2 as a control. To improve the infection efficiency, 0.01% Silwet L-77 was added into the Agrobacterium mixer. Four to six-leaf-stage plants were selected for the infection. The infected plants were kept in darkness for 48 h, and then grown at an open field at the campus of the Shanghai University, China. The phenotype was then evaluated after 6–8 weeks. The same procedure was applied to generate the Pa24ISO-silenced plants.For down-regulating the expression of -ΔΔCt method.The Real-time PCR (qRT-PCR) was performed on Bio-Rad CFX96™ Real-Time PCR instrument  using TransStart Green qPCR SuperMix (Transgen). Primers used in this article are listed in P. angulata samples was extracted with 1.7 ml of 90% methanol by sonication for 30 min, and this procedure was repeated three times. To normalize variations between the different extractions, isofraxidin at a final concentration of 1.5 μg/ml was added as an internal standard. The solvent extracts were collected by centrifugation at 12,000 g for 20 min, and passed through a membrane filter (0.22 mm) prior to the Ultra Performance Liquid Chromatography–Tandem Mass Spectrometry (UPLC–MS) analysis.Every 1 g of powdered One microliter of the extracts was injected for the UPLC-MS analysis. LC-MS analysis was performed using a Q-Exactive Focus mass spectrometer, coupled with a VanquishTM UPLC system  and a HESI source (Thermo Fisher Scientific). The column  was used to separate the sample, the column temperature was 40°C, and the flow rate was 0.35 ml/min. The mobile phases contain 0.1% formic acid (solvent A) and acetonitrile (solvent B), and the solvent gradient is set as follows: 15% solution B (0–1.0 min), 20–25% solution B (2.0–13.0 min), 25–40% solution B (13.0–25.0 min), 40–90% solution B (25–26 min), 90% solution B (26.0–29.0 min), and 15% solution B (29.0–33.0 min). The MS detection was performed with a full MS mode. The parameters of the mass spectrometers were as follows: spray voltage, 3.2 kV; source capillary temperature, 320°C; sheath gas flow rate (nitrogen), 25 ml/min; Aux gas flow rate (nitrogen), 8 ml/min; Aux gas heater temperature, 30°C, Scan range 150.0–700.0 m/z.Pa24ISO in yeast, the full-length sequences of Pa24ISO were manually synthesized following the S. cerevisiae codon usage preference  at 90°C for 2 h. When the saponified sample was cooled down, it was extracted with 2 ml of hexane for 30 min, and then was repeatedly extracted two times with another 1 ml hexane. The pooled hexane extracts were then dried in a vacuum freeze drier, and derivatized in 50 μl of N,O-bis(trimethylsilyl)-trifluoroacetamide (BSTFA) at 60°C for at least 1 h for sterol analysis by GC–MS.2O and 500 μl of hexane were added, and extracted with shaking for 30 min. After centrifugation, the upper clear hexane extract was collected. The lower aqueous phase was then extracted twice with an additional 500 μl of hexane. The pooled hexane extracts were evaporated to dryness, and treated with 50 μl of the silylating reagent BSTFA [bis(trimethylsilyl)trifluoroacetamide]. The trimethylsilyl esters were dissolved in 80 μl of hexane, and a 1-μl sample was used for GC–MS analysis. For quantifying the sterol content, 40 μg of cholesterol  was added to each sample as an internal standard. GC–MS analysis was performed using a Shimadzu 2010 Plus GC machine coupled to a QP2020 Ultra mass spectrometer . Samples were separated using a RTX-5 MS column (30 m × 0.25 mm × 0.25 mm) with helium being used as a carrier gas at a flow rate of 1.0 ml/min. The injection temperature was 280°C. The GC oven temperature program was as follows: 90°C for 1 min, then increased by 30°C/min to 300°C and hold at 300°C for 15 min. The mass of sterol compounds was detected in a select ion-scan mode with electronic ionization (70 eV), and monitored at multiple ions of m/z 343, 382, and 472 for campesterol, m/z 341, 365, and 470 for 24-methylenecholesterol, m/z 365, 386, and 470 for 24-methyldesmosterol, and m/z 329, 368, and 458 for the internal standard cholesterol.Phytosterol was extracted as described previously  with smaP. angulata , hereafter refer to as Pa24ISO, was found to show the highest amino acid identity (96.0%) with Ws24ISO. A phylogenetic analysis shows that Pa24ISO closely clustered with Ws24ISO in the tree  system for P. angulata, as there have been no reports of VIGS in this plant. For this purpose, the tobacco rattle virus  was retrieved from the calyx transcriptome, which is available in our laboratory, and a 382 bp-fragment of PaPDS was selected as a target sequence. Six-to-eight week-old seedlings were infected with the Agrobacterium slurry containing the pTRV2-PaPDS vector or the empty pTRV2 vector as a control. At around 4 weeks post infiltration, a greenish–whitish variegated phenotype was observed in the upper leaves. With the growing of the infected seedlings, some of the newly emerging leaves exhibited a completely white phenotype  in the content of 24-methyldesmosterol, which was accompanied with increased accumulation of 24-methylenecholesterol and campesterol . Based on its mass spectra  from P. angulata. Heterologous expression of Pa24ISO in yeast suggested that Pa24ISO is able to convert 24-methylenecholesterol to 24-methyldesmosterol. Silencing of Pa24ISO in P. angulata by the VIGS approach revealed a direct role of Pa24ISO in biosynthesis of physalin, such as physalin B at this case.This study reports the isolation and functional analysis of a 24ISO-encoding gene  and the start-up funding grant to YZ from Shanghai University (N.13-G210-20-327).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.


