To obtain metabolite concentrations specific to the putamen, we investigated a correction method previously proposed to account for the tissue composition of the volume of interest. We compared the method with the conventional approach, which a priori assumes equal metabolite concentrations in GM and WM. Finally, we compared the concentrations acquired at 3 Tesla (T) and 7 T MRI scanners. Spectra were acquired from 15 subjects (age: 67.7 ± 8.3 years) at 3 T and 7 T, using an ultra-short echo time, stimulated echo acquisition mode sequence. To robustly estimate the WM-to-GM metabolite concentration ratio, five additional subjects were measured for whom the MRS voxel was deliberately shifted from the putamen in order to increase the covered amount of surrounding WM. The concentration and WM-to-GM concentration ratio for 16 metabolites were reliably estimated. These ratios ranged from ~0.3 for γ-aminobutyric acid to ~4 for N-acetylaspartylglutamate. The investigated correction method led to significant changes in concentrations compared to the conventional method, provided that the ratio significantly differed from unity. Finally, we demonstrated that differences in tissue voxel composition cannot fully account for the observed concentration difference between field strengths. We provide not only a fully comprehensive quantification of the neurochemical profile of the putamen in elderly subjects, but also a quantification of the WM-to-GM concentration ratio. This knowledge may serve as a basis for future studies with varying tissue voxel composition, either due to tissue atrophy, inconsistent voxel positioning or simply when pooling data from different voxel locations.The aim of this work is to quantify the metabolic profile of the human putamen  It has proven to be a valuable tool for the assessment of several pathologies and disorders of the human brain, such as tumours  , ƒi are the tissue volume fractions within the VOI, and ciw, are the tissue water concentrations . Here we assume cw,WM = 36100 mM, cw,GM = 43300 mM, and cw,CSF = 53800 mM  and cGM and cWM are free parameters. Clearly, a sufficiently broad φGM range needs to be covered to achieve a robust estimation of cGM and cWM. However, most studies seek to position the VOI in a consistent manner throughout the subjects, inevitably leading to a rather small φGM range. Consequently, some works have opted to pool the data acquired from different voxel locations to achieve a more robust estimation of αm [The pure WM and GM metabolite concentrations, and consequently the concentration ratio, ƒGM+ƒWM) . The linon of αm , 33.αm, we evaluated the relative change, Δαm, as followsb can be either T1, T2, cw,WM or cw,GM and the subscript “0” denotes the unbiased counterpart. For the sake of simplicity, an equal bias in the water relaxation times for both WM and GM was assumed. Notice that due to the intrinsic assumption of non-distinct metabolite relaxation times between WM and GM in αm.In order to consider the effect of a potential bias in the water relaxation times or tissue water content used in Eqs  and 2)  2) in αm2, we defined the error, ε, in the absolute concentration if the concentration ratio used, i.e. αm, differs from the actual concentration ratio, αm,0, as follows:Finally, to investigate the way in which a possible bias in the estimated concentration ratio propagates into the metabolite concentration obtained following MφGM in the MRS voxels of that study, five more age-matched subjects (three VOIs each) were measured. For these subjects, the VOI was deliberately shifted to include higher fractions of WM (see Section 2.4 MR protocol). The Parkinson’s disease study was approved by the Ethics Committee of the Medical Faculty of the University of Cologne, Germany. The five extra subjects were measured under the approval of the Ethics Committee of the Medical Faculty of the RWTH Aachen University, Germany. All participants gave written informed consent according to the Declaration of Helsinki.MRS spectra were acquired from the putamen of 15 healthy subjects  in the framework of a case-control Parkinson’s disease study . Due to 3; matrix-size = 176 × 232 × 256; flip angle, 7º. At 7 T, the MP2RAGE sequence [3; matrix-size = 208 × 300 × 320; flip angle, 5º.For all subjects, spectra were initially acquired on a Siemens 3 T hybrid PET/MR Tim Trio scanner  and then on a Siemens 7 T Terra scanner. The 3 T scanner was equipped with a birdcage transmit coil and an 8-channel receive coil provided by the vendor, whereas the 7 T scanner used a single transmit coil with a 32-channel receive coil . The MPRAGE sequence  was usedsequence  was util3; receive-bandwidth, 2000 Hz; vector-size = 2048; flip angle = 90°; RF pulse duration = 1920 μs. At 7 T, the protocol parameters were: TE / TR / TM = 4 / 8000 / 28 ms, 72 averages; voxel-size = 14 (left-right) × 32 (anterior-posterior) × 17  mm3; receive-bandwidth, 6000 Hz; vector-size = 2048; flip angle = 90°; RF pulse duration = 1920 μs. One extra complete phase cycle was measured without the water suppression RF pulses for eddy-current correction and absolute quantification. The same 3 T and 7 T MRS protocols were also used for the five additional subjects, for whom three different voxel positions were measured, namely occipital (WM1), parietal (WM2), and frontal (WM3) WM  WM .myo-inositol (Ins), lactate (Lac), NAA, N-acetylaspartylglutamate (NAAG), phosphocreatine (PCr), phosphorylcholine (PCh), phosphorylethanolamine (PE), scyllo-inositol (Scyllo), taurine (Tau). Macromolecular spectra measured using the inversion recovery STEAM sequence at 3 T (TE = 8 ms) and 7 T (TE = 6 ms) available at MM Consensus Data Collection (https://github.com/mrshub/mm-consensus-data-collection), were additionally used in the basis sets [All data sets from the 3 T and 7 T scanners were processed following the same pipeline with the help of the FID-A package , availabsis sets , 46. Thesis sets , 48. Onlsis sets , 48.αm-ratio, is the tissue segmentation approach [T1-weighted images in this work, due to its limited contrast with WM. Therefore, given that the putamen VOI in our study contains both cortical and subcortical GM, in order to assess the tissue volume fractions, we utilised a combined approach in which FAST was used for the segmentation of cortical GM, WM, and CSF, and FIRST [An important issue in metabolite quantification, as well as in the estimation of the approach . In contapproach . Traditiapproach , face prnd FIRST  was usedαm-ratio, the distributions of metabolite concentrations evaluated using M1  were compared between the scanners. If cm,1 concentrations at 3 T vs. 7 T were not significantly different, then the pooled cm,1 concentrations (3 T and 7 T) were used for the estimation of the αm-ratio. Otherwise, 3 T and 7 T cm,1 were considered separately, and the αm-ratio was estimated for each field strength. A one‐way analysis of covariance (ANCOVA) was used to compare the cm,1 concentrations between the scanners, where φGM at each scanner was used as the covariate. This was performed with the help of the SPSS software (28.0.1.0 (142)).Prior to the estimation of the 1 and M2 were compared for each field strength separately. To this end, a two-sample t-test was performed for each group of subjects. Additionally, a scanner comparison was performed for each of the correction methods utilising a two-sample t-test.The metabolite concentrations obtained using the correction methods Mαm-ratio method (M2) compared to the case of neglecting it (M1) was performed by calculating the Pearson’s correlation coefficient of the concentrations cm,1  vs. φGM. While a significant correlation of cm,1 vs. φGM for a given metabolite denotes that αm ≠ 1 (i.e. different WM and GM concentrations), a lack of a correlation of cm,2 vs. φGM for the same metabolite signifies that the use of method M2 can indeed remove the dependence of the absolute metabolite concentration on the tissue voxel composition.A further evaluation of the effect of using the r = 0.83; p < 10−7. The mean linewidth values for the putamen, averaged across subjects, were 0.077 ± 0.008 ppm (3 T) and 0.07 ± 0.01 ppm (7 T), 0.052 ± 0.005 ppm (3 T) and 0.05 ± 0.01 ppm (7 T) for occipital WM, 0.064 ± 0.008 ppm (3 T) and 0.054 ± 0.009 ppm (7 T) for parietal WM, and 0.057 ± 0.004 ppm (3 T) and 0.05 ± 0.01 ppm (7 T) for frontal WM.A mean of 6.9 averages (5.4% of the number of averages) were discarded from the total number of averages at 3 T, whereas at 7 T, a mean of 2.4 (3.3%) averages were discarded. The spectral characteristics of the putamen VOI at 3 T (left column) and 7 T (right column) are depicted in 1  between field strengths. Therefore, the αm-ratio values for these metabolites were estimated separately for each field strength . The putamen mean and standard error of the mean (SEM) metabolite concentrations calculated with the correction method M2 are also summarised in αm evaluated using b were evaluated according to Δb = (100% × (b − b0) /b0. The parameter Δαm was assessed for several values of ΔT1, ΔT2, Δcw,WM and Δcw,GM ranging from -20% to +20%. For the case of the water relaxation times, Δαm was evaluated for a range of TE, TR, and TM values  and p-values of cm,1 and cm,2 vs. φGM for each metabolite. As expected, concentrations cm,1 show a high correlation (p < 0.05) with φGM, for metabolites whose WM and GM concentrations are notoriously different. However, cm,2 concentrations show no correlation with φGM (p > 0.05) in any metabolites, highlighting the ability of M2 to remove the dependence of the concentration on φGM.ε , 1.0 (b) and 2.0 (c) are assumed. The error is shown for relative GM fractions with φGM ranging from 0.5 to 0.95. ε as a function of φGM assuming actual concentration ratios of αm,0 = 0.3 (d), 1.0 (e) and 2.0 (f). Several values of αm are considered, ranging from an underestimation equal to -50% to an overestimation of +50% of the actual concentration ratio αm,0. We observed that ε increases as the relative fraction φGM decreases. It is also clear that, for a given αm,0, the error tends to be smaller if αm > αm,0 than if αm < αm,0 by the same amount. In other words, it is preferable to overestimate the concentration ratio used in M2 than to underestimate it . Note aαm,0 > 1  comparedαm,0 < 1 . That is1 and M2, for 3 T (red) and 7 T (blue) scanners. In the case of metabolites where αm differed significantly from unity, we observed that M2 led to significantly different concentrations  compared to M1. It is, however, important to note that another factor influencing the significance of the difference between M1 and M2 is the inter-subject variability of the metabolite concentration. Notice that the M2 method for the case of Gln, tCr, and Glx was evaluated using the corresponding separate αm-ratio values (either 3 T or 7 T), based on the observed significant difference of the cm,1 concentration distributions between field strengths. For the rest of the metabolites, the αm-ratio values obtained using pooled data were used. Regarding the comparison between the scanners, Asp, tNAA, tCr, Glx, and Gln showed a significant difference between the field strengths  for both correction methods. The relative difference between the concentrations calculated with M2 and M1, Δcm = 100% × /cm,1, is plotted in αm-ratio values for the whole metabolic profile.1 method have already been broadly discussed in the literature [αm-ratio method has only been investigated for the case of GABA in a few works [αm-ratio method, with the putamen being the anatomical target. Finally, concentrations quantified at field strengths of 3 T and 7 T were compared as a means of validation.We have provided a thorough quantification of the metabolic profile of the human putamen in a cohort of healthy, elderly subjects. Although the features covered by the Mterature , 33, 58,ew works , 22, 59.The data quality for the 3 T and 7 T scanners, as assessed via the linewidth of the unsuppressed water peaks, reflected a strong, positive correlation between the scanners . Furtherin vivo experiments [in vivo experiments [The use of STEAM enabled the data acquisition at ultra-short TE, which in turn has the advantage of providing i) high SNR spectra and ii) low spectral distortion of multiplets. The combination of these features facilitated the reliable detection and quantification of 12 metabolites and four total concentrations, as reflected by the low CRLB values , 24, 39.eriments , 64. Fureriments  and in veriments , 64. Fineriments , 65 and Conversely, a drawback of short-TE sequences is the presence of strong macromolecular background peaks that complicate spectral fitting . To overαm due to any potential inaccuracy in the relaxation times used for water and metabolites in Eqs ..