The current study aimed to analyze, using accelerometer-based activity, acute workload, chronic workload, acutechronic workloads ratio, training-monotony and training-strain throughout a competitive soccer-season and to compare these variables between players from different playing positions. Twenty-one professional soccer-players were monitored during the 48 weeks of the season. Players were grouped according to their position. Four lateral-defenders and four winger-players formed LDW group, four central-defenders and four forwards formed CDF group, and six midfielder-players formed MDF group. Accelerometer-based variables were collected during training and match contexts and were used to generate indicators of weekly acute and chronic workload, training monotony, training strain and metabolic power. A one-way ANOVA compared all dependent variables between groups, and effect sizes for pairwise comparisons were calculated. Results revealed variations in the weekly load throughout the season, which demands caution from coaches to avoid injuries. There were no differences in weekly-loads for all dependent variables . We conclude that the weekly-load is not constant during a competitive season and players from different positions have similar weekly-loads. Therefore, previously reported in the literature, possible match-related positional differences might be compensated by differences in training-related loads, leading to a similar profile when considering the whole week. However, the competitive schedule requires frequent adjustments in load prescription because players are differently demanded during the matches3. For example, players from different playing positions differ in internal and external loads during a match5, leading to other weekly training loads. Hence, assessing training loads during matches and training is essential for an adequate load prescription.Stimulating players with adequate training loads challenges coaches in elite soccer. Recent approaches to load monitoring proposed that variations in weekly training load reduce monotony and increase the potential for positive adaptations7. Data recorded with Global Positioning Systems (GPS) allows coaches to calculate different variables associated with the external training load. Traditionally, external load data collection and recording has focused mainly on variables such as volume, intensity, and occasionally frenquency and density. However, since training load variability may condition the type of adaptations that can be attained, it is essential to consider parameters such as training monotony, training strain8, metabolic power, and acute-chronic workload ratio (ACWR)10, which were previously calculated using rates of perceived exertion only. Training monotony is the mean of the training loads performed during the week divided by the standard deviation of these training loads11; training strain is the sum of the training loads performed in all training sessions and matches during a week multiplied by training monotony11. The metabolic power is the overall energy cost of activity12, and the ACWR compares the acute workload (the workload players were exposed in the current week) with the chronic workload . Together, these variables describe the variation in the weekly training load and can be used to adjust the load prescription according to players’ responses. This multidimensional approach to load monitoring seems crucial in training13 and competition since isolated variables may not capture the whole phenomenon.The external training load is defined as the work performed by an athlete during training or competition4 showed increased match demands among all playing positions in the past decade, mainly the high-intensity efforts. Full-backs and wing forwards perform the highest number of sprints during the match15, which may result in a higher external load in these players compared to the other playing positions. Hence, differences in the weekly training load in players from different playing positions are expected, although this hypothesis still needs to be tested. The limited knowledge about the possible differences in the weekly training loads in players from different playing positions limits the evidence-based load prescription and must be investigated.The impact of the playing position on match demands is well established. Bush et al.Considering the abovementioned rationale, this study aimed to (1) analyze the acute and chronic training loads, the ACWR, the training monotony and the training strain throughout a competitive season in professional soccer players and (2) compare these variables between players from different playing positions. Furthermore, based on the match data, we expected that external players  would present higher weekly training loads than the other playing positions.16, so 4 Lateral Defender and 4 Wingers players formed the LDW group, another 4 Central Defender and 3 Forwards constituted the CDF group and 6 Midfielder players formed the MDF group. Players were monitored and controlled by GPS throughout the season .This study was a cohort of an entire season at the highest level of the Iranian Premier League, the Persian Gulf Premier League and the Hazfi cup in 2018–2019. In this study, based on the availability of one team, we divided of players into three groups based on similar activity needs of playing positionThe monitoring was performed daily for each training session and competition. Finally, we explored two specific goals, which are: (i) to describe (mean/standard deviation (SD)), weekly average acute (wAW), chronic (wCW), wACWR, training monotony (wTM) and training strain (wTS) based on body load (BL) variations across the full season by play position; (ii) to analyze the variations in pairwise comparisons by play position of wAW, wCW, wACWR, wTM, wTS, weekly metabolic power average (wMPA), weekly average of Accelerations zone 1 (wAccZ1), Accelerations zone 2 (wAccZ2), Accelerations zone 3 (wAccZ3), Decelerations zone 1 (wDecZ1), Decelerations zone 2 (wDecZ2), Decelerations zone 3 (wDecZ3), Ratio between wMPA/wAW, Ratio between wAccZ1/wDecZ1,Ratio between wAccZ2/wDecZ2, Ratio between wAccZ3/wDecZ3 in the full season.2) who were monitored for 48 weeks during a full season in the Iranian Premier League. For the players' information to be calculated, they had to attend at least three weekly sessions. The exclusion criteria of this study were defined as follows (i) If a player were absent for two consecutive weeks, he would be excluded from the study for analysis. (ii) Goalkeepers were not included in the study for analysis. The current study was approved by the University of Mohaghegh Ardabili (1395.10.20). As well as the club's official license and the players' informed consent were also obtained for study. We followed the Helsinki Declaration on Human Research at all stages of the study.The study included 21 professional soccer players  GPS with 15 Hz through the accelerometer variables (Acc & Dec); (ii) BL with 16G, 100 Hz, and Tri-Axial; (iii) All transfer of information is done by the infrared; (iv) It has the smallest size and is waterproof on the market; (v) It weighs about fifty grams and has very high battery power. Data collected during the season had good weather states regarding the satellite. GPS unit has high validity and reliability for measuring external load variables17. We set the GPS default to SPI IQ Absolutes throughout the season. The variables used in this study were as follows among the external loaded variables: 1. BL, considered the newest workload training, calculates the training load marker as well as the rate of training load (BL/min); 2. MPA calculates the average rate of energy consumed per second (W/kg) for the athlete according to the specifications entered in the device default (i.e. height and weight), and the previous report showed that it has high reliability of 3–5%19; 3. AccZ1 (< 2 m/s2); 4. AccZ2 (2 to 4 m/s2); 5. AccZ3 (> 4 m/s2); 6. DecZ1 (< − 2 m/s2); 7. DecZ2 (− 2 to − 4 m/s2); 8. DecZ3 (> − 4 m/s220).To ensure accurate data collection by GPS, we pursued the following procedure. First, before starting the training, we put the GPS unit in the special belt position of the device and then we had to check the light before the start of the workout. At the end of the workout, we removed the GPS unit from the belt and entered the docking station to store information. Data storage was done by updated specialized AMS software 21 was used to reduce the re-porting error in this study. For this reason, in the first four weeks of the season, two variables (wCW and wACWR), were not reported in the study; wTM 22; wTS ; Ultimately, for other accelerometer variables calculations by form the weekly in during a throughout the season for 48 weeks.From the BL we obtained, respectively; wAW ; wCW ; wACWR , uncoupled methodpost-hoc test was used to detect the pairwise comparisons between different workload parameters and accelerometry variables by playing position. Cohen's d effect size with 95% CI was used for variables in this study. The Hopkins’ thresholds for Cohen’s d effect size statistics were used as follows: ≤ 0.2, trivial; > 0.2, small; > 0.6, moderate; > 1.2, large; > 2.0, very large; and > 4.0, nearly perfect23. Significant differences were considered for p ≤ 0.05. All statistical analyzes were considered by SPSS  at a significance level of p ≤ 0.05. Prism software  was used to draw the diagrams.Shapiro–Wilk test was used for normality, and Levene’s tests for homogeneity. The information was used for descriptive (mean and SD), and the information analysis was inferential tests. One-way ANOVA was used to find the differences between groups, and then the Bonferroni Figure Figure Table d = 1.79) and MDF . However, no significant differences were observed when compared to CDF and MDF. Also, there were higher significant wAccZ3 in LDW compared to CDF  and MDF ; no differences were observed when comparing CDF and MDF.Between-group comparisons for derived-GPS variables of acceleration and deceleration in the full season were presented in Table hypothesis.We aimed to analyze the weekly training loads throughout an entire season in a professional soccer team and also to compare these loads between players from different positions. Our results revealed that the weekly training loads varied throughout the season, and peaks of ACWR, training monotony, and training strain were more frequent in the pre-season and the midseason. However, no significant changes were observed between positions for all dependent variables. Therefore, we rejected our main 15 in official matches15. Also, when analyzing training contents, players from different positions showed different responses, even when the training scenario was the same25. The matches may represent the highest load within the training process and play a major role in biochemical and neuromuscular responses related to fatigue27. It is possible that coaches adopted tapering strategies to decrease the other training loads for these players, to reduce the stress and maximize performance, as suggested in previous studies28. This reduction probably compensated for the higher match load performed by external players leading to similar weekly loads among playing positions. Nevertheless, since we did not distinguish match-related load from the load imposed by other training activities, this hypothesis could not be confirmed.In general, the results of the present study showed that players from different positions were exposed to similar external loads throughout the season. Therefore, we expected higher weekly training loads for external players compared to the other playing positions because these players have shown the highest physical demand28. The individualization of these strategies is also important to match the needs of each player, including the specificities related to playing position. Finally, the wACWR ranged between 1.05 and 1.08 arbitrary units for all playing positions, falling within the zone recommended to prevent injuries and reach positive adaptations29. We also observed recommended values of training monotony11 (below 2.00) in all playing positions. Both ACWR and training monotony indicate similar information when they control the training loads in soccer. However, we must be aware of the controversial aspects of ACWR30, such as the lack of a proper causal effect between ACWR and the injury rate. Therefore, future studies should seek clarification of these issues and comprehend the possible relationship between these variables.The load monitoring over the whole season showed an oscillatory behavior of the external loads and a drastic increase in many variables between weeks 26 and 34. The congested fixture observed in many teams during the season highlights the importance of load monitoring and illustrates the difficulty of controlling the weekly loads over the competitive period. These sudden increases in the external loads require adequate strategists to reduce the injury risk, such as taperingThis study provides an example of load distribution over the weeks of an entire season in professional soccer and highlights the importance of monitoring the external load throughout the season. Nevertheless, the participants of the present study were players from one professional team, which requires further investigation into the impact of playing positions on weekly training loads in different contexts or teams. Furthermore, we neither evaluate the training load of each day of the week, which could reveal the impact of each training activity  on the weekly loads. Finally, it remains unclear whether the external load changes impact the players’ physiological responses and injury rate in soccer since most studies on load monitoring in soccer comprise only short periods. Providing further validity of the variables investigated in this study  would help sports scientists better understand the impact of load distribution over the season on players’ responses.More frequent peaks of ACWR, training monotony, and training strain were found in pre and mid season Moreover, despite an oscillatory training load dynamic was observed, these changes across the time are not position-dependent, as players from different positions showed similar training loads over the whole period. The absence of significant differences between positions could be related to the use of tapering strategies to increase sports performance in soccer competitions.


