Tetralophozia filiformis s.l. is known from a number of localities mostly in amphi-oceanic areas in Northern Hemisphere, including Atlantic Europe, amphi-Pacific Asia, South Siberia, and western North America. The newly obtained collections of this ‘species’ show strong variation in morphology of the taxon across amphi-Pacific Asia although connected by some ‘intergrading’ modifications. This implies the genetic diversity within this unit earlier recognized as a single taxon. Authors used molecular-genetic, morphological, and chorological methods to understand if the geographically correlated morphological variation also correlates with genetic differences and if it is possible to distinguish some additional taxa within the series of specimens originating from the various areas in amphi-Pacific Asia. It was found that Tetralophozia filiformis is a complex of at least three morphologically similar species, including one long forgotten name (Chandonanthus pusillus) that should be reinstated as separate species and one taxon  that should be described as new. Tetralophozia filiformis and Chandonanthus pusillus are lectotypified, and the new combination is provided for the latter. The three accepted taxa distinctly differ one from another in distribution patterns, preferable climate characteristics, and genetic distances, besides minor differentiations in morphology. The main morphological distinguishing features are the leaf cell size, height of undivided part in leaf lamina, and leaf dentation characteristics. Taking into account the robust correlation between the climate-based and molecular-genetic-based clusters, one more (fourth) taxon could be probably segregated from Tetralophozia filiformis. Tetralophozia (R.M.Schust.) Schljakov is the broadly distributed Arctic-Montane circumpolar T. setiformis (Ehrh.) Schljakov. Other taxa of the genus are locally distributed or rare. One of the lesser known is the predominantly East Asian Tetralophozia filiformis (Steph.) Urmi, originally described from Yunnan Province of China as Chandonanthus filiformis Steph.  … T. setiformis *Plants commonly more than 0.7–0.8 mm wide 3–4 as long as wide, terminating by 1-several uniseriate cells, leaf cuticle papillose-verrucose, sometimes scarcely or coarsely so … 2.2.T. sibiricaPlants 0.3–0.5 mm wide, cells in the lobe base 12–30 × 12–15 µm, cuticle papillose, sometimes obscurely so, leaf basal teeth less 10 cells long [hemiboreal Asian mainland with continental climate] … 2.Plants wider, 0.6–0.75 mm wide, or almost the same size (0.4–0.6 mm wide), but then with cells in the lobe base are distinctly smaller (10–20 × 10–12 µm), leaf basal teeth less than 10 cells long or to 20 cells long  … 33.T. filiformisPlants 0.6–0.75 mm wide, leaf undivided part 30–50 µm (2–5 cells) high, cells in the leaf lobe base 12–25 × (7–)10–12(–15) µm, cuticle distinctly papillose-verrucose, sometimes coarsely so [warm-temperate to tropical East and SE Asia] … 3.T. pusillaPlants 0.4–0.6 mm wide, leaf undivided part 50–90 µm (3–6 cells) high, cells in the lobe base 10–20 × 10–12 µm, cuticle distinctly papillose-verrucose, sometimes scarcely so (Korean-Japanese taxon) … * The most recent and comprehensive treatments are in Damsholt , KonstanFrullania-type (looking as dichotomous); stem cross section slightly transversely elliptic, 100–110 × 120–130 µm, with loosely defined cortex, external wall distinctly striolate, outer cells with thick walls, irregular in shape, ca. 10–12 µm in diameter, inner cells 10–20 µm in diameter, with thickened to thin walls and large, mostly convex trigones. Leaves transversely inserted, mainly subimbricate or  obliquely spreading; subimbricate leaves distinctly concave, with lower 1/4–1/3 erect spreading, then suddenly curved and lobes subparallel to the stem; (3–)4-lobed, lobes subequal , (250–)400–450 µm long and (at the level of leaf lamina) (200–)250–300 µm wide, undivided part 50–90 µm (4–5 cells) high, lobe apices prominently acuminate, with straight axis and 1–3-celled uniseriate ends, sinus strongly recurved near its base, each lobe with (1–)2(–3) acute 1–3-celled teeth near base, besides the lateral sides of the leaf have one more additional, commonly curved, to 10 cells long teeth. Underleaves bilobed, 250–350 µm long, 120–150 µm wide (at the level of lamina), undivided part 30–50 µm (2–3 cells) high, lobes prominently acuminate with straight or distinctly curved axes and 2–4-celled uniseriate ends, sinus margin recurved in the base, lobe bases in the sinus with 2–4 teeth, while underleaf side bases with 3–5 teeth, with the basal tooth largest. Cells in the lobe base 12–30 × 12–15 µm, oblong to nearly isodiameteric, walls strongly vermiculately thickened, with prominent, large and convex trigones; cuticle papillose, sometimes obscurely so. Generative structures unknown. , 780 m a.s.l., N.A. Konstantinova 13-24-01, 04 Aug. 2001 . Other specimens examined are in Tetralophozia filiformis (Steph.) Urmi. J. Bryol. 12 (3): 394. 1983.Chandonanthus filiformis Steph., Sp. Hepat. [Basionym: . Hepat.  3: 644.Lectotype (selected here): Ma Cul Chan, Delavay s.n. (G00112121/3698!)Note on the lectotypification: Franz Stephani (with very limited exceptions) did not designate holotypes in the materials he studied. Moreover, the vast majority of the collections he studied were sent to him on loan and were returned to the sender after Stephani separated a small part of the specimen for his own herbarium in Leipzig (now is in G). Therefore, at least two type specimens for each taxon are presumed to exist in the majority of cases, and the formal lectotype should be designated. A discussion on this issue, including also the questionable lectotypification of Stephani’s taxa by Bonner, is provided by Engel and Merrill  with theFrullania-type , rarely ventral intercalary; stem cross section slightly transversely elliptic with smooth external wall, 130–150 × 160–180 µm, with loosely defined cortex, outer cells with thick walls, irregular in shape, ca. 7–13 µm in diameter, inner cells to 25 µm in diameter, with thin walls and large to moderate in size, concave trigones. Leaves transversely inserted, mainly subimbricate or  obliquely spreading, somewhat turned dorsally; subimbricate leaves distinctly concave, with lower 1/4–1/3 erect spreading, then suddenly curved and going subparallel to the stem; (3–)4-lobed, lobes subequal , 600–800 µm long and (at the level of leaf lamina) 250–350 µm wide, undivided part 30–50 µm (2–5 cells) high, lobe apices prominently acuminate, with straight axis and 2–4(–5)-celled uniseriate ends, sinus strongly recurved in the base, each lobe with 2–4 acute 3–12-celled teeth in lower 1/3–1/2, besides each lateral side of the leaf have one more additional, commonly curved and branched, to 20 cells long tooth. Underleaves bilobed, 400–500 µm long, 150–250 µm wide (at the level of lamina), undivided part 20–40 µm (2–3 cells) high, lobes prominently acuminate with straight or distinctly recurved axes and 3–6-celled uniseriate ends, if uniseriate end shorter then the lobe ends by biseriate end to 8–10 cell-pairs long, sinus margin recurved in the base, lobe lower half in the sinus side with 2–4 teeth, leaf lateral bases with 3–5 teeth, the basal tooth largest. Cells in the lobe base 12–25 × (7–)10–12(–15) µm, oblong to nearly isodiametric, walls strongly vermiculately thickened, with prominent, large, and convex trigones; cuticle distinctly papillose-verrucose, sometimes coarsely so (Generative structures in molecularly studied specimens are absent) (Description (based on specimens examined placed in the  absent) .Comment. The type specimen of the taxon is somewhat smaller than other specimens from Yunnan and Sichuan. It has shorter but the same in width leaves and somewhat wider underleaves . Moreover, the plants in the lectotype are slightly crumbled, which gives the impression of smaller and shorter plants than commonly in the species. The old, dried herbarium specimens commonly have a less pronounced leaf surface armature, including papillae. The latter may be the consequence of the partial collapsing of the cells and indenting of surface elements inside. The latter is the reason the cuticle elements are not as coarse as in relatively fresh material.Tetralophozia pusilla (Steph.) Bakalin et Vilnet comb. nov.Chandonanthus pusillus Steph., Sp. Hepat. [Basionym: . Hepat.  3: 645.Lectotype (selected here): Komagadake Mt., Kai, No. 35, Aug. 1903 Coll. K. Tamura G00283305/11030! Stephani [Frullania-type (looking as dichotomous) and as subfloral innovations (1–)2 per gynoecium; stem cross section slightly transversely elliptic, 115–125 × 150–175 µm, with loosely defined cortex, distinctly striolate, outer cells with thick walls, with nearly rounded lumens, ca. 7–10 µm in diameter, inner cells 8–15 µm in diameter, with thin walls and large, mostly concave trigones. Leaves transversely inserted, mainly subimbricate or  obliquely spreading; subimbricate leaves distinctly concave, with lower 1/4–1/3 erect spreading, then suddenly curved and going subparallel to the stem; (3–)4-lobed, lobes subequal , 400–500 µm long and (at the level of leaf lamina) 250–450 µm wide, undivided part 50–90 µm (3–6 cells) high, lobe apices prominently acuminate, with straight axis and 1–3-celled uniseriate ends, sinus strongly recurved in the base, each lobe with 1–3 acute 1–6-celled teeth in lower third or basal teeth completely absent (as in the type specimen), each lateral side of the leaf have one more additional, commonly curved and sometimes branched, long (to 12 cells long) tooth. Underleaves bilobed, 300–400 µm long, 120–150 µm wide (at the level of lamina), undivided part 30–50 µm (2–3 cells) high, lobes prominently acuminate with straight or distinctly curved axes and 3–4-celled uniseriate ends, sinus recurved in the base, lobe bases in the sinus with 2–4 teeth, lateral underleaf bases with 3–5 teeth, the basal tooth the largest, sometimes branched and strongly curved, 1–2 basal teeth sometimes terminating with slime papilla. Cells in the lobe base 10–20 × 10–12 µm, oblong to nearly isodiameteric, strongly vermiculately thickened, with prominent, large, and convex trigones; cuticle papillose, sometimes scarcely so. Generative structures unknown  but far less so in North America and Asia, where the southernmost localities lie at 43° N in the Russian Far East mainland. The species is also illustrated here  an oro-temperate-oro-boreal amphioceanic range, and all records are from rocky substrates. Tetralophozia cavalli (Gola) Váňa is confined to the Central African high mountains . The species occurs on the bark of trees from middle to high elevations [Tetralophozia pilifera (Steph.) R.M. Schust. is a New Guinean endemic species growing mostly in epiphytic habitats and occurring much more rarely in epixylous habitats [The genus chljakov , who firbination . Seven ybination , Urmi [2Schuster —with Cham et al. , includeted here , based ohabitats . All recChandonanthus pusillus Steph. was treated as the synonym of Tetralophozia filiformis, although Schuster [T. filiformis is more strongly armed with spinescent teeth and cilia, which may occur far up the lobe margins; in C. pusillus, lobes are entire or bear 1–2 small teeth just above their