Heavy metal concentrations  in children and adolescents were approximately half that of adults, while some organic substances  were high. BPA showed higher levels than in the US or Canada, whereas BPF and BPS showed lower detection rates in this cycle; however, as these are increasingly used as a substitute for BPA, further research is necessary. As environmental chemicals may affect childhood health and development, additional analyses should assess exposure sources and routes through continuous observations.The Korean National Environmental Health Survey (KoNEHS) program provides useful information on chemical exposure, serves as the basis for environmental health policies, and suggests appropriate measures to protect public health. Initiated on a three-year cycle in 2009, it reports the concentrations of major environmental chemicals among the representative Korean population. KoNEHS Cycle 3 introduced children and adolescents into the analysis, where the blood and urine samples of 6167 participants were measured for major metals, phthalates, phenolics, and other organic compounds. Lead, mercury, cadmium, metabolites of DEHP and DnBP, and 3-phenoxybenzoic acid levels of the Korean adult population tended to decrease compared to previous survey cycles but remained higher than those observed in the US or Canada. Both bisphenol A (BPA) and  Human biomonitoring is an important tool for assessing internal exposure to environmental chemicals. Its history extends back to the 1890s, when exposure levels among occupational workers were measured in the workplace ,3,4,5. IThe Korean National Environmental Health Survey (KoNEHS) has been conducted in three-year cycles since its implementation in 2009. Three cycles have thus been conducted up to 2017, providing baseline information on major chemical exposure profiles. Cycles 1 and 2 were focused on the adult population n = 12,789), measuring a total of 16 and 21 chemicals, respectively, including metals, bisphenol A (BPA), and metabolites of phthalates ,12. ,789, meaKoNEHS Cycle 3 2015–2017) extended the population coverage to include children and adolescents. Many studies have reported that environmental chemicals are related to delayed growth and development in children and adolescents, who are particularly vulnerable to environmental pollution and related chronic diseases ,14,15,16–2017 extThis study provides the updated exposure profiles for major environmental chemicals among the representative Korean population, thus helping to prioritize chemicals of concern that warrant further environmental health management efforts.To ensure representativeness of the adult population, the results of the 2010 Population and Housing Census (Statistics Korea) were used, and a stratified multi-stage sampling process was conducted. Through a household visit in the sampled survey district, participation was asked, and the survey was conducted with those who consented.These sampling and field survey methods were the same as in Cycles 1 and 2 ,12, the Considering the participant accessibility and ease of sample collection, children and adolescents were extracted in units of kindergartens, childcare facilities, and educational institutions. Using households as sampling units enabled comprehensive sampling across a range of populations; however, children and adolescents who did not attend childcare or educational institutions may be omitted. However, the proportion of that those who attending such institutions was higher than >90%, which would be representative of the same age groups in Koreans .Sampling institutions and sample sizes by age were selected based on the status of daycare centers and educational institutions published by the Ministry of Education and the Ministry of Health and Welfare The country was divided into five regions to be used as variables: Seoul, Gyeonggi/Incheon/Gangwon, Chungcheong, Honam/Jeju, and Yeongnam. In total, 127 schools and 56 kindergartens/childcare facilities were selected across all regions following the sampling process for the KorEHS-C .This survey was approved by the Institutional Review Board of the National Institute of Environmental Research (NIER), Korea (NIER-2015-BR-006-01) and carried out only for those providing prior consent.Through a field survey, 18 mL of blood and 60 mL of spot urine were collected from 6167 participants. We sent a message to each subjects notifying them of relevant information 2 days before the survey began. On the day of a field survey, we guided them to the location by mobile phone and finally requested their participation. For adults, samples were taken at the survey site. For preschoolers and children, urine samples were directly collected in a specimen cup at their homes on the day of survey. For adolescents, urine samples were similarly collected upon visiting a pre-negotiated hospital near the investigation. In adults, 26 substances in their blood and urine were analyzed , whereasQuestionnaire items were continuous with Cycles 1 and 2, focusing on demographic and socioeconomic characteristics, living environments, recent diet, and lifestyle to identify exposure factors and routes of environmental chemicals. A single household member representing preschoolers and elementary school students completed both the household and individual questionnaire surveys. Completed questionnaires were submitted to kindergartens, childcare facilities, and schools to be collected later. For middle and high school students, one household member representing a student completed the common household questionnaire, and individual questions were conducted face-to-face by a trained interviewer. For adults, survey questionnaires were completed solely through face-to-face interviews.The biological samples collected in the field survey were transported under cold conditions (2–6 °C), with temperatures checked using a temperature logger . The samples were aliquoted from the center within 24 h, and kept frozen at −20 °C until analysis. All processes were performed according to the “Biological sample management guidelines of the KoNEHS Cycle 3 ”.Of the 26 target chemicals selected , nine weThe target chemicals were analyzed according to the “Analysis manual of the KoNEHS Cycle 3” ,28, and Measured values were log-transformed and analyzed using the survey means procedure, after accounting for stratification variables and survey sample weight. Measurements below the LOD were assigned a value equal to LOD/√2 for the calculation of geometric means ,29. WhenThe study population consisted of preschoolers , children , adolescents , and adults , for a total of participants of 6167 . Within the adults, the proportion of older subjects > 50 years was high, especially higher number of females present among participants >50 years as well. Many more female adult contacts than young people or elderly male for academic or work reasons were made when visiting households for recruitment. Furthermore, the voluntary participation rate was higher for women than men. Children and adolescents were recruited through educational institutions; the gender ratios of the final participants were maintained by adjusting for age and gender when recruiting. −1) were approximately half those of adults  concentration of blood lead in adolescents  phthalate (MEOHP): 25.5 μg·L−1, 19.2 μg·L−1, and 9.24 μg·L−1; and Mono(2-ethyl-5-hydroxyhexyl) phthalate (MECPP): 45.3 μg·L−1, 44.5 μg·L−1, and 28.4 μg·L−1, respectively. Accordingly, phthalate metabolite concentrations in preschoolers were approximately 2–3 times higher than those of adults: MEHHP, 13.2 μg·L−1; MEOHP, 9.88 μg·L−1; MECPP, 23.2 μg·L−1. For MCNP, MCOP, and MCPP added in Cycle 3, the GM concentrations were similar in preschoolers, children, adolescents, and adults: MCNP: 0.491 μg·L−1, 0.533 μg·L−1, 0.452 μg·L−1, and 0.441 μg·L−1; MCOP: 1.62 μg·L−1, 2.24 μg·L−1, 1.71 μg·L−1, and 1.07 μg·L−1; and MCPP: 1.80 μg·L−1, 1.56 μg·L−1, 1.48 μg·L−1, and 1.13 μg·L−1, respectively  phthalate (MEHHP): 34.6 μg·Lectively . −1, 1.70 μg·L−1, 1.39 μg·L−1, and 1.18 μg·L−1 in preschoolers, children, adolescents, and adults, respectively . Notably, the 95th percentile of urinary methyl paraben in preschoolers was 3445 μg·L−1, ≤10 times higher than those of other age groups. The preschooler concentrations of propyl paraben showed a similar pattern, with a 95th percentile of 699 μg·L−1, ≤6 times greater than other age group populations. The GM concentrations of urinary ethyl paraben in preschoolers, children, and adolescents were 14.2 μg·L−1, 11.4 μg·L−1, 19.1 μg·L−1, respectively : 1.08 μg·L5 μg·L−1  for pres−1, 1.20 μg·L−1, 3.04 μg·L−1, and 5.59 μg·L−1 for preschoolers, children, adolescents, and adults, respectively (−1) was significantly higher than that of non-smokers (1.87 μg·L−1). The GM concentrations of urinary cotinine in adolescents were significantly higher than for preschoolers and children: 1.05 μg·Lectively . For adu−1, 0.163 μg·L−1, and 0.130 μg·L−1  were notably higher for preschoolers compared to children, adolescents, and adults: 0.495 μg·L−1, 0.209 μg·L−1, 0.263 μg·L−1, and 0.321 μg·L−1, respectively.The GM concentration of urinary 1-hydroxypyrene (1-OH-Pyr) in children, adolescents, and adults was 0.108 μg·L0 μg·L−1 . The conchoolers . The GM trans,trans-muconic acid  in preschoolers, children, adolescents, and adults were 82.2 μg·L−1, 91.2 μg·L−1, 80.4 μg·L−1, and 86.2 μg·L−1, respectively; whereas those of urinary N-Acetyl-S-(benzyl)-L-cysteine (BMA) were 10.6 μg·L−1, 7.39 μg·L−1, 5.66 μg·L−1, and 4.63 μg·L−1, respectively.The GM concentrations of urinary Comparisons were made between the results of KoNEHS Cycle 3 (2015–2017), the United States National Health and Nutrition Examination Survey , and the Canadian Health Measures