αm due t0% in WM , whereasearly 8% . Regardiy 8% [T2 , 71. On ghly 18%  while otghly 18%  or even ghly 18% . Given ob et al.  reportedt et al.  also obsThe impact of the intrinsic assumption made in Eqs  and 2)  2) that αm-ratio values, we first utilised ANCOVA to compare the cm,1 vs. φGM distributions at 3 T and 7 T. A lack of significance in the difference indicated that the cm,1 vs. φGM distributions could be pooled for the estimation of a single αm-ratio. On the contrary, a significant difference in the cm,1 vs. φGM distributions suggests that the αm-ratios at 3 T and 7 T could diverge due to reasons other than just differences in the tissue voxel composition. However, even though the cm,1 vs. φGM distributions were significantly different for Gln, tCr, and Glx, the αm-ratio values were comparable. The αm-ratios measured here were generally in line with the αm-ratio values calculated using literature WM and GM metabolite concentrations . Given that the observed changes in metabolite concentrations due to activation primarily take place in GM, it is expected that, as per the definition of αm, changes in αm will also occur as a result of brain activity. Starting from the derivation of c = fGM ΔcGM and therefore |Δc| ≤ |ΔcGM|. Hence, the change in the metabolite concentration between activation and resting state, as assessed using αm-ratio. However, studies have shown that in the presence of some stimuli, the concentration of metabolites, e.g. Lac, can fluctuate as much as 30% within the VOI [αm-ratio within the framework of fMRS is worth considering for future studies.A context in which the  the VOI . Therefoαm-ratio for the whole metabolic profile for the different brain regions becomes paramount and emphasises the importance of our results.It is worth mentioning that we have not made use of the tissue-correction approach normalised to the study-specific voxel composition, as originally proposed by Harris et al. (Eq (6) in reference ). The reA limitation of this work is the fact that the metabolite basis sets used in LCModel were simulated using ideal RF pulses. However, while real RF pulses with localisation gradients are preferable, the improvement is not expected to be substantial for the case of STEAM, as shown by Kaiser et al. .αm-ratios, as shown in A further important limitation of this work is that the tissue water concentration was assumed based on literature values, which normally correspond to the case of healthy, young volunteers. However, it has been shown that the tissue water concentration can be altered, not only in the case of brain pathologies, e.g. hepatic encephalopathy , stroke αm significantly differs from unity. Furthermore, the investigated method was able to remove any dependence of the putamen metabolite concentration on the tissue voxel composition. Finally, not only have we provided a quantification of the neurochemical profile of the human putamen, but also the αm-ratio of the metabolite concentration profile with the surrounding WM. Our data may potentially serve as a reference for the classification of the degree of metabolic changes in the putamen within the framework of neurodegenerative diseases.We have investigated a method for the quantification of the neurochemical profile of the brain using MRS, which accounts for differences in the metabolite concentrations of the different tissues comprised in the voxel. The method was applied to quantify the neurochemical profile of the human putamen in a cohort of healthy elderly subjects using STEAM MRS at 3 T and 7 T. The data were then compared to the metabolite concentrations obtained using the conventional correction approach that assumes equal concentration in all tissues. We demonstrate that accounting for differences in the metabolite concentrations between WM and GM leads to significant differences in the estimated metabolite concentration provided that S1 Fig(DOCX)Click here for additional data file.S2 FigDotted lines denote the subjects for whom the spectra were skipped due to a linewidth greater than 0.07 ppm.(DOCX)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.


The emergence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and subsequent Coronavirus Disease 2019 (COVID-19) pandemic has resulted in a significant global public health burden, leading to an urgent need for effective therapeutic strategies. Monoclonal antibodies (mAbs) are a potentially effective therapeutic option. We identified a potent antibody JMB2002 against the SARS-CoV-2 receptor binding domain. JMB2002 has demonstrated therapeutic efficacy in a SARS-CoV-2 infected rhesus macaque model.We conducted a randomized, double-blind, phase 1 trial to evaluate the JMB2002’s safety, tolerability, pharmacokinetics, and immunogenicity in healthy Chinese adults. Participants were randomly assigned to one of four cohorts with sequential dose, administrated intravenously with JMB2002 or placebo, and followed up for 85 ± 5 days.0−∞, AUC0 − t of JMB2002 infusion increased dose-dependently from 5 mg/kg to 50 mg/kg while there is also a linear trend between doses and Cmax.40 participants were recruited and completed in the study. Eight (25.0%) participants experienced 13 treatment emergent adverse events (TEAEs) that were drug-related. No serious adverse events (SAEs), dose limiting events (DLTs), or adverse events of special interest (AESIs), such as infusion related/allergic reactions, were observed, and no drop out due to adverse events (AEs) occurred. There was no significant safety difference observed between JMB2002 and the placebo, suggesting it was well tolerated. The AUCTherefore, JMB2002 was well tolerated after administration of a single dose in the range of 5 mg/kg to 50 mg/kg in healthy Chinese adults.https://www.chictr.org.cn/searchproj.aspx (14/01/2021).ChiCTR2100042150 at  In late 2019, the novel beta corona virus SARS-CoV-2 emerged from an animal reservoir into humans, causing an acute respiratory infection known as COVID-19 , such asOne approach to combating pathogen outbreak, such as SARS-CoV-2, would be to use plasma from the convalescent patients , 7, e.g.D=0.47 nM), inhibited the binding of ACE2 to the Omicron spike protein, and blocked the entry of Omicron pseudo virus into human ACE2 expressing cells [JMB2002–a human monoclonal antibody specifically binds to the RBD in S protein of SARS-CoV-2, was developed by Shanghai JiYu Pharmaceutical Technology Co. LTD, through a self-created FeiTai platform. It is a fully human IgG1 antibody containing N297A mutation in its constant region to attenuate Fc function . There ing cells . The pseng cells , 21. Furng cells   to four escalation dose cohorts of 5, 15, 30 to 50 mg/kg.This study was conducted in accordance with the principles of Declaration of Helsinki and Good Clinical Practice. The protocol and informed consent were confirmed by the Clinical Trial Ethics Committee of our Hospital. All participants signed informed consents before receiving the screening. This trial was registered with Chinese Clinical Trial Registry, ChiCTR2100042150.https://www.chictr.org.cn/searchproj.aspx).Participants aged 18 ~ 45 years in good health were eligible to enrolled. Exclusion criteria included those confirmed cases of previous coronavirus infection or carriers, suspected coronavirus disease, and high-risk exposure within 14 days. A complete list of inclusion and exclusion criteria was written in the study protocol ChiCTR2100042150 at Chinese Clinical Trial Registry and placebo, both were colorless to light yellow and transparent to micro-opalescent liquid, were identical in the appearance, viscosity and color.Based on preclinical studies, this study applied 4 dose levels, from 5, 15, 30 to 50 mg/kg. The study started from the lowest dose group and proceeded, to the next dose level only after safety assessment was confirmed not reaching the below-listed criteria for dose escalation termination. For the safety assessment, dose limit toxicity (DLT) was observed for at least 7 days. Blindness was kept when assessing the data of all subjects of the current dose group. No subject participated in two or more dose groups.The dose escalation termination criteria were: (1) more than 1/2 participants had drug-related adverse events (AEs) ≥ grade 2 (CTCAE 5.0), or (2) more than 1/4 participants had drug-related AEs ≥ grade 3, or (3) 1 drug-related serious adverse event (SAE). Termination criteria for sentinels was that at least 1 participant had drug-related AEs ≥ grade 3 or 1 drug-related SAE. All AEs were graded using CTCAEv5.0 (Common Terminology Criteria for Adverse Event) and coded with MedDRA23.1 .Both JMB2002 and placebo were diluted in a 0.9% sodium chloride (250 mL), and then were perfused intravenously for 60 min. First 10 participants were randomized (4:1) to receive 5 mg/kg of JMB2002 or placebo in cohort 1, and monitored for adverse events. After a successful safety evaluation on Day 7, the cohort 2 was proceeded with another 10 randomly-selected participants receiving 15 mg/kg of JMB2002 or placebo (4:1), and so on. Particularly in cohort 3 (30 mg/kg) and 4 (50 mg/kg), two sentinels were first randomly selected and assigned (1:1) to receive JMB2002 or placebo respectively; and only if there were no drug-related AEs ≥ graded 3 after day 1, the remaining 8 participants were proceeded.Vital signs, such as blood pressure, pulse, respiration and ear temperature, were taken at 0 h as baseline, then 1, 3, 8, 24, 48, and 72 h after administration and on days 8, 15, 29, 43, 57, and 85 or early termination day. Physical examination, including skin, mucous membrane, lymph node and so on, was taken on days 8, 29,57 and 85 or early termination day. Laboratory examination, including blood routine, urine routine, blood biochemistry and coagulation function, and 12-lead electrocardiogram were carried out on days 4,8,29,57 and 85 or early termination day.0–t) or time to infinity (AUC0−∞), maximum concentration (Cmax), elimination half-life (t1/2), volume of distribution (Vz), plasma clearance (CLz), time to reach Cmax (Tmax), the terminal phase elimination rate constant (λz), and mean residence time (MRT), were calculated accordingly [max, AUC0–t, or AUC0−∞ respectively was performed after log-transformation, using a power model first described by Gough et al. [The primary outcome was the safety and tolerability of the JMB2002 in healthy Chinese adults, which was quantified by the number and