bases”. The type localities of Chandonanthus filiformis and C. pusillus are, although in East Asia, situated at a strong distance from one another. The type locality of Chandonanthus filiformis is in Ma’ershan (‘Ma Cul Chan’ by Delavay), which is in Yunnan Province of China . The bryofloristic richness of this region is also obvious [As it was admitted starting from Urmi , ChandonSchuster  called tf China . Where possible, the specimens were processed via molecular-genetic analysis. The basic principle that was accepted in this work was not only to study quite old-type materials  but also to try to collect specimens in type localities or nearby—to reveal their real morphological variability in classic localities and to obtain material suitable for molecular-genetic analysis. Therefore, we visited Komagadake Mt. in Japan and the Ma’ershan area  in Yunnan Province of China. In addition, other areas in southern China (Sichuan Province), the Korean Peninsula and Russian Siberia were explored. In total, 17 specimens  were studied: 4 from Russian Siberia, 1 from the Russian Far East, 6 from the Korean Peninsula, 1 from Japan, 1 from North Vietnam and 3 from China. Additionally, three specimens of T. setiformis were included for molecular-genetic comparison. All specimens are listed in The studied specimens of the Tetralophozia remains questionable since two species from the genus Plicanthus R.M. Schust. were subsequently found within it [Plicanthus, we were not able to clarify the generic concept of both genera and concentrated here only on affinity among known Tetralophozia species. For molecular estimation, we selected 11 specimens of Tetralophozia and 11 species from the family Anastrophyllaceae, and Lophozia ascendens (Warnst.) R.M. Schust. from Lophoziaceae was chosen as an outgroup. In total, ITS1–2 nrDNA and trnL–F cpDNA sequence data for 14 samples were taken from our previous studies, GenBank accession numbers for them are provided in The monophyly of the genus ithin it ,24. Due trnL–F and Shaw et al. [trnG-intron were used for amplification and sequencing. Polymerase chain reaction (PCR) was carried out in 20 µL volumes with the following protocol: 3 min at 94 °C, 30 cycles , 2 min of final extension at 72 °C. Amplified fragments were visualized on 1% agarose TAE gels by EthBr staining, purified using the QIAquick Gel Extraction Kit , and used as a template in sequencing reactions with the ABI Prism BigDye Terminator v. 3.1 Ready Reaction Kit  following the standard protocol provided for the 3730 DNA Analyser .DNA was extracted with a DNeasy Plant Mini Kit . The primers suggested by White et al.  for ITS1w et al.  for trnGtrnL–F were produced manually; all positions were considered. Due to the absence of appropriate data, the trnG-intron dataset was not produced. Preliminary phylogenetic estimation revealed congruent results from both datasets; thus, they were combined in a single dataset ITS1–2+trnL–F for subsequent analyses by the maximum parsimony (MP) method with TNT v.1.5 [Newly generated sequences were assembled and aligned with previously obtained sequences in BioEdit 7.0.1 . AlignmeNT v.1.5  and the NT v.1.5 . The MP NT v.1.5  selectedNT v.1.5 . Thus, Mp-distances for the genus Tetralophozia were calculated in Mega 11 [The average pairwise  Mega 11  based onTetralophozia filiformis complex, we obtained the bioclimate variables for collecting localities of the specimens: 1) studied by molecular and/or morphological methods, and 2) randomly selected from the GBIF database  and for two specimens from Spain on which the first report of the taxon for Europe was based [https://www.worldclim.org/): BIO1 = Annual Mean Temperature, BIO2 = Mean Diurnal Range (Mean of monthly (max temp − min temp))m BIO3 = Isothermality (BIO2/BIO7) (×100), BIO4 = Temperature Seasonality (standard deviation ×100), BIO5 = Max Temperature of Warmest Month, BIO6 = Min Temperature of Coldest Month, BIO7 = Temperature Annual Range (BIO5–BIO6), BIO8 = Mean Temperature of Wettest Quarter, BIO9 = Mean Temperature of Driest Quarter, BIO10 = Mean Temperature of Warmest Quarter, BIO11 = Mean Temperature of Coldest Quarter, BIO12 = Annual Precipitation, BIO13 = Precipitation of Wettest Month, BIO14 = Precipitation of Driest Month, BIO15 = Precipitation Seasonality (Coefficient of Variation), BIO16 = Precipitation of Wettest Quarter, BIO17 = Precipitation of Driest Quarter, BIO18 = Precipitation of Warmest Quarter, BIO19 = Precipitation of Coldest Quarter. The obtained data were then tested using multivariate analysis (using Past ver. 4.03c [Since the climate is obviously changing in the distribution range of the r. 4.03c ). The hir. 4.03c , and EucTetralophozia filiformis is in hand because all our attention was attracted to the robust difference of collected material from T. setiformis . The involvement of additional material from type localities (including types) derived from other regions in East Asia revealed noticeable variation within widely treated T. filiformis in four main groups of species features: molecular genetics, morphology, ecology (including climate characteristics), and geography. The situation is somewhat similar to the recently published Ptilidium himalayanum, the species molecularly more different from the pair P. ciliare–P. pulcherrimum than the taxa constructing the pair and occupying a distinctly defined area in the Sino-Himalaya [The study illustrates that when only a few materials and from limited regions are available, the distinctive traits of the regional ‘populations’ may be overlooked, and a potentially new taxa could be neglected. In our case, when we had an experience restricted to the specimens collected in South Siberia, we certainly could not recognize that a taxon different from Himalaya .Tetralophozia sibirica as the terminal link in the adaptation process to the cold and dry climate within T. filiformis s. lat. complex, then it could be stressed the main morphological pathways of this ‘adaptation’  seem imaginable in regard to the northerly distributed Arctic-alpine T. setiformis (showing the same features in much more pronounced manner). This regularity to possess shorter divided leaves northward may be compared with trends observed in other genera, e.g., (1) Scapania with deeply divided leaves are not present in Arctic-distributed taxa , (2) the wide distribution of Gymnomitrion taxa with shallowly-lobed leaves in the Arctic and the absence taxa with deeply-divided leaves there.Treating


The unit cell parameter estimated from X-ray diffraction spectra exhibited a decreasing trend with an increase in wt % of Si content in both alloy systems. The particle size of powders estimated using bright field transmission electron microscopy images for ferritic (3 wt % Si) and austenitic (3 wt % Si) SS powders was found to be 65 ± 5 nm and 18 ± 3 nm, respectively. In case of the ferritic system, 3 wt % Si exhibited the highest densification (~98%) and micro-hardness of about 350.6 ± 11.2 HV, respectively. Similarly, for the austenitic system (3 wt % Si), maximum densification and micro-hardness values were about 99% and 476.6 ± 15.2 HV, respectively. Comparative analysis of potentiodynamic polarization, linear polarization, and electrochemical impedance spectroscopy results indicates an increase in electrochemical performance of both alloy systems as the wt % Si was increased. The increase in electrochemical performance is directly related to the increase in densification owing to Si addition in these alloys.Nanostructured ferritic (Fe Stainless steels (SSs) are the most diversely employed Fe-based alloys—finding their applications in oil and gas, power generation setups, desalination plants, nuclear reactors etc.—owing to their excellent corrosion resistance and mechanical properties. The development of powder metallurgy (PM) has revolutionized the near-net shape bulk synthesis of complex geometries with a variety of metallic powders . SS compIt is generally believed that powder metallurgy (PM) alloys exhibit lower corrosion and oxidation resistance as compared to wrought alloys of similar chemical compositions ,13,14. TAmong the different available routes of preparing nano-crystalline alloy powders, mechanical alloying (MA) is considered one of the promising techniques for such applications ,24,25,262 layer at the interface of outer Cr2O3 (passive film) and the base metal, thus providing additional protection to the base metal. Although there are few studies on the mechanical alloying and corrosion performance of Fe-Cr based alloys [(73−x)-Cr18-Ni9-Six, x = 0, 1, 2, 3 wt %) and ferritic (Fe(82−x)-Cr18-Six, x = 0, 1, 2, 3 wt %) alloys prepared by MA and SPS is not well documented in the literature. Therefore, the main objective of this research work was to synthesize nano-structured austenitic and ferritic stainless steel powders with varying Si content via mechanical alloying, followed by SPS and the conducting of an in-depth study on their electrochemical performance. The presence of nanostructure will help in achieving better densification and so will improve the corrosion resistance of the alloys along with the presence of Si in these alloys.The commercially available SSs do have a small amount of Si and the main reason for such an addition is oxygen absorption along with an increased oxidation resistance of such alloys, irrespective of ferritic and austenitic SSs ,35,36,37d alloys ,22, the (73−x)-Cr18-Ni9-Six  and Fe(82−x)-Cr18-Six  alloys. Ferritic powders were milled for 100 h and austenitic powders for 160 h respectively and the details are reported elsewhere [High purity (>99.99%) Fe, Cr, Ni, and Si powders—with particle sizes of <45 μm, <140 μm, <100 μm, and <50 μm respectively—were used to prepare the nominal compositions of FeAfter mechanical alloying, ball-milled powders were sintered in an automated, spark plasma sintering machine . A 20 mm graphite die (current was passed through it) was used to hold the powders. The alloys were sintered in vacuum to avoid oxidation and a thermocouple was used for temperature measurement. To ease the specimen removal once the sintering process was completed, a sheet of graphite was employed which also helped in reducing the friction issues between the die and powder. Sintering was performed at 1100 °C for 15 min with an applied pressure of 60 MPa and a heating rate of 100 °C/min under vacuum and sintering parameter optimization  is explained elsewhere . SintereA JOEL field emission scanning electron microscope (FE-SEM) equipped with energy dispersive spectroscopy (EDX) was leveraged to confirm the chemical composition of ball-milled powders as well as after sintering. A JOEL FE-transmission electron microscope (FE-TEM) was utilized to confirm the XRD data collected from MA powders. For TEM microscopy, the powders were dispersed in acetone in a sealed glass container and ultrasonicated for 5 min to remove their stickiness. For particle size analysis of milled powders, low magnification TEM images were taken, and the average of at least 20 particles is reported here.2 was selected for accuracy and repeatability, and experiments were performed in 0.2 M NaCl solution at room temperature. A Gamry Reference 3000 potentiostat was used for these experiments, which