Survey  and paraben (methyl paraben: 46.3 μg·L−1 and propyl paraben: 4.36 μg·L−1) GM concentrations were 3–4 times higher than in Canada . In children, paraben (methyl paraben: 28.9 μg·L−1 and propyl paraben: 1.83 μg·L−1) GM concentrations were 2–3 times higher than in Canada (methyl paraben: 7.5 μg·L−1 and propyl paraben: 0.96 μg·L−1). Methyl paraben GM concentrations were similar to that of children in the US (methyl paraben: 28.9 μg·L−1), but two times lower in adolescents (KoNEHS: 26.1 μg·L−1 and US: 40.5 μg·L−1) and adults (KoNEHS: 35.2 μg·L−1 and US: 52.2 μg·L−1). In preschoolers, BPA (2.41 μg·L−1 and 0.289 μg·L−1) concentrations were four times higher than in the US (0.057 μg·L−1 and 0.055 μg·L−1), where levels were highest in the Asian ethnic group compared to others [−1) GM concentrations were four times higher than in the US (0.395 μg·L−1).In children and adolescents, cadmium  were 2–3 times higher than in the US  and Canada (Hg: 0.72 μg·L−1 and Cd: 0.22 μg·L−1). Moreover, DEHP metabolites, and MnBP  were 2–3 times higher than in the US  and Canada .The DEHP metabolite and MnBP, 3-PBA GM concentrations were 2–3 times higher in all age than in the US and Canada. The concentrations are presented in detailed in p < 0.001). A previous study showed that it has been shown that alcohol consumption in adults accounted for the largest proportion of changes in lead concentrations, followed by smoking [Blood lead concentrations in adults were approximately twice those observed in adolescents, similar to the findings of other research ,32. Both smoking . Additio smoking . Blood l smoking . −1) showed higher blood mercury levels than those in urban (GM 2.76 μg·L−1) or rural areas . We confirmed similar results in previous studies. It has been demonstrated that very high blood mercury levels detected in Yeongnam region of Korea were attributable to a local culture of consuming shark meat [In adolescents, blood mercury concentrations were half those of adults. Overall mercury levels were positively associated with fish, egg, meat, and vegetable intake, similar to findings that fish are a potential cause of total mercury exposure among Swedish adolescents . Other sark meat . Analyziark meat . Similarark meat ,43. p = 0.004) and soybean  intake increased with age, with a further positive correlation revealed between GM cadmium concentrations and soybean intake . Monitoring cadmium exposure in the Iranian population indicated that cadmium contamination occurred in food groups, such as rice, cereals, legumes, and vegetables. Specifically, 75% of the consumed rice samples showed cadmium concentrations higher than the maximum cadmium limit (0.06 mg⸱kg−1) allowed by the Institute of Standards and Industrial Research of Iran (ISIRI) [−1) and tuna (mean: 0.022 μg·g−1) samples showed high cadmium concentrations (ISIRI: 0.05 mg⸱kg−1) [The concentration of urinary cadmium increased with age, similar to the findings from other studies ,45. The  (ISIRI) . In addimg⸱kg−1) ,53. TherThe concentrations of phthalate, BPA, and TCS in this analysis were higher in preschoolers and children. Children’s ratio of body area to body weight is higher than that of adults, so any correlated exposure to harmful substances through the skin is higher ,55.p < 0.05) than on a common diet. Several findings have confirmed that childhood phthalate exposure is associated with obesity [p < 0.001), and the correlated MEHHP and MEOHP GM concentrations were significantly high .Several studies have similarly shown that the younger the age, the higher the concentration of phthalates in one’s body . Estimat obesity ,60,61, w obesity . The youHigher BPA levels in younger ages have also been observed in several other studies ,63,64. Bp = 0.021) lead to higher concentrations of TCS in non-adults, whereas adult concentrations were significantly correlated to frequency of body wash use (p = 0.041).When investigating the association between TCS and the use of personal care products, the former was detected in >70% of children, with notable higher concentrations in the group using hand soap, increasing with the frequency of hand washing. Additionally, the use of toothpaste by children was also positively related to TCS, further explaining its high correlation with personal care products . In the −1, p < 0.001) and propyl  parabens were higher in women, highlighting their association with cosmetic and personal care product use. For both methyl and propyl paraben, males were higher in infancy, whereas females were far higher in adolescence and adulthood. Methyl paraben is one of the most frequently used preservatives in cosmetic products such as lipsticks, perfumes, and personal care products [In the case of methyl and propyl paraben (commonly used together), concentrations in preschoolers were relatively higher. Parabens are often used as preservatives in medicines, such as pills and liquid antipyretics , both ofproducts , thus poproducts . Ethyl pproducts . An analproducts . Therefop = 0.0006) than outdoors, and coffee intake was also found to have an effect (p = 0.007) [The GM concentrations of urinary 3-PBA in children was slightly higher than that of other age groups. Further, children < 9 years had greater 3-PBA concentrations when living in rural residential apartments compared to similar urban locations, indicating the effects of landscaping pesticides on nearby playgrounds and public areas . Various= 0.007) . Furtherp < 0.001), or when more time was spent exposed to secondhand smoke (p < 0.001). Adults also showed higher cotinine concentrations with family smokers present (p < 0.001), and secondhand smoke exposure (p = 0.001).Adolescent males showed significantly higher cotinine concentrations than females. As age increased, the number of male adolescents who started smoking increased while the number of female adolescents decreased . Accordip = 0.005), whereas increased frequencies of charcoal grilling was correlated with higher 1-OH-Phe concentrations in non-adults (p = 0.032). PAHs metabolites were detected in over 78% of urine samples from 522 children, aged 5–12 years of Valencia, Spain, and 2-NAP was detection rate in 100%. In addition to the consumption of legumes and packaged foods, the education level, demographic characteristics, socioeconomic characteristics were previously identified as the most relevant factors influencing PAHs exposure levels in children [p = 0.001, p < 0.001, p < 0.001, and p = 0.001, respectively). Elsewhere, studies have indicated that diet, smoking, and air pollution were major determinants of internal PAHs exposure [The GM concentrations of urinary 2-NAP and 2-OH-Flu in preschoolers were significantly higher than the other age ranges. When food was cooked using gas, 2-NAP concentrations were significantly higher in non-adults . According to the results of the National Health and Nutrition Survey in the US, the BMA concentrations in children were higher than in adults. This can be explained either by higher childhood levels of exposure to chemicals, such as ethylbenzene-styrene and toluene, or slower excretion rates than that of adults [The GM concentrations of urinary BMA in preschoolers were 10.6 μg·Lf adults . A previf adults . The KoNEHS is the largest biomonitoring survey capable of identifying environmental chemical exposure levels across a nationally representative population. The most current exposure profiles for major environmental chemicals among the Korean population, KoNEHS Cycle 3 (2015–2017), is summarized here. In Korean children and adolescents, concentrations of mercury and cadmium were higher than in other countries. Although these levels are not of great concern when compared with Human Biomonitoring Commission recommendations, continuous observations are required to ensure they are not deleteriously affecting children’s growth, development, or health. BPA and paraben levels were also high; further studies are needed due to the increased use of bisphenol substitutes  and parabens, and there were different trends according to gender and age. In addition, the levels of phthalate biomarkers measured in the KoNEHS were higher in children and adolescents than in adults. Accordingly, the results indicated the need for exposure reduction measures throughout basic lifestyle and daily life, while continuously monitoring children’s exposure in particular. Among Korean adults, lead, mercury, cadmium, metabolites of DEHP and DnBP, and 3-PBA levels were higher than those reported in the US or Canada; however, their concentrations have been decreasing with time. Urinary BPA and t,t-MA concentrations showed an increasing trend, both warranting further studies on their exposure pathways. For children and adolescents, the Cycle 3 survey represented the first national biomonitoring results. Although it was not possible to explain the exact mechanism of substances with a large concentration difference compared to foreign standards, it is believed that the effect will be due to differences in dietary habits and the resulting basic body shape and living environment. Additional research is needed on the effects of exposure to environmental chemical substances. Moving forward, if data are secured through continuous surveys, it will be possible to identify various exposure factors by comparing data over time and analyzing connections within the survey results. Furthermore, The Ministry of Environment of Korea has established a “concentration standard in the human body” and is conducting a detailed investigation by reviewing whether these are exceeded through the results of the KoNEHS. Findings from these efforts will be used to develop environmental health policies, and appropriate mitigation measures for protecting the health of the people.