proportion of treatment emergent adverse events (TEAEs). Any anomaly in the exam results and occurred adverse medical events were also recorded. Secondary outcomes were pharmacokinetic characteristics and immunogenicity. Pharmacokinetic parameters were calculated and analyzed using non-compartmental analysis (Phoenix WinNonlin version 6.4 or above), by measuring the concentration of JMB2002 in serum of participants at 0 h as baseline, then 1, 3, 8, 24, 48 and 72 h, and on days 8, 15, 29, 43, 57 and 85 after receiving a single dose of JMB2002 by intravenous infusion. The area under the serum concentration–time curve from time zero to the time of the last quantifiable concentration , median, Q1, Q3, minimum and maximum. Pharmacokinetic variables were additionally described with coefficient of variation and geometric mean. Count variable was described with frequency and percentage. Unless otherwise stated, all statistical tests were two-sided with α = 0.05, and two-sided 95% confidence intervals were calculated. If the P value was ≥ 0.001, it was rounded to 3 decimal places; if the P value was < 0.001, it was reported as “<0.001”.A total of 109 screened subjects, 40 participants were eligible and enrolled in the study. Ten participants were randomly assigned into 5, 15, 30, and 50 mg/kg dose group respectively, of which 8 received JMB2002 and 2 received placebo  participants developed 27 cases of TEAEs, including 13 (40.6%) in the test groups and 4 (50%) in the control group. Among them, 11 (27.5%) participants developed 17 drug-related TEAEs, including 8 (25.0%) in the test groups and 3 (37.5%) in control group  was 141.5 ± 49.4, 310.7 ± 166.8, 841.6 ± 149.6, and 1340.6 ± 226.1 ug/mL in the 5, 15, 30, and 50 mg/kg groups respectively; and the AUC0 − t (mean ± SD) was 39984.0 ± 5594.5, 104043.0 ± 26631.5, 265375.9 ± 43010.3, and 366394.5 ± 45374.6 h*ug/mL respectively. A dose proportionality analysis revealed that AUC0−∞, AUC0 − t exhibited linear relationship with the dose of the study drug while there is also a linear trend between dose and Cmax.  in the 50 mg/kg dose group was found to have ADA prior to administration (day 0) and tested positive for antibody titers at day 85. The similar results have been reported in other clinical trials . It was The primary objective of this study was to evaluate the safety and tolerability of single dose of JMB2002 by intravenous infusion in healthy participants. The incidence of drug-related TEAEs at 50 mg/kg dose level was higher than that at other dose levels, suggesting that increased JMB2002 may increase the safety risk. Nevertheless, it remained tolerable in all the participants, with only 1 grade 2 and no grade ≥ 3 or worser TEAEs.Our results are comparable to similar drugs reported in previous literatures. In a phase I clinical study, Etesevimab , a recombinant neutralizing human IgG1 monoclonal antibody developed by Top Alliance Biosciences in China, triggered 173 TEAEs in all 40 (100%) participants and 22 drug-related TEAEs in 17 (42.5%) participants . With a max, AUC0 − t, AUC0−∞ of JMB2002 increased along with dose over a range of 5 to 50 mg/kg, in a similar manner reported previously in the study of Etesevimab and LY-CovMab [1/2) across dose levels. Therefore, the JMB2002 had similar PK profile with the other antibodies under development. On the other hand, only 1 (2.5%) participant tested positive for ADAs, which was unrelated to the antibody administration. In comparison, 3 (7.5%) participants tested positive for ADA with Etesevimab in the test groups throughout the study, and 5 (12.5%) participants were tested ADA positive caused by LY- CovMab. Therefore, the JMB2002 had similar or lower immunogenicity risk to the similar antibodies under development, correlated with the prediction by in silico analysis and somatic hypermutation rate [The pharmacokinetic parameters CY-CovMab , 23, whiion rate .In contrast to many NAbs showing low affinity or efficacy to the most recent Omicron variant , 17, ourThere are several limitations in this study. First, as a phase 1 clinical trial, the sample size is small. Second, participants were recruited from healthy Chinese adults aged 21–45 years old. It remains undetermined whether JMB2002 is safe for high-risk population, such as older adults and those with underlying comorbidities including but not limit to hypertension, diabetes, or obesity , or mildIn conclusion, the result of this study confirmed the safety and tolerability of JMB2002 in healthy participants, and warrants the phase II clinical trial to explore its efficacy and safety in patients with novel coronavirus pneumonia.


Dioscorea villosa) was evaluated for its intrinsic physicochemical properties. From the results, essential metals such as K, Ca, P, and Fe were detected along with some nonessential heavy metals below the WHO permissible limits. Bulk density was 0.13–0.63 g/mL. The water absorption capacity, oil absorption capacity, swelling power, and solubility of the starch were pH-responsive. Thermal profiles showed onset temperature, To (59.21 °C), peak temperature, Tp (60.22 °C), endset temperature, Tc (63.12 °C), gelatinization enthalpy, ΔHgel (0.54 J/g), temperature range of gelatinization, R (3.91 °C), and peak height index, PHI (1.87 J/g °C). Exhibiting a crystallite size of 0.03 nm, absorption peaks of 15.3119°, 24.4120°, and 18.4170°, corresponding to interplanar d-spacings of 3.7500 Ǻ, 5.14000 Ǻ, and 4.954610 Ǻ, were obtained. Evidence of C–H at 1338.1 cm−1, C–O at 640.0 cm−1, C–H stretch at 2829.7 cm−1, and a strong and broad O–H group at 3291.2 cm−1 were obtained. The starch granules had low particle sizes, were homogeneous, and were aggregates of irregular shapes. At a lower pH (2-4), the wild yam starch studied could be a potential absorbent material in the production of disposable diapers and female napkins and as biodegradable films due to its high hydrophobicity at a high pH (8-12).Native starch extracted from wild yam ( Dioscorea , is a major home meal in West Africa, especially Nigeria [Dioscorea villosa).Yam, a species in the genus  Nigeria , 2. Alth Nigeria , only se Nigeria . HoweverWild yam is a perennial crop with nonfleshy but dried, narrow, and crocked roots, bearing horizontal branches of long creeping runners . Wild yaThe inedibility of wild yam species is due to the presence of antinutrients such as phenols, tannins, hydrogen cyanide, oxalate, amylase inhibitor activity, and trypsin inhibitor, which however, this can be eliminated through soaking, cooking, autoclaving, baking, and oil-frying, resulting in a good dietary source with high proportions of carbohydrate, fibres, essential amino acids, and low-level fats and protein –11.Starch is a highly versatile biopolymer that is widely used in both food and nonfood applications due to its exceptional bioavailability, biocompatibility, biodegradability, and low cost. It is composed of amylose and amylopectin chains and is readily available in various plant storage organs such as roots, tubers, corms, cormels, and legume grains. The functionality of starch in industrial products is influenced by its source and intrinsic physicochemical properties, including chemical composition, functional properties, morphology, thermal properties, and rheological properties –14.This research paper emphasizes the importance of analyzing the intrinsic physicochemical properties of wild yam starch to determine its potential for increased use as the demand for starch continues to grow.Dioscorea villosa) were obtained from a farm in Etsako-West Local Government Area, Edo State, Nigeria. The tubers were identified by the herbarium curator of the University of Medical Sciences, Ondo, Nigeria, having voucher details, UNIMED PBTH No. 012, and were deposited in the herbarium of the university. The reagents used in this investigation were of Analar quality and were utilized in their original form. All the methods were carried out following local and international institutional guidelines.Freshly harvested tubers of wild yam tubers of the wild yam species . After drying, grinding, and sieving, a 200 mg, 2.5 cm diameter pellet was formed with a pellet-pressing machine. The pellet was then exposed to radiation from a Cd-109 radioactive source for 2500 s while placed in the sample compartment. To correct for absorption, two measurements were taken using a liquid nitrogen-cooled Si (Li) detector. Different filters were used to ensure optimal detection of minerals , 16.The starch sample (1 g) was measured into a 25 mL graduated measuring cylinder and gently tapped on the bench top ten times from a height of 5 cm. After tapping, the volume of the sample was recorded. This process was repeated for 2, 3, 4, and 5 g weights of the same starch sample. Bulk density (BD) was calculated as the ratio of the weight of the sample to the volume of the sample after tapping .One (1) g of the starch sample was mixed with 10 mL of distilled water/oil using a Thermolyne Maxi mix II mixer (type 37600) for 30 seconds to obtain a slurry. The pH of the slurry was then adjusted to 2, 4, 6, 8, 10, and 12 by adding 0.1 M HCl and 0.1 M NaOH while stirring. After allowing the slurries to stand at 21 °C for 30 min, they were centrifuged at 5000 rpm for 30 min, and the volume of the supernatant was measured in a 10 mL graduated cylinder. The results were expressed on a dry weight basis, using densities of 1 g/mL and 0.89 g/mL for distilled water and Devon King's Vegetable Oil, respectively , 19.Six preweighed test tubes were taken, and 0.1 g of starch was dispersed in 50 mL of distilled water in each one. The pH of each solution was then adjusted to 2, 4, 6, 8, 10, and 12 by adding 0.1 M HCl or 0.1 M NaOH while stirring. The resulting slurries were heated in a water bath at 90 °C for 30 min and then allowed to cool down to 28 ± 2 °C. After centrifuging the slurries at 2500 rpm for 15 minutes, the supernatants were poured off. Finally, the swelling power was calculated using a specific equation . (1)SwelFor solubility, the supernatants were spread onto clean, preweighed Petri dishes and dried in an oven at 100 °C until a constant weight was achieved . (2)SoluThe thermal properties of the starch were analyzed using a differential scanning calorimeter . A sample of 3.00 ± 0.01 mg starch (mixed with water in a ratio of 1 : 3) was placed in an aluminium sample pan, which was then sealed and weighed. The pan was allowed to equilibrate at room temperature for an hour. The machine was conditioned at 75 °C for 2 hours, and the reference and indium pans were conditioned at a rate of 10 °C/min between 30-180 °C for 20 min. The sample was then tested for thermal properties by replacing the indium pan with the sample pans and heating at a rate of 5 °C/min from 20 to 100 °C. The DSC cell was purged with nitrogen gas at a flow rate of 100 mL/min, and the thermal profiles were generated using TA Instruments Universal Analysis software .