were repeated thrice to confirm the authenticity of the reported data. A scan rate of 0.2 mV/s was selected for potentiodynamic polarization (PDP) experiments using DC105 software, which was subsequently used for data analysis as well. The common practice of cathodic cleaning (for removing any traces of surface oxides) was employed at −1.0 VSCE for 180 s, before corrosion experiments. A sinusoidal voltage perturbation signal of 10 mVSCE with a frequency range of 100,000 to 0.2 Hz, was used for electrochemical impedance spectroscopy (EIS) tests. To evaluate the electrochemical impedance response of the alloys, first an oxide film on the surface of these alloys. The potential selected for this film formation was 0.05 VSCE in deaerated 0.2 M NaCl solution, based on PDP data. The duration for film formation was 30 min under potentiostatic conditions. Linear polarization resistance (LPR) experiments were carried out over a range of potentials (±20 mVSCE) around the open circuit potential (OCP) using a scan rate of 0.3 mV/s. Before performing the LPR experiments versus open circuit potential (OCP), its value was measured, after the samples were kept inside the solution for 30 min.A three-electrode corrosion cell  was used for electrochemical tests. Grinding was carried out up to 2000 grade of SiC abrasive paper before applying 0.5 μm alumina suspension for mirror polishing. Finally, the samples were ultrasonically cleaned in acetone before performing the corrosion experiments. For corrosion tests, an exposed surface area of 0.1 cm(82−x)-Cr18-Six, x = 3 wt %) SS powders milled for 100 h and austenitic (Fe(73−x)-Cr18-Ni9-Six, x = 3 wt %) SS powders milled for 160 h followed by annealing is presented in XRD spectra collected from ferritic (Fe(73−x)-Cr18-Ni9-Six (x = 2) milled for 5, 10, 160 h, and the 160 h annealed sample presents phase evolution during ball milling. XRD spectra from pure Fe, Cr, Si, and Ni powder are also presented before milling. The first specimen was collected after 5 h of ball milling and the XRD spectrum is presented in The XRD spectrum collected from 160 h milled powder indicates that the powders are still amorphous and the transformation of α to γ has not occurred yet. After 160 h of milling, the authors decided to anneal the powders at 1100 °C for 1 h holding time under argon. The XRD data collected from annealed powder are presented in FE-TEM was leveraged to confirm the XRD results—i.e., formation of α-phase at 5 h milling, amorphous phase at 20 h, and evolution of γ-phase after annealing—and the data are presented in (82−x)-Cr18-Six, x = 3 wt %) and austenitic (Fe(73−x)-Cr18-Ni9-Six, x = 3 wt %) specimens are presented in (82−x)-Cr18-Six, x = 0, 1, and 2 wt %) austenitic (Fe(73−x)-Cr18-Ni9-Six, x = 0, 1, and 2 wt %). The analysis of the XRD data confirmed the BCC and FCC crystal structures of ferritic and austenitic SS specimens, respectively. The inset in The alloys were sintered under optimized conditions of 1100 °C for 10 min, applied pressure of 60 MPa, and at a heating rate of 100 °C/min under vacuum. The XRD spectra collected from the spark plasma sintered ferritic (Fe(82−x)-Cr18-Six, x = 0, 1, 2, and 3 wt %) SS sintered specimens, the maximum densification achieved was lesser than 98% while in case of austenitic specimens (Fe(73−x)-Cr18-Ni9-Six, x = 0, 1, 2, and 3 wt %), at the same SPS conditions, ~99% relative density was observed. Micro-hardness (HV) values show an increasing trend with the increase in % relative density. Starting with the α-BCC system, a minimum and maximum hardness was calculated at 209.6 ± 18.2 HV and 350.6 ± 11.2 HV for 0 wt % and 3 wt % Si, respectively. Similarly, for the γ-FCC system, minimum and maximum hardness was calculated at 280.8 ± 13.2 HV and 476.6 ± 15.2 HV for 0 wt % and 3 wt % Si, respectively.Micro-hardness and relative densification data calculated from both BCC (ferritic SS) and FCC (austenitic SS) are presented in (82−x)-Cr18-Six, x = 0, 1, 2, and 3 wt %) and austenitic (Fe(73−x)-Cr18-Ni9-Six, x = 0, 1, 2, and 3 wt %) SS sintered specimens in a deaerated 0.2 M NaCl solution at room temperature are presented in pitE), passive current density (passi), and corrosion potential (corrE) etc.) are presented in corrE, but that corri monotonically decreases with the increase in wt % Si, indicating the higher propensity of the 0 wt % Si sample to corrosion. Furthermore, to estimate passi, a potential at −0.55 VSCE (in the passive region) was selected and the data indicate that passive current density is not changing with wt % change in Si. However, the electrochemical performance significantly changes; i.e., the width of passive region increases. pitE values in pitE is indicated as N/A; and for the 3 wt % Si, pitE is calculated at 0.128 VSCE. The difference in pitE values from the 0 wt % Si to the 3 wt % Si sample clearly indicate an increase in corrosion resistance due to the formation of more protective passive film with an increase in Si content (reasons are presented in the Discussion section). Similar analysis of kinetic parameters obtained from the austenitic SS sintered specimens is represented in corrE and highest corri estimated at −0.231.6 VSCE and 0.886 pA/cm2 respectively. The sample with 3 wt % Si exhibits the highest corrE and lowest corri values estimated at −104.4 VSCE and 739.2 pA/cm2 respectively. The visual comparison of PDP curves indicates that the 0 wt % Si sample does not exhibit a clear passive region and the 3 wt % Si sample has a wider passive region. The comparative analysis of pitE values indicates a monotonic increase in corrosion resistance with an increase in Si content. To conclude, increase in the width of a passive region in both systems with an increase in Si content is directly related to the improvement in electrochemical performance. The electrochemical performance comparison between ferritic and austenitic SSs is presented in the Discussion section.The PDP scans collected from both ferritic (Fe (82−x)-Cr18-Six, x = 0, 1, 2, and 3 wt %) and austenitic (Fe(73−x)-Cr18-Ni9-Six, x = 0, 1, 2, and 3 wt %) sintered specimens  and −0.05 VSCE (for austenitic SS) in deaerated 0.2 M NaCl at room temperature. The potential was selected from the PDP data in (82−x)-Cr18-Six, x = 0, 1, 2, and 3 wt %) austenitic (Fe(73−x)-Cr18-Ni9-Six, x = 0, 1, 2, and 3 wt %) are presented in Rs represents the solution/electrolyte resistance, while Rp is the polarization resistance of the passive film. The data collected after fitting the experimental EIS data with the EEC model are presented in Rp values estimated after the EEC model fit indicates a monotonic increase with wt % Si in both ferritic and austenitic SS samples.EIS experiments were performed at −0.55 V(82−x)-Cr18-Six, x = 0, 1, 2, and 3 wt %) and austenitic (Fe(73−x)-Cr18-Ni9-Six, x = 0, 1, 2, and 3 wt %) SS are synthesized under similar ball milling conditions  and sintered under the same conditions. The examination of the XRD spectrum of austenitic stainless steel powders milled for 160 h revealed a characteristic amorphous spectrum. Ferritic powders also showed a similar trend for up to 50 h of milling; however, continued milling up to 85–100 h resulted in a crystallized alpha phase [(82−x)-Cr18-Six, x = 3 wt %) and austenitic SS (Fe(73−x)-Cr18-Ni9-Six, x = 3 wt %) indicates that austenitic alloy exhibited the higher density. A possible reason for the monotonic increase in relative density in both ferritic and austenitic SS can be the presence of Si in these alloys. Many studies [The mechanical properties—especially corrosion resistance—of powder metallurgy (PM) steels is strongly dependent on the density of the sintered alloys. Many sintering parameters such as sintering temperature , holdingha phase . On the ha phase . Similarha phase  observedha phase . Obviousha phase . As repoha phase  highlighha phase  favored  studies ,57 have  studies  reportedThe comparison of the data presented above in (82−x)-Cr18-Six, x = 3 wt %) and austenitic (Fe(73−x)-Cr18-Ni9-Six, x = 3 wt %) SS samples indicates that austenitic SSs are more corrosion resistant than ferritic SSs ((82−x)-Cr18-Six, x = 3 wt %) is around 98% and austenitic (Fe(73−x)-Cr18-Ni9-Six, x = 3 wt %) SS is around 99%, but the corrosion response is dramatically different. Therefore, it is safe to conclude that the dramatic increase in corrosion resistance is not due to the relative densification. For better comparison, a PDP plot for ferritic SS (Fe (82−x)-Cr18-Six, x = 3 wt %, density = 98%) is plotted against austenitic SS (Fe(73−x)-Cr18-Ni9-Six, x = 2 wt. %, density~99) The comparative analysis of ferritic SS (Feitic SSs a. It is (82−x)-Cr18-Six, x = 0, 1, 2, and 3 wt %) and austenitic (Fe(73−x)-Cr18-Ni9-Six, x = 0, 1, 2, and 3 wt %) stainless steel powder alloys were prepared by mechanical alloying (MA). Various techniques such as X-ray diffraction (XRD), scanning electron microscopy (SEM), and field emission transmission electron microscopy (FE-TEM) were leveraged for microstructural characterizations. Ferritic powders exhibiting a body centered cubic (BCC) structure were obtained after 100 h of milling while austenitic powders were milled for 160 h followed by annealing at 1100 °C for 60 min in an argon atmosphere. The particle size after ball milling was estimated from low magnification FE-TEM images. Ferritic (Fe(82−x)-Cr18-Six, x = 3 wt %) SS powders have an average particle size of 65 ± 5 nm and austenitic (Fe(73−x)-Cr18-Ni9-Six, x = 3 wt %) powders average 18 ± 3 nm. Spark plasma sintering (SPS) was performed at 1100 °C for 10 min under an applied pressure of 60 MPa and at a heating rate of 100 °C/min under vacuum. XRD data confirm the BCC and face centered cubic (FCC) crystal structure of ferritic and austenitic SSs, respectively. The percentage of densification was estimated by Archimedes’ principle and the data indicate that both alloy systems exhibit an increasing trend of densification with the increase in wt % Si content. In both ferritic and austenitic systems maximum densifications of ~98% and ~99%, respectively, were observed for 3 wt % Si. A similar trend was observed for micro-hardness: a 3 wt % system exhibited the highest micro-hardness. The comparative analysis of corrosion kinetic parameters for the ferritic system (Fe(82−x)-Cr18-Six, x = 0, 1, 2, and 3 wt %) showed that there was no significant difference in corrosion potential (corrE) values ; however, pitting potential was significantly improved for 3 wt % Si alloys. Visual comparison of PDP scans shows that a 0 wt % Si system does not exhibit a clear passive region while a 3 wt % Si system exhibited a wide passive region. The maximum pitE was estimated to be 125 mV for a 3 wt % Si system. A similar trend of kinetic parameter variation was observed for an austenitic system at 0 wt %. Si has no clear passive region and 3 wt % Si exhibited a wide passive region. Maximum pitE was estimated to be 259 mV. LPR values indicated a monotonic increase with the wt % Si content in both alloy systems. An electrical equivalent circuit (EEC) with a constant phase element was used to fit the EIS data, and the results showed an increase in electrochemical impedance performance of the alloys with an increase in Si content of the alloy systems.Nanostructured ferritic (Fe