Graphium  confucius Hu, Duan & Cotton, 2018 is a recently discovered, wide ranging, multivoltine swordtail butterfly in China and Vietnam. The present study reports the complete mitochondrial genome of this butterfly, which is the fifth mitochondrial genome record for subgenus Pazala Moore, 1888. The mitochondrial genome of G. (P.) confucius is circular and 15,212 bp in length, and consists of 37 genes, including 13 PCGs, 22 tRNAs, and two rRNAs. The Bayesian phylogenetic tree containing the focal species and 33 other Papilioninae members clusters G. (P.) confucius with other Pazala taxa inside tribe Leptocircini, which agrees with its taxonomic position. The findings of this study added data to the complex subgenus Pazala and are beneficial to future understanding and conservation planning of butterfly diversity. Graphium  confucius Hu, Duan & Cotton, 2018 is among several recently discovered species of the subgenus Pazala Moore, 1888 (Lepidoptera: Papilionidae), a complex Sino-Himalayan group of butterflies (Racheli and Cotton G. (P.) confucius occupies a wide distribution range from Southwest to Central China, as well as North Vietnam  confucius reported herein would add new data to facilitate future understanding and conservation planning for butterflies.Phylogenetic diversity has been increasingly accepted as a tool in conservation planning (Lu and Bullock 2 (25 mM), 2 μl dNTPs (2.5 mM each), 0.5 μl each of forward and reverse primers , 0.25 μl Taq DNA polymerase , and 1 μl genomic DNA. The PCR thermal profile consisted of an initially denaturation at 95 °C for 3 min; followed by 30 cycles of 94 °C denaturation for 60 s, 50 °C annealing for 60 s, and 72 °C extension for 90 s; then an external extension at 72 °C for 5 min. All PCR products were sequenced on an ABI 3730xl automatic sequencer . Resultant gene fragments were assembled using DNAStar (https://www.dnastar.com/) with G. (P.) parus (MT198821), G. (P.) mullah chungianus (MW549197), and G. (P.) eurous asakurae (MW549198) as the reference genomes  .The butterfly specimen used in this study was collected from Da Moyu , Kunming, Yunnan, China. The specimen was deposited in the Zoological Museum (insect collection) of Yunnan University, Kunming, China . Genomic DNA was extracted from the thoracic muscle tissue of a single male adult using the TianGen TIANamp Genome DNA Kit . The PCR amplification was performed using in a 25 μl system containing 2.5 μl 10× PCR buffer, 2 μL MgClG. (P.) confucius is circular and 15,212 bp in length (GenBank accession number: OK136253). The base composition is 39.84% for A, 39.28% for T, 8.13% for G, and 12.75% for C. This mitochondrial genome contains 37 genes, including 13 PCGs, 22 tRNAs, and two rRNAs, plus a non-coding control region. The plus (+) strand encodes nine PCGs , while the minus (−) strand encodes four PCGs .The gene arrangement and character of this genome fit those of ditrysian Lepidoptera mitochondrial genomes   confucius clusters with G. (P.) parus first and then with other Pazala species. All Pazala taxa are related to other Graphium species within Leptocircini, forming a monophyletic clade, supported by the maximal support values (To validate this mitochondrial genome, a Bayesian phylogenetic tree was reconstructed by PhyloSuite 1.2.2 (Zhang et al. t values .Click here for additional data file.


Scylla spp.)—from megalopa to several, further crablet stagesneeds to be carried out in optimal environmental conditions, until they reach a larger size suitable to being stocked to a pond. This study observed the behavioral thermoregulation of crablet instar phases, and determined the effects of several levels of water temperature on growth, survival, molting cycle, gill condition, and sex ratio in the nursery phase of S. paramamosain. In this study, optimal temperature for the long-term nursery phase of S. paramamosain, beginning with megalopa stage, was found to be 28 to 30 °C, with the possibility that water temperature could affect the sex ratio of mud crabs. Findings of this study could result in increased crablet production from hatcheries, and assist in realizing potential of monosex seed production of S. paramamosain, through temperature treatment.The nursery phase of mud crab , critical thermal minimum (17.33 ± SD 0.58 °C), critical thermal maximum (40 ± SD 0.00 °C), and thermal tolerance interval (22.67 ± SD 0.58 °C), were checked for Crablet 1 stage only (with ambient temperature as acclimation temperature).Both low (24 °C) and high (32 °C) temperatures were associated with lower growth performance, and survival rate (p < 0.05), in comparison with both 28 °C and ambient temperature treatments.Male ratio at low temperaturetreatment (24 °C) was higher (80.09 ± SD 18.86%) than for other treatments (p < 0.05), observed as 44.81 ± D 10.50%, 41.94 ± SD 19.44%, and 76.30 ± SD 5.13% for 28 °C, 32 °C and ambient temperature treatments, respectively. However, there was no significant difference observed between 24 °C, 28 °C, and ambient temperature treatments. Anatomical alterations of gill lamellae of S. paramamosain crablet for both 32 °C, and 24 °C treatments, appeared thinner and paler than at both 28 °C, and ambient temperature treatments. Based on this study, temperature of 28 to 30 °C was recommended as the optimal temperature for the long-term nursery phase of S. paramamosain.This study was carried out to determine the physiological changes  of mud crab,  Scylla paramamosain is one of four species of mud crabs from the genus Scylla, based on revision of Keenan et al. [S. serrata has the widest distribution area in comparison with other species [n et al. . It is an et al. ,3, and mn et al. . Each sp species . Tempera species ,7,8,9,10 species ,12, to m species ,14.In aquaculture, it is very important to understand the thermoregulatory behavior of cultured organisms. Thermoregulatory behaviour is used to inform about the tolerance limits, and the preferred temperature of aquatic species , and thePorcellionides pruinosus (Isopod) [Tigriopus californicus (Copepod) [Neocaridina davidi (Caridea) [Scylla. A study of the impact of water temperature on sex ratio of mud crab is expected to provide opportunities to produce monosex seeds, especially for aquaculture purposes.Temperature is also known as the most important environmental, sex determining factor for aquatic species . Some st(Isopod) , TigriopCopepod) , and NeoCaridea) . KnowledCaridea) . To dateS. paramamosain, for long term periods, is still scarce. Previous research conducted by Gong et al. [S. paramamosain.Studies on the effect of temperature on the nursery phase of g et al.  only loog et al.  only froS. paramamosain) in this experiment were produced at the Institute of Tropical Aquaculture and Fisheries, Universiti Malaysia Terengganu hatchery, using local female broodstock from Setiu Wetland, Terengganu, Malaysia. Larval production started from broodstock maintenance, through spawning, incubation, and larval rearing. Female broodstock of mean BW (bodyweight) 348 ± 70.88 g, and mean CW/CL (carapace width/carapace length) 13.33 ± 0.65/8.83 ± 0.61 mm, were maintained in a 7.5 m2 square fiberglass tank, at a density of about 2 ind/m2. Broodstock were fed with Scad fish, Decapterus macrosoma, and occasionally (2–4 times a month) with shellfish , as often as once a day, at a dose of about 3% of biomass. Spawned females were allowed to remain in the broodstock tank until eggs started to turn black (estimated to hatch in 2–3 days), then were transferred to the incubation tank. The incubation tank was a 500 L round fiberglass tank (d = 1.1 m). After the eggs had hatched completely, aeration in the incubation tank was turned off for several minutes, then larvae that had clustered and actively swam on the surface were collected by gentle scooping.Samples of mud crab megalopa  with a density of about 2 to 5 ind/mL. In Zoea 1 and Zoea 2, larvae were given both live feed and frozen Artemia nauplii to facilitate feed capture. In the Zoea 3 to Megalopa (M) phase, larvae were only given live Artemia nauplii. Rearing time from Zoea 1 to M phase was 16 days, and on the 22nd day M was collected for experiment.Zoea 1 larvae were maintained in larval rearing tanks (500&1000 L round fiberglass tanks) at a density >100 ind/L. Larvae were fed with ®) at one end, whereas at the other end, extreme cold water was provided by adding frozen sea water (by plastic cup). The gradient in temperature ranged between 9 to 38 °C. In each virtual segment, a tube diffuser/aeration stone was placed along the gradient, to provide a very gentle aeration, and avoid stratification in the water column. The temperature was measured in each virtual segment with a glass thermometer. Crablets were not fed for 24 h before testing, to avoid interference from their diet. Within the formed gradient, one crablet was introduced at the virtual segment which had temperature as same as of the