ObjectiveThis study aims to compare clinical and laboratory features between Lyme arthritis (LA) and oligoarticular juvenile idiopathic arthritis (oligoarticular JIA) by examining several potential predictors in pediatric patients. This study also aims to improve and increase awareness of ways to detect LA and oligoarticular JIA in pediatric patients who present with clinical features of joint pain.MethodsA medical chart review was conducted among pediatric patients diagnosed with LA or oligoarticular JIA at John R. Oishei Children’s Hospital of Buffalo between January 2014 and September 2018. Patients’ diagnoses were identified using the International Classification of Disease 10th Revision code for LA (ICD 10 code A69.23) and oligoarticular JIA (ICD 10 code M08.40).Borrelia burgdorferi proteins by IgG antibodies or at least two of three B. burgdorferi proteins by IgM antibodies, and (4) at the age of 16 or below at the time of diagnosis. Patients with oligoarticular JIA were included in this study if they (1) exhibited arthritis affecting one to four joints for at least six weeks in the first six months of diagnosis and (2) are at the age of 16 or below at the time of diagnosis after ruling out LA and reactive arthritis.Patients with LA were only included in this study if they (1) exhibited arthritis, (2) tested positive for Lyme antibodies, (3) indicated a positive western blot (WB) of five or more out of 10 B. burgdorferi DNA were also collected and analyzed.In this study, clinical presentations, physical exam findings during initial healthcare visits, and demographics including age, sex, and race of patients were obtained. In addition, laboratory results including white blood cells (WBCs), hemoglobin (Hgb), platelet count, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) level, Lyme antibodies through enzyme-linked immunosorbent assay (ELISA) and WB, synovial fluid analysis for red blood cells (RBCs), nucleated cells, and polymerase chain reaction (PCR) for ResultsIn our data, ESR and CRP were significantly higher in LA compared to oligoarticular JIA . The mean WBC in the synovial joint fluid was significantly higher in LA compared to oligoarticular JIA (P=0.002).ConclusionLA shares features with oligoarticular JIA. This overlap prevents the creation of a clinically useful predictive model for LA. Therefore, Lyme testing should be performed on all patients presenting with monoarticular and oligoarticular arthritis. In addition, ESR, CRP, and WBC in the synovial joint fluid were significantly higher in LA compared to oligoarticular JIA in our findings. While this difference is not definitive by any means, it may help distinguish between the two in cases where the diagnosis is not clear-cut, and the values of ESR, CRP, and WBC in the joint aspirate may help guide clinical judgment in cases that lack a definitive diagnosis. Borrelia burgdorferi through the transmission of tick bites [Lyme disease results from the infection of the spirochete ck bites ,2. An esck bites . The click bites ,5. Lyme ck bites . The click bites .In children, LA is often misdiagnosed as the oligoarticular subtype of juvenile idiopathic arthritis (JIA) . JIA is Due to the high incidence of Lyme disease in the Western New York (WNY) region, we instituted a mandatory Lyme disease screening protocol for all patients under the age of 16 years and presenting with oligoarticular JIA symptoms in 2016, and immediately noticed a significant increase in the incidence of LA . In thisIn this study, medical records of pediatric patients diagnosed with LA or oligoarticular JIA at John R. Oishei Children’s Hospital  between January 2014 and September 2018, were retrospectively reviewed. Approval was obtained from the State University of New York (SUNY) and the University at Buffalo Institutional Review Board (UBIRB) (reference numbers: STUDY00002006 and STUDY00003014).Patients’ diagnoses were identified using the International Classification of Disease 10th Revision Code for LA (ICD 10 code A69.23) and oligoarticular JIA (ICD 10 code M08.40).B. burgdorferi proteins by IgG antibodies or at least two of three B. burgdorferi proteins by IgM antibodies, and at the age of 16 or below at the time of diagnosis. Patients with oligoarticular JIA were included in this study if they exhibited arthritis affecting one to four joints in the first six months of diagnosis, at the age of 16 or below at the time of diagnosis, and had arthritis that lasted more than six weeks. Patients with LA were included in this study if they exhibited arthritis, tested positive for Lyme antibodies, indicated a positive western blot (WB) of at least five of 10 B. burgdorferi DNA were collected and analyzed. Clinical presentations, physical exam findings during initial healthcare visits, and demographics including age, sex, and race of patients in this study were reviewed. In addition, laboratory results including white blood cell count (WBC), hemoglobin (Hgb), platelet count, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) level, Lyme antibodies through enzyme-linked immunosorbent assay (ELISA) and WB, synovial fluid analysis for nucleated cells, and polymerase chain reaction (PCR) for Descriptive statistics were computed to characterize the study sample. Categorical variables  were reported as proportions in percentage, and continuous-level variables  as medians and interquartile ranges. Separate Mann-Whitney U tests were used to assess differences between patients with Lyme arthritis and oligoarticular juvenile idiopathic arthritis for continuous-level variables. Separate chi-square tests were used to assess differences for categorical variables between the two diseases, with Fisher’s exact test used for contingency tables having any cell less than five observations. All statistical tests were conducted as two-tailed, based on an alpha of 0.05, and with SYSTAT 13 .Baseline demographicsAmong the 61 patients included in this study, 44 patients were diagnosed with oligoarticular JIA, and 17 patients were diagnosed with LA  presented with involvement of a single joint  and LA (82.4%). Patients presented with only knee joint involvement in 65.6% of patients with oligoarticular JIA and 59.1% of patients with LA. The next most commonly affected joints were the ankle (10.6%) and wrist (10.6%). In an uncommon presentation for LA, arthritis was present bilaterally at the interphalangeal joint in two patients (11.7%) (Table A majority (77.3%) of patients in the cohort, arthritis was the only clinical presentation at the time of diagnosis. Within the LA group, 59.1% of patients did not have documentation or recollection of any symptoms during the early stages of Lyme disease, including no rashes, no history of a tick bite, and no flu-like symptoms during the summer, and arthritis was the initial presenting symptom. In addition to arthritis, three (13.6%) patients with LA had neurologic symptoms, including unilateral seventh cranial nerve palsy (n=1), numbness in the lower extremities (n=1), and headaches (n=1). Patients with neurological symptoms showed no symptoms of nuchal rigidity or photophobia to warranty lumber puncture. Moreover, 3 (13.6%) patients also reported the presence of a rash, but they did not seek medical care, and another 3 (13.6%) patients reported a previous history of a rash or bug bite three to four months before the presentation that had resolved. In addition, 4 (18.2%) patients reported a history of camping or residing in an area at high risk for Lyme disease, such as a wooded area. 3/μL (IQR 2.93 × 103/μL) in the oligoarticular JIA group, compared to 8.9 × 103/μL (IQR 4.20 × 103/μL) in the LA group, p=0.3  positivity in serum, and WBC counts in synovial fluid. A significantly higher proportion of patients with LA had elevated ESR levels compared to patients with JIA . Similarly, a higher frequency of patients with LA had an elevated CRP level compared to patients with JIA . We compared the median ESR and CRP levels in patients with LA and oligoarticular JIA who had elevated levels of these inflammatory markers. While the median ESR was numerically higher in patients with LA (59 mm/h) compared to oligoarticular JIA (44.5 mm/h), this did not achieve statistical significance, p=0.051. Similar results were noted in patients with elevated CRP levels. There was no significant difference in the proportion of patients with a positive ANA between the two groups  . The synovial fluid WBC count was >10,000/uL in eight patients . However, a higher proportion of patients with synovial fluid WBC <10,000/uL had oligoarticular JIA   were more frequently noted in patients with LA  compared to oligoarticular JIA . However, the IQR varied considerably among patients. In addition, not all patients with LA had elevated inflammatory markers, and similarly, significant ESR and CRP elevations were seen in some patients with oligoarticular JIA. These findings are consistent with the reported literature, where ESR and CRP elevations are often but not always seen in patients with LA, highlighting the importance of not excluding this diagnosis in the presence of normal ESR and CRP levels ,16. The We also noted that patients with LA had a higher median synovial fluid WBC count and more frequent elevations in the synovial fluid WBC count compared to patients with oligoarticular JIA, another potentially important marker to suggest the diagnosis of LA. There are other studies in the literature comparing synovial fluid markers in Lyme disease versus septic arthritis or Lyme disease among pediatric and adult populations, but none with such a direct comparison between Lyme disease and oligoarticular JIA ,10.Our study has a few limitations. Firstly, our sample size is small. Second, our data are based on a retrospective chart review at a single institution. Despite this, our study highlights the significant overlap in clinical presentation between patients with oligoarticular JIA and LA and highlights the importance of Lyme antibody testing in all patients presenting with mono- or oligoarticular arthritis in endemic regions. With the continued rise in LA cases in Western New York, it is essential for providers across the state to be aware of the distinct clinical and laboratory features of Lyme arthritis and oligoarticular JIA and maintain a high suspicion for both in pediatric patients presenting with mono or oligoarticular joint arthritis.LA shares features with oligoarticular JIA. This overlap prevents the creation of a clinically useful predictive model for LA. Therefore, Lyme testing should be performed on all patients presenting with mono- and oligoarticular arthritis. In addition, ESR, CRP, and WBC in the synovial joint fluid were significantly higher in LA compared to oligoarticular JIA in our findings. While this difference is not definitive by any means, it may help distinguish between the two in cases where the diagnosis is not clear-cut, and the values of ESR, CRP, and WBC in the joint aspirate may help guide clinical judgment in cases that lack a definitive diagnosis.