α radiation. The resulting X-ray patterns were analyzed using peak positions to determine the starch sample's crystalline nature [k is the Scherrer constant (0.84), λ is 0.154 nm, B(hkl) = FWHM , and θ is the corresponding Bragg's angle to FWHM.The starch sample was equilibrated in a desiccator for three days above distilled water. It was then subjected to X-ray diffraction analysis using a D-5000 Siemens diffractometer with Ni-filtered Cu-Ke nature . (3)Crys−1 was used, and 32 scans were recorded for the sample [The functional groups present in the starch sample were identified using a Nicolet AVATAR 360 Fourier infrared spectrometer with KBR disks. Prior to testing, the sample was pressed into a pellet and dispersed in a KBR matrix of 100 mg, with a total weight of 1.0 mg. A spectral resolution of 4 cme sample .Gold-coated starch granules were positioned on an aluminium specimen stub and observed using a scanning electron microscope , which was equipped with a sputter coater . The sample morphology was analyzed at various magnifications, including 150 K, 500 K, and 1000 K .The statistical analysis of the data collected for mean comparison was conducted with SPSS 26.0 software. Duncan's least significant test and one-way analysis of variance (ANOVA) were utilized at a significance level of 5%.Bioaccumulation of K, Ca, P, Fe, Cr, S, Al, Si, Cl, Ti, and Mn in wild yam starch was studied . Fe, Cr,The bulk density is in a constant ratio of 1 : 1 with the weight of the starch sample studied . Thus, ip < 0.05) values as pH changes from 2 to 12  of 3.50 ± 0.01% is obtained at pH 10. As the pH increases from 2 to 10, OAC increases because the intermolecular bonds of the starch granules are weak; thus, the pores are enlarged, oil molecules are absorbed and retained in the interstitial network, and the starch granules behave oleophilic in nature . The impDioscorea villosa) starch studied at different pH of 2, 4, 6, 8, 10, and 12. The swelling and solubility of wild yam starch are pH-sensitive and vary significantly at different pH levels (p < 0.05). Swelling power (SP) is an indicator of the ability of the starch to hold water and form gels at temperatures close to or equal to its gelatinization temperature [Figures perature . The SP perature , 32. Staperature . Wild yaperature . As the perature . This phperature .To), peak temperature (Tp), and endset temperature (Tc) of gelatinization of the wild yam starch are 59.21, 60.22, and 63.12 °C, respectively  [Hgel) of 0.54 J/g compared to 17.0 J/g reported for potato and taro [R) obtained for the starch sample is 3.91 °C. R is the difference between To and Tc and significantly indicates the homogeneity or heterogeneity of starch granules. Homogeneity increases as R decreases [The onset temperature  . The sta59.8 °C) . This maand taro , 41. Theecreases . The peaB-type starch [The three major peaks used in characterizing the wild yam starch sample show absorption bands at 15.3119°, 24.4120°, and 18.4170°, which correspond to the interplanar d-spacings of 3.7500 Ǻ, 5.14000 Ǻ, and 4.954610 Ǻ, respectively . The date starch . The crye starch . Highly Discorea villosa) as a function of transmittance and wave number is depicted in −1 corresponding to transmittance values of 63.907, 68.330, 72.907, 68.095, 28.763, 56.052, 67.672, 80.119, 84.157, 86.432, 80.757, and 70.189%, respectively. The fingerprint representations at 928.1 cm−1 and 998.9 cm−1 are evidence of the presence of vinyl C–H out-of-plane bending in the starch sample studied. The transmittance peak at 1338.1 cm−1 represents CH in-plane bending, 1640.0 cm−1 is the carbonyl group, C–O, which can be oxidized to a carboxyl group, 2829.7 cm−1 is the C–H stretch of alkanes, and 3291.2 cm−1 represents a strong and broad O–H group, which is indicative of a strong hydrogen bonding between the starch molecules [The FTIR pattern of wild yam (olecules –47. The olecules . This unR) and PHI, as previously reported in studies [As shown in  studies , 21. The studies , 22.Discorea villosa) exhibits intrinsic physicochemical properties. The wild yam starch studied bioaccumulates some macro and heavy metals  in nonlethal proportions. The bulk density is not altered by a change in the weight of the starch sample, whereas the water absorption capacity, oil absorption capacity, swelling power, and solubility are pH-responsive. Swelling power is elevated in an acidic medium, while solubility is elevated in an alkaline medium. The granules of wild yam starch are thermally stable and homogeneous. Strong hydrogen bonding is identified between the molecules of the starch granules. Due to its intrinsic properties, the wild yam starch studied can serve as a suitable recipe for snack foods, food emulsifiers, and products such as mayonnaise. Its crystallite size is advantageous where penetration of the treatment agent into a starch matrix is required. At a lower pH (2-4), the wild yam starch studied could be a potential absorbent material in the production of disposable diapers and female napkins and as biodegradable films due to its high hydrophobicity at a high pH (8-12). At a pH level of 8, which is similar to the natural pH level of the body, wild yam starch is safe for human use as a diagnostic fluid in MRI and MPI recipes. The granules are shaped irregularly but are arranged in separate clusters.The native starch isolated from wild yam (


The aim this study is to determine anterior chamber parameters variations induced by PreserFlo MicroShunt implantation, in the early post-operative days.n = 30) or combined with phacoemulsification (n = 18). Anterior chamber depth (ACD) and volume (ACV), central corneal thickness (CCT) and total corneal astigmatism (TCA) were evaluated pre-operatively, post-operatively at day-1 and at 1 week with the Pentacam tomography.This is a prospective observational study on 48 eyes undergoing PreserFlo MicroShunt implantation alone (p < 0.0001) and to 10.8 ± 3.7 mmHg (p = 0.0001) at day-1 and week-1, respectively. TCA varied significantly from baseline (1.5 ± 1.2 D) to both day 1 follow up  and week 1 follow up . Nevertheless, only K1 showed a transient flattening at day 1, while K2 value didn’t show any statistical variation in the early post-operative period. CCT value rose significantly at day 1 , but then returned toward pre-operative values at week 1 . In contrast, ACD values changed insignificantly from 3.3 ± 0.9 to 3.7 ± 1.0 mm at day 1 (p = 0.21), and then stabilized at 3.4 ± 0.9 mm (p = 0.82) at week 1 follow up. ACV changed from 150.0 ± 36.2 to 159.5 ± 42.1 mm3 at day 1 (p = 0.58), and successively to 153.9 ± 37.9 mm3 at week 1 follow up (p = 0.96). The subgroup analysis in eyes undergoing standalone PreserFlo implantation didn’t show significant changes in both ACD and ACV.Intraocular pressure decreased significantly from 20.9 ± 4.0 to 8.0 ± 2.8 mmHg (PreserFlo implantation minimizes the anterior chamber modifications generated by traditional filtering surgery, inducing low and transient corneal and biometric changes only in the very early postoperative period and insignificant changes to ACD and ACV, label of its safety and minimal invasiveness. Glaucoma filtering surgeries have the aim to bypass the trabecular meshwork by shunting aqueous humor (AH) to the subconjunctival or sub-Tenon space.  The goldMIGS  refers to a series of relatively novel surgical techniques for lowering intraocular pressure (IOP), with the goal of a speedier recovery, decreased risk of complications, and less refractive and visual alterations than traditional trabeculectomy.  New ab iThe 8.5-mm PreserFlo MicroShunt  is made of the stable and flexible polymer ‘SIBS’ (poly[styrene-blockisobutylene-block-styrene]), which showed its biocompatibility in cardiac stents.  It has aThe aim of our study is to determine anterior chamber (AC) variations, such as ACD, ACV, and corneal curvature parameters, induced by PreserFlo MicroShunt implantation, in the early post-operative days, to check whether the minimal invasiveness and reported safety of this device is also evident in the maintenance of anterior segment ocular parameters.In this prospective observational study, we included patients with primary open-angle glaucoma (POAG) who underwent Microshunt PreserFlo implantation at Fondazione Policlinico Universitario A. Gemelli—Rome between November 27, 2019 and December 8, 2021. POAG was characterized by the presence of an open iridocorneal angle, evidence of glaucomatous optic nerve injury, and visual field (VF) deficits. The research comprised individuals who were on the most tolerated medication treatment and had increasing vision loss. This research adhered to the tenets of the Declaration of Helsinki and was approved by Catholic University of the Sacred Heart Ethical Committee in Rome, Italy. An informed and written consent was obtained from all enrolled patients.We established the following inclusion criteria: uncontrolled glaucoma on maximum tolerated medication with an intraocular pressure (IOP) of 12–45 mmHg; best-corrected visual acuity (BCVA) of 20/200 or better; phakic or pseudophakic patients; and individuals with significant worsening of visual field parameters during a follow-up of at least 1 one year . In the event of bilateral surgical indication, both eyes may be treated, if necessary, at least 1 month apart.Exclusion criteria included prior filtering treatments (trabeculectomy), angle closure, pigmentary, pseudoexfoliative, uveitic, and neovascular glaucoma. In addition, the following exclusion criteria were considered: age less than 18, a history of intraocular inflammation or retinal abnormalities, a history of non-glaucomatous optic neuropathy, optic nerve drusen, poor image quality due to hypermature cataract, and unstable fixation. In situations with concomitant cataracts, implantation of PreserFlo together with phacoemulsification and IOL implantation was suggested. Consequently, our cohort was split in two subgroups: PreserFlo MicroShunt implantation alone or combined with phacoemulsification. All individuals provided written informed permission in accordance with the Declaration of Helsinki for this research. A total of 46 eyes who underwent PreserFlo implantation was included.All patients’ demographics , type of surgery , and ocular parameters, were collected at the preoperative visit, including BCVA assessed with the Snellen chart, IOP at which the decision for surgery was made (measured with Goldmann applanation tonometer) and number of glaucoma medications at the preoperative visit. The post-operative IOP was analyzed at post-operative day 1 and week 1. At every follow-up, patients underwent three consecutive IOP measurements, 5 min apart from each other, and an average value from the three was collected. The number of patients with early adverse effects were also gathered.All surgeries were performed by expert surgeons  under peribulbar anesthesia. A traction suture on the superior cornea was used to expose the upper nasal conjunctiva in order to perform conjunctival peritomy and careful Tenon dissection, in order to create a posterior pocket in the supero-nasal quadrant. To reduce bleeding and provide a clean surgical area, a diathermy probe was used on the sclera.All patients were treated with Mitomycin C (MMC) 0.2 mg/mL by placing three wet surgical sponges supplied by the manufacturer beneath Tenon’s layer for 2 min, avoiding limbus, and then abundantly washing with a balanced salt solution (BSS). With the tip of the caliper 3 mm away from the limbus, a trypan blue mark was made, and a microknife-made 1-mm wide scleral preincision was performede. A 25-G needle was inserted into the AC at the trabecular meshwork to form a scleral tunnel parallel to the sclera’s surface. The PreserFlo MicroShunt, just after priming with BSS to ensure its patency, was then put with the bevel up in the tunnel until reaching the AC, where it was visually verified to control that it was not too near to the iris or endothelium.A planar attachment mechanism, like the fins of an arrow, is located halfway down the tube and secures the device in the pocket, avoiding tube migration. Flow through the implant was verified visualizing drop-by-drop flow from the end of the tube with a surgical sponge. Tenon’s layer was moved ahead of the conjunctiva to ensure that the implant did not become stuck in it, and the conjunctiva was then sutured watertight over Tenon’s layer using a 10–0 nylon fornix-based removable chain suture.  