The RD was paclitaxel 80 mg/m2 on Day (D)1,D8 plus lurbinectedin 2.2 mg/m2 on D1. At this RD, myelotoxicity was reversible and manageable, and most non-hematological toxicities were mild/moderate. Adding BEV did not notably change tolerability. Twenty-five confirmed responses were observed: 20/51 evaluable patients in Group A , and 5/10 evaluable patients in Group B (ORR = 50%). Most responders had breast (n = 7/12 patients), small cell lung (SCLC) (n = 5/7), epithelial ovarian (n = 3/9) and endometrial cancer (n = 3/11) in Group A, and epithelial ovarian (n = 3/4) and NSCLC (n = 2/6) in Group B. Clinical benefit rate was 61% in Group A (58% at the RD), and 90% in Group B. No major pharmacokinetic drug-drug interactions were observed. Paclitaxel/lurbinectedin and paclitaxel/lurbinectedin/BEV are feasible combinations. Further development is warranted of paclitaxel/lurbinectedin in SCLC, breast, and endometrial cancer, and of paclitaxel/lurbinectedin/BEV in epithelial ovarian cancer.Lurbinectedin and paclitaxel showed synergism in preclinical studies and have non-completely overlapping toxicity profiles. This phase I trial evaluated a combination of paclitaxel and lurbinectedin with/without bevacizumab in advanced tumors. This trial was divided into Group A, which evaluated weekly paclitaxel (60 or 80 mg) plus lurbinectedin (3.0–5.0 mg flat dose [FD] or 2.2 mg/mThe online version contains supplementary material available at 10.1007/s10637-022-01281-z. It inhibits oncogenic transcription primarily through binding to the exocyclic amino group of guanine-rich DNA sequences around promoters of protein-coding genes, thereby altering the 3D DNA structure and evicting oncogenic transcription factors from their binding sites, thus halting their aberrant transcription programs ) were due to disease progression. One discontinuation was due to treatment-related thrombocytopenia. The median time on treatment was 7.9 months. The median DIs were 50.3 mg/m2/week for paclitaxel, 0.7 mg/m2/week for lurbinectedin and 5.0 mg/m2/week for BEV; the median relative DIs were 94%, 100% and 99%, respectively. Three patients (25%) had cycle delays and 2 patients (17%) had study drug dose reductions owing to treatment-related reasons .In Group B, 12 of the 14 enrolled patients received a total of 111 treatment cycles at a dose of paclitaxel 80 mg/m2, lurbinectedin 5.0 mg FD) had delayed DLTs (grade 3 or 4 neutropenia after Cycle 1). After DL3, the paclitaxel schedule was changed to D1 and D8 q3wk owing to the finding of a high incidence (4 of 6 patients) of paclitaxel dose omissions due to treatment-related neutropenia during Cycle 1 at DL3. At DL4 , 3 of 6 patients had DLTs ; as a result, this dose was defined as the MTD. None of the first 6 patients at DL5  had DLTs. The DL5 cohort was then expanded, converting lurbinectedin to a BSA-based dose of 2.2 mg/m2 . Six of 28 patients in the expanded DL5 cohort  had DLTs  [n = 1], and grade 3 vomiting [n = 1]), thereby confirming paclitaxel 80 mg/m2 on D1 and D8 plus lurbinectedin 2.2 mg/m2 on D1 as the RD in Group A, with prolonged severe neutropenia as the most common DLT. No episodes of febrile neutropenia occurred as DLTs in Group A.Fifty-two treated patients in Group A were evaluable for DLTs . Three patients were non-evaluable because they did not receive a complete Cycle 1 due to disease-related events (n = 2), or because of lack of laboratory assessment for DLT evaluation during Cycle 1 (n = 1). Paclitaxel was initially administered at a schedule of D1, D8 and D15 q3wk. No DLTs occurred at the first two dose levels. Two of 6 patients at DL3 . Three patients had DLTs: grade 3 febrile neutropenia (n = 1), grade 4 neutropenia > 3 days (n = 1), and grade 4 large intestine perforation . No treatment-related deaths occurred in Group B. The percentage of patients with DLTs in Group B  in the absence of treatment-related mortality was below the threshold of one third of patients defined in the study protocol, thereby confirming the feasibility of adding BEV to the RD defined in Group A.All 12 patients treated with paclitaxel 80 mg/mAll treated patients were evaluable for safety. Treatment-related AEs and laboratory abnormalities at the RD in Group A and in Group B are shown in Table Thirty-four of 37 patients (92%) treated at the RD in Group A had at least one treatment-related AE. Most of these AEs were grade 1/2, with the most common being fatigue (59% of patients), nausea (54%), vomiting (46%), peripheral sensory neuropathy (27%), diarrhea and alopecia (24% each). Grade ≥ 3 treatment-related AEs comprised catheter site cellulitis, diarrhea, fatigue, maculopapular rash, nausea, sepsis, and vomiting in one patient each (3%). Of these, only sepsis reached grade 4. Hematological abnormalities consisted of anemia , neutropenia  and thrombocytopenia ; one patient (3%) had treatment-related grade 3 febrile neutropenia. The most frequent biochemical abnormalities were increases in creatinine (81%), alanine aminotransferase (ALT) , aspartate aminotransferase (AST)  and AP . Seven patients (18.9%) required red blood cell (RBC) transfusions and four patients (10.8%) required secondary granulocyte colony-stimulating factor (G-CSF) support. No treatment discontinuations or deaths occurred due to toxicity.All 12 treated patients in Group B had at least one treatment-related AE. Most of these AEs were also grade 1/2, with the most frequent being fatigue (58%), nausea (50%), alopecia (42%) and diarrhea (33%). Grade ≥ 3 treatment-related AEs comprised abdominal abscess and pulmonary embolism in two patients each (17%), and colonic fistula, fatigue, gastrointestinal fistula, large intestine perforation, and septic shock in one patient each (8%); of these, only the intestinal perforation was grade 4. Hematological abnormalities consisted of anemia , neutropenia  and thrombocytopenia . Two patients (17%) had grade 3 febrile neutropenia. All biochemical abnormalities were grade 1/2, with the most common being increases in creatinine and ALT (75% each), AST and AP (42% each). Three patients (25.0%) required RBC transfusions and three (25.0%) were given G-CSF support. One patient discontinued treatment due to toxicity (grade 4 large intestine perforation). No deaths occurred due to toxicity.Fifty-one patients in Group A and 10 patients in Group B were evaluable for efficacy. Four patients in Group A and 2 in Group B were not evaluable because no tumor evaluations were conducted after baseline imaging.In Group A, 20 patients had confirmed responses, including one complete response (CR)  and 11 patients had stable disease (SD) ≥ 3 months  at all dose levels, with objective tumor shrinkage in 35 (71%) of 49 evaluable patients  and 4 patients had SD ≥ 3 months   at the RD in Group A and 6 of 10 patients (60%) in Group B showed greater antitumor activity with paclitaxel and lurbinectedin with or without BEV compared to the last prior therapy . Among patients pretreated with taxanes, 8 confirmed responses and 5 SD ≥ 3 months occurred at the RD in Group A , and 5 confirmed responses and one SD ≥ 3 months in Group B .Clinical benefit for > 12 months was observed in 5 patients: 4 in Group A (3 at the RD) and one in Group B. These patients had SCLC (n = 2), epithelial ovarian, endometrial cancer, and NSCLC (n = 1 each), had received 1–3 prior chemotherapy lines, and were given 18–35 cycles of study treatment each . Four of these patients showed no signs of disease progression prior to study termination.max) and area under the concentration–time curve (AUC). Potential drug-drug interactions (DDIs) between paclitaxel and lurbinectedin could not be fully ruled out, since a slight decrease in the CL of each drug was observed at the high AUC of the other drug. No statistically significant differences were observed in the PK parameters of paclitaxel and lurbinectedin in the presence or absence of BEV. Additional details are provided in Supplemental Information.All patients were sampled for PK analysis and were suitable for non-compartmental analysis (NCA). Parameters obtained for paclitaxel and lurbinectedin at each dose level are shown in Supplemental Information. Wide variability was observed in lurbinectedin and paclitaxel total clearance (CL). No dose linearity could be established for lurbinectedin maximum concentration  [The safety profile of the paclitaxel plus lurbinectedin combination was predictable. Myelotoxicity at the RD was common but reversible and manageable, and most non-hematological toxicities were mild/moderate. As expected, severe myelotoxicity and some non-hematological toxicities  were slightly more frequent at this RD compared to the established dose for lurbinectedin monotherapy (3.2 mg/m2 q3wk) , 17. Thi2 q3wk) –20. No pvs. chemotherapy alone in patients with solid tumors [As expected, the addition of BEV 15 mg/kg q3wk to this RD increased the incidence of some treatment-related events , and severe neutropenia. Similar increases have been reported in clinical trials comparing chemotherapy plus BEV d tumors , 21, 22.vs. 35%) [BRCA-unselected breast cancer (50% vs. 9%) [vs. 14–23%) [vs. 20–27%) [vs. 22–42%) [vs. 21–60%) [2/week). These effects were achieved at a lower paclitaxel dose intensity (50.3 mg/m2/week vs. 72–78 mg/m2/week) [vs. 50–100%; grade 3/4, 0% vs. 3–21%) [Encouraging antitumor activity was observed herein with the paclitaxel plus lurbinectedin combination in several indications. Recognizing the limitations of cross trial comparisons, the response rates at the RD without BEV were higher than those reported for lurbinectedin monotherapy in second-line treatment of patients with SCLC (80% vs. 35%) , BRCA-un vs. 9%) , and ova 14–23%) , 24. Wit 20–27%)  and meta 22–42%) , 27, and. 3–21%) , 27–31. . 3–21%)  and ixab. 3–21%) , 34.vs. chemotherapy alone in patients with recurrent ovarian cancer [The addition of BEV improved response rate to the combination at the RD in patients with epithelial ovarian cancer (from 33 to 75%) and NSCLC (from 25 to 33%). This is in line with the finding, in previous studies, of a 15–20% improvement in ORR with the addition of BEV to chemotherapy n cancer , 22, 35 n cancer , 36, 37.max and AUC was probably due to the dose levels explored being very close. DDIs between paclitaxel and lurbinectedin could not be ruled out, although their clinical relevance would be marginal, based on the slight changes in CL and the large variability of PK parameters observed. BEV had no significantly effects on the PK profile of either lurbinectedin or paclitaxel.The PK parameters of paclitaxel and lurbinectedin in the present study were generally similar to those reported elsewhere , 38. The2 on D1 and D8 combined with lurbinectedin 2.2 mg/m2 on D1 q3wk, with or without the addition of BEV 15 mg/kg on D1, showed a manageable overall safety profile and promising antitumor activity in patients with selected advanced solid tumors. These results support further development of this combination without BEV in the treatment of SCLC, breast, and endometrial cancer, and with added BEV in the treatment of epithelial ovarian cancer.In conclusion, weekly paclitaxel 80 mg/mSupplementary file1 (DOCX 163 KB)Below is the link to the electronic supplementary material.