Brassica napus and the mechanism. To shed light on the effect of TDZ on the seed size of B. napus, four different concentrations of TDZ were applied to B. napus. Results indicated that TDZ treatment could increase the seed diameter and silique length of B. napus to varying degrees and 100 and 200 μmol/L TDZ treatments were the most effective with a 3.6 and 4.6% increase in seed diameter, respectively. In addition, the yield of B. napus was also substantially increased under TDZ treatment. On the other hand, confocal micrographs of embryos and cotyledon cells suggested that embryos and their cotyledon epidermal cells treated with 200 μmol/L TDZ were obviously larger in size than the control. Furthermore, TDZ promoted the upregulation of some key maternal tissue growth-related genes, including two G-protein signaling genes (AGG3 and RGA1) and two transcriptional regulators (ANT and GS2). The expression analysis of genes related to the auxin metabolic pathways, G-protein signaling, endosperm growth and transcriptional regulators confirmed that treatment with TDZ negatively regulated the key genes ABI5, AGB1, AP2, ARF2, and ARF18 during bud development stage and florescence. The results strongly suggested that TDZ might regulate the transcriptional levels of key genes involved in auxin metabolic pathways, G-protein signaling, endosperm growth and transcriptional regulators, which resulted in bigger cotyledon epidermal cells and seed size in B. napus. This study explored the mechanism of TDZ treatment on the seed size of B. napus and provided an important reference for improving rapeseed yield.Thidiazuron (TDZ) is a novel and efficient cytokinin commonly used in tissue culture, and numerous studies have demonstrated that TDZ can increase berry size. However, no study to date has explored the effect of TDZ on seed size of  Brassica napus  is not only an important oilseed but also a valuable biofuel. It is one of the main sources of natural edible vegetable oils in people’s daily life and has gradually become the green energy that people urgently need now. Nevertheless, owning to the dramatically increase of world population and sharply decrease in arable land area, although rapeseed has been widely cultivated all over the world, the supply of oilseed crop still can’t adequate to the production demand.B. napus, there are still few commercially available chemical reagents that can promote seed size, and the underlying mechanisms by which it regulates seed size remain unclear.Three direct determinants of oilseed rape yield include seed number per silique, silique number per plant and seed weight. Whether rapeseed productivity can be improved or not is greatly influenced by these components . In addi3 and TDZ treatments promoted fruit maturity and significantly increased berry cracking rate compared to the control treatment (Thidiazuron (TDZ) is a highly effective synthetic phenyl urea plant growth regulator, which not only has auxin-like activity, but also a cytokinin analogue . Nowadayreatment . TDZ hasArabidopsis was discovered to produce smaller seeds and organs. In contrast, AGG3 overexpression resulted in increased seed size, seed weight and seed number per plant (RGA1) in the heterotrimeric G protein complex has been found to be involved in the regulation of grain weight and cell function during plant development by coupling extracellular signals sensed by receptors  transcriptional repressors and the AUXIN RESPONSE FACTOR (ARF) are jointly involved in the auxin signaling process  is a transcription factor of the AP2-domain family members that mediates cell proliferation and growth control (ANT gene generated larger seeds (ANT promoter combined with ARF2 could directly regulate the expression of COLD-REGULATED15A (COR15a), and the mutants after knocking out cor15a displayed a smaller seed size phenotype, indicating that COR15A positively regulated seed size  is considered to be an important seed growth regulator through maternal tissues , APETALA2 (AP2) and TRANSPARENT TESTA GLABRA2 (TTG2) (AP2 members in the APETALA2/ethylene response element binding protein (AP2/EREBP) transcription factor superfamily, which are mainly expressed in floral organs, ovules and seed coat development , and seed weight was also predominantly reduced relative to the WT, the above results demonstrated that AfAP2-2 might be negatively correlated with seed size and weight  . There aelopment , 2009. Ad weight . In summg OsGRF4 . MicroRNin yield .B. napus was treated with different concentration TDZ in this research, and the morphology and physiology of treated plants were determined. Laser confocal microscopy was used to observe embryos and cotyledon cells. Quantitative Real-time PCR (qRT-PCR) was used to measure the transcriptional levels of genes associated with maternal tissue growth  and seed size . The results provided the evidence that a molecular mechanism by which TDZ regulated seed size in Brassica napus, which greatly benefitted to the improvement of rapeseed yield.To study the effect of TDZ on seeds, °36′N 112°08′E). The area belongs to the subtropical monsoon humid climate zone, with an average annual temperature of 15.9–16.6°C, annual rainfall of 2600–3100 mm, and annual sunshine hours of 1800–2000 h. Changyou No. 1 cultivar of B. napus was used material in this study, which was a hybrid between zhongshuang 11 and a material from Czechia. It is a winter-hardy variety with high yield which is extremely sensitive to light and temperature. Completely randomized block method was used with 3 replicates and the area of each block was 18 m2, with a planting density of 18 cm between the plant individuals and 25 cm between rows. Field management and pest and disease prevention were conducted based on local standard practices.The field trials were conducted during growing season of 2020–2021 in Agricultural Science and Technology Industrial Park of Yangtze University, Hubei province, China , silique length (cm) and seed number per silique were recorded. Thirty uniformly growing plants from rape treated with different concentrations of TDZ were selected for the determination of growth and yield indicators. Meanwhile all siliques of the selected plants were measured. In order to analyze the effect of TDZ on seed size, the main stem of plants was divided into three parts: top, middle and bottom, according to the branching characteristics of rape. Total number of seeds per plant was added up by using a knife to open all siliques from each plant and counting the number of seeds in it. Additionally, the number of existing seeds and empty shells number were counted. Seed setting ratio was determined as the number of existing seeds divided by the sum of the number of existing seeds and the empty shells number. Seed diameter was measured with vernier calipers. Twenty siliques were selected at the optimal concentration after TDZ treatment, and their silique lengths were measured with a ruler and photographed for comparison with the control. The seeds collected from each part of the plant were evenly mixed together and randomly selected 30 seeds from them to measure their seed diameter, then divided by the number of seeds to get the average seed diameter per plant. To calculate thousand-seed weight (g), 1,000 seeds per plant were randomly selected and weighed. Poured the seeds into the test tube to its 5 ml mark and weighed the seeds at this moment. Seed volume weight (g/ml) was determined as the seed weight divided by the volume of tick mark. The all seeds of each block was harvested separately and its yield was measured.A dissecting needle was used to dissect CK and seeds treated with 200 μmol/L TDZ under a microscope to isolate the embryos. Seeds of 10 days after flowering and mature seeds are treated in this way. Then, the embryos placed in DIC buffer  were incubated in 37°C constant temperature incubator overnight (at least 12 h), fixed with FAA fixative . The fixed embryos were dehydrated with 30, 50, 70, 80, 90, 95, and 100% ethanol (1 h for each stage) and then treated in Hoyer’s solution  for 1 h , 2009. T® II Q RT SuperMix for qPCR Kit  according to the manufacturer’s protocol.RNA prep Pure Plant Plus Kit  was used to extract the total RNA from flowers and buds under different concentrations of TDZ treatment according to the manual. In short, samples were ground with a grinder (MB-24) after freezing about 100 mg of flowers and buds in liquid nitrogen. The concentration and purity of RNA was evaluated using Ultramicro ultraviolet spectrophotometer  by determining absorption at 260 and 280 nm. Subsequently, 1 μg RNA was reverse transcribed into cDNA using HiScriptABI5), three transcriptional regulatory factors , three G-protein signaling genes  and two auxin metabolic relative genes (BnARF18 and ARF2). The nine genes were searched directly or BLAST in the BnPIR genome browser by using the protein sequences of Arabidopsis as a query. Then, Vector NTI software was used to design the specific primers for qRT-PCR accord to the genes sequences related seed development  was used to draw figures.The experiment was carried out following a completely randomized devise with four replications at different concentrations of TDZ treatment. The phenotypic data of seed diameters, silique number per plant, seed number per silique, thousand-seed weight and seed volume weight were subjected to the mean ± standard error. The statistical analysis of total data was determined by applying Duncan’s Test in DPS 8.50 . A significant difference was indicated when the B. napus at 40th day after full maturity is shown in p ≤ 0.05) (p ≤ 0.05) (p ≤ 0.05)  . Regardi ≤ 0.05) . Meanwhi ≤ 0.05) . For the ≤ 0.05) . AdditioPlants treated with TDZ showed basically consistent changes in the seed yield-related traits, including increases in thousand-seed weight, seed volume weight, the silique number per plant, yield and the seed number per silique relative to control. Thousand-seed weight is one of the important components of yield. The thousand-seed weight after treatment with 200 μmol/L TDZ was remarkably higher than control and the other concentrations of TDZ treatments. Additionally, as compared to control, the thousand-seed weight also significantly increased in the other TDZ treatment. As for seed volume weight, like thousand-seed weight, it also reached a maximum under the treatment of 200 μmol/L TDZ, while the effect of TDZ (400 μmol/L) treatment was statistically comparable to TDZ (10 and 100 μmol/L), but all of the seed volume weight treated with them was greater than that of control .p ≤ 0.05). Moreover, the 400 μmol/L TDZ treatment outperformed the other TDZ treatments (except for the 200 μmol/L TDZ treatment) both in terms of silique number per