The surgTomographic analysis were performed the day before surgery, and then repeated 24 h after surgical procedure and at the 7-days follow-up. The Pentacam HR (Oculus Inc.) is an eye tomography device exploiting a 360-degree rotating Scheimpflug camera and a monochromatic slit-light source  to obtain cross-sectional pictures of the anterior segment. This instrument can capture up to 50 photos in 2 s. Data regarding ACV, ACD, central corneal thickness (CCT), anterior corneal keratometries (Ks) and total corneal astigmatism (TCA) were collected Fig. . The datp > 0.05 was utilized to confirm the null hypothesis. We performed Analysis of Variance (ANOVA) and Dunnett’s multiple comparison test with the Geisser–Greenhouse adjustment for matched pairs. To compare the difference between each pair of non-matched means, the Tukey test, which computes confidence intervals, was used. For contingency analysis, Chi-square and Fisher’s exact tests were utilized. In addition, correlation studies were performed on continuous variables. The quantitative results were represented as the mean standard deviation, and a p value 0.05 was deemed statistically significant. The 95 percent confidence interval (CI) of the mean was used.The statistical analysis was conducted using GraphPad PRISM Software, . Our sample’s normality was determined using the Shapiro–Wilk test, and A total of 48 eyes with POAG were included in the study. The study population’s mean age at the time of surgery was 72.2 ± 5.7 years; laterality (RE/ LE) was 30/18; sex (M/F) was 20/28. Mean preoperative IOP treated with anti-glaucoma drugs was 20.9 ± 4.0 , on a mean number of topical drugs of 2.8 ± 0.7; mean BCVA was 0.61 ± 0.3 and mean CCT was 529 ± 32 µm. A total of 28 patients (58%) were already pseudophakic, while 18 patients (37.5%) underwent combined surgery. Two eyes underwent standalone procedure without the need for cataract surgery. A total of 30 eyes underwent a standalone PreserFlo implantation (62.5%). Demographic charachteristics are visible in Table p < 0.0001) at the first post-operative visit after 24 h, and to 10.8 ± 3.7 mmHg at 1 week (p = 0.0001).PreserFlo MicroShunt decreased IOP from a mean baseline of 20.9 ± 4.0 to 8.0 ± 2.8 mmHg (n = 9) at day 1 and 8% (n = 4) at week 1, but resolved in all instances at the successive follow ups. In the first week, one of the patients developed choroidal effusion, which was successfully managed with conservative therapy (occlusion and 1% atropine eye drops), and resolved within the first month. Early self-limiting hyphema was often documented  but fully resolved at week 1 in all instances.In accordance with the World Glaucoma Association’s guidelines, hypotony was defined as an intraocular pressure (IOP) of 5 mmHg following surgery.  The inciThe findings of the change in anterior chamber parameters as well as anterior astigmatism are shown in Table p = 0.005) in corneal K1 at day 1, from 43.8 ± 1.6 D (CI 43.3–44.2) to 42.5 ± 1.7 D (CI 42.0–43.0), which was not significant at week 1 . On the other hand, no statistical variations were found in corneal K2, neither at day 1 , nor at week 1 . Consequently, a statistical difference was found when comparing TCA from baseline (1.5 ± 1.2 D) with both day 1 follow up  and week 1 follow up . Nevertheless, mean K value didn’t show any statistical variation in the early post-operative period .Compared to the pre-operative period, we found a statistical difference (p = 0.04), but then returned toward pre-operative values at week 1 . In contrast, ACD and ACV didn’t show any significant variation at both follow ups. ACD values changed from 3.3 ± 0.9 mm, CI 2.9–3.5, to 3.7 ± 1.0, CI 3.3–4.0 at day 1 (p = 0.21), and then stabilized at 3.4 ± 0.9, CI 3.1–3.7 (p = 0.82) at week 1 follow up. ACV changed from 150.0 ± 36.2 mm3, CI 139.2–160.7, to 159.5 ± 42.1 mm3, CI 146.7–172.4, at day 1 (p = 0.58), and successively to 153.9 ± 37.9 mm3, CI 142.5 to 165.2, at week 1 follow up (p = 0.96) . As a result, ACD values didn’t show significant variations when comparing pre-operative values (3.1 ± 0.7 mm) to day-1  and week-1  follow ups, respectively. Similarly, ACV remained statistically stable when comparing baseline values (155.0 ± 24.2 mm3) to post-operative day 1  ad week 1 .A subgroup analysis was conducted in order to exclude the effect of lens extraction as a bias for ACV and ACD changes after surgery. In particular, ANOVA tests were applied specifically to the standalone PreserFlo surgery group (p = 0.02) at day 1, which decreased towards the first week . In particular, we found that the group of combined surgery showed a significant flattening in K1, which was linked to the site of the 2.2 mm clear corneal incision, at both day 1  and day 7 . On the other side, K2 showed no significant changes (p = 0.36). In the standalone group, we reported no significant variation in both K1 and K2 .When comparing post-operative TCA variations between standalone PreserFlo surgery and combined surgery groups, we found higher astigmatism changes in the latter  revealed no statistically significant association, as visible in Table Previous studies reported that trabeculectomy caused considerable alterations in anterior segment parameters in the first postoperative week, which generally reverted to baseline in the first postoperative month. , 26 In fMoreover, a widespread interest was focused on the impact of glaucoma surgery on astigmatism. Initially, Hugkulstone, using keratometry, observed a statistically significant decrease in vertical corneal radius from 7.71 mm preoperatively to 7.36 mm on day 1 postoperatively, which persisted at all time periods up to 28-days follow-up.  ClaridgeOvertight scleral flap sutures around 12 o’clock, according to Dietze et al., might induce local tissue compression, steepening the vertical meridian.  AstigmatTzu et al. assessed the refractive results of cataract-only and combined cataract with trabeculectomy (using 0.4 mg/mL MMC) or Ahmed  or Baerveldt  implants, reporting a difference in cylinder of 1.31 ± 0.86 in the combination group against 0.99 ± 0.72 in the cataract-only group, which was not statistically significant. Moreover, no difference between the combined trabeculectomy and the glaucoma draining devices (GDD) groups was found. Ioannidis et al. investigated refraction, visual acuity, and manifest astigmatism, using a vector-based analysis, in 106 eyes undergoing combined femtosecond laser-assisted cataract surgery and the implantation of two iStent inject trabecular microbypass devices . As a result, the mean cylinder dropped from 0.91 D preoperatively to 0.41 D after 4 weeks, with 73.8% of eyes having 0.5 D or less residual refractive astigmatism following the procedure.  Hammel ep = 0.002).The use of Pentacam Scheimpflug tomography helped us to reduce possible corneal astigmatism underestimation, given its high accuracy in analyzing anterior and posterior corneal surfaces.  A recentThe advantage of the PreserFlo MicroShunt implant relies on the fact that scleral flap and sutures are not needed, eliminating two of the risk factors for the induction of astigmatism. Furthermore, some recent anterior segment optical coherence tomography (AS-OCT) analysis showed that the most common filtering bleb morphologies determined by PreserFlo MicroShunt are the microcystic multiform and the multiple internal layer ones, located posteriorly in relation to the limbus, with no cases of large or avascular blebs linked to changes in corneal curvature reported to date. , 33 To tOn the other side, several works have neen published regarding the variation of ACD after glaucoma surgery. The first investigations were conducted with the Haag–Streit pachymetry, indicating that ACD achieves its shortest value on the second and third days after trabeculectomy, then deepens and reaches its preoperative value by the 14th day following surgery, as reported by Cunliffe et al.  Husain eThe introduction of Scheimpflug imaging offered novel results in this setting, with a recent research showing how ACD levels rose from the first postoperative week to the first postoperative month in this research, with no significant difference between preoperative and first postoperative-month ACD values. In the same research, TCA and ACV values fell in the same way as the ACD value in the first postoperative week, but showed no statistical difference after 1 month when compared to the baseline values.  SimilarlPrevious research on the influence of trabeculectomy on CCT levels has shown mixed findings, with Moschos et al. showing a slow but substantial decline in these values in the first, sixth, and 12 months following trabeculectomy as compared to preoperative values,  while HuMore studies were conducted focusing on anterior chamber parameters variations after MIGS surgery. Hammel et al. reported the first Pentacam Scheimpflug analysis after Ex-PRESS Miniature Glaucoma Implant surgery in 2013, showing that surgery had a temporary influence on anterior segment characteristics such as posterior corneal astigmatism, ACD, and ACV.  Differen3 preoperatively vs. 159.5 mm3 at day 1), allowing for more reproducible surgical results and increased safety. These results were comparable in both the standalone and the phaco-combined surgical approach and appeared to stabilize early, with the 1-week follow up parameters reported to be similar to baseline. As far as we know, this is the first research to analyze ACV variations after PreserFlo MicroShunt implantation, confirming the minimal invasiveness of this approach which consistently reduced the risk of shallow anterior chamber in the first post-operative days.Based on our findings, we hypothesize that the PreserFlo MicroShunt structure is able to keep a steady aqueous outflow via the tube’s lumen even from the very first hours after surgery, since we found a non-significant variation in both ACD (3.3 mm preoperatively vs. 3.7 mm the day after surgery) and ACV (150.0 mmn = 48) than other researches in this field, allowing for a better statistical reliability. In conclusion, the PreserFlo MicroShunt approach seems to restrain the anterior chamber modifications generated by traditional filtering surgery. In comparison to trabeculectomy, which has been reported to cause visually significant changes in the ACD, ACV and keratometric parameters that can last for more than a year in some cases, our research suggest that the implantation of this device determines slight modifications to ocular biometrics, inducing low and transient corneal and biometric changes only in the very early postoperative period and, even more important, insignificant changes to ACD and ACV, which is a label of its safety and minimal invasiveness. More research regarding AC changes, also combined with AS-OCT analysis, as well as a longer follow-up time, are certainly required to understand the long-term effects of the PreserFlo MicroShunt implantation on the anterior segment parameters.This research has several limitations, primarily due to the short-term follow up period, but has an higher sample size (


