Let  Dedicated to the memory of our friend and colleague Yahya ould Hamidoune1Let Long sets of lengths have a well-defined structure: they are AAMPs  with a universal bound for all parameters There are two atoms  Thus Little is known about short sets of lengths. If Theorem ALetbe a Krull monoid with finite class group, and suppose that every class contains a prime divisor. ThenOur first main result  shows thher rank , and theher rank .It is well known that all questions on sets of lengths in a Krull monoid translate into zero-sum problems in its class group. Thus, after applying well-studied transfer machinery , all the2sumset, and by set of (successive) distances of Let independent if all elements are non-zero and basis if total rank of Let Monoids and factorizations. By a monoid, we always mean a commutative semigroup with identity which satisfies the cancellation law ≤2 see  .Lemma 2.1Letbe a Krull monoid,a divisor theory with some nonempty setits class group, andthe set of classes containing prime divisors. Letdenote the unique homomorphism defined byfor all. Thenis called the block monoid associated to, and the homomorphismhas the following property:This implies thatandfor every.ProofSee  The following simple technical lemma will be used without further mention.Lemma 2.2Letbe a finite abelian group and.1., andif and only ifand.2.If, then. In particular, ifwith, then.ProofSee  3The following characterization of minimal zero-sum sequences of maximal length over groups of rank two—formulated in Theorem 3.1Letwithand. A sequenceoverof lengthis a minimal zero-sum sequence if and only if it has one of the following two forms:•where(a)is a basis of,(b)and.In this case, we say thatis of type  I.•where(a)is a basis ofwithand,(b), and,(c)with,(d), and(e)eitheror.In this case, we say thatis of type  II.ProofSee the corollary in  If If Lemma 3.2Letwithand.1.We have.2.If, then.Proof1. This follows immediately from the special case in 2. For Case 1. We set Case 2. Let If If Suppose that Lemma 3.3Let, whereis a possibly trivial subgroup and,, such that. Then there exists somewith.ProofLet The following proposition is one of the more lengthy and difficult portions of the paper.Proposition 3.4Letwithand, and letwith. Then.ProofPer Case 1. Then there is a basis of A.Let Proof of ASince Since Since If, for some Assume by contradiction that Now assume by contradiction that Case 1.1. Some  with If Case 1.2. Then A to conclude Now, since Since But now, since there exists Suppose In this case, Case 2. In this case, we have a basis Assume by contradiction that we have a factorization We begin by handling the case when Case 2.1. Suppose Since Case 2.1.1. Suppose Suppose If there is an  However, in the former case, Thus, if If Case 2.1.2. Suppose In this case, Suppose As a result, if Suppose In this case, we either have Case 2.2. In this case, we conclude that the atoms Suppose Case 2.2.1. Let Suppose However, Case 2.2.2. Let Suppose If From As in Case 2.2.1, if all terms of Suppose Since Let us next examine the atom In view of 1)e2 see . TherefoLet us next examine the atom In view of Case 2.2.2.1. Suppose Then Case 2.2.2.2. Suppose Since But now, if Since Theorem 3.5Letbe a Krull monoid with class group, whereand, and suppose that every class contains a prime divisor. ThenProofBy Case 1. Case 2.  We have Case 3.  We have Case 4.  We have  We have For Case 5.  We have  We have Next we show that  We have For 4We start with a simple technical lemma.Lemma 4.1Letwithnon-trivial subgroups satisfyingand suppose thatwithand. Then there exists awithsuch that. In particular,ProofBy hypothesis, there are We choose a basis Theorem 4.2Letbe a Krull monoid with class group, whereand, and suppose that every class contains a prime divisor. Then.ProofBy A1.Let A1 holds. We set A1, and Suppose that A1. To do so, we need two auxiliary assertions.Thus it remains to prove A2.••Any zero-sum subsequence of Let A3.•••Any subsequence from Let Proof of A2We choose an element Case 1. We set We find Case 2. Note that  If We find Proof of A3Let Proof of A1If Let Case 1.  and we find In this particular case, we set Case 2.  and we find Let Case 3.  and we find Let 5Two reduced Krull monoids ers see  . In this two see .cross number of cross number of To introduce the necessary concepts, let almost arithmetical progression (AAP) with differencelengthboundLet  For every  The sets We set A subset Lemma 5.1Letbe a finite abelian group with.1..2..ProofFor item 1, see  Lemma 5.2Letandbe finite abelian groups withsuch that. Then we haveand. Moreover, we haveProofThe first three statements are proved in  Lemma 5.3Let, whereandis a subgroup which has no direct summand isomorphic to. Then, and equality holds if and only ifis an elementary  2-group or an elementary  3-group.ProofBy  Now suppose that Suppose that It is easy to verify that Proposition 5.4Letandbe finite abelian groups such thatand suppose thatand. If, then.ProofIf First, suppose that the maximum on the right hand side equals  Hence, using  Second, suppose that the maximum on the right hand side equals There is a recent result due to Schmid Lemma 5.5Let.1.If, then.2.If, then.Proof1. Let 2. Let We will use Therefore, if Thus If Let Suppose that But now Theorem 5.6Letbe a finite abelian group with,with, and suppose that.1.If, then.2.Ifis a power of a prime, then.ProofIf tion see . If m=2,Since Now we suppose that Suppose that


RH) has great potential to increase atmospheric CO2 concentrations, it is important to understand warming effects on RH for a better prediction of carbon–climate feedbacks. However, it remains unclear how RH responds to warming in subtropical forests. Here, we carried out trenching alone and trenching with warming treatments to test the climate warming effect on RH in a subtropical forest in southwestern China. During the measurement period, warming increased annual soil temperature by 2.1 °C, and increased annual mean RH by 22.9%. Warming effect on soil temperature (WET) showed very similar pattern with warming effect on RH (WERH), decreasing yearly. Regression analyses suggest that WERH was controlled by WET and also regulated by the soil water content. These results showed that the decrease of WERH was not caused by acclimation to the warmer temperature, but was instead due to decrease of WET. We therefore suggest that global warming will accelerate soil carbon efflux to the atmosphere, regulated by the change in soil water content in subtropical forests.As heterotrophic respiration ( Forests cover only about 30% of the terrestrial ecosystems, but contain about 45% of terrestrial carbonGlobal soil CORH) and autotrophic respiration (RA). Responses of heterotrophic respiration (RH) and autotrophic respiration (RA) to warming are not always the same. For example, Schindlbacher, et al.RH and RA responded similarly to temperature increase; both increasing with warming. However, Zhou, et al.RH and RA. Other studies showed that warming increased RH, but decreased RA13RH component of soil respiration has potential to increase atmospheric CO2RH from soil respiration and to examine how warming may affect RH1617Many previous studies focused on the warming effects on soil respiration6789WERH were carried out in grassland ecosystems12131419201122Forest soil organic carbon decomposition plays an important role in the carbon cycle, but most previous studies about RH exhibited seasonal variation and had a significant positive relationship with soil temperature28RH in this subtropical forest.The subtropical evergreen broad-leaved forest in the Ailao Mountains in Yunnan of southwestern China has been considered a carbon sinkWERH. A few studies suggested that warming decreased RH12RH which can be sustained for 5–10 years18308221619333536371117RH acclimated to continuous warming in this forest. To test this hypothesis, we conducted ‘trenching alone’ and ‘trenching with warming’ treatments in this forest.Previous studies