plant and yield (p ≤ 0.05)  . On the  ≤ 0.05) . The yieB. napus, the control had the most empty shells number and was significantly more than 10 μmol/L TDZ treatment. Additionally, in the siliques located in the bottom and various portion, the empty shells number in control was significantly higher than in 100 and 400 μmol/L TDZ treatments (p ≤ 0.05)  . NeverthB. napus . For theIn order to explore the reasons for the larger seeds, a cytological analysis of the seeds was performed. The embryos were isolated and observed. The size of embryo and cotyledon cells in the siliques at 20th day after TDZ treatment is shown in Epidermal cells in the middle of cotyledons treated with TDZ and control were selected respectively to measure the cell size. The results proved that TDZ treatments enlarged the epidermal cells of cotyledons. For the 20 days seeds after treatments, the average area of cotyledon epidermal cells was 1.24 times larger than the area of the corresponding control cells . The cotABI5 gene related to endosperm formation was significantly decreased in buds  in buds. In flowers, the transcriptional levels of AGG3 and ANT also reached a maximum at moderate concentrations (100 or 200 μmol/L) of TDZ treatment. In contrast, compared to that of control, the transcriptional level of the genes BnaA05.ABI5, AGB1 (except BnaC01.AGB1), AP2 (except BnaA03.AP2) and ARF18 reached a minimum when treated with TDZ at the same concentrations (100 or 200 μmol/L). Combined with the previous phenotypic and morphological results, it could be speculated that a medium concentration (100 or 200 μmol/L) of TDZ treatment may be the optimum concentration for increasing seed size. To summarize, TDZ might affect seed growth and regulate seed size by promoting or repressing the transcription of genes related to seed size.As shown in  in buds . Compareame buds . By cont in buds . Simulta in buds . OverallThidiazuron is considered as a substituted phenyl urea compound with strong auxin- and cytokinin-like activities. It has been proved to promote seed germination and stomatal formation on floral parts . What’s –1 TDZ increases berry width (–1) on small-fruited “Spadona” and “Coscia” pears resulted in a significant increase in fruit size  were identified in G-protein signaling, two genes (BnARF18 and ARF2) in auxin metabolic pathway, three genes  in transcriptional regulatory factors and an endosperm growth-related gene (ABI5). In order to more intuitively reflect the difference in the overall expression level of each gene between different tissues, we summed up the expression levels of genes located on different chromosomes. ARF2 and BnARF18. ARF2 was a suppressor of cell division, tissue development and organ growth, and it determined seed size and weight by regulating integument growth and development. Subsequently, ARF2 was shown to negatively regulate the expression of the homologous structural domain gene HB33, thereby controlling ABA-mediated primary root growth and seed germination (ARF2 could negatively regulate the expression of ANT after binding to the ANT promoter region, which in turn regulated seed weight and drought resistance by promoting cell proliferation (ARF18 was showed as a transcriptional repressor, which can activate downstream auxin genes to increase cell size, thereby regulating SL and SW (ABI5, AP2, and AGB1 (ABI5 has the effect of attenuating or terminating ABA signaling during seed germination, hence ABI5 can regulate seed dormancy, germination and seedling growth (Arabidopsis, ABA downstream signaling component ABI5 have been proposed to decrease the expression of SHB1 by combining with the SHB1 promoter region during early seed development, which caused the seed size increased in abi5 (AGB1 has been shown previously to function as a negative regulator in ABA responses (SIDREB3 also negatively regulated ABA responses in tomato and its over-expression reduced final fruit size (Arabidopsis, AP2 was shown to be a negative regulator of seed size and weight. When AP2 gene expression decreased, ap2 mutants produced abnormally shaped large seeds in which not only the seed size and weight increased, but also the number and size of embryo cells increased simultaneously due to endosperm cellularization and outward growth of the central endosperm vacuole (AP2 has a negative regulatory effect on seed size and weight. Additionally, AP2 genes have been found to have similar effects in other plant species, such as Larix Aechmea fasciata (AGG3, RGA1, ANT, GS2 were up-regulated by TDZ treatment. Prior research in rice highlighted the importance of RGA1 in dwarfism and set small seed, and this study identified RGA1 as a positive regulator of cellular proliferation and dwarfism in d1 was caused by a decrease in the number of cells (AGG3 in the control of seed and organ size in Arabidopsis, AGG3 overexpression resulted in increased seed size, fruit length, and number of seeds per fruit. Instead, AGG3 loss-of-function mutants had smaller seeds and reduced number of seeds per fruit (ANT gene is a key transcription factor for ovule development and organ growth in Arabidopsis. It has been proved that after overexpression of the ANT gene driven by the 35s promoter, ant mutants exhibit a larger seed phenotype due to cell expansion (GS2/OsGRF4 directly interacted with the transcriptional co-activator OsGIF1, the elevated expression levels of both resulted in increased cell size and number, which positively regulated grain size and grain weight, and significantly improved yield (A previous review showed that seed size is controlled by a combination of several different signaling pathways that regulate maternal tissues growth . Figure mination . A recenferation . In addiL and SW . Our resand AGB1 . ABI5 hag growth . In Arab in abi5 . The Gβ  in abi5 . AGB1 haesponses . The plaesponses . Interesuit size . In Arab vacuole , 2009. Tfasciata . Two G pof cells . Consister fruit . The ANTxpansion . Recentled yield .B. napus to varying degrees and greatly improve rapeseed yield. TDZ increased seed size by promoting cell elongation, cell division and regulating gene expression in several signaling pathways associated with maternal tissue growth. This study preliminarily explored the mechanism of TDZ treatment on the seed size of B. napus and provided an important reference for improving rapeseed yield.Thidiazuron treatment could increase the seed diameter and silique length of The original contributions presented in this study are included in the article/BX, JD, and XZ: designed the research. LZ: conducted the research and analyzed the data. JX: supplied the materials. LZ, LX, BX, JX, JD, and XZ: wrote and edited the manuscript. All authors scrutinized and corrected the manuscript.


Raw material analyses of prehistoric stone tool assemblages can reveal insight into mobility and exchange patterns in hunter-gatherer populations by reconstructing the circulation of stone throughout ancient landscapes. In Pleistocene Southeast Asia, stone tools are generally thought to have been fashioned from easily accessible local raw materials. However, despite the consistent presence of stone tools made of igneous raw material at prehistoric sites throughout the Tràng An Landscape Complex in northern Vietnam, there are no sources of igneous raw material in the immediate vicinity. This paper presents the results of geochemical sourcing analysis of late Pleistocene igneous stone tools from Tràng An: the first analysis of its type in mainland Southeast Asia. The results shed light on mobility and raw material provisioning strategies in Pleistocene mainland Southeast Asian hunter-gatherer populations and raise questions surrounding the relationship between technological organization, raw material, and expediency in Southeast Asian stone tool assemblages. Late Pleistocene stone tool assemblages recovered from sites in mainland Southeast Asia have a history of being discussed in the context of a prevailing archaeological technocomplex called the ‘Hoabinhian’ –6. Whileth century research suggested that rainforests were generally unsuitable for human occupation without the aid of agriculture [The analytical objectives of the paper are underpinned by theoretical considerations regarding our understanding of prehistoric tropical forager mobility. One of the primary goals of this research has been to gain insight into the mobility strategies of ancient hunter-gatherers at Tràng An and their potential wider implications. Two broad types of prehistoric hunter-gatherer mobility strategies have been prevalent in the literature since the 1980’s. In this brief opening review I explore archaeological correlates associated with each of them . The firiculture , 27. Howiculture –32. TherIn seasonal tropical mainland Southeast Asia, Shoocongdej  suggestschaîne opératoire for each assemblage. The application of pXRF represents a novel approach to the study of raw material sourcing and the still poorly understood nature of tropical forager mobility in Southeast Asia.Assemblage-wide investigations into tropical forager mobility in mainland Southeast Asia remain a rarity. With few exceptions, such as Marwick’s 2013 study tracking the relationship between technological variation and environmental change to test the validity of behavioral ecological models of forager behavior at Tham Lod and Ban Rai rockshelters in Thailand, the majority of studies , 15, 34 The Tràng An World Heritage Complex is located at the southern margin of the Red River (Song Hong) Delta, approximately 85 km south of Hanoi in Ninh Bình Province, Vietnam Figs  and 2. TPrevious analyses of lithic assemblages from Tràng An , 41–46 hIn order to investigate the source of this raw material, potential locales would need to fulfill two major criteria. The first is the presence of igneous stone. There are only two geological zones that are likely to have been a primary geological source in this case, as they encompass the only two igneous deposits that are accessible in the Red River Delta. The first is the Song Ma zone and the second is the Song Da rift structure . The sec2 wt% < 1.0) and high-Ti basalt (TiO2 wt% > 1.0). High-Ti basalt is considered to originate from enriched mantle sources whereas low-Ti basalts are thought to originate from depleted mantle sources [Geological studies of the southwestern Red River Delta divide igneous stone found there into two main categories based on geological formation. The Song Ma zone, created by the collision of the South China and Indochina blocks during the Indosinian orogeny in the middle Triassic , 48 and  sources .2 wt% > 1.0) basalt. Other candidate formations include the Song Ma formation (part of the Song Ma zone), accessible as close as 50 km west of Tràng