The suspensory strategy, a method for controlling postural balance in the vertical direction of the center of mass (COM), is considered by the elderly as a means of balance control. The vertical COM control might alter the sensory integration and regularity of postural sway, which in turn impacts balance. However, to date, this was not confirmed. Thus, this study aimed at investigating the influence of the suspensory strategy achieved through knee flexion on the static standing balance.Nineteen participants were monitored at knee flexion angles of 0°, 15°, and 65°. Time-frequency analysis and sample entropy were employed to analyze the COM data. Time-frequency analysis was utilized to assess the energy content across various frequency bands and corresponding percentage of energy within each frequency band. The outcomes of time-frequency are hypothesized to reflect the balance-related sensory input and sensory weights. Sample entropy was applied to evaluate the regularity of the COM displacement patterns.Knee flexion led to a decreased COM height. The highest energy content was observed at 65° knee flexion, in contrast with the lowest energy observed at 0° in both the anterior–posterior (AP) and medial-lateral (ML) directions. Additionally, the ultra-low-frequency band was more pronounced at 65° than that at 0° or 15° in the ML direction. Furthermore, the COM amplitudes were notably higher at 65° than those at 0° and 15° in the AP and ML directions, respectively. The sample entropy values were lower at 65° and 15° than those at 0° in the ML direction, with the lowest value observed at 65° in the vertical direction.The suspensory strategy could enhance the sensory input and cause sensory reweighting, culminating in a more regular balance control. Such suspensory strategy-induced postural control modifications may potentially provide balance benefits for people with declining balance-related sensory, central processing, and musculoskeletal system functions. In tThe rationale of the suspensory strategy is that a flexed knee combined with flexion of the ankle and hip joints can lower the COM height and absorb internal and external perturbations; thus, effectively increasing stability and reducing the likelihood of falling . PreviouTime-frequency analysis and nonlinear analysis of COP or COM data have been demonstrated as crucial tools for evaluating the overall posture system to estimate the sensory component and regularity of postural sway . Time-frIn addition, a review has reported that the COM vertical control is crucial for the standing balance and requires integration of multisensory cues that are critical for balance in vertical direct estimation, vertical perception, and stability . However2.2.1.f = 0.4, alpha = 0.05, and power = 0.8  was used with an F test. The sample size calculation revealed that a minimum of 12 samples were necessary for the experiment, with an effect size of er = 0.8 . NineteeThree-dimensional motion capture kinematic data were acquired using seven high-speed cameras . The collected data were then analyzed using a motion analysis system . Thirty-five retroreflective markers were placed at the specific anatomical points , 25. Mar2.2.To manipulate the suspensory strategy, participants were instructed to adopt three different postures with 0° (knee is locked by screw home mechanism) , 26, 15°2.3.Kinematic data, including knee joint angles and COM position, were analyzed using Visual3D software  and processed with MATLAB . A fourth-order, zero-lag Butterworth filter with a 10-Hz cutoff frequency was applied to the kinematic data to obtain a low-pass filtered signal . For theIn accordance with prior studies , 13, a tThe Gaussian window with a width controlled by parameter σ in The total energy content was determined by adding the energy in the frequency range of 0–6.25 Hz . To assem(r), and the probability that sequences match for m point is denoted by Bm(r). However, there is no consensus on the parameter selection, but for balance control studies, the parameter settings are commonly set to m = 2 or 3 and r between 0.1 and 0.25 standard deviation (SD)  . For thiion (SD) . Generalion (SD) , 36.2.4.Post hoc analyses between the two postures with different knee flexions were conducted using the Bonferroni paired comparison method or pairwise comparisons. For one-way repeated-measures ANOVA, Cohen suggested that small, medium, and large effects would be reflected in the partial eta squared values (η2) to 0.01, 0.06, and 0.14 . The Shapiro–Wilk test was conducted to test normality. One-way repeated-measures analysis of variance (ANOVA) or Friedman’s test was used to examine the effects of posture (three different degrees of knee flexion) on the outcome measure. and 0.14 . Accordiand 0.14 . The sig3.2(2) = 38.00, p < 0.001, Kendall’s W = 1.00]. Post hoc analyses revealed a significant difference in the COM height among the three knee flexion angles . Specifically, the median (Q1–Q3) of the COM height were 51.62% (50.47–52.68), 50.65% (49.33–51.35), and 41.47% (39.03–43.22) at 0°, 15°, and 65° knee flexion, respectively.Friedman’s test indicated a significant effect of the knee flexion angle on the COM height , ML , and vertical  direction in the COM sway among the three knee flexion angles. Post hoc analyses were performed to further investigate specific pairwise differences. COM sway amplitude at 0° knee flexion in the AP direction was significantly smaller than that at 65° (p < 0.05), and the sway amplitude at the 0° knee flexion in the ML direction angle was significantly smaller than that at both 15° and 65°  = 14.632, p < 0.001, Kendall’s W = 0.39] were revealed. Post hoc analyses indicated that the total energy content at 65° knee flexion was significantly lower than that at 0° and 15°  = 5.71, p < 0.05, η2 = 0.24]. Post hoc analysis showed that the energy percentage of the very-low-frequency band was significantly lower at the 65° knee flexion angle than those at 0° and 15° knee flexion angles  = 20.632, p < 0.001, Kendall’s W = 0.543]. Post hoc analyses indicated that the total energy content at the 65° knee flexion was significantly lower than those at 0° and 15° = 9.58, p < 0.05, Kendall’s W = 0.25] and very-low-frequency band  among the three knee flexion angles. Post hoc analysis showed that the energy percentage of the ultra-low-frequency band was significantly lower at a 65° knee flexion angle than those at 0° and 15°  = 4.11, p = 0.13, Kendall’s W = 0.23]. A significant difference was observed in the COM ML direction , with the 0° knee flexion angle exhibiting a significantly higher sample entropy than those at 15° and 65°  = 21.72, p < 0.001, Kendall’s W = 0.87]. Post hoc analysis revealed that the 65° knee flexion angle displayed a significantly lower sample entropy than those at 0° and 15° , time-frequency analysis, and non-linear analysis to investigate the effect of the suspensory strategy on the static standing balance by knee flexion. In the classical parameter analysis, the COM sway amplitude in the AP, ML, and vertical directions showed an increase during knee flexion. In terms of the time-frequency analysis, the results showed that the full-band energy content increased and the sensory weights changed while the knee was flexed. Thus, this work demonstrated that knee flexion alters the postural control in the vertical direction and this suspensory strategy alters the balance-related sensory integration and postural sway regularity.4.1.During static standing, the COM sway amplitude is typically inversely related to the balance ability; thus, smaller amplitudes indicate better balance , 37. How4.2.One of the aims of this study was to examine the effects of suspensory strategy of bending the knee on sensory integration. Time-frequency analysis of the COM or COP is a frequently employed method to explore sensory input and measure sensory reweighting , 16, 31.4.3.Typically, a decrease in sample entropy increases the regularity of postural sway, reflecting a shift toward more attention and less automatic control required to control balance. However, an increase in sample entropy decreases the regularity of postural sway, which in turn corresponds to less attention and more automatic control required for balance , 17, 19.4.4.This study has several limitations. First, the study sample consisted of only young individuals, limiting the generalizability of the findings to other older age groups. Future research should include participants across a wider age range to examine the potential age-related differences in balance control during squatting. Second, this study focused solely on sensory input and sample entropy under open-eye conditions. Future investigations could consider incorporating other sensory conditions, such as eyes closed or altered visual feedback, to provide a more comprehensive understanding of the sensory contributions to balance control during squatting. This study examined three specific knee flexion angles  but did not include intermediate angles between 15° and 65°. Investigating a broader range of knee flexion angles would provide a more detailed understanding of how different angles affect balance control and the associated sensory input. Finally, this study did not collect electromyograms, and a more detailed analysis of the conclusions or muscle contributions may require support from these data. Addressing these limitations in future studies will enhance our knowledge of balance control during squatting and provide a more comprehensive understanding of the underlying mechanisms.In conclusion, this study revealed that suspensory strategy by knee flexion led to an increased COM amplitude in healthy adults, and the suspensory strategy may be able to increase the functionality and attentional control of postural sway to some extent. Balance gains from suspensory strategy may be more appropriate for older adults with declining balance-related sensory, central processing, and musculoskeletal system function. However, it should be noted that this study population did not include older adults and more complex dynamic balances. Further research should be conducted to fully analyze the suspensory strategy. From a clinical perspective, this study provides new insights into a rehabilitative approach to balance control in older adults using a suspensory strategy. This work could also offer pertinent insights for the design of knee joint exoskeletons, particularly concerning the interplay between the use of knee joint exoskeletons, overall postural sway control, the potential effects of exoskeleton movement on the integration of sensory functions, and the attention of the user toward postural control.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving humans were approved by Faculty of Health Sciences at Hokkaido University. The studies were conducted in accordance with the local legislation and institutional requirements. The participants provided their written informed consent to participate in this study.LJ: Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Software, Visualization, Writing – original draft, Writing – review & editing, Funding acquisition. SK: Conceptualization, Data curation, Investigation, Methodology, Project administration, Supervision, Writing – original draft, Writing – review & editing, Funding acquisition. TI: Conceptualization, Methodology, Project administration, Supervision, Writing – review & editing. YK: Conceptualization, Methodology, Supervision, Writing – review & editing. AC: Data curation, Supervision, Validation, Writing – review & editing. KY: Data curation, Supervision, Writing – review & editing. YW: Supervision, Writing – review & editing. MS: Conceptualization, Investigation, Methodology, Supervision, Writing – review & editing. HT: Conceptualization, Investigation, Methodology, Supervision, Writing – review & editing.