A randomized, blank-controlled clinical trial included 68 T2D patients and 10 controls. Based on the urinary albumin-to-creatinine ratio (UACR), 68 diabetic patients were stratified into three levels, UACR < 30 mg/g, UACR ≧ 30 mg/g to ≦ 300 mg/g and UACR ˃ 300 mg/g, who were randomized (1:1:1) to receive SGLT2i treatment for 12 weeks. The concentrations of NPH and TGF-β1 in urine were measured as indications of podocyte injury and renal fibrosis. Low-grade inflammation was assessed by the levels of IL-6, TNFα and hsCRP. After 12 weeks of SGLT2i treatment, the levels of UACR and NPH decreased, UTGF-β1 increased in the T2D with microalbuminuria and macroalbuminuria groups, NPH  and , UTGF-β1  and , respectively. The changes in NPH were positively correlated with the UACR and negatively correlated with UTGF-β1 in T2D with albuminuria. SGLT2i alleviate nephrin loss and enhance TGF-β1 excretion in urine in T2DM with albuminuria. The anti-albuminuric effect of SGLT2i could be attributed to mitigating podocyte apoptosis and attenuating renal fibrosis.To evaluate the effect of SGLT2 inhibitor (SGLT2i) on albuminuria, nephrin (NPH) and transforming-growth-factor-betaTrial registration This clinical trial was registered on 15/10/2019, in ClinicalTrials.gov, and the registry number is NCT04127084. Therefore, effectively reducing albuminuria is extremely important and can reduce the risk of ESRD due to DN. Current standard albuminuria intervention strategies include sodium restriction, weight loss, blood pressure control, and inhibition or blocking of RASS activation2. However, these traditional methods did not take into account the effects of high sugar and high fat toxicity on local renal hemodynamics. Albuminuria may not be well controlled by simply following the above strategies. Sodium-glucose cotransporter 2 inhibitors (SGLT2i) are a newer class of hypoglycemic drugs that act on sodium-glucose cotransporter 2 in the curvature of the proximal tubule of the kidney, lower the renal glucose threshold, inhibit glucose and sodium reabsorption in proximal tubules, excrete excess urinary glucose and urinary sodium, transport urinary sodium to the distal convoluted tubule, stimulate tubuloglomerular feedback, contract afferent arterioles, and reduce the pressure of the glomerulus3. Therefore, SGLT2 inhibitors reduce glomerular hyperfiltration, effectively improve early glomerular hemodynamic abnormalities and reduce albuminuria. Animal experiments and human studies have shown that SGLT2i can reduce glucose, weight, and systolic blood pressure while reducing albuminuria6. Nevertheless, the mechanisms behind the renal protective effect of SGLT2 inhibitors are not fully understood. In the current trial, the concentrations of NPH and TGF-β1 in urine were measured as indications of podocyte injury and renal fibrosis. Low-grade inflammation was assessed by the levels of IL-6, TNFα and hsCRP. The purpose of our research was to observe the intervention of SGLT2i in type 2 diabetic patients with different levels of albuminuria for 12 weeks and to investigate the effect of SGLT2i on nephrin and TGF-β1 in urine, glucose and lipid metabolism, and low-grade inflammation in type 2 diabetes patients.Diabetic nephropathy (DN) is one of the most important chronic microvascular complications of type 2 diabetes and is also the most common cause of end-stage renal disease (ESRD) on a global scale. Early diagnosis and effective intervention for albuminuria are the core strategies to prevent the progression from DN to ESRD. The presence and deterioration of albuminuria and the increase in serum creatinine are still the main indicators for clinical evaluation of the progression of DN2), women in pregnancy or lactation, alcoholics. The study was approved by the ethics committee of Zhongshan Hospital of Xiamen University and conducted in accordance with the Helsinki Declaration. Written informed consent was obtained from each participant.Based on the urinary albumin-to-creatinine ratio (UACR), type 2 diabetes patients were stratified into three levels, UACR < 30 mg/g, UACR ≧ 30 mg/g to ≦ 300 mg/g and UACR > 300 mg/g. Participants with T2D patients and healthy controls were allocated (2:2:2:1) to UACR < 30 mg/g, UACR ≧ 30 mg/g to ≦ 300 mg/g and UACR ˃ 300 mg/g or blank-controlled group. Considering the lost follow-up rate of at least 20%, 68 T2D patients and 10 normal controls were ultimately recruited between October 2019 and October 2020 at Zhongshan Hospital of Xiamen University in the Department of Endocrinology and Metabolism and Nephrology, which included 21 T2D with nonalbuminuria (UACR < 30 mg/g), 20 T2D with microalbuminuria (UACR30 ~ 300 mg/g), and 27 T2D with macroalbuminuria (UACR > 300 mg/g). Inclusion criteria for type 2 diabetes participants were as follows: the patients were initially diagnosed with type 2 diabetes according to World Health Organization (WHO) criteria, were 18–80 years of age, had HbA1c between 7 and 11%, received antidiabetic agents except SGLT2i and GLP-1, and had normal liver function and renal function. Exclusion criteria were set as follows: acute or chronic urinary tract infection, genital tract infection, type 1 diabetes, special type of diabetes, recent acute complications including diabetic ketoacidosis and hyperglycaemic hyperosmolar coma, acute or chronic glomerulonephritis, Long term use of corticosteroids or immunosuppressants, impaired liver function, impaired renal function . In this clinical trial, 68 subjects with T2D were divided into three groups according to UACR levels: T2DM with nonalbuminuria (UACR < 30 mg/g), T2DM with microalbuminuria (UACR30 ~ 300 mg/g), and T2DM with macroalbuminuria (UACR > 300 mg/g), who were randomized (1:1:1) by computer-generated numbers to treated with dapagliflozin 10 mg per day , empagliflozin 10 mg per day  or canagliflozin 100 mg per day  for 12 weeks. Considering the beneficial effects of RAS receptor blockers (ACEIs or ARBs) on albuminuria7, the subjects were asked to stop using ACEIs or ARBs for at least 4 weeks. If the patients’ SBP exceeded 140 mmHg or DBP exceeded 90 mmHg, CCB, β1 receptor blocker, and α receptor blocker were prescribed. We collected fasting blood and lean-catch 24-h urine samples for each participant before and after 12 weeks of treatment. All patients received standard care, including diet and exercise education, by professional nurses. The patients received follow-up every two weeks to take a routine urine test, measure blood glucose, systolic blood pressure (SBP), diastolic blood pressure (DBP), body mass index (BMI), body weight (BW), waist circumference (WC), hip circumference (HC), and WHR, and monitor the adverse events of drugs, including diabetic ketoacidosis, acute genital and urinary tract infection, bone fracture, leg and foot amputation, cancer.This was a 12-week, randomized, blank-controlled clinical trial included 68 T2D patients and 10 controls , total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were assayed using standard methods . The estimated glomerular filtration rate (eGFR) was based on the CKD-EPI creatinine equation8. Fasting blood glucose (FPG) was measured by the hexokinase method, high-sensitivity C-reactive protein (hsCRP) concentrations were measured using immune turbidimetry, uric acid (UA) was measured by the uricase-peroxidase method, and serum creatinine (Scr) was measured by the picric acid method. All measurements were performed using an automated method . Interleukin-6 (IL-6) and tumor necrosis factor α (TNFα) were measured by electrochemiluminescence . HbA1c was measured by high-performance liquid chromatography (HPLC) . Urinary albumin was determined by immunoturbidimetry, and creatinine was measured by the picric acid method . Urinary TGF-β1 concentrations were assayed using a dual-antibody sandwich enzyme-linked immunosorbent assay (ELISA) kit . Nephrin excretion was measured in the urinary samples of patients using quantitative ELISA kits in indirect competitive mode .The waist-hip ratio (WHR) was calculated as the ratio of waist circumference to hip circumference. The enrolled patients fasted for at least 8 h before fasting cubital venous blood was drawn to determine HbA1c, FPG, Scr, UA, CRP, IL-6, and TNFα. On the other hand, 24 h of urine was collected to determine UACR, nephrin and TGF-βχ2) was used to analyze the proportion of SGLT2i initiation, the percentage of metformin and DPP-IV inhibitors in baseline medications in this study. The comparisons of basic characteristics among multiple groups were made using one-way analysis of variance. Comparisons of baseline and posttreatment data in the same group were made using a paired t test or Wilcoxon signed rank test. The Wilcoxon signed rank test was used to determine the changes in NPH, TGF-β1, hsCRP, IL-6 and TNFα before and after 12 weeks of SGLT2i treatment. Spearman correlation coefficients were calculated to explore the relationships between baseline UACR and NPH, eGFR, Scr, Age, HbA1c, SBP, hsCRP, and IL-6, to explore the relationship between changes in urinary nephrin and those in urinary TGFβ1, UACR, eGFR, and weight before and after 12 weeks of treatment with SGLT2i in type 2 diabetes with albuminuria. Two-tailed significance was set at P < 0.05.SPSS version 21  and GraphPad Prism version 9.0  were utilized for statistical analysis and the construction of graphs. Normally distributed data are presented as the mean±standard deviation. The skewed distribution variables are expressed as the median . The chi square test  were more likely to be younger and have a higher baseline BMI. There were significant differences in the values of serum creatinine (Scr) and LDL-C in all four groups (P < 0.05), as shown in Table The clinical and laboratory characteristics are shown in Table The percentage of metformin and DPP-IV inhibitors in the baseline analysis were not statistically significant between different UACR levels, as shown in Table Three SGLT2 inhibitors, including dapagliflozin, empagliflozin, and canagliflozin, of which 22 (32.4%) used dapagliflozin, 27 (39.7%) used empagliflozin, and 19 (27.9%) used canagliflozin, were randomized to administer throughout the study. The percentage of SGLT2 inhibitor initiation was not statistically significant, as shown in Table P < 0.001) gradually increased in the three diabetic groups. Patients with higher baseline UACR (UACR ≥ 300 mg/g) categories were more likely to have lower eGFR and higher Scr (P < 0.05). Spearman rank correlations showed that NPH , eGFR , Scr , age , HbA1c , SBP , hsCRP  and IL-6  were highly relevant to the baseline UACR, as shown in Table The median values of baseline UACR  showed significant differences among the three diabetic groups, in which nephrin and Scr were highest and GFR was lowest at baseline UACR ≥ 300 mg/g. After 12 weeks of SGLT2 inhibitor treatment, the UACR decreased in the microalbuminuria and macroalbuminuria groups compared to baseline, which were 87  vs. 75  mg/g, P = 0.003 and 861  vs. 738  mg/g, P = 0.037, respectively. The values of NPH were significantly lower in the microalbuminuria and macroalbuminuria groups than at baseline   were comparable across the three diabetic groups. After 12 weeks of SGLT2i treatment, TGF-β1 secretion in urine was significantly enhanced in the three groups compared with baseline, which was 4.34 ± 1.52 versus 6.78 ± 1.36 pg/mL, P < 0.001, 4.88 ± 1.31 versus 7.27 ± 1.21 pg/mL, P < 0.001 and 4.30 ± 1.34 versus 6.78 ± 2.59 pg/mL, P < 0.001, respectively . The values of UA  and WHR  were significantly lowered, and HDL-C was significantly increased  compared with baseline in the UACR < 30 mg/g group, while TG, HbA1c, HC, WHR and DBP were markedly reduced in the microalbuminuria group after 12 weeks of SGLT2i treatment  , which was negatively correlated with UTGF-β1 , eGFR , BMI , WC , WHR  after 12 weeks of SGLT2i treatment in T2D patients with albuminuria.Spearman’s correlation analysis showed that the changes in NPH were positively correlated with the UACR . The slit diaphragm is the final barrier preventing passage of proteins into the urinary filtrate. Mutations or downregulation of nephrin is associated with loss of defective filtration barrier, causing albuminuria13. Our results indicated that baseline nephrin in urine gradually increased with the progression of albuminuria, nephrin loss into urine was highest and eGFR was slowest in the macroalbuminuria subgroup.Our findings show that baseline UACR was positively correlated with nephrin expression. After 12 weeks of SGLT2i treatment, UACR and NPH remarkably decreased in the T2D with microalbuminuria and macroalbuminuria groups. The pathogenesis of albuminuria in diabetic nethropathy has been linked to podocyte foot