An, and other outcrops of the Song Da rift structure that include low-Ti basalt  Figs , with a The Tràng An excavated material selected for geochemical analysis comprised 33 stone tools from Hang Trong (HTC), 87 from Hang Thung Binh 1 (TB1), and 37 from Hang Thung Binh 3 (TB3). These sites were selected because they contain the highest proportion of igneous stone tools and cover a wide range of dates. Radiocarbon dates presented in this section were calibrated with IntCal20 through the CALIB 8.20 program  unless oArchaeological material analyzed in this study is currently being stored between facilities at the Tràng An Visitor’s Center, Ninh Binh province, Vietnam and at the University of Cambridge, United Kingdom on authority of an agreement between the SUNDASIA Project, the Ninh Binh Provincial People’s Committee and the Tràng An Management Board. Each specimen has a unique identifier that is available in the data supplement . All arcCyclophorus shell midden context in trench 1, dated to 18,757–15,736 cal. BP (lab codes UBA-14886 and UBA-14884). Previous analyses of archaeological material from HTC suggests that it might have been used as a temporary stopping point or as a corridor between valleys in the complex [HTC is located in the central area of the Tràng An massif  and lies approximately 142 meters above sea level. It was first excavated in 2009–2010 and 2014 by the Tràng An Archaeological Project. There are three excavated trenches at HTC, and material from this study includes 32 specimens from trench 1 (center of the cave) and 1 from trench 3 . Specimens from trench 1 were excavated from contexts radiocarbon dated to between 37,676–18,115 cal. BP (lab codes UBA-35063 and UBA-14884). The specimen from trench 3 was excavated from a cyclophorid shell midden assumed to be contiguous with a  complex . The abuCyclophorus-dominated shell midden dated to between 18,634–12,739 cal. BP (lab codes UBA-40554 and UB-38671). Notable faunal evidence from TB1 includes Cervidae, Hystricidae, Panthera, and Geoemydidae [Cyclophorus shells paired with faunal evidence suggests that TB1 may again have been primarily occupied during the wet season, and that it served as a temporary hunting campsite. Igneous stone tools from TB1  is situated in an isolated karstic formation in the northwest corner of the complex, approximately 1 km west of the main massif and c. 27 meters above sea level. It was first excavated by the Vietnam Institute of Archaeology in 2012 and subsequently by the SUNDASIA Project from 2017–19 . Materiafrom TB1  are geneBrotia sp. shell, for which a calibration curve is currently unavailable. Specimens in this study were excavated from a Cyclophorus shell midden rich with other faunal elements, including marine and freshwater mollusks and larger mammalian fauna, notably Cervidae and Bovidae [TB3  is situated in the same karstic formation as TB1 and was excavated by the Vietnam Institute of Archaeology in 2012. Radiocarbon dates from TB3 indicate occupation from 15.5–13.5 uncal. BP. . These d Bovidae . The zooPortable X-Ray Fluorescence Spectroscopy (pXRF) is a technique used in geochemical composition analysis. It is particularly attractive for archaeological applications because it is non-destructive. While other techniques such as thin sectioning or LA-ICP-MS are more accurate, they inflict irreparable damage to specimens , 65. XRF2 and Fe2O3. Trace elements were calibrated as metals in parts per million (ppm) and included Ni, As, Rb, Sr, Y, Zr, Nb, Sn, Sb, Ba, and Th. A table of elemental readings is available in the data supplement . These readings were discarded prior to quantitative analysis.Major elements were characterized by recording three readings from different locations on each specimen for 180 seconds each. XRF machine settings were 15 volts/25 μA (microampere) with no filter inserted. For trace elements, three readings were taken from different locations for 60 seconds each. The XRF machine was set at 40 volts/12.7 μA with a 12 mm Al/1 mm Ti (yellow) filter to reduce background noise. The XRF machine was controlled by Bruker pXRF software, including X-Ray Ops (version 1.2.21) to manage instrument settings and S1PXRF (version 3.8.3.0) for spectrum collection, storage, and export. Raw readings were calibrated using USGS standards . Viable readings were then averaged into a single point for each specimen.Data analysis sought firstly to categorize specimens based on geochemical composition. Previous archaeological analyses of stone tools from Tràng An have classified igneous stone as rhyolite, basalt, and andesite, but macroscopic identification methods are rarely adequate for classifying raw material. When comparing raw materials for the purpose of geological source discrimination, it is important to compare the same types of raw material . The second goal was to identify patterns within each geological subgroup using cluster analysis. The final goal was to assess the hypothesis that igneous archaeological specimens could be attributed to the Cam Thuy formation.2 and K2O against the weight of SiO2. However, these elements are not reliably measured by pXRF, and are strongly affected by long-term weathering [For the first goal, a geochemical discriminant diagram was used to plot diagnostic elemental concentrations against each other to elementally categorize specimens in the study sample into distinct rock types. The most common geochemical discriminant diagram for the classification of igneous stone is the total alkali silica (TAS) diagram, which compares the combined weight of NaOathering . Insteadathering , 79 was athering . For thiPrincipal Component Analysis (PCA) is statistical clustering method used to simultaneously minimize information loss and increase interpretability of multivariate datasets by transforming large sets of variables into smaller components . It has For the third goal, boxplots and biplots were constructed to examine variability within geochemical subgroups at more specific resolution, and to compare elemental data from the study sample against known values (e.g. titanium wt%). Boxplots were used to present variability in single elements, and biplots were used to show clustering based on elemental ratios.Statistical analysis for this study was performed in R . Graphs The following sections will present 1) the geochemical classification analysis, 2) cluster analysis including all elemental data, and 3)specific diagnostic elemental data.2 ratios, placing them into the andesitic category. Specimens from the TB1 and TB3 sites are similar to one another, and largely cluster between the subalkaline basalt and alkaline-basalt categories. A chi-square test with Monte Carlo p-value simulation to adjust for low cell counts [When plotted on a Pearce diagram , specimel counts  indicateStudy specimens were separated into categories based on the Pearce W-F diagram, and further cluster analysis was conducted on elemental data within the alkali basalt, andesite basalt, and trachyandesite subgroups Figs –14. Whil2, Nb, and Zr  is potentially indicative of self-provisioning of raw material, though the relationship between non-local raw material and seasonal mobility patterns remains for the moment unclear. It is possible, for example, that non-local raw materials are indicative of logistic foraging expeditions to areas where igneous raw materials are available, or of large-scale residential mobility strategies during the wet season. As noted, subtler seasonally related mobility may also factor. Reasons for the investment involved in the transport of igneous cobbles and/or tools around the landscape remain unknown. However, it is possible that the use of rafts or small boats  significIt is also important to note that the observed technological differences between assemblages are not necessarily driven by deliberate behavioral decisions in the reduction process. Limestone at Tràng An is relatively hard and suitable for stone tool manufacture but frequently has substructural inclusions and is prone to fracturing unpredictably. Therefore, otherwise identical stone tool reduction methods might result in different observed outcomes for limestone and igneous cobbles. Further experimental research into the knapping characteristics of limestone and basalt, matched with the analysis of zooarchaeological and botanical remains will be necessary to help further explain the archaeological record in Tràng An and the impact that seasonality had more generally on the kinds of tropical mobility strategies hinted at in this study.There is a considerable degree of elemental variability between igneous specimens from Hang Trong and the Thung Binh caves, which is also reflected in macroscopic differences in color and texture. The former is older than both TB1 and TB3, and the results of this study might indicate changes in the circulation of stone, by whatever means, before and after the Last Glacial Maximum  , 46. HowPortable XRF has limited analytical resolution, and some lighter diagnostic elements that might further narrow down the geological source  are not detected well by this method . The appHistorical typological approaches defined mainland Southeast Asian stone tool assemblages by a series of diagnostic retouched core types. However, a growing body of evidence suggests that the small, expedient, and unretouched elements that dominate these assemblages are at least as important as the larger, retouched elements. Therefore, research into the lesser studied components of these assemblages has the potential to contribute significantly to a more comprehensive understanding of behavioral variability in prehistoric mainland Southeast Asia.This study has revealed that, despite the ubiquity of locally available technologically suitable raw material, the prehistoric inhabitants of Tràng An invested in obtaining igneous stone from a relatively long distance: perhaps as far as 85 km away. Despite this investment, they seemingly did not reduce this non-local stone in any significantly different manner than from local limestone. The reasoning behind this series of technological choices is unknown, though this study provides clear evidence that a notable level of behavioral complexity underlies the apparent simplicity of these tool kits. Further application of geochemical techniques will help identify more precisely the geographic locations of exploited igneous outcrops (and if these changed through time) and help to build a more detailed picture of forager mobility and technological decision-making that will have wider relevance for our understanding of Palaeolithic societies in this part of the world.S1 File(CSV)Click here for additional data file.S2 File(R)Click here for additional data file.