In healthy subjects without diabetes the associations between OBG dose and glucose iAUC and iPeak were linear . MinDoses for glucose iAUC for high-MW, medium-MW and low-MW OBG, respectively, were estimated to be 0.2 g, 2.2 g and 3.2 g per 30 g avCHO; MinDoses for glucose iPeak were less than those for iAUC. Insufficient data were available to assess MinDose for insulin, however, there was no evidence of a disproportionate increase in insulin. More high-quality trials are needed to establish MinDose in individuals with diabetes.To determine the minimum amount of oat β-glucan (OBG) required to reduce glycaemic responses (MinDose), we conducted a systematic review and meta-regression analysis of acute, crossover, single-meal feeding trials that examined the effects of adding OBG or oat bran to a carbohydrate-containing test-meal versus a control test-meal containing an equivalent amount of available-carbohydrate (avCHO) from the same or similar source. Medline, Embase, and Cochrane Library were searched up to 18 August 2021. The primary outcome was glucose incremental-area-under-the-curve (iAUC). Secondary outcomes included insulin iAUC, and glucose and insulin incremental peak-rise (iPeak). Two independent reviewers extracted data. Results were expressed as ratio-of-means (RoM) with 95% confidence intervals (CIs). Linear associations were assessed by random effects meta-regression. MinDose was defined as the dose at which the upper 95% CI of the regression line cut the line of no effect . Fifty-nine comparisons ( A totalThis SRMRA was conducted according to the Cochrane Handbook for Systematic Reviews of Interventions . Data weMEDLINE, EMBASE, and the Cochrane Central Register of Controlled Trials were searched through August 18, 2021. Electronic searches were supplemented with manual searches of references from included studies. The detailed search strategy is outlined in Supplementary Table We included randomized and non-randomized acute, crossover, single-meal, controlled feeding trials that examined the effects of adding OBG or oat bran (high in OBG) to a carbohydrate-containing meal in humans regardless of age, sex, or health condition. For a trial comparison to be included, the comparator (control) test meal had to contain an equivalent amount of avCHO from the same or similar source as the OBG test-meals . To minimize bias and heterogeneity trial comparisons where the comparator (control) test-meals had different sources of avCHO  were excluded. Protein, fat and glycaemic index (GI) have independent effects on glucose and insulin responses . Thus, dn = 8–10 by as much as ±28% of the mean ) were: 4.2 g (0.65 [0.44–0.94]) for low-MW, 1.8 g (0.63 [0.43–0.91]) for medium-MW, and 1.2 g (0.75 [0.59–0.96]) for high-MW .Figure When trials with 120–180 min follow-up duration assessing the effect of OBG on glucose iPeak in individuals with and without T2D were included, the MinDose for low MW-OBG could not be determined because the upper 95% CI of the regression line was >1 across the entire dose range studied, but the MinDoses for medium-MW (1.6 g) and low-MW (<0.2 g) were similar to those for 120 min studies in subjects without diabetes , were 2.8 g, 4.0 and 6.6 g per 30 g avCHO for medium-MW OBG and <0.2 g, 0.8 g and 1.7 g per 30 g avCHO for high-MW OBG, but could not be determined for low-MW OBG because the upper 95% CI was ROM ≥ 0.9. The lowest dose (g/30 g avCHO) used in any single study that elicited a statistically significant reduction in glucose iPeak (RoM [95% CI]) was: 2.4 g (0.65 [0.46–0.93]) for medium-MW, and 0.8 g (0.72 [0.59–0.87]) for high-MW; none of the low-MW studies showed a statistically significant reduction in glucose iPeak  and did not differ from each other (P = 0.55). Similarly, mean ± SEM RoM for glucose iPeak and insulin iPeak, respectively, for the n = 8 comparisons in subjects without diabetes, 0.79 ± 0.045 and 0.75 ± 0.052, respectively, were similar to those for iAUC, were both less than 1 (P = 0.002) and did not differ from each other (P = 0.32).Due to insufficient data , the MinDose for insulin iAUC and iPeak could not be determined. However, nine publications reported data for 16 comparisons of insulin iAUC  were 0.2 g per 30 g avCHO for high-MW OBG, 2.2 g per 30 g avCHO for medium-MW OBG and 3.2 g for low-MW OBG per 30 g avCHO. The respective MinDoses for glucose iPeak were <0.2 g, 1.8 g and 2.3 g per 30 g avCHO. Not enough data were available to determine MinDose for insulin iAUC and iPeak. However, in the available trials, reductions in glucose iAUC or iPeak occurred without a disproportional increase in insulin iAUC or iPeak.In the context of our analysis, MinDose means that doses of OBG ≥ MinDose will be associated with a RoM <1 with 95% certainty, or, in other words, that the food with OBG will elicit a lower mean glucose iAUC or iPeak than the food without OBG with ≥95% certainty. This does not necessarily mean that the difference in glucose response will be either statistically or physiologically significant. This approach could be justified because, to make a claim, EFSA did not indicate the measure of glycaemic response to be used, the effect-size required nor whether the difference had to be statistically significant . Neverthwithin) of glucose iAUC varies widely in different laboratories. In a study involving 28 labs around the world, lab mean CVwithin varied from 12 to 40% [within of 12–40%, respectively, n = 10 provides 80% power detect significant differences of 17% or 56%; if n = 90, differences of 5% or 17% can be detected. The factors that determine the magnitude of CVwithin are not well understood. An early study suggested that CVwithin of glucose iAUC is highest in type 1 diabetes, intermediate in subjects without diabetes and lowest in subjects with T2D [within is: high with avCHO intakes <20 g [within is not always seen [The ability to demonstrate a statistically significant difference in a cross-over trial depends on the number of subjects and the magnitude of their within-individual variation. Within-individual coefficient of variation  [The ability to demonstrate a statistically significant reduction of PPGR also depends on the % reduction targeted. It is not clear how big a difference in PPGR is physiologically relevant. Health Canada opined that the minimum physiologically relevant difference in glucose iAUC is 20% . However< 0.001) , an effe< 0.001)  and the < 0.001) . Thus, iOur previous SRMA  includedThe mechanism by which viscous fibres such as OBG reduce PPGR is thought to be related to their ability to increase the viscosity of the contents of the gastrointestinal tract (GIT) as has been demonstrated in in-vitro –61, animNevertheless, the effect of OBG on PPGR also depends on its solubility, or bioavailability, which, in turn, may vary depending on the food matrix  , 41 and Our analysis has several limitations. Nearly all the trial comparisons (95%) were in subjects without diabetes. There were two trials in subjects with T2D. Although our findings did not change materially with the addition of these trial comparisons, we cannot conclude with a high degree of certainty that the MinDoses estimated in subjects without diabetes would apply to those with diabetes. In addition, there were insufficient data to conduct reliable meta-regression analyses for insulin outcomes. However, the results demonstrate that reductions in PPIR occurred in parallel to those for PPGR and, therefore, indicate there is no disproportionate increase in insulin. Other potential limitations were the inclusion of non-randomized studies and the need for using Plot Digitizer. However, of the 20 studies included, there was concern about the randomization process for 1 study  and PlotFinally, the primary method we used to determine the MinDose, i.e., the smallest amount that elicits a mean reduction >0% (RoM < 1) with 95% certainty, may not be considered appropriate by other scientists or regulatory authorities. For this reason, we provided results for some other methods of determining MinDose. What is very clear from these results, however, is that whatever method is used to establish MinDose, the amount is far less than the 4 g/30 g avCHO currently required by EFSA . FurtherWe conclude that the estimated minimum amounts of OBG per 30 g avCHO required to reduce mean glucose iAUC (or iPeak) in healthy subjects without diabetes with 95% certainty are: 0.2 g (<0.2 g) for high-MW OBG, 2.2 g (1.8 g) for medium-MW OBG and 3.2 g (2.3 g) for low-MW OBG. For high-MW OBG, 1.4 g (<0.2 g), 2.1 g (0.8 g) and 2.8 g (1.7 g) per 30 g avCHO, respectively, are required to obtain at least 10%, 15% or 20% reductions in iAUC (iPeak). More high-quality trials are needed to determine the MinDose for insulin responses and MinDose for glucose and insulin responses in subjects with T2D.Supplemental Material


The density and biomass of phytoplankton were higher in summer ((14.64 ± 20.34) × 107 cells/L, 10.61 ± 13.16 mg/L) and autumn ((6.79 ± 3.97) × 107 cells/L, 5.57 ± 2.40 mg/L), with the M and H2 of dominant functional groups. The dominant functional groups were N, C, D, J, MP, H2, and M in spring, whereas functional groups C, N, T, and Y dominated in winter. The distribution of phytoplankton community structure and dominant functional groups exhibited significant spatial heterogeneity in the lake, which was consistent with the environmental heterogeneity of the lake and could be classified into four locations. Location I had higher phytoplankton density and biomass than the other three locations. Additionally, dominant functional groups M, C, and H2 were present throughout the lake, and all 13 dominant functional groups were observed in Location II. Our findings suggest that environmental heterogeneity is a key factor influencing the spatial distribution of phytoplankton functional groups in Lake Chaohu.The growth and development of phytoplankton are influenced by physico-chemical parameters, which can also affect the spatial distribution of phytoplankton community structure. However, it is unclear whether environmental heterogeneity caused by multiple physico-chemical factors can affect the spatial distribution of phytoplankton and its functional groups. In this study, we investigated the seasonal variation and spatial distribution of phytoplankton community structure and its relationships with environmental factors in Lake Chaohu from August 2020 to July 2021. We recorded a total of 190 species from 8 phyla, which were divided into 30 functional groups, including 13 dominating functional groups. The average annual phytoplankton density and biomass were (5.46 ± 7.17) × 10The online version contains supplementary material available at 10.1007/s11356-023-28043-5. Phytoplankton is the primary producer in aquatic ecosystem, and their growth and reproduction are limited