process effacement (FPE), reduced nephrin-positive areas in glomeruli, mesangial expansion, glomerular basement membrane thickening, and increased area of TGF-β1 staining in glomeruli, which result in podocyte apoptosis, tubulointerstitial collagen accumulation, glomerular fibrosis and glomerulosclerosis. Therefore, podocyte apoptosis plays a central and critical role in the disruption of the structural and functional integrity of the glomerular filtration barrier14. Our research showed that the change in NPH was positively correlated with the UACR and negatively correlated with UTGF-β1 after 12 weeks of SGLT2i treatment in T2D patients with albuminuria. Nephrin loss is a sensitive marker of podocyte apoptosis. Common antidiabetic agent, metformin and DPP-4 inhibitors, can also ameliorate podocyte damage by restoring renal tissue nephrin expression16. We have compared the application of metformin and DPP-IVinhibitors between different groups in the baseline analysis to eliminate their effects on the nephrin expression. Our result showed that the percentage of metformin and DPP-IV inhibitors between different UACR levels groups were not statistically significant. Staining for the presence of nephrin showed a significant decrease in Western diet-fed C57BL/6J mice compared with Low fat diet-fed mice. Dapagliflozin prevented loss of podocytes as determined by Immunofluorescence staining for nephrin17.Nephrin, a podocyte marker, is not only an essential protein in the SD complex but also involved in podocyte survival. Its extracellular domain forms the SD zipper-like structure, while its intracellular domain acts as a signaling molecule18. Dapagliflozin prevented glomerular pathology and renal fibrosis in Western diet -fed mice, which could relieve the increases in extracellular matrix accumulation in the tubular interstitium as determined by type IV collagen and fibronectin, mitigate mesangial expansion, reduce albuminuria17. Second, TGFβ1 but not HG increased SGLT2 expression via phosphorylated smad3 and inhibited cell survival signaling NF-κB, resulting in amplification of TGF-β–mediated podocyte apoptosis19. Canagliflozin (CAN) transcriptionally inhibited NLRP3 inflammasome-related proteins by inhibiting the transduction of the nuclear factor κB (NFκB) signal20. Third, TGF-β1 is known to activate the Notch pathway22, which is correlated with albuminuria and glomeruloesclerosis23. The Notch activation induced by TGF-β1 has been involved in epithelial mesenchymal transition, augmented vascular endothelial growth factor (VEGF) expression, decreased nephrin expression and podocyte number. Blockade of Notch-1 signaling significantly abrogated VEGF activation and nephrin repression in HG-stressed cells and ameliorated albuminuria in the diabetic kidney24. SGLT2 inhibitors blocked TGF-β1-induced mRNA expression of thrombospondin 1 (THBS1), tenascin C (TNC), and platelet-derived growth factor subunit B (PDGF-B), which are key mediators of renal fibrosis and kidney disease progression in two human proximal tubular (PT) cell lines25. Dapagliflozin significantly reduced effluent TGF-β concentrations, peritoneal thickening and fibrosis, resulting in improved ultrafiltration in a mouse model of chronic peritoneal exposure to high-glucose dialysate26. Empagliflozin treatment or downregulation of SGLT-2 expression significantly ameliorated peritoneal fibrosis by suppressing TGF-β/Smad signaling-associated proteins, such as TGF-β1 and phosphorylated Smad3 (p-Smad3)27.TGF-β1, a potent fibrogenic cytokine, inhibits the expression of the slit-diaphragm protein nephrin and results in podocyte apoptosis in several ways. First, TGF-β1 induces podocyte apoptosis by activating caspase3 via p38 mitogen-activated protein kinase (MAPK) and Smad7. In TGF-β1 transgenic mice, TGF-β induces apoptosis by activating p38MAPK and the classic effector caspase-328. Therefore, SGLT2i should be considered one of the most promising drugs to mitigate podocyte apoptosis and alleviate renal fibrosis.We have demonstrated for the first time that SGLT2 inhibitors alleviate nephrin loss and enhance TGF-β1 excretion in urine at the same time in type 2 diabetic patients with albuminuria. This means that administration of SGLT2 inhibitors in diabetic nethropathy could restore nephrin expression in glomeruli, attenuate nephrin loss in the urine and relieve podocyte apoptosis, inhibit TGF-β1 expression in glomeruli, enhance TGF-β1 secretion by the renal pathway, and ameliorate the progression of renal fibrosis, which is aligned with the results in db/db diabetic mice30. SGLT2i treatment had a beneficial effect on weight loss, which was attributed to improved primary glucose transporters in proximal tubular cells (PTCs), blocked reabsorption of filtered glucose, and led to massive glycosuria and deprivation of energy. The decrease in body weight is the result of two major effects of SGLT2 inhibition: caloric loss due to glucose excretion and loss of body water due to osmotic diuresis, which reduce glomerular pressure and inhibit albuminuria excretion32.Significant body weight loss from baseline was observed after 12 weeks of SGLT2i treatment, which caused a mean weight loss of approximately 2–3 kg regardless of the baseline UACR. Furthermore, Spearman’s correlation analysis showed that the changes in NPH were negatively correlated with BMI, WC, and WHR after 12 weeks of SGLT2i treatment in T2D patients with albuminuria. Obesity is associated with an increased risk of CKD. There are potential mechanisms that can explain the association between obesity and renal injury, including hemodynamic effects, inflammation and renal lipotoxicityThe strengths of this study are that it is the first to verify that SGLT2 inhibitors alleviate nephrin loss and enhance TGF-β1 secretion in urine at the same time in type 2 diabetic patients with albuminuria. Moreover. The changes in NPH were negatively correlated with UTGF-β1 after 12 weeks of SGLT2i treatment in T2D with albuminuria. However, this study has certain limitations. First, this was a randomized, blank-controlled clinical trial study with a relatively small sample size and a comparatively short study period, lack of active-controlled group. Second, further studies are needed to explore the internal mechanisms by which SGLT2i attenuate podocyte apoptosis and reduce the secretion of nephrin with respect to changes in glomerular ultrastructure, inflammatory pathway transcription and translation via histological results of renal biopsy.In conclusion, this study demonstrated that SGLT2 inhibitors can reduce albuminuria, decrease nephrin loss and enhance TGF-β1 secretion in urine. Furthermore, the changes in NPH were positively correlated with the UACR and negatively correlated with UTGF-β1 after 12 weeks of SGLT2i treatment in T2D patients with albuminuria. Significant weight loss was also observed after 12 weeks of SGLT2i treatment and was negatively correlated with NPH. Our findings suggest that the anti-albuminuric effect of SGLT2 inhibitors could be attributed to mitigating podocyte apoptosis, attenuating renal fibrosis and promoting weight loss.Supplementary Information 1.Supplementary Information 2.Supplementary Information 3.Supplementary Information 4.


Clinical and experimental evidence shows lung fluid volume as a modulator of fetal lung growth with important value in treating fetal lung hypoplasia. Thus, understanding the mechanisms underlying these morphological dynamics has been the topic of multiple investigations with, however, limited results, partially due to the difficulty of capturing or recapitulating these movements in the lab. In this sense, this study aims to establish an ex vivo model allowing the study of lung fluid function in branching morphogenesis and identify the subsequent molecular/ cellular mechanisms.−) concentrations  or Cl− channels inhibitors  were injected into lung lumen at two timepoints, day0 (D0) and D2. At D4, morphological and molecular analyses were performed in terms of branching morphogenesis, spatial distribution (immunofluorescence), and protein quantification (western blot) of mechanoreceptors (PIEZO1 and PIEZO2), neuroendocrine  and smooth muscle [alpha-smooth muscle actin (α-SMA) and myosin light chain 2 (MLC2)] markers.Ex vivo lung explant culture was selected as a model to study branching morphogenesis, and intraluminal injections were performed to change the composition of lung fluid. Distinct chloride (Cl−], 715 mM Cl−, through the overexpression of PIEZO1, PIEZO2, ghrelin, bombesin, MLC2, and α-SMA. In contrast, intraluminal injection of CFTRinh or CaCCinh decreases fetal lung growth and the expression of PIEZO1, PIEZO2, ghrelin, bombesin, MLC2, and α-SMA. Finally, the inhibition of PIEZO1/PIEZO2 by GsMTx4 decreases branching morphogenesis and ghrelin, bombesin, MLC2, and α-SMA expression in an intraluminal injection-independent manner.For the first time, we described effective intraluminal injections at D0 and D2 and demonstrated intraluminal movements at D4 in ex vivo lung explant cultures. Through immunofluorescence assay in in vivo and ex vivo branching morphogenesis, we show that PGP9.5 colocalizes with PIEZO1 and PIEZO2 receptors. Fetal lung growth is increased at higher [ClOur results identify PIEZO1/PIEZO2 expressed in neuroendocrine cells as a regulator of fetal lung growth induced by lung fluid.The online version contains supplementary material available at 10.1186/s12931-023-02328-2. The physiological circulation of lung fluid filling the air spaces is critical to lung development. In fact, if it is disturbed, lung growth and maturation are impaired. For instance, excess fluid drainage during fetal life or a decrease of fluid pressure due to premature rupture of the membranes or oligohydramnios are associated with lung hypoplasia with underbranched lungs, which is a major cause of respiratory insufficiency and mortality in newborns ) that favors the movement of sodium and water into the lumen and promotes lung liquid formation and the consequent pulmonary expansion  . The intraluminal [Cl−] manipulation was achieved using three experimental concentrations: 5.8, 29, and 715 mM Cl−. Lungs were randomly assigned to one of four experimental groups (n ≥ 12 per condition). After intraluminal injections at day 0 (D0) and D2, morphological and molecular dynamics were determined at D4.According to in vivo  and ex v−] maintaining a similar concentration of the remaining ions, the following chemical compounds were used: potassium chloride , magnesium chloride , calcium chloride , potassium d-Gluconate , MgSO4  and calcium d-gluconate . Specifically, KCl, MgCl2, and CaCl2 were used as donors of Cl−, K+, Mg2+, and Ca2+, while potassium d-gluconate, MgSO4, and calcium d-gluconate worked as replace compounds as demonstrated in the Additional file −] were as follow , MgCl2 (0.049), CaCl2 (0.041), d-glucose , HEPES , potassium d-gluconate (698.290), MgSO4 , Calcium d-gluconate ; 715 mM Cl−: KCl (704.000), MgCl2 (6.000), CaCl2 (5.000), d-glucose (10.000), HEPES (5.000) as shown in Additional file To strictly manipulate the  , and Cl− channels inhibitors . Two-way ANOVA was used in the analysis of both morphological  and the molecular (protein expression levels) effect after GsMTx4 exposure. The parametric test assumptions were previously verified, and an additional LSD test was used for post-test analysis. Statistical analysis was performed using the statistical software IBM SPSS Statistics 26.0. Statistical significance was confirmed at p < µ0.05, γ0.01, β0.001, and α0.0001.All quantitative data are presented as mean ± standard deviation (SD). One-way ANOVA was performed for the number of peripheral airway buds, epithelial perimeter, and protein expression levels on : 5.8, 29, 143, 715 mM Cl− or Cl− channels inhibitors: SS, A9C, CFTRinh, CaCCinh.To study the role of intraluminal composition in branching morphogenesis, ex vivo lung explants were cultured for 4 days after injection of distinct  activates PNECs/NEBs through PIEZO1/PIEZO2 mechanoreceptors that, in turn, regulate the expression of ghrelin, bombesin, α-SMA, and MLC2 thus regulating fetal lung growth  and crescent chloride concentrations, [Cl−]: 5.8, 29, 143, and 715 mM. b demonstrates the ionic composition in terms of chloride (Cl−), potassium (K+), magnesium (Mg2+) and calcium (Ca2+) in SS, 5.8, 29, 143 and 715 mM Cl−.Additional file 2: Movie S1. Injection of standard solution into the lumen of ex vivo lung explant cultures at day0 (D0).Additional file 3: Movie S2. Injection of standard solution into the lumen of ex vivo lung explant cultures at day2 (D2).Additional file 4: Movie S3. Dynamic intraluminal movement in ex vivo lung explant cultures at day4 (D4).