Olea europaea L.) reproductive buds as affected by cold accumulation, tree bearing status, and budburst temperature was studied under natural and controlled conditions, using both cuttings and container- and field-grown plants. The chilling necessary for dormancy release was acquired at different times within the bud population, presenting a progressive pattern of reproductive budburst. Once sufficient chilling is accumulated, 20 °C is a suitable temperature for reproductive budburst, although higher temperature, e.g., 30 °C, during dormancy release can inhibit budburst. While the bearing status of trees determined the amount of return bloom, dormancy release followed a similar pattern for previously bearing and non-bearing trees. Concurrent with investigating budburst factors, the use of shoot cuttings was tested as a method for olive dormancy release studies by contrasting with results from whole trees. It was found it to be valid for studying reproductive budburst, thus providing a useful method to screen chilling requirements in cultivar evaluation and the breeding programs currently ongoing in this species. However, the method was not valid for vegetative budburst, with varying results between cuttings and the whole plant.Dormancy release dynamics in olive tree ( Olea europaea L., reproductive development and its well-known biennial bearing cycle is axillary bud dormancy, so understanding the processes of olive bud dormancy onset, maintenance of dormancy, and dormancy release is critical for crop management, selection, and breeding. Furthermore, it is imperative to develop simple and efficient measurement tools to achieve these objectives. This knowledge is especially relevant in current times, with the spread of olive growing to new and climatically diverse areas of cultivation, as well as in considering the global threat of climate change [A key factor in olive tree, e change .Bud dormancy of deciduous fruit trees has been extensively studied for many years ,3,4,5. LA major difference with deciduous trees, in which reproductive differentiation occurs within the bud prior to the onset of winter dormancy, is that olive tree bud morphology remains undifferentiated as either vegetative or reproductive until dormancy release the following year, when bud reproductive induction and initiation are evident ,8. That Forcing, the placing of plants under beneficial growth conditions, is a useful technique to determine whether winter dormancy has been overcome. This procedure is consistent with the hypothesized entrance of the buds in ecodormancy , during Cuttings provide a valuable experimental system for forcing, as their smaller size in comparison to whole trees facilitates manipulation and repeatability within a relatively small space, especially desirable for trees, where the alternative of using potted trees in growth chambers requires substantial time, space, and financial inversion. In deciduous trees, forcing of cuttings has long been used to test whether winter dormancy has been overcome . In deciIn agreement with the overall hypothesis that olive tree reproductive budburst requires sufficient winter chilling followed by adequate warm temperature, the experiments presented here intend to characterize the temporal dormancy release pattern of olive tree reproductive buds. Different factors in overcoming this physiological stage, including chilling accumulation, tree bearing status, and the response of budburst to different spring temperatures are considered. Additionally, this study uses both cuttings and potted trees, providing the opportunity to test the use of cuttings as representing the behavior of whole trees.p = 0.0000), but less significant in A (p = 0.1191). For the ON trees, means comparisons of budburst for all dates of each treatment by the Tukey test showed a unique homogeneous group of means for As winter chilling increased under natural conditions in the field, progressively greater reproductive budburst occurred for the buds from all treatments, although with different timing and in different amounts. Reproductive budburst started at an earlier field-sample date for the buds on cuttings forced at 20 °C . Moreover, for each forcing temperature, reproductive budburst from ON and OFF trees started on the same sampling date, but under natural conditions appeared to be slightly delayed for ON trees compared to OFF. Still, although budburst percentage was much lower for ON than OFF trees, within each of the three types of observations  the overall temporal patterns were similar for ON and OFF  bud size for ON trees and OFF trees, and, consequently, no growth occurring between the two dates .In addition, no reproductive differentiation was visible at those times, and the first reproductive differentiation was seen at 11 February (ON) or 1 February (OFF) in the field, evidenced by the d0 observations . In a siForcing for 21 days at 30 °C did not yield reproductive budburst until the 11 February field samples C,F. ThesWhen potted trees underwent chilling in growth chambers, the results of forcing using cuttings and whole trees were similar for reproductive budburst, but not for vegetative budburst . That isIn contrast to flowering, results for vegetative budburst showed highly significant differences between the two forcing formats. In ON potted trees vegetative shoots formed from approximately 55% of the buds at 20 °C forcing and 25% at 30 °C forcing, after both periods of cold accumulation A. For thOur data illustrate the progression in the percentage of inflorescence emergence as cold accumulated during winter  and the The percentage of flowering buds was much higher for trees which had borne few or no fruit in the fall (OFF) than for trees which had born fruits (ON). Approximately 65% and 25% of buds formed inflorescences for OFF and ON, respectively, in the field study , and 60%When the buds were forced at 20 °C, reproductive budburst started in the 9 Jan samples, while at 30 °C, it was not observed until approximately one month later, in the 11 Feb samples . NonetheUnder the 30 °C forcing conditions, once budburst started A, bud grIn this study, no reproductive differentiation of the buds was observed until budburst conditions were fulfilled , consistThe results for progressive sampling dates  and inflIn the scheme presented in There is a period, between 21 January and 11 February in the conditions of this study, where dormancy release and budburst overlap. That overlap occurs due to the coinciding dormancy-release response to chilling unit accumulation and budburst response to accumulation of heat units, as may occur under natural temperature fluctuation and is demonstrated in modeling these processes ,28,29.In the upper part of Shoot-derived cuttings, by assuming dormancy release progresses in the cuttings as in the whole tree, provide a valid organ-level plant structure for testing temperature requirements of dormancy phases of olive reproductive buds. They have been used in dormancy release studies in deciduous fruit trees ,4,5,30, To best standardize the cutting methodology for studying olive reproductive budburst, our results highly recommend obtaining the cuttings from trees in OFF (non-bearing). Even though the temporal pattern for dormancy release was similar for ON and OFF trees, reproductive budburst levels were much lower or even absent in ON trees , while tIn contrast to reproductive budburst, the cutting system was not found to be consistent for determining vegetative budburst, as very large differences were observed compared to the buds on whole potted trees . VegetatThis study comprises two different experiments that consisted of an initial cold treatment of variable duration that was supplied either naturally  or artificially . Following different periods of cold treatment, the buds, present either on cuttings (both experiments) or on the potted trees (experiment 2), were forced in climatic chambers at 20° (standard temperature for bud growth once dormancy has been overcome) or 30 °C (a representative high Spring temperature). Temperatures and 21-day total forcing period were indicated by results of Ramos et al. . The expIn fall–winter season, six field-grown adult (seven-years-old) olive trees cv. Arbequina were chosen at the IFAPA  at the Alameda del Obispo Center in Córdoba, Spain, three in ON and four in OFF bearing status. Temperatures registered in the IFAPA weather station during the experiment were typical for the experimental site, ranging from 0.5 °C minimum to 20.8 °C maximum in January, and from 13.1 °C to 20.3 °C in February.v/v/v); Jensen [Testing budburst by forcing cuttings was carried out as follows. The olive-tree buds are located in the axils of the two opposite leaves at each node of current-year shoots . At succ; Jensen ) for mic; Jensen  were mad4) at 200 ppm for fungal control and placed in plastic trays containing perlite in the climatic chambers at 20 °C and 30 °C, with 12/12 h day/night photoperiod, and photon flux density of 200 μmol m−2 s−1. Ten cuttings per tree were evaluated for each of the six  sampling dates and two spring forcing temperatures (20 °C and 30 °C). In addition, fifteen similar shoots were tagged on each tree as controls at the beginning of the experiment to observe budburst under natural conditions.The cuttings were sprayed with copper sulfate  buds for 21 December and 21 January were measured in the central longitudinal sections, using an optical microscope equipped with image analysis . Eighty buds (20 buds per tree) for the four trees of each bearing status were measured as by . Width oThe experiment employed 32 three-year-old olive trees cv. Arbequina planted in 14 L pots with abundant flowering were selected. In spring, all inflorescences were removed from half of them for their use as non-bearing trees, while the inflorescences of the other 16 trees were left to set fruit and the trees used as bearing trees. On 1 September, all trees were placed in a climatic chamber at constant 10–12 °C temperature of for cold accumulation and the trees were split in two groups for two cold-treatment durations (8 weeks or 12 weeks). At the end of the cold treatments, ten shoots at least eight nodes long were selected from the central canopy zone of each tree. Five of these shoots were used to prepare 3-node cuttings as described for experiment 1 , and theExperiments 1 and 2 evaluated the reproductive budburst response to chilling accumulation in trees with either ON or OFF bearing status. In experiment 1, there were three (ON trees) and four (OFF trees) replications for each of the six sampling dates. In experiment 2, there were four trees and four groups of five cuttings for each of the eight forcing groups: two bearing levels, (ON or OFF), two periods of treatments (8 or 12 weeks), and two forcing temperatures (20 and 30 °C).For statistical analysis, we used the Statistix Program for Windows (Version 9.0). To determine the progression of reproductive budburst, we tested the data for normality using the Shapiro–Wilk test and then ANOVA carried out as data distribution was normal. In experiment 1, when differences were significant for sampling dates, separation of means between budburst dates was performed using the Tukey test, while in experiment 2, comparison of means was used.The results characterize the temporal pattern of dormancy release of olive tree reproductive buds, following chilling accumulation, and expand knowledge regarding factors involved. Using both natural (field) conditions and forcing at different temperatures in controlled chambers, the percentage of flowering buds was much higher for nonbearing than bearing trees but the pattern of reproductive budbreak over time was consistent. Once sufficient chilling has been accumulated, 20 °C forcing was confirmed suitable for normal reproductive budburst; when buds had just barely overcome dormancy, however, high (30 °C) temperature inhibited budburst, but that did not occur on later dates, after buds had initiated reproductive differentiation. We present an illustrative scheme to summarize the pattern of release from dormancy and reproductive budburst in olive buds.It is important to determine chilling requirements for dormancy release and subsequent flowering for the selection and breeding of appropriate olive genotypes, particularly to identify those with low requirements, and, thus, be able to maintain productivity under climate warming and in new, mild-winter growing areas. By comparison with forcing whole trees and with budburst in the field, forcing of cuttings was verified as suitable for testing olive reproductive but not vegetative budburst. The use of cuttings, representing valuable savings in time, space and financial inversion, can now be solidly recommended for olive tree cultivar selection and breeding. Furthermore, the results presented here not only provide specific information regarding experimental procedures, but also indicate relevant parameters, such as dates of onset and completion of budburst, and the length of the budburst period.