acclimation temperature [Thermal preference was determined by the acute method, with a horizontal gradient of temperature, using Crablet 1 (C1) stage. Crablets were acclimated at ambient temperature 27 to 30 °C), prior to assessment. Apparatus consisted of a PVC pipe 330 cm long, and 10.16 cm in diameter, with 21 virtual segments of 15 cm each. The depth of the water column was 8 cm, and a gradient was formed by placing one 300 W heater  were used to monitor temperature during the experiments.Two aeration stones placed in the aquarium to maintain oxygen saturation, and to avoid thermal stratification in the water column. A 1500 W immersion heater was placed in the bath for CTmax determination. To determine CTmin, this bath was conditioned as a cold chamber, using ice flakes. The heating and cooling rate were approximately 1 °C min−1. Visual monitoring was performed to describe the behavioral stress responses, shown by crablets exposed to increases and decreases in temperature. Observation of CTmax and CTmin ended when the crablets showed loss of righting position, i.e., when crablets were on their back and could not recover an upright posture (about 2 min). When the juveniles reached this point, they were returned to the acclimation temperature  for a 96 h recovery period. Data of crablets that did not recover after 96 h was discarded. Determination of CTmin and CTmax of C1 (crablet instar) were performed three times each, with individual crablets used only once. A total of 40 and 35 crablets were used for CTmax and CTmin experiments, respectively. A thermal tolerance interval (TTI) was calculated as the difference between CTmax and CTmin mean values.A 40-litre aquarium was used as a thermo-regulated bath, containing one experimental chamber . Both a glass thermometer, and DO (dissolved oxygen) meter (YSI® Chiller was used to control temperature in Treatments A (24 °C) and B (28 °C), while Treatment C (32 °C) used a 300 W (Eheim®) heater. Crablets were maintained individually in plastic cups with 6–9 cm of bottom diameter. The cups were placed in a rectangular fiber tank (117 cm length × 70 cm width) for each treatment, with approximately 7 cm of water depth. A recirculation aquaculture system (RAS) with gentle aeration was used to ensure mixing of the water bath. Water was exchanged every two days, as much as 20 to 30%. The used water salinity ranged from 25 to 30 ppt. During 45 days of nursery period, crablets were fed with frozen adult Artemia once a day (ad libitum). The observed parameters were growth performance, survival, molting cycle , and sex ratio. Based on the molting cycle data, the results from each crablet stage, in each experiment, were determined at the end of experiment. Carapace measurements were performed using digital calipers (0.01 mm), and weight measurements using a digital scale (0.01 g). Crablet stages, as described with C1, C2, C3 and so on, were determined based on molting activity observed daily [A completely randomized design was used for this experiment, with four temperature condition treatments: water temperatures of 24, 28 and 32 °C, and ambient temperature (27 to 30 °C) as a control. The daily water temperature fluctuation of the 24, 28 and 32 °C treatments was <1 °C. Each treatment had three replicates, and each replication consisted of 10 samples. A Resun−1), which was determined both in terms of BW (SGRBW) and CW (SGRCW), molting interval, and percentage of molt increment [BW (% day−1) = /days of culture) × 100SGRCW (% day−1) = /days of culture × 100SGRPercentage of molting increment (%) = ((postmolt CW—premolt CW)/premolt CW) × 100Molting interval (days) = time period between two molt eventsSurvival rate (%) =  × 100Sex ratio (%) = ) × 100Growth performance was evaluated using growth indices: specific growth rate (SGR)  software.Samples of C5 stage from each treatment were fixed in Davidson solution for 24–48 h, stored in a 70% ethanol solution until further normal histological processing. Parafin wax sections (5μm thick) were stained with H&E and were observed using NikonData of carapace size and ratio, survival, and molting interval were processed first with normality (Shapiro–Wilk test), and homogeneity test (Levene test). The data were transformed using square root transformation prior to analysis, if the original data did not meet standards of normality and homogeneity. Data were analyzed using analysis of variance (ANOVA), and post hoc Tukey’s test, to determine statistical significance. If the data did not meet parametric analysis standards, even though the data had been transformed, the data were then analyzed using non-parametric statistics, namely the Krusskal–Wallis ANOVA, and post hoc Dunn test. Statistical analyses were run on R-program, whereas tables and graphs were generated using Microsoft Excel program. Data of CTmax, CTmin, and preferred temperature, were presented as mean values with standard deviation.The mean value of CTmin, CTmax, preferred temperature, and thermal tolerance interval (TTI) of C1, were 17.33 ± 0.58 °C, 40 ± 0.00 °C, 29.83 ± 2.47 °C, and 22.67 ± 0.58 °C, respectively, as listed in detail in p < 0.05), but not significantly different than those kept at both 32 °C and ambient temperature. Means of CL and final BW were not significantly different (p > 0.05) between treatments, but 28 °C and ambient temperature treatment showed higher values than for both 24 and 32 °C treatments. SGRBW observed for 24 °C was significantly different than for 28 °C (p < 0.05), but neither were significantly different from 32 °C, or ambient temperature. SGRCW value observed for 24 °C was the lowest, and significantly different from both 28 °C and ambient temperature, but not significantly different from 32 °C. Highest survival rates were obtained at 28 °C, followed by ambient temperature, 24 °C, and 32 °C treatments, in order. Survival for 28 °C was significantly higher than 32 °C, but it was not significantly different from either 24 °C or ambient temperature. The daily survival rate in p < 0.05) which was not significantly different from either of the 28 °C or ambient temperature treatments  between stages M to C5, and C1 to C5 for the 28 °C treatment, while it was longest for the 24 °C treatment (p < 0.05). Molt increment in 32 °C treatment showed the lowest value, particularly in the last three stages (C3 until °C6) (p < 0.05), eventhough the 32 °C treatment carapace had been of similar size to size in the other treatments during the C1–C3 stages  mean was shortest (reatment . MI was til °C6) . The car3 stages .The color of the gill lamella at the treatment of low (24 °C) and high (32 °C) temperature treatments seemed paler, compared to the treatment temperature of 28 °C and ambient temperature (27–30 °C). In addition, the color, size, and shape of the gill lamella, at low and high temperature treatments also appeared different from both treatments 28 °C and ambient, while both the gill size and shape of treatments 28 °C and ambient were similar. The gill lamella size at high C and lowS. paramamosain was 22.67 ± 0.58 °C , which demonstrated that thermal tolerance can change depending on age and life cycle stages [In this study, TTI of the crablet instar (C1) of  0.58 °C , which i26.04 °C . These pe stages ,35.S. paramamosain C1, in this present study, showed that C1 had a wide temperature tolerance, making it suitable to be stocked into a safe, predator-free pond. Although survival rate of M to C1 was high in this study (>80%), stocking of M directly into a pond is not recommended [TTI, and preferred temperature values of ommended , becauseS. Paramamosain, during nursery phase. 