Released Cu2+ played a critical role in the toxicity mechanism of Cu(OH)2 nanowires in lettuce seedlings, as evidenced by the substantial accumulation of Cu in the seedling roots (p< 0.01) rather than in the leaves. Polystyrene (PS) MPs (1 mg/L) stimulated lettuce seedling growth, as shown by the (highly) significant increase in root and leaf length and in the seed vigor index (p< 0.01 or 0.05). Notably, PS MPs (1 mg/L) neutralized the hazardous effects of 1 mg/L Cu(OH)2 nanowire treatment on lettuce growth, as reflected by the vitality and root length of the seedlings returning to normal levels. The PS MPs (1 mg/L) absorbed on middle root surfaces and strongly hindered Cu accumulation in lettuce roots, which was the predominant mechanism by which PS MPs suppressed the hazardous effects of the Cu(OH)2 nanowires. This study strengthens the understanding of the toxicity and toxicity mechanisms of Cu(OH)2 nanowires with or without PS MPs in the environment.Copper-based nanopesticides are released into the environment during foliar spray application, and they could, on their own or in combination with microplastics (MPs), pose threats to environmental safety and human health. In this study, Cu(OH) Inevitan plants , and thun health . Corresp species . For exaea mays) . In addima cells . Notablyn humans . Howeverrecently .Triticum aestivum)  , on growita pepo , and on ita pepo . From thita pepo . This plita pepo  and imprita pepo . Due to ita pepo , Cd  pepo , which is a model plant and is widely applied in toxicity assays 2 nanowires could induce hazardous effects on L. sativa seed germination and seedling growth after Cu(OH)2 nanopesticides enter the environment. Furthermore, we proposed that PS MPs that have been widely detected in realistic environments  determine the hazardous effects of Cu(OH)2 nanowires on seed germination and seedling growth of lettuce and carry out a biological analysis, and (ii) explore the effects of PS MPs (1 mg/L) on stimulating lettuce seedling growth and suppressing Cu(OH)2 nanowire toxicity to lettuce.To bridge these gaps, Lettuce  were obtained from Beijing Dk Nano Technology Co., Ltd. Scanning electron microscopy  was employed to determine the sizes and morphologies of the PS MPs and Cu(OH)2 nanowires. The hydrodynamic diameters and zeta potentials of the PS MPs and/or Cu(OH)2 nanowires were determined with a Zetasizer Nano instrument . Lettuce (Lactuca sativa) seeds were purchased from Nanyang Seed Inc. . Peroxidase (POD), superoxide dismutase (SOD), malondialdehyde (MDA) and catalase (CAT) activity assay kits were purchased from Nanjing Jiancheng Bioengineering Institute, Nanjing, China.PS MPs (0.1 µm) and fluorescent PS MPs were purchased from Tianjin Baseline Chromtech Research Centre, China. Cu(OH)2 nanowire group, a 10 mg/L Cu(OH)2 nanowire group, a 10 mg/L Cu2+ group, a PS (1 mg/L) group, a PS (1 mg/L) + 1 mg/L Cu(OH)2 nanowire group, a PS (1 mg/L) + 10 mg/L Cu(OH)2 nanowire group, and a PS (1 mg/L) + 10 mg/L Cu2+ group. The 10 mg of Cu(OH)2 nanowire were added into a 100 glass measuring flask and sonicated on an ultrasonic water bath (0.5 min) (ready-to-use). Certain of Cu(OH)2 nanowire/Cu2+(CuSO4) were added to certain of 100 mg/L of PS MPs solution or DI water to obtain the above different exposure groups. Four replicates for these groups were performed, and 100 seeds were placed in each glass Petri dish (⌀ 90 mm). Depending on the treatment group to which the seeds were randomly assigned, the seeds were incubated with 7 mL of the assigned Cu(OH)2 nanowires or Cu2+ with or without PS solution and were kept in the dark at 25°C for seed germination.The lettuce seeds were surface disinfected in hydrogen peroxide  for 30 min with continuous stirring, rinsed 3 times with deionized water (DI water), and immersed in DI water for 24 h. The seeds were randomly divided into eight different groups (100 seeds), including a control group (CK), a 1 mg/L Cu(OH)The germination rate was observed and recorded for three days. Accordingly, the germination rate, germination index, vigor index and germination energy of the seeds were calculated by equations (1) to (4). The biomass of the lettuce seedlings was measured, as well as the root length and shoot height, and the fresh weights of shoots and roots were recorded.Where GR and GI represent the germination rate and germination index, respectively. Gt represents the germination number at t days, and Dt represents the corresponding germination time (days). VI and GE represent the vigor index and germination energy, respectively.Antioxidant enzyme levels, including SOD, CAT and POD activity levels and MDA levels, were measured with commercial assay kits obtained from Nanjing Jiancheng Bioengineering Research Institute during the seed germination (3 d) and seedling growth (7 d) stages. The measurement processes for the antioxidant enzyme activities were in accordance with the manufacturer’s instructions. Chlorophyll a (Chl a) and chlorophyll b (Chl b) were measured at the seedling growth stage (7 d). A specific weight (m) of freeze-dried cotyledons of lettuce samples was ground in a mortar with quartz sand and then extracted with 80% acetone to a specific volume (V). The obtained supernatant was centrifuged (5000 r/min for 10 min at 4 °C), and the absorbance was measured at 663 and 645 nm with UV−Vis spectrometry. Finally, Chl a and Chl b were calculated according to equations (5) and (6), respectively.Where m is the specific weight of the lettuce cotyledon sample and V is the specific volume created by adding 80% acetone to the ground cotyledons.Surface-disinfected lettuce seeds were immersed in DI water for 24 h, after which 100 of these seeds were selected and exposed to 1 mg/L fluorescent PS MPs. The seeds were cultured with fluorescent PS MPs  for seed germination. After 3 d of exposure in the dark at 25°C, the lettuce seed and seedling samples were washed three times with DI water to exclude fluorescent PS MPs adsorbed on the root surface. Then, the uptake of fluorescent PS MPs by the lettuce seeds and seedlings was examined using fluorescence microscopy  with a photomicrography system . The excitation wavelength of the fluorescence microscope was 488 nm, and the emission wavelength was 518 nm.2+ or Cu(OH)2 on the seed and seedling surfaces. Then, the sample tissues were washed three times with DI water and oven-dried to constant weight for digestion (by adding 4 mL of a mixture (1:3) of H2O2 and HNO3 and heating at 80 °C for 2 h and then at 160°C for 4 h) and Cu accumulation quantification (by inductively coupled plasma−mass spectrometry).Ten of lettuce seeds (at 3 d) and seedlings (at 7 d) samples were taken and washed with EDTA-2Na  to exclude extracellular Cut test and/or one-way ANOVA to explore any significant differences between treatment groups. Significant differences are marked with asterisks (* (p< 0.05) and ** (p< 0.01), denoting statistically significant and highly statistically significant differences, respectively).The experimental data were for four replicates were analyzed with SigmaPlot 12.5. The data are presented as the means ± standard deviations. These data were normally distributed and were evaluated with a 2 nanowires exhibited a typical wire shape with a nanosized diameter (40-80 nm) and a microsized length (1-2 µm) (2 nanowires tended to aggregate slightly in ultrapure water (1 mg/L), based on their average hydrodynamic diameter of 1584.97 (± 674.29) nm and on their average zeta potential of –21.58 (± 1.81) mV. Interestingly, the Cu(OH)2 nanowires (1 mg/L) with PS MPs (1 mg/L) presented a large hydrodynamic diameter of 2365.25 (± 856.37) nm and a low zeta potential of –35.12 (± 5.47) mV and tended to aggregate in ultrapure water.The PS MPs exhibited a spherical shape with quite a smooth surface and had an average size of ~ 100 nm 2 nanowires induce toxic effects in plants (2+ stress treatment induced adverse effects comparable to those induced by the 10 mg/L Cu(OH)2 nanowire stress treatment, suggesting that released Cu ions played a critical role in the toxicity effects induced by Cu(OH)2 nanowires.Based on the seed germination period (3 d) data, the Cu(OH)n plants . In this2 nanowire and Cu2+ treatments (highly) significantly inhibited the average root length 2 nanowire treatments displayed similar effects, (highly) significantly inhibiting the average root length and weight, as well as adversely affecting the vitality of the lettuce seedlings, which again emphasizes the important role of Cu2+ release in the hazardous effects induced by Cu(OH)2 nanowires. Additionally, the antioxidant defence system and MDA content were significantly modified under Cu stress at 7 d of lettuce seedling growth. For example, compared with the SOD activity and MDA levels in seedling roots or leaves from the water treatment groups, those in the 10 mg/L Cu(OH)2 nanowire or Cu2+ treatments were (highly) significantly increased  significantly decreased 2 nanopesticides may be a nonnegligible toxicity mechanism of the nanopesticides. Comparably, Cu was shown to predominantly accumulate in plant leaves but not in roots when a Cu(OH)2 nanopesticide was applied through foliar spraying (Based on the seedling growth period (7 d) data, the Cu(OH)spraying . It seemspraying .p > 0.05; p< 0.05; p > 0.05; P< 0.05) and bottom-to-top transportation of PS MPs have been confirmed in wheat roots , the PS MPs treatment helped SOD and CAT activities return to the control level (p > 0.05), and these activities were the dominant mechanisms for protection of lettuce seed germination against Cu(OH)2 nanowire or Cu2+ stress (as mentioned in Section 3.2). Furthermore, when PS MPs were present, the highly decreased vigor index that was induced by the 1 mg/L Cu(OH)2 nanowire treatment was reversed, and the vigor index returned to the control level 2 nanowire treatment was significantly restrained by PS MPs treatment, as shown in 2 nanowire or Cu2+ treatments on seedling root growth, although the average seedling root length in these treatment groups was still much shorter than that in the water treatment groups 2 nanowires and Cu2+ with PS MPs were (highly) significantly decreased. Furthermore, the MDA level in the 10 mg/L Cu(OH)2 nanowire treatment group was significantly decreased by the presence of PS MPs, although it was still much higher than that in the control  by the 1 mg/L PS MPs treatment. For example, the Cu concentration from the Cu(OH)2 nanowire (1 mg/L) treatment decreased from 0.093 (± 0.017) to 0.023 (± 0.008) mg/g with PS MPs + Cu(OH)2 nanowire (1 mg/L) treatment, and thus, Cu2+ bioavailability was decreased by PS MPs treatment in this study. Furthermore, the PS MPs were likely adsorbed on the surface of the middle root section  are possible (2 nanowires or Cu2+ from entering the lettuce by binding to the lettuce root surface.Based on the seedling growth data (7 d), the decrease in lettuce seedling root length caused by the 1 mg/L Cu(OH)eviously , and it possible . Howeverpossible Figure 62+ toxicity on macrophyte growth 2 nanowires in lettuce seedlings. Interestingly, PS MPs (1 mg/L) stimulated lettuce seedling growth. Notably, treatment with PS MPs (1 mg/L) strongly hindered Cu accumulation in lettuce roots and thus neutralized the hazardous effects of the 1 mg/L Cu(OH)2 nanowire treatment on lettuce growth. Therefore, PS MPs in the environment could protect lettuce from the hazardous effects of Cu(OH)2 nanopesticides and decrease Cu risk in the food chain.The protective effects of the MPs treatment on other plants have also been previously confirmed. For example, the presence of PE MPs reduced the accumulation of phenanthrene in wheat roots and leaves , decrease growth . PS MPs e growth  and to me growth . The unde growth . It is p2 nanowires on lettuce seed germination and seedling growth were confirmed. These effects were highly related to the lettuce antioxidant defence system and MDA content. Released Cu2+ was a nonnegligible toxicity mechanism for the hazardous effects of Cu(OH)2 nanowires on lettuce. PS MPs (1 mg/L) increased the lettuce seed vigor index and stimulated lettuce seedling growth. Furthermore, the PS MPs treatment (1 mg/L) partially suppressed the hazardous effects of Cu(OH)2 nanowires on lettuce growth. This study provides new insights into the toxicity and toxicity mechanisms of Cu(OH)2 nanowires and/or PS MPs to plants.In this study, the hazardous effects of Cu(OH)The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.GY, data curation, investigation, and writing of the original draft. YS and LY, data curation and investigation. YZ, resources and visualization. WZ, funding acquisition, formal analysis, writing-review, and editing. All authors contributed to the article and approved the submitted version.