Pregnant women represent a potentially high‐risk population in the COVID‐19 pandemic.To summarize clinical characteristics and outcomes among pregnant women hospitalized with COVID‐19.Relevant databases were searched up until May 29, 2020.Case series/reports of hospitalized pregnant women with laboratory‐confirmed COVID‐19.PRISMA guidelines were followed. Methodologic quality was assessed via NIH assessment tools.Overall, 63 observational studies of 637 women (84.6% in third trimester) with laboratory‐confirmed SARS‐CoV‐2 infection were included. Most (76.5%) women experienced mild disease. Maternal fatality, stillbirth, and neonatal fatality rates were 1.6%, 1.4%, and 1.0%, respectively. Older age, obesity, diabetes mellitus, and raised serum D‐dimer and interleukin‐6 were predictive of poor outcomes. Overall, 33.7% of live births were preterm, of which half were iatrogenic among women with mild COVID‐19 and no complications. Most women underwent cesarean despite lacking a clear indication. Eight (2.0%) neonates had positive nasopharyngeal swabs after delivery and developed chest infection within 48 hours.Advanced gestation, maternal age, obesity, diabetes mellitus, and a combination of elevated D‐dimer and interleukin‐6 levels are predictive of poor pregnancy outcomes in COVID‐19. The rate of iatrogenic preterm birth and cesarean delivery is high; vertical transmission may be possible but has not been proved. Perinatal and maternal morbidity and mortality are increased in severe/critical COVID‐19 disease; there is a high rate of iatrogenic preterm birth and cesarean delivery. The infection has spread swiftly across the globe to 188 countries, affecting over thirteen million individuals with a case fatality rate of 5%, and was declared a pandemic by the WHO on March 11, 2020.The initial data on COVID‐19 outcomes in pregnancy were derived from small numbers of patients, first from China and then from Italy, with uncertainty regarding the extent to which these findings might be extrapolated to inform obstetric practice elsewhere in the world. Small international data sets have subsequently emerged, but these are often from different institutions with heterogeneous testing policies, obstetric management, and patient populations. Population‐level studies are therefore urgently required to provide robust data on the incidence of COVID‐19 in pregnancy and its effects on the pregnant woman, developing fetus, and newborn.The objective of the present review was to evaluate clinical characteristics and maternal, fetal, and neonatal outcomes among pregnant women admitted to hospital with laboratory‐confirmed SARS‐CoV‐2 infection by conducting a large global comprehensive review of data from various epicenters. By addressing practical issues surrounding the parameters that predict prognosis across the disease spectrum, the review aims to inform clinical practice and guide policy in maternity services for this and future global pandemics affecting pregnant women and those wishing to conceive.2The present systematic review was conducted in accordance with Preferred Reporting Items for Systematic Reviews and Meta‐Analyses (PRISMA) guidelines.The review included case series and case reports of pregnant women with SARS‐CoV‐2 infection confirmed by either quantitative real‐time polymerase chain reaction (RT‐PCR) or dual fluorescence PCR assessment. Unpublished reports, articles in which the date and location of the study were not specified, women with suspected COVID‐19 that was not confirmed by laboratory tests, and studies that did not report maternal, fetal, or neonatal outcomes were excluded.Studies were independently retrieved and reviewed for eligibility by two authors (OT and RK). The following data were extracted: Severity of COVID‐19, gestational age at presentation, presenting symptoms, laboratory parameters, admission to intensive care unit (ICU), need for Invasive mechanical ventilation (IMV), gestational age at delivery, mode of delivery, admission to NICU, stillbirth, neonatal death and maternal death.Data analysis, including prevalence and incidence levels, was performed by using SPSS version 26.0 . Categoric variables were expressed as number (percentage).The methodologic quality of studies was assessed by using National Institutes of Health quality assessment tools.33.1In total, 261 articles were identified in the initial search from March 26 to May 29, 2020; of these, 63 observational studies met the inclusion criteria  across all case series; 4 (14%) and 8 (29%) of case series reported high or moderate risk of publication bias, respectively. Subgroup analysis indicated that a high percentage of case series  adequately described the research questions and study population, and ensured that study subjects were comparable. Only 12 (43%) and 15 (54%) of case series fully described statistical methods and patient results, respectively. Consequently, reporting of statistical methods and full analysis of study data posed the greatest risk of publication bias.3.2The clinical characteristics of all 63 observational studies are documented in File Overall, 487 (76.5%), 101 (15.9%), and 49 (7.7%) were diagnosed on admission as having, respectively, mild, severe, and critical COVID‐19 disease, according to clinical criteria defined by the Chinese Centre for Disease Control and Prevention.Serum biochemistry results were reported in 37 studies encompassing 381 women respectively.Chest imaging findings were reported in 42 studies (n=274 women), and comprised 267 computed tomography (CT) scans of the chest, 15 plain chest X‐rays, and 5 CT pulmonary angiogram (CTPA) examinations.3.3The incidence of adverse maternal outcomes was assessed for all 637 pregnant women Table . The ratOverall, the incidence of maternal mortality was 1.6% (10/637); all of these women were admitted to the ICU with severe or critical COVID‐19 requiring IMV.Hemorrhagic complications were observed in 10 cases  of the 637 study women had delivered by the end of the study period, comprising 479 (98.6%) live births and 7 (1.4%) stillbirths, and 135 women were still pregnant. Six stillbirths occurred preterm between 24+0–36+6 weeks) and 318 (66.4%) term deliveries (≥37 weeks).+0–33+6 weeks) and 71 (59.7%) late preterm deliveries (34+0–36+6 weeks).Among the 479 women who had a live birth, there were 161 (33.7%) preterm deliveries  of the total number of cesareans performed. Among 119 preterm deliveries where information about mode of delivery and disease severity were available, the indication for delivery was ‘maternal SARS‐CoV‐2 infection’ in 59/119 (49.6%) of preterm deliveries, involving 10 early and 49 late preterm pregnancies among women with mild symptoms and no fetal or maternal complications. The remaining 60/119 (50.4%) preterm deliveries were performed owing to medical indications to aid maternal resuscitation among those with severe or critical COVID‐19. Of these 60 women, 28 (46.7%) had evidence of concurrent fetal distress. Among the preterm deliveries, spontaneous preterm labor complicated 25 pregnancies , of which 8 reported preterm prelabor rupture of membranes. The spectrum of severity of COVID‐19 among women who presented with spontaneous preterm labor included 14 (56.0%) mild and 11 (44.0%) severe or critical infections. All women who experienced preterm labor delivered by cesarean.Among the 318 term neonates, 76 were delivered vaginally and 242 were delivered by cesarean. The indication for cesarean was deteriorating maternal respiratory condition for 25 women, all of whom had severe or critical disease; fetal distress for 22 women (9 with severe and 13 mild disease); failure to progress in labor for 19 women; previous cesarean for 15 women; prolonged rupture of membranes for one woman; and pre‐eclampsia for one woman. In 159 (65.7%) cases, cesarean was performed because of COVID‐19 among women with mild disease and no fetal and maternal compromise.Adverse neonatal outcomes, including admission to the neonatal ICU (NICU), neonatal death rate, and vertical transmission were assessed among 479 live births; in addition, Apgar scores were reported in 29 studies comprising 361 newborns  of newborns were admitted to the NICU. Of these, 3 were delivered by mothers with mild COVID‐19, and 51 were delivered by mothers with severe or critical infection.+5 gestational weeks. Another two deaths were twins who were delivered at 28 gestational weeks due to signs of fetal distress. They died on day three of life owing to complications of prematurity, despite normal blood parameters and chest radiographs. Their mothers also died from complications of critical COVID‐19 infection. The last two neonates were born with Apgar scores of 1 at 1 and 5 minutes after cesarean delivery due to signs of fetal distress among women with critical disease. The neonates died on days 0 and 1 of life, respectively. All five neonates who died had negative nasopharyngeal COVID‐19 swabs.Among the 479 live births, the incidence of neonatal death was 1.0% (n=5).Of the 479 live newborns, 405 were tested for COVID‐19 via naso‐ and oropharyngeal swabs and 8 (2.0%) positive swabs were detected, obtained between 16 and 48 hours of life.4To our knowledge, the present review is one of the largest international systematic reviews of published studies on COVID‐19 infection in pregnancy, stratified by risk according to severity of COVID‐19 disease and timing in pregnancy. The review has assessed outcomes across a latitude of geographic locations, including Asia (mainly China), Europe, and the United States, from the start of epidemic in Hubei Province to its eventual reach as a global pandemic. The review provides sufficient data to support previous findings that pregnancy is not associated with a more severe course of infection overall. However, women in their third trimester and those with co‐morbidities are at particular risk of developing critical infection. This review also provides lessons from the high rate of obstetric interventions due to concerns of worsening maternal condition, potentially resulting both in cesaerean deliveries that are not indicated and in iatrogenic preterm birth with associated adverse neonatal outcomes among women with only mild COVID‐19.Most pregnant women admitted to hospital had mild COVID‐19 with good outcomes, and only approximately 24% had severe or critical disease. The latter two subgroups accounted for the majority of adverse maternal, fetal, and neonatal outcomes. All maternal deaths, consumptive coagulopathies, and thrombotic complications occurred among women with severe and critical disease. Similarly, all instances of neonatal death, and NICU admissions and six of seven stillbirths occurred among neonates of mothers with severe or critical COVID‐19. These findings support a policy of safely managing pregnant women with mild infection in an outpatient setting to avoid unnecessary hospital admission during the flux of a pandemic with stretched healthcare services. Nevertheless, given that 3% of women with mild COVID‐19 in the present study deteriorated to a more severe infection, it is imperative that these women continue to be regularly reviewed and have access to acute medical care.The review found that 9.6% of women were admitted to ICU and the overall maternal mortality was 1.6%. In a recent report from the US Center of Disease Control and Prevention of COVID‐19 among women aged 15–44years, 31.5% of pregnant women were hospitalized as compared with 5.8% of non‐pregnant women.COVID‐19 is a novel infection with rapidly evolving scientific data regarding its management. This has a significant impact on the psychosocial wellbeing of women during pregnancy. In a study from Ireland, 71 pregnant women expressed anxiety about the wellbeing of their family and 63% were concerned about the wellbeing of their unborn baby.Coagulopathy and thromboembolism are now recognized complications of severe COVID‐19 and are associated with poor outcomes.There remains a paucity of data on the impact of COVID‐19 on pregnancy in the first and second trimester. The present review, in line with UK Obstetric Surveillance data,The data reveal that a high proportion of severe (~60%) and critically unwell (~80%) pregnant women had both raised D‐dimer and raised IL‐6 levels, which are biomarkers of abnormal coagulation and molecular inflammation, respectively. This may have prognostic value in predicting COVID‐19 severity and poor outcomes, especially as these factors have been implicated in orchestrating the cytokine storm observed in critically unwell non‐pregnant patients with the infection.During the early surge of COVID‐19, there was a lack of information on the vertical transmission potential of SARS‐CoV‐2, resulting in understandable anxiety among women and obstetricians. This is reflected in the high rates of preterm birth  and cesarean delivery in the review. Although deteriorating maternal condition and fetal distress accounted for ~50% of all preterm births, the remainder seemed to be iatrogenic among women with mild COVID‐19 with no maternal or fetal compromise. It seems unlikely that COVID‐19 itself increases the risk of spontaneous preterm labor; the rate in the review was approximately 4%  as compared with a rate of 5%–8% reported for the general population.The review demonstrated that COVID‐19 increases perinatal mortality with an overall stillbirth rate of 1.4% and neonatal death of 1.0%. Stillbirth and neonatal mortality rates are 0.42%The possibility of vertical transmission of SARS‐CoV‐2 virus has been widely debated and the picture remains unclear. In the present study, only 8 (2%) of 405 neonates who underwent naso‐ and oropharyngeal PCR testing for SARS‐CoV‐2 virus had a positive test. Some neonates were also found to be positive for throat swabs immediately after cesarean delivery, despite measures to isolate the neonate from the mother on delivery. Recently, the visualization of SARS‐CoV‐2 virions in syncytiotrophoblasts and microvilli of the placenta suggests that the placenta, which is known to express the ACE2 receptor (the putative receptor for SARS‐CoV‐2), may be a target for virion entry.A major strength of the review is the inclusion of data from a large number of pregnant women with laboratory‐confirmed SARS‐CoV‐2 infection from various regions of the world, including Asia, Europe, and the United States. However, the analysis also has limitations because COVID‐19 management decisions, as well as variations in healthcare resources, differ from country to country. In addition, datasets were obtained from retrospective observational studies that are prone to recall and/or misclassification bias, and limited the ability to explore risk factors. However, the overall risk for publication bias was moderate and more than 85% of the case series in the review adequately described the research questions and study population, and ensured that study women were comparable.In conclusion, as our collective understanding grows, we believe that obstetricians will become less interventional, especially in preterm gestations, when managing mild COVID‐19 disease, thereby reducing the burden of iatrogenic neonatal morbidity. The data support previously documented findings of reassuring maternal outcomes for mild COVID‐19 infection, with poor maternal and fetal outcomes among those with severe or critical disease, predisposed by obesity, diabetes mellitus, advanced maternal age, and advanced pregnancy. This, we believe, should inform the counselling of women diagnosed with COVID‐19 in pregnancy.OT was responsible for conceptualization, project administration, investigation, methodology, validation, data curation, formal analysis, and drafting and revising the manuscript. AH was responsible for conceptualization, methodology, formal analysis, data curation, and drafting and revising the manuscript. PD was responsible for investigation, validation, and drafting and revising the manuscript. WJL was responsible for investigation, validation, and drafting and revising the manuscript. AW was responsible for and drafting and revising the manuscript. RA‐K was responsible for conceptualization, methodology, validation, supervision, and revision of the manuscript.The authors have no conflicts of interest.File S1. Clinical characteristics of pregnant women at initial presentation of COVID‐19 disease.Click here for additional data file.