28 °C treatment produced the highest mean of final CW, SGRCW, SGRBW, and survival rate (p < 0.05). The higher values of growth performance and survival rate at this temperature, compared to other treatments, showed that the optimal temperature for the nursery of S. paramamosain  was around 28 °C. The current findings are almost similar to the optimal temperature of 30 °C recommended for juveniles of S. Serrata, during nursery phase from C2 to C3 [Eriocheir sinensis, for which temperatures of 28 and 30 °C were the optimal water temperatures for growth and molting in the juvenile phase [This study showed the effect of long-term exposure (chronic exposure) of water temperature on both growth, and survival rate of BW and SGRCW), and stage composition, was influenced by different temperature regimes, where low (24 °C) and high temperatures (32 °C) tended to have lower values than for temperature regimes of both 28 °C and ambient temperature (27 to 30 °C). Temperature is one of the external factors that can affect the growth of crustaceans, where increasing temperature from optimal temperature can result in reduction of molt increment [S. serrata [Callinectes sapidus [Growth performance in this study, as measured by CW, and BW at harvest, MI , molting increment, SGR in terms of BW and CW , during the molting phase from C1 to C2, although its value was not significantly different from either the 28 °C or ambient temperature treatments. This result indicated that high temperature had an effect on accelerationof molting cycle at C1 to C2, but in subsequent phases, the 28 °C treatment had improved MI, even though the values were not significantly different from the 32 °C and ambient temperature treatments. Furthermore, MI from C5 to C6 for the32 °C treatment had the shortest MI, and its value was significantly different (p < 0.05) from both 28 °C and ambient temperature treatments. The shorter MI periods at high temperatures are common in crustaceans [In this study, 32 °C treatment had the shortest MI (2.96 ± 1.22 days) compared to other treatments , which was thought to be related to a lower expression of Ecdysone receptor (Ecr) transcript in the crablet at salinity of 40 ppt. The results obtained by Ruscoe et al. [S. Serrata, from C2 to C3, was longer at salinity 35 ppt in comparison to salinities of 5, 10, 20, and 30 ppt, at temperature conditions of 25, 30 and 35 °C.In this study, the presence of daily salinity fluctuations (>1 ppt) at 32 °C, due to evaporation, may have had an effect on MI fluctuations at 32 °C. Future experiments need to minimize such salinity fluctuations for higher temperature treatments. Although it was known that mud crab have a broad tolerance to temperature and salinity ,40,41, te et al.  also shoS. Paramamosain, and at C2 to C3 of S. Serrata, respectively. In the current study, exposure of juveniles of S. Paramamosain at non-optimal temperatures for long-term, likely caused loss ofenergy, and triggered mortality. Optimization of physiological and biochemical processes that provide maximal efficiency in energy use can occur within the optimal temperature range [S. serrata. Further, Matozzo et al. [Carcinus aestuarii, modulated its cellular and biochemical parameters  in order to cope with temperature. In the crablet phase, S. paramamosain had a better resistance to low temperature, which could be attributed to the increasing thickness of the integument with growth, that provides increasing protective, mechanical protection against abrupt changes in temperature [S. Paramamosain can reach 86.7%, from C1 to C2, under water temperature of 20 °C , but at water temperature of 14 °C, C1 becomes inactive with a substantial decrease in feed consumption, and C1 also did not molt to C2, during the current study. In C2 to C3 phase, Ruscoe et al. [S. serrata could only molt as much as 10 and 20%, at salinity conditions of 5 and 20 ppt, respectively. Generally, the optimal temperature range for tropical and subtropical crab species is above 25 °C. Temperatures below 20 °C cause increase in larval mortality [Survival rate has been useful in identifying the effect of inadequate diets on the crab larvae ,44, whilre range ,12, whilre range . Paital re range  supposedo et al.  concludeperature . Gong ete et al.  found thortality , particuortality ,41.S. paramamosain crablets. Although the reason for changes in the size of gill lamella remain unknownin this study, they are likely to be either a form of adaptation to temperature changes, or to result from negative impact of long-term exposure to temperatures above or below the optimal temperature. For example, one of the ways in which intertidal decapod crustaceans adapt to an environment, which is sometimes without water, is the specialization of the structure of their branchia apparatus, which functions to prevent the gill lamella from adhering together, and reduces availablesurface area for gas exchange [Histological observationof the gill lamella section showed that various temperature treatments had a direct effect on the gills of exchange . The chaexchange . The obsexchange , where cexchange .p < 0.05), which indicated a possibility of water temperature having an affect on sex ratio in mud crab, such as has been commonly reported in teleostei fish [Male ratio was higher at low temperature treatment 24 °C) than for the other temperature treatments  of S.paramamosain was 28 to 30 °C. Below or above optimal temperature range could have negative impacts on growth performance, survival, or gill lamella structure of S. paramamosain crablet during long-term nursery phase. Findings of this study could increase crablet hatchery productivity, and reduce dependence on seeds taken from the wild. In addition, the potential of monosex seed production of S. paramamosain through temperature treatment needs to be studied further in future.TTI and preferred temperature values observed in the current study showed that C1 of


Tribulus terrestris L. (TT) supplementation have been shown to enhance sports performance in many but not all studies. Moreover, data regarding the potential impact of TT supplementation on CrossFit® endurance is limited. This study aimed to determine whether TT supplementation improve body composition, hormonal response, and performance among CrossFit® athletes. In a randomized, single-blind, placebo-controlled trial, a total of 30 healthy CrossFit®-trained males were randomly allocated to receive either 770 mg of TT supplementation or a placebo daily for 6 weeks. Body mass, fat mass, fat composition, testosterone and cortisol levels, and CrossFit® performance  were assessed before and after intervention. There were no significant group x time interactions for the outcomes of the study except for testosterone levels and bench press performance (p < 0.05). TT supplementation did not impact enhance performance or body composition in CrossFit® male athletes. However, TT supplementation may act as a testosterone booster helping the recovery after physical loads and mitigating fatigue. Dietary supplements are substances that can be used to add nutrients to one’s diet and that have a potential nutritional or physiological effect . In 2020Tribulus terrestris L. (TT), originally from India. TT is a famous traditional Chinese medicine that has been widely used in many countries for thousands of years  [Total testosterone was determined by enzyme-linked immunosorbent assay (ELISA) . Cortisol concentration was assessed by enzyme-linked fluorescence assay (ELFA)-based technology by ready-to-use reagents with a compact multiparametric immunoanalyzer MinividasV)/100)] ,33. ® warm-up, strength and/or CrossFit® skill technique as well as a structured strength or conditioning workout between 10 to 30 minutes, followed by stretching and/or cool down. Each workout was performed in the participants’ own gym, supervised by a certified CrossFit® Degree 1- or 2-certified instructor (at each). All subjects completed the same physical activity routines to ensure that they performed the same prescribed workout during the study.During the 6 weeks of the study, training involved 3 weekly sessions on alternate days . Each session was one-hour long and was divided into a CrossFit® athletes were briefed on two diet-tracking systems  for CG and IG. Effect size among participants were calculated using eta-squared (η2p). The η2p is a measure of effector size association. Because this measure often overestimates effect size, values were interpreted as the following: where no effect is indicated, if 0 ≤ η2p < 0.05; a minimal effect is shown if 0.05 ≤ η2p < 0.26; a moderate effect is indicated if 0.26 ≤ η2p < 0.64; and a strong effect is indicated if η2p ≥ 0.64. The treatment was randomly assigned utilizing a sequence generated by date (or day) of admission in the study by Random Sequence Generator. Random Sequence Generator is an application, which was accessed through the Internet. The data were presented as means and standard deviations. Analyses was performed using SPSS version 4.0 software . Statistical significance was indicated when p > 0.05) for energy, proteins, fats, carbohydrates, micronutrients, and vitamins (There was no significant difference between groups (CG and IG) (vitamins .p < 0.05). However, the group-by-time interaction between CG and IG did not reveal significant differences (p > 0.05).In ® Total and Grace) for both GC and IG at the beginning and end of the study . Within the CrossFit® Total tests , only a significant increase (p < 0.05) was observed in the bench press in GI (T1: 69.17 ± 11.58 vs. T2: 80.00 ± 11.40). In addition, significant differences (p < 0.05) were only observed in group-time for the bench press . In both groups, a shorter execution time was observed, without statistical significance (p > 0.05), in the Grace test in both GI (T1: 28.00 ± 6.23 22.67 ± 5.92 vs. T2: 22.67 ± 5.92 28.00 ± 6.23) and CG (T1:30.17 ± 5.08 vs. T2: 24.33 ± 4.46). No significant group-time differences were observed for the Grace test .p > 0.05) were observed between baseline and at the end of the six-week study in testosterone, cortisol, and testosterone/cortisol ratio. Nor were there significant differences in group-by-time for cortisol  