Low-cost and high-efficiency iodine removal materials are urgently needed. In this study, we synthesized two aniline-based hypercrosslinked polymers (AHCPs), AHCP-1 and AHCP-2, for iodine capture in both aqueous and gaseous phases. They are obtained by aniline polymerization through Friedel–Crafts alkylation and Scholl coupling reaction, respectively, with high chemical and thermal stability. Notably, AHCP-1 exhibits record-high static iodine adsorption (250 wt%) in aqueous solution. In the iodine vapor adsorption, AHCP-2 presents an excellent total iodine capture (596 wt%), surpassing the most reported amorphous polymer adsorbents. The rich primary amine groups of AHCPs promote the rapid physical capture of iodine from iodine water and iodine vapor. Intrinsic features such as low-cost preparation, good recyclability, as well as excellent performance in iodine capture indicate that the AHCPs can be used as potential candidates for the removal of iodine from radioactive wastewater and gas mixtures.Effective capture and safe disposal of radioactive iodine ( Thus, development of novel materials for treating contaminated water and gas containing radioactive iodine with high efficiency and good durability is of primary importance but is still challenging [Nuclear energy is considered a type of renewable and clean energy; however, the challenges and threats of radioactive substances are inevitable in nuclear fuel reprocessing ,2,3. Amo systems . The othllenging ,7,8. To remove the iodine radioisotopes, vast adsorbents including activated carbon ,10, zeol3, –NH2, –(OH)2, –COOH, –(COOH)2, –NO2, –Cl, and –Br) and found that the best sorption uptake (60%) was obtained when the ligand was decorated by the electro-donor group –NH2. Tesfay Reda et al. [2-Mel) with high iodine adsorption (128.1 wt%), which may be attributed to the N–H⋯I hydrogen bonding and N⋯I halogen bonding interactions. Kobinata et al. [Due to the electron-deficient inherency, the iodine molecule is a superior electron-acceptor and, therefore, the affinity of the adsorbent binding sites to iodine also plays an important role . Volkrina et al.  prepareda et al.  measured2 from readily available raw materials. Herein, aniline was chosen to prepare two aniline-based hypercrosslinked polymers (AHCPs), AHCP-1 and AHCP-2, containing rich primary amine groups for effective uptake of iodine. Then, their iodine uptake capacities and adsorption kinetics from the vapor and aqueous solutions were systematically evaluated. The adsorption mechanism was comprehensively described by combining theoretical calculation and these characterizations such as Raman spectra, X-ray photoelectron spectroscopy (XPS), Fourier-transform infrared spectroscopy (FT-IR), and powder X-ray diffraction (PXRD). In this context, we aim to develop a new type of adsorbent with rich affinity sites such as –NH3 and AlCl3, respectively  at 3500–3300 cm−1 and bending vibrations (δN-H) near 1620 cm−1 were preserved compared with the monomer aniline [vas C–H) at 2930 cm−1 and (vs C–H) at 2845 cm−1 and scissor-like vibrations (δC-H) at 1456 cm−1 were preserved compared with dimethoxymethane. The benzene ring possesses stretching vibrations occurring near 1600 cm−1, 1560 cm−1, 1500 cm−1, and 1450 cm−1 [2 sorption were 14.4 m2∙g−1 and 75.2 m2∙g−1, respectively  spectra . Solid-s spectra a of AHCP spectra  of AHCP-450 cm−1 . Powder ectively b, which ectively c, which To evaluate the iodine capture performance of these two aniline-based hypercrosslinked polymers with rich primary amine groups, abundant π-electrons and high water stability, time-dependent UV–vis spectroscopy was carried out to study the iodine uptake of the AHCPs from water. The adsorption amount of iodine in AHCPs was monitored by recording sample mass as a function of time after the sample was completely dried a. The AHFurthermore, the iodine adsorption performance of AHCP-1 and AHCP-2 in iodine vapor was measured. The iodine adsorption amounts of AHCPs in iodine vapor were monitored by recording sample mass as a function of time at 75 °C or 100 °C. As shown in 2 adsorption in aqueous solutions. This model based on the adsorption capacity indicated that the rate controlling step was dependent on a chemisorption phenomenon. The mechanism of chemisorption involved strong interactions between the surface and the adsorbate, confirming the formation of charge transfer complexes between iodine and the AHCPs. It can be seen from Iodine aqueous solutions sorption kinetics were investigated by considering different equations: pseudo-first- and pseudo-second-order kinetic models . The resNext, a time-dependent UV–vis spectroscopy experiment was performed to study the iodine release of iodine@AHCPs, as shown in 2 vapor uptake capacity. The partial loss was most likely due to the chemical reaction between iodine and AHCPs at 75 °C and 100 °C [2∙g−1, 7.1 m2∙g−1, 63.2 m2∙g−1, and 46.7 m2∙g−1, respectively, exhibiting various decline.In order to test the recyclability of AHCPs for iodine capture from water and vapor phases, the iodine re-adsorption and desorption experiments for five times using a gravimetric method were carried out. As shown in d 100 °C , namely,−1 and 109 cm−1; nevertheless, the pristine AHCPs do not. The peaks at ~165 cm−1 and ~109 cm−1 are assigned to the stretching vibrational modes of polyiodine anions −1 are greater than those of iodine@AHCPs(vapor) at ~165 cm−1, which may imply that the content of The outstanding performance of AHCPs on iodine uptake from water solution and from vapor phase encouraged us to investigate the possible mechanism of the iodine adsorption. Various characterization methods, such as Raman spectra, XPS, FT-IR, and PXRD, were performed to detect the interactions between AHCPs and iodine. As shown in ectively ,41. Comp5/2 and I3d3/2 peaks at ~620 eV and ~631 eV. I3d5/2 demonstrated the split peaks at 619.0 eV and 620.7 eV in a different ratio. The former can be attributed to Moreover, the XPS spectra were used to analyze the interactions between guest iodine species and AHCPs. As shown in −1 and 11 cm−1 in iodine@AHCPs (water), and blue-shifted by 28 cm−1 and 37 cm−1 in iodine@AHCPs (vapor) upon complexation with iodine [−1) of the benzene ring in iodine@AHCPs were red-shifted by 5–22 cm−1 compared with AHCPs. All these FT-IR results indicated obvious interactions between AHCPs and iodine [FT-IR and PXRD were used to study the structural changes of AHCPs before and after iodine adsorption as well. As shown in h iodine . In addid iodine ,43. In a2 (iodine vapor) and aniline unit/2 (iodine vapor), the bond distances of N⋯I and N–H⋯I are, in turn, 2.751 Å and 2.930 Å, and the corresponding binding energy is −0.835 eV and −0.584 eV. The results demonstrated that the binding modes of N⋯I were more stable than that of N–H⋯I. For the structural optimization model of aniline unit/A theoretical calculation can also provide great insights into the adsorption properties in polymers ,45. To f for I3− c. These 2 flowing gas. Then, FeCl3  and 1,2-dichloroethane (20 mL) were added. The reaction was then heated to 80 °C for 10 h. After cooling down to room temperature, the precipitate was removed by filtration and extracted extensively by Soxhlet extraction with methanol for 3 h. Finally, the solid was dried under vacuum and at 60 °C overnight. A dark brown powder was obtained (AHCP-1). Yields: 280.7 mg (94%). Elemental analysis of AHCP-1, calcd: C, 80.21%, N, 9.99 %, and H, 9.80%. Found: C, 78.91%, N, 10.6 %, and H, 8.14%.AHCP-1 was prepared according to the previous reported procedure with a slight modification ,33. In a3 , and CHCl3 (25 mL) were mixed together under N2 flowing gas and then refluxed for 10 h. The precipitate was obtained by filtration and subsequently washed with CHCl3. The product was further washed with 10% NaOH aqueous solution and distilled water until the pH reached 7. Finally, the product was washed with CHCl3 at 70 °C for 3 h. The black solid was dried in a vacuum oven at 60 °C overnight. Yields: 165.6 mg (90%). Elemental analysis of AHCP-2, calcd: C, 77.42%, N, 15.05%, and H, 7.53%. Found: C, 75.78%, N, 13.11%, and H, 6.57%. In a 50 mL round-bottom flask, aniline , AlCl1H NMR spectrum was recorded on a Bruker Avance III 400 MHz instrument. Solid-state NMR spectrum was recorded on a Bruker Avance III 400 MHz spectrometer by conventional double-resonance 4 mm CPMAS probe at 12 kHz MAS and 298 K with sample packed into a 4 mm MAS zirconia rotor. Scanning electron microscopy (SEM) imaging was performed on a Bruker-BioSpin S-3000N SEM. Low-pressure N2 adsorption and desorption isotherms were measured at 77.3 K. The cumulative apparent surface areas for N2 were calculated using the Brunauer–Emmett–Teller (BET) model range from 0 to 1.0 bar for all samples. Micropore volumes were calculated using the T-plot method, while the total porous volumes were obtained from the N2 isotherm at P/P0 = 0.99 and 0.10. Pore size distributions were derived from the N2 adsorption isotherms using H-K methods. Powder X-ray diffraction (PXRD) was measured using Bruker-BioSpin D8 Advance-PXRD. X-ray photoelectron emission microscopy (XPS) analysis was performed on a spectrometer . AHCPs were characterized with Raman spectroscopy having laser excitation energy of 514 nm in inVia Reflex laser Raman spectroscopy.The successful preparation of AHCPs adsorbents was confirmed by the following instruments. TENSOR27 Fourier-transform infrared (FT-IR) spectra were collected in transmission on a VARIAN 1000 FT-IR spectrometer using KBr disks. Thermo-gravimetric analysis (TGA) was performed at a heating rate of 10 °C/min under air atmosphere. 2 − m1)/m1, where m1 and m2 are the mass of AHCPs samples before and after iodine uptake. Iodine adsorption experiments from water were performed based on the gravimetric method. An amount of 20 mg of AHCPs adsorbents were soaked in 0.1 M (20 mL) of iodine aqueous solution. After iodine adsorption of AHCPs for a given time, the iodine-loaded solid sample was filtered and washed by DI water until the filtrate was colorless, and then dried in vacuum at room temperature for 48 h. The same adsorption experiments were performed in parallel for three times. The iodine uptake of AHCPs samples was calculated according the following equation (mIodine vapor adsorption experiments based on gravimetric measurements were performed as the following procedure. An open glass bottle (1.5 mL) containing 25 mg of AHCPs and excess iodine solids were placed in a sealed glass bottle (25 mL), which were then heated at 348.15 K and 373.15 K, respectively. After adsorption of AHCPs in iodine vapor for a while, the bottle was taken out and cooled down to room temperature and then weighted. The same adsorption experiments were performed in parallel three times. The iodine uptake capacity of adsorbents was calculated according to the same equation as the adsorption in iodine aqueous solution. After absorbing iodine of determined mass, place the material in a vial and soak it in methanol at room temperature. Replace the eluent every 2 h until the eluent is colorless, indicating that the desorption is completed.Herein, two aniline-based hypercrosslinked polymers (AHCPs) were successfully developed for industrially cost-effective, efficient, and safe disposal of radioactive iodine from aqueous solution or from their gas vapor. It was notable that AHCPs showed excellent performance in iodine capture including high adsorption capacity, fast rate, and good recycle. Our results suggest that a primary amine group in these aniline-based polymers are crucial for strong iodine adsorption. Considering the low-cost aniline and the facile synthesis, we believe that such aniline-based adsorbents have great potential for iodine removal in nuclear power industry.