by environmental factors including water temperature, nutrient levels, and light availability Vanni . The fluThe composition of phytoplankton communities is influenced by the abiotic factors  and biotic factors  together, which is able to respond quickly to water environmental changes  in Lake Chaohu experienced a complex and long-term process. It initially appeared in the western zone of the lake and subsequently spread to other regions before occurring throughout the entire lake. However, the western zone of the lake has experienced stronger cyanobacterial blooms than the eastern zone for more than 30 years   are rare in warm seasons  in Lake Chaohu, affecting the succession of phytoplankton functional groups. However, fishing ban at the whole lake has been executed since January 2020 in Lake Chaohu. This way of comprehensive fishing ban in a large lake (with an area of 780 km2) is rare in the world, and its impact mechanism on plankton remains unclear.Furthermore, there have been significant alterations in the fish yield and composition of Lake Chaohu. In 1952, the annual fish yield was recorded at 3500 t, with 41.1% of phytophagous fish species such as 02) Wang . At presThe aim of this study is to assess the impact of environmental heterogeneity on the temporal fluctuations and spatial patterns of phytoplankton functional groups in a persistent high-phosphorus lake and to explore the response mechanism of phytoplankton community structure and dominant functional groups under high-intensity fish predation in Lake Chaohu.2. It is a subtropical shallow lake and has the functions of climate regulation, recreation, and fishery resources  is located in the middle and lower reaches of the Yangtze River, China, with an area of 780 kmpH was measured by a portable pH meter ; dissolved oxygen (DO) and water temperature (WT) were determined by a portable dissolved oxygen meter  in the field. Water depth (WD) and transparency (SD) were determined by the Secchi disk at each sampling station.Water samples for nutrient measurements were a mixture of sub-samples from the surface to the bottom at 1-m or 2-m intervals. One-liter mixed water sample was taken to determine the nitrogen and phosphorus concentrations in the laboratory. Total nitrogen (TN) was analyzed with the alkaline persulfate oxidation method Huang . Total pCrustacean zooplankton samples were collected with a 5-L modified Patalas bottle sampler. A total of 30-L mixed water samples at each sampling site were obtained from the surface to the bottom at 0.5-m or 1-m intervals. Cladocerans and copepods were filtered with a 25# plankton net (64 μm) and then fixed with 4% formaldehyde solution. The crustacean zooplankton were identified according to the methods of Jiang and Du  and ShenEach phytoplankton sample was a mixture of sub-samples taken from the surface to bottom with a 5-L modified Patalas bottle sampler at 1-m or 0.5-m intervals. Phytoplankton sample (1 L) was collected from a mixed sub-sample at each station and then was fixed with 10 mL of Lugol’s iodine solution and sedimented for 48 h prior to counting under a microscope. The remains were stored in a 50-mL plastic bottle and preserved in the dark in the laboratory. The concentrated solution (0.1 mL) was absorbed after thoroughly shaking and placed in a counting box under a microscope (Olympus CX31) and then observed and counted. For each sample, it was counted twice, and about 100 fields were observed each time. The average value twice was regarded as the final result. It was valid of the error twice which was within 15% , where N is the cell density of a species in a sample (× 105 cells/L); A is the total number of fields in the microscope; Ac is the number of fields observed; Va is the volume of the counting box (mL); Vs is the volume (mL) concentrated by 1-L original water sample precipitation; and n is the cell number of the counted phytoplankton.The cell density of each phytoplankton species was calculated in a formula: Phytoplankton species were identified according to the methods of Hu , Hu and Pi = ni/N, in which ni is the biomass of the ith functional group, and N is total biomass of phytoplankton.The biomass of phytoplankton functional groups can explain the dynamics of phytoplankton community structure and response to environmental characteristics  of each species. The formula is Y = (ni/N)*fi, in which ni stand for the individual number of i species, N is total individual number of phytoplankton, and fi is the frequency of occurrence of i species in the sample. When Y ≧ 0.02, phytoplankton is regarded as dominant species  is performed to analyze the relationships between dominant functional groups and environmental factors, and the maximum gradient length is found to be less than 3. Therefore, redundancy analysis (RDA) and variation partitioning analysis (VPA) are determined to analyze the relationships between dominant functional groups of phytoplankton and environmental factors using Canoco 5.0 software  before analysis except pH. Two-way ANOVA was employed to examine the influence of locations , seasons , and their combination on the physico-chemical factors, phytoplankton density, and biomass in Lake Chaohu using SPSS 26.0 software.During statistic analysis, the data are transformed by Log (The maximum water temperature appeared in August (30.95 ± 0.62 °C) whereas the minimum value was in January (2.67 ± 0.34 °C) Table . The maxNon-metric multidimensional scaling analysis (NMDS) shows there is a clear spatial heterogeneity in the physico-chemical properties of the lake, and it can be divided into four locations Fig. . Both LoBosmina longirostris, Ceriodaphnia cornuta, Chydorus sphaericus, Diaphanosoma brachyurum, Moina micrura, Thermocyclops sp., and Mesocyclops leuckarti, and the minimum value (6.94 ± 3.59 ind./L) occurs in April, in which the dominant species are Sinocalanus dorrii, Limnoithona sinensis, and B. longirostris. Spatially, the maximum (134.98 ± 32.47 ind./L) of crustacean zooplankton occurs in Location I in summer, in which B. longirostris, C. cornuta, C. sphaericus, L. sinensis, and Thermocyclops sp. dominate, and the minimum density (16.54 ± 7.17 ind./L) appears in Location IV in spring, in which B. longirostris, S. dorrii, Bosmina fatalis, and L. sinensis dominated.In Lake Chaohu, the density of crustacean zooplankton shows obvious tempo-spatial variations. The annual average density of crustacean zooplankton is 58.91 ± 63.72 ind./L, with the range of monthly average density from 6.94 ± 3.59 ind./L to 236.15 ± 98.82 ind./L. The maximum value (236.15 ± 98.82 ind./L) appears in August, in which the dominant species are 7 cells/L to 20.91 × 107 cells/L, with an annual mean value of (5.46 ± 7.42) × 107 cells/L in Lake Chaohu. Phytoplankton density shows a significant seasonal difference (P < 0.001), in which it increases in spring, reaches the peak in summer, and declines in autumn and winter  and station 11 ((16.79 ± 0.24) × 107 cells/L), and the lowest density appears at station 20 ((1.46 ± 0.16) × 107 cells/L). The relative density of Cyanophyta in each sampling station is more than 65% among total phytoplankton density, followed by Chlorophyta and Bacillariophyta. The highest relative density of Cyanophyta occurs at station 16 (97.47%). The peak of relative density of Chlorophyta and Bacillariophyta both appear at station 18 (20.60%) and station 20 (15.31%), respectively. Two-way ANOVA shows that the effects of seasonal variation  and spatial distribution  on phytoplankton density are all very significant (Supplementary Table The density of phytoplankton vary from 0.43 × 10Cyclotella meneghiniana (Y = 0.175) is the dominant species. It reached the peak (10.61 ± 13.16 mg/L) in summer, in which the proportion of Cyanophyta is 81%, and Microcystis sp. (Y = 0.566) and Dolichospermum sp. (Y = 0.147) are the dominant species. In autumn, it is 5.57 ± 2.40 mg/L, with a relative biomass of 69% Cyanophyta, and then, it is 1.69 ± 0.48 mg/L in winter, with the proportion of 44% Bacillariophyta and 22% Chlorophyta. Spatially, phytoplankton biomass in the western zone of the lake is higher than that in the eastern zone in four seasons , spatial distribution , and their combination  on phytoplankton biomass are all very significant  occurs in July whereas the minimum value (1.27 ± 0.47 mg/L) appears in December. Phytoplankton biomass shows obvious temporal and spatial variations , H2 (27.70%), and C 6.89%). Functional groups M (29.19%) and C (16.97%) dominate in Location II. Functional groups J, C, H2, and M are the dominant functional groups in Location III, accounting for 68.26% of the total relative biomass. The dominant function groups in Location IV are M (43.95%), H2 (23.38%), and C (9.20%), respectively. In summary, dominant function groups M, H2, and C remain at a high level in space, and their maximum relative biomass is in Location I (52.01%), Location III (27.87%), and Location IV (23.38%), respectively , whereas functional groups X3 and Y are positively correlated with TN, N/P, and SD (P < 0.05). In addition, functional group M is positively correlated with DTP, WD, pH, and zooplankton (P < 0.05), whereas functional group C is positively correlated with TN and N/P (P < 0.01).The discriminant correspondence analysis (DCA) conducted on the biomass data of the dominant functional groups of phytoplankton reveals that the sorting axis length is below 3 in Lake Chaohu. Consequently, RDA is performed to investigate the relationship between the dominant functional groups of phytoplankton and environmental factors. The results indicate that the eigenvalues of the first two axes of the dominant functional group of phytoplankton are 0.21 and 0.12, respectively. These two axes account for 76.03% of the variance in the relationship between the dominant functional group of phytoplankton and environmental factors (Supplementary Table P < 0.01) and dominate in summer and autumn, showing a positive correlation with water temperature and dissolved total phosphorus. Functional group C dominates in spring and winter. Functional groups C, D, W1, X3, and N dominate significantly in Locations I, II, and III (P < 0.001), which are all positively correlated with TN, N/P, and DO. In Location III, J is the dominant functional group in winter and spring. VPA shows that the explanation rate of nutrients to phytoplankton functional groups is 11.5%, whereas environmental factors and zooplankton account for 40.7% and 2.2%, respectively , 7.3% , 6.2% , and 4.6% , respectively.Spatially, functional groups M, H2, and G at the biomass level of Location I are significantly higher than those of the other three locations  Varol . ConsequH. molitrix and A. nobilis may directly affect phytoplankton abundance and biomass Below is the link to the electronic supplementary material.