and ratio testosterone/cortisol ratio . Significant differences (p < 0.05) can be seen in the group-by-time for testosterone .In addition, p > 0.05) in the perceived fatigue between T1 vs. T2 in both groups. Additionally, there were no significant differences in the group-by-time for RPE .Finally, p < 0.05) in the bench press test between the IG vs. CG. There were no significant differences (p > 0.05) in dead lift (IG 4.64 ± 2.55 vs. CG 3.32 ± 4.17) and CrossFit® Total (IG 6.19 ± 3.19 vs. CG 5.07 ± 4.61), but there was an improving trend in the IG group. However, in the CG the percentage change was greater for squat than IG (CG 5.70 ± 8.303 vs. IG 3.01 ± 4.96). The results for the CrossFit® Total test were similar for both groups (CG −24.73 ± 10.58 vs. IG −25.00 ± 10.74). p < 0.05) were observed in the testosterone levels (IG −0.20 ± 13.63 vs. CG −21.85 ± 11.10). There were no significant differences (p > 0.05) in cortisol (IG 4.64 ± 2.55 vs. CG 28.65 ± 43.19) and testosterone/cortisol ratio (IG 22.43 ± 69.60 vs. CG −29.07 ± 37.38), but there was an improving trend in the IG group. In addition, −1) on body composition, hormonal response, RPE, and CrossFit® performance in CrossFit®-trained males. We hypothesized that TT supplementation would improve CrossFit® performance and testosterone levels since previous studies in athletes demonstrated that TT enhanced sports performance [® Total and Grace test), body composition parameters , cortisol, and testosterone/cortisol ratio.The purpose of our investigation was to evaluate the impact of six-weeks TT supplementation  and with no significant difference in the group-time interaction, indicating that the CrossFitathletes ,27,36,37athletes . Howeverathletes . ® training program, and/or the absence of TT could have led to a lower level of adaptation in CG. Therefore, the information from bodybuilding and fitness websites on the activity of TT compound supplements in decreasing body fat and increasing lean muscle mass is questionable [These results would suggest that the increase in bench press performance in IG was not mediated by changes in body composition or increased muscle mass. This could be because of the adequate adaptability of the IG athletes to the CrossFittionable ,11. −1 of TT [−1 for 8 weeks) [® Total exercise, squat and dead lift showed no significant gains. The squat showed a slightly better percentage improvement in IG than in CG. However, in dead lift, the opposite was true. Squat and dead lift employ muscle groups, mainly of the posterior chain, involved in many other CrossFit® movements . This could indicate that habitual training is responsible for the individual adaptations of each athlete [We did not observe significant improvements in WODs, which evaluates global strength by the sum of the measurement of 1 repetition maximum (RM) of bench press, squat, and dead lift. These results are similar to those reported in a study conducted in rugby players, where no significant improvement in total strength was observed after supplementing for five weeks with 450 mg · day−1 of TT . In addi8 weeks) , researc athlete , which i athlete ,18. Alternatively, the bench press showed significant improvements in the IG between T1 and T2, twice as much as in the CG. Our results are similar to those obtained in 1-RM bench press after three weeks of TT supplementation in powerlifters ,21. The TT-mediated inhibition of cyclooxygenase-2 (COX-2)  could alTT is recommended as an herbal supplement that optimizes and increases mood ,41, prob−1 · day−1) in young adults [−1) for three months in adults with idiopathic infertility [Continuous, intense exercise directly influences an inadequate functioning of the hypothalamus-pituitary-testes axis . This alg adults . Similarertility . To our ertility . In thisertility ,26. Addiertility . Other mertility , and theertility .−1 increased, but this change was not statistically significant [® trained vs. athletes belonging to a physical education program). These results could indicate that the possible efficacy of TT on hormonal status would be conditioned in relation to the sport modality and/or the physical condition of the athletes.Cortisol is released from the adrenal cortex in response to stressors linked to exercise and catabolic actions; it is also directly influenced by duration and intensity . The cornificant . Althougp > 0.05) in the testosterone/cortisol ratio concerning CG at the end of the study, which could translate into a potential better response to training load and volume and establish a positive trend in the recovery process in athletes. In this way, improvements in muscle recovery after high-intensity endurance exercises by reduction of creatine phosphokinase (CPK) and lactate dehydrogenase (LDH) have been reported after supplementation with 500 mg · day of TT [Testosterone/cortisol ratio is indicative of the anabolism/catabolism ratio in the body due to their role in protein synthesis and degradation. It can be used as a biological indicator to supervise fatigue and establish optimal training loads for athletes . Testostay of TT . Nutritional supplements contaminated with banned substances have been reported . TaintedThe present study has certain limitations. The sample size is small; a larger one would produce more robust results. Moreover, the realization in professional athletes of different sports modalities would also give us the accurate dimension of the potential properties of TT supplementation because the nature of each sport has demands that differ between disciplines, such as aerobic or anaerobic capacity, the type of strength used, the predominant energy systems, and other factors that would justify the individualization of TT testing. Additionally, the design exclusively focuses on males, which may reduce generalizability. Nor were biochemical tests included, especially determination of muscle damage marker levels, such as lactate dehydrogenase, creatine kinase, and myoglobin. Thus, it would be necessary to include such as analysis in future studies.−1 of TT used in CrossFit®-trained men does not appear to influence body composition, RPE, total strength, and anaerobic performance. However, our results suggest that it could potentially intervene in the recovery process by a tendency, although not significant, to increase serum testosterone and maintain a favorable trend in the testosterone/cortisol ratio, which would mean that it could potentially influence less fatigue and catabolism associated with exercise. The dose 770 mg · dayTT supplementation could be recommended for athletes playing sports at levels where the competition rules prevent adequate recovery, such as play-offs in team sports  or tournaments in individual sports  and in training stages characterized by high loads and volumes. In addition, since the beginning of the COVID-19 pandemic, more males have been affected as well as a higher rate of intensive care unit admissions and deaths. Perhaps a lower bioavailability of testosterone or a decrease in its receptors could be the cause. In this regard, low total testosterone levels are associated with a hyperinflammatory state in hospitalized men with COVID-19 and represent a risk factor for hospital mortality in COVID-19 patients. Therefore, inadequate total testosterone levels may serve as markers of poor prognosis in COVID-19 . Thus, g


Freelite and N-Latex methods are two common monitoring methods at present. But the two meanings are not completely equivalent, especially for patients with renal insufficiency. We analyzed the changes of serum and urine FLC in renal insufficiency patients without monoclonal gammopathy and the clinical significance of these changes.  Free light chains κ, serum FLC λ, SCr, serum β2-microglobulin, urinary FLC κ, urinary FLC λ, urinary α1-microglobulin, and urinary β2-microglobulin.  This study is an observational study. Patients ≥ 18 years old, who met the diagnostic criteria of chronic kidney disease (CKD), excluding monoclonal gammopathy, were selected. Fasting serum and 24-hour urine were taken to detect serum FLC κ/λ remained stable, which was basically within the reference range of healthy people and was not affected by renal function. There was a good correlation between FLC detected by N-Latex and microglobulin in serum and urine.  