Sepsis is a severe disease with high mortality, and liver injury is an independent risk factor for sepsis morbidity and mortality. We analyzed co-differentially expressed genes (co-DEGs) to explore potential biomarkers and therapeutic targets for sepsis-related liver injury. Three gene expression datasets  were downloaded from the Gene Expression Omnibus (GEO). DEGs were screened between sepsis and control samples using GEO2R. The association of these DEGs with infection and liver disease was analyzed by using the CTD database. GO functional analysis, KEGG pathway enrichment analysis, and protein-protein interaction (PPI) network analysis were performed to elucidate the potential molecular mechanism of DEGs. DEGs of different tissues in GSE60088 were analyzed again to obtain specific markers of septic liver injury. Mouse model of sepsis was also established by cecal ligation and puncture (CLP), and the expression of specific markers in liver, lung, and kidney tissues was analyzed using Western blot. Here, we identified 21 DEGs in three datasets with 8 hub genes, all of which showed higher inference scores in liver diseases than bacterial infections. Among them, only TNFRSF1A had a liver-specific differential expression. TNFRSF1A was also confirmed to be specifically reduced in septic liver tissues in mice. Therefore, TNFRSF1A may serve as a potential biomarker for septic liver injury. Sepsis is an infection-induced systemic inflammatory response syndrome. It is a common complication of patients with severe trauma, shock, and critical diseases and a leading cause of death for patients in the intensive care unit (ICU). The occurrence of sepsis is associated with the hypersensitivity cascade and excessive inflammatory mediators –3. IdentMicroarray technology and bioinformatics analysis have been widely used to screen for genetic alterations at the genome level, contributing to the identification of differentially expressed genes (DEGs) and functional pathways in disease models. Analysis of microarray data in a septic liver injury model can build gene networks and screen for potential key molecular targets to provide a new understanding of the pathogenesis of septic liver injury and a potential strategy for clinical treatment , 17. In https://www.ncbi.nlm.nih.gov/geo) is a public functional genomics repository containing global gene expression data and microarrays [GEO  was used to screen significant DEGs between sepsis and control liver samples. The application of P-values and Benjamini and Hochberg false discovery rates provided a balance between the discovery of statistically significant genes and false positive limits. The probe sets without any gene symbols or genes with multiple probe sets were excluded or averaged, respectively. A fold change (logFC) >1 and P-value <0.05 were considered statistically significant.GEO2R (https://www.string-db.org) to retrieve gene interactions [PPI networks were predicted using an online database searching tool  is an online bioinformatics database that integrates biological data and analysis tools to provide complete annotation information of functional genes and proteins for users [P < 0.05 was considered statistically significant.The Database for Annotation, Visualization, and Integrated Discovery 6.7  is a public resource that describes the interactions between environmental chemicals and gene products and their relationship to disease [The Comparative Toxicogenomics Database  and CLP (n = 3) groups. The sepsis model was induced by CLP as previously described [μm thickness, and stained with hematoxylin and eosin. The pathological sections were randomly observed by pathologists to analyze the pathological damage in each group. The animal study was performed following the Guide for the Care and Use of Laboratory Animals and approved by the ethics committee of the Xijing Hospital .Male C57BL/6 mice  from the Fourth Military Medical University  were housed under standard laboratory conditions. Mice were randomly divided into sham . BCA protein assay kit  was used for protein quantification. Certain amounts of protein were loaded onto the SDS-PAGE gels and then transferred to PVDF membranes for Western blot analysis. After blocking, the membranes were incubated with the anti-TNFRSF1A primary antibody at 4°C overnight, followed by incubation with diluted HRP-conjugated secondary antibody  for 1 h at room temperature. Blots were visualized with ECL-Plus reagent . t-test was used. All animal experiments had at least three replicates. A value of P < 0.05 was considered statistically significant.Data analysis was performed using GraphPad Prism software . Student's After normalization of the microarray results, there were 364, 1,030, and 665 DEGs identified in GSE60088, GSE23767, and GSE71530, respectively . The oveWe further analyzed the functional and pathway enrichment of the eight DEGs through DAVID. Results from GO functional analysis indicated that the alteration of BP was mainly on the inflammatory and immune responses . The chaα, mainly participates in the mTOR, MAPK, caspase 3, and NFκB pathways that were related to the regulation of inflammatory responses and cell apoptosis (The eight DEGs were compared with the DEGs in the septic liver, kidney, and lung tissues of GSE60088, and it was found that only TNFRSF1A had a specific expression change in the liver . Combinepoptosis .Despite extensive research in sepsis, there are still few biomarkers that can be used to effectively detect and treat sepsis . Liver hMicroarray assay is an effective method to screen novel biomarkers of disease and find genetic alterations in disease progression, which has been proved to be applicable in the study of septic biomarkers and organ damage . In the α can be exerted through two different receptors belonging to the TNF receptor superfamily. The type I receptor is TNFRSF1A, also known as p55, p60, CD120a, or TNFR1. TNFRSF1A is a 60 kDa transmembrane glycoprotein and expressed in almost all cells except for erythrocytes, but type II receptor mainly exists in immune cells, endothelial cells, and cells of the hematopoietic lineage. Both receptors can be activated by transmembrane TNF-α, but TNFRSF1A can also have functions by soluble TNF-α [Staphylococcus aureus infections and is associated with the bacterial clearance from the spleen [However, these DEGs were not liver-specific. Through further analysis of dataset GSE60088 that contained DEGs of the septic liver, lung, and kidney, it was found that only TNFRSF1A was differentially liver-specific expressed among the 4 key genes. The effects of TNF-le TNF-α . The actle TNF-α , 35. Sevle TNF-α . In addile TNF-α . In sepse spleen . The sige spleen .−/− mice in sepsis was comparable with wildtype mice [κB pathways to control cell apoptosis, which maybe the inner mechanism of TNFRSF1A regulating liver immune defense and immunity adjustment. Our study indicated that not only the TNFRSF1A membrane protein in septic liver cells significantly decreased but also its gene expression had a specific decline. Its gene regulation mechanism remains to be investigated. Only by further exploring the duration, concentration, and related inflammatory factors of TNFRSF1A in the immunoregulation of sepsis, can we better understand its role in the occurrence and development of septic liver injury and provide potential insights and targets for the diagnosis and treatment.In our experimental study, TNFRSF1A expression was detected in the liver, lung, and kidney tissues of sepsis mice, and only TNFRSF1A in the liver was significantly downregulated, which was consistent with the bioinformatics analysis. Hepatocytes from septic mice developed extensive vacuolar degeneration and nuclear rupture, which were closely related to the cytokine storm and inflammatory cascade triggered by sepsis. Membrane TNFRSF1A induces cellular inflammatory damage and apoptosis by participating in mTOR, JNK, IKK, caspase 3, MAPK, and NF-kB pathways. Therefore, the decreased TNFRSF1A protein expression in the liver tissue of the sepsis model combined with the downregulation of gene expression verified that TNFRSF1A may serve as a specific biomarker of septic liver damage and liver immunoregulation. Although the mortality of TNFRSF1Aype mice , it is sype mice , so inteIn conclusion, our study showed that TNFRSF1A is closely associated with sepsis-induced liver injury, which provides a potential diagnostic signature for septic liver injury and a basis for exploring the roles the liver plays in defense homeostasis during sepsis. TNFRSF1A may serve as an intervention target to alleviate and treat sepsis in the future.