There was a good correlation between the two methods for determining serum/urinary FLC. No matter serum or urine, FLC showed a good correlation with renal function by the N-Latex method, but not by the Freelite method. Under the N-Latex method, FLC κ/λ determined by the N-Latex method remained stable within the recommended range. When the concentration of FLC is low, the N-Latex method is more recommended to monitor FLC. The FLC measured by the N-Latex method is more closely related to renal function. The ratio of FLC  The i[μmol/L) . ImpaireIn 2001, a method for the detection of FLC was developed for the first time : FreelitThrough this study, we intend to understand the changes of serum and urine FLC in patients with renal insufficiency without monoclonal gammopathy and the clinical significance of these changes.2; stage 2: 60 mL/min/1.73 m2 ≤ eGFR < 90 mL/min/1.73 m2; stage 3: 30 mL/min/1.73 m2 ≤ eGFR < 60 mL/min/1.73 m2; stage 4: 15 mL/min/1.73 m2 ≤ eGFR < 30 mL/min/1.73 m2; and stage 5: eGFR < 15 mL/min/1.73 m2). There were 50 patients in each group, and the number of patients in the group was stopped after the number of patients reached the standard.This study is an observational study. Since May 1, 2019, patients were continuously enrolled in the group according to the order in which they were admitted to the ward of the Department of Nephrology of Beijing Hospital. According to K/DOQI guidelines , patient2 for ≥3months, with or without kidney damage [μmol/L) within 48 h or an increase in serum creatinine to ≥1.5 times baseline within the previous 7 days or urine volume ≤ 0.5 mL/kg/h for 6 h [The inclusion criteria were as follows: (1) age ≥ 18 years old; (2) it meets the diagnostic criteria of chronic kidney disease (CKD) which was diagnosed by kidney damage for ≥3 months as defined by structural or functional abnormalities of the kidney, with or without decreased GFR, or GFR < 60 mL/min/1.73 mThis study has been approved by the Ethics Committee of Beijing Hospital .κ, serum FLC λ, SCr, serum β2-microglobulin, urinary FLC κ, urinary FLC λ, urinary α1-microglobulin, and urinary β2-microglobulin. Serum levels of creatinine were determined by an enzymatic method . Other items were tested by Siemens BN II. FLC in serum and urine were detected by the Freelite™ assay  and the N-Latex FLC assay  at the same time.Fasting serum and 24-hour urine were taken from each patient in the same period. The following tests were performed: serum FLC κ6.7-22.4 mg/L; FLC λ 8.3-27.0 mg/L, FLC κ/λ0.31-1.56; Freelite: FLC κ 3.3.0-19.40 mg/L; FLC λ 5.71-26.30 mg/L, FLC κ/λ 0.26-1.65 (0.37-3.10 mg/L if eGFR < 60 mL/min/1.73 m2).We chose to use the reference ranges proposed in the literature , 6, 9: NIBM SPSS 22.0 were used for data analysis. First, we verified whether the continuous variables belong to normal distribution. For normal distribution variables, mean ± standard deviation was used; single-factor ANOVA was used to compare different groups; Pearson was used to test the correlation between data.A total of 250 patients were selected, including 145 males (58.0%) and 105 females (42.0%). The average age was 58.93 ± 15.75 years old, of which 58.18 ± 16.59 years old in males and 60.10 ± 14.35 years old in females. 95 patients (38.0%) suffered from primary glomerulopathy , 73 patients (29.2%) with diabetic nephropathy, 23 patients (9.2%) with hypertensive nephropathy, 14 patients (5.6%) with secondary glomerulonephritis, 8 patients (3.2%) with chronic interstitial glomerulonephritis, 7 patients (3.5%) with obstructive nephropathy, 6 patients (2.4%) with renal artery stenosis, and 24 patients (9.6%) with renal artery stenosis.κ: R = 0.703, P ≤ 0.001; FLC λ: R = 0.505, P = 0.001, κ: R = 0.463, P = 0.071; FLC λ: R = 0.500, P = 0.051, κ/λ was within the reference intervals , while under the Freelite method, the ratio was greatly affected by renal function, and the ratio was significantly higher than the normal range in stage 4 (0.37-3.10) .Using N-Latex to detect FLC, with the aggravation of renal function injury, the concentration of serum FLC increased step by step  . There wκ: R = 0.188, P = 0.003; λ: R = 0.325, P ≤ 0.001). There was no clear correlation by the Freelite method. The level of urinary FLC κ/λ obtained by the N-Latex method was relatively stable, but only phase 2 and stage 3 were within the recommended range of the manual . However, the ratio obtained by the Freelite method was greatly affected by renal function, in which stage 2 and stage 3 were significantly higher than the reference range (0.461-4.0) .As can be seen from Figures 461-4.0) . There wMicroglobulin can freely pass through the glomerulus and reabsorb through the renal tubule. Therefore, serum microglobulin reflects the ability of glomerular filtration, and urinary microglobulin reflects the state of early renal tubular injury. As can be seen from κ or λ light chains. After forming a complete immunoglobulin molecule, about 40% of the light chains remain into the circulation. The production speed of κ chain is about twice as fast as that of λ chain, and FLC κ is often a monomer, while FLC λ is easy to form a dimer. In normal individuals, serum FLC is rapidly cleared and metabolized by the kidney. Monomer FLC (κ chain) is cleared at 40% glomerular filtration rate within 2-4 hours. The dimer FLC (λ chain) is cleared at 20% glomerular filtration rate within 3-6 hours. FLC is completely through the glomeruli, degraded in the proximal renal tubules, and then absorbed or excreted in the form of fragments. The scavenging rate of FLC κ is 3 times faster than that of dimer FLC λ, so the concentration in serum is only about half of that of FLC λ [Each plasma cell produces one of the five heavy chains and of FLC λ .κ/λ changes only in a small range—it can be used to distinguish between monoclonal and polyclonal diseases. However, with the aggravation of renal failure, the ratio of κ/λ will increase, even as high as 3.1 [κ/λ is greater. Therefore, it can reflect small changes in renal processing function, urine dilution function, and FLC changes.In patients with CKD, the clearance rate of polyclonal FLC decreases and the serum concentration increases. Mild to moderate renal failure prolongs the half-life of both FLC from hours to days, so the concentration of FLC in serum increases by 20 times or more, but h as 3.1 . The conWith the availability of the Freelite method, the determination of FLC has been incorporated into guidelines for multiple myeloma and related diseases –3. HowevThe Freelite method is established using sheep polyclonal antibodies to detect only FLC wavelength dimers, while the N-Latex test includes several mouse monoclonal antibodies that only detected FLC monomers . In the κ and λ is slightly different, but this difference does not seem to affect the prognosis of the patients. Therefore, researchers believe that the two detection methods are limited in clinical application and cannot be replaced with each other [Many researchers have compared the two different detection methods and found that the results of the two kinds of FLC have good correlation and consistency are even in patients with kidney failure. This study confirmed this finding. However, the accuracy of the two methods for FLC ch other –17. Our There may be another reason for this. When testing different reagent batches at low concentrations , the Freelite method will result in a large error in the results , whereasκ and λ is of greater significance [κ/λ was determined by Freelite and N-Latex FLC, respectively. The regression line almost overlapped with the same line in the reference range of healthy control, and the best consistency was observed. In patients with highly abnormal FLC ratios , greater measurement uncertainty was observed, resulting in significant differences in FLC ratios between the two methods [κ/λ measured by the N-Latex method had little change in patients with impaired renal function, and no FLC κ/λ exceeded the N-Latex reference limit of healthy controls. However, the FLC κ/λ determined by the Freelite method increased significantly with the increase of renal function damage [κ/λ obtained by the Freelite method was negatively correlated with eGFR, while N-Latex was positively correlated, but they were all within the reference range of healthy people [κ/λ was within the recommended range, while under the Freelite method, the ratio was significantly higher than the normal range. The expression of urinary FLC κ/λ was less than that of N-Latex, and it was more stable than that of urine κ/λ.In view of the influence of renal function on FLC, the ratio of urinary FLC ificance . The rat methods . In 2014n damage . It has n damage . The laty people . Our resIn view of the fact that FLC passes freely through the glomerular filtration membrane, most of it is reabsorbed by renal tubules, which is similar to microglobulin. In our study, there was a high correlation between FLC and microglobulin measured by the N-Latex method, while the correlation was poor by the Freelite method. It also showed that the FLC measured by the N-Latex method can better reflect the state of renal disease in patients with renal insufficiency.κ/λ determined by the N-Latex method is not affected by renal function and remained stable within the recommended range.The metabolism of FLC depends on renal function. When the concentration of FLC is low, the N-Latex method is more recommended to monitor FLC. The FLC measured by the N-Latex method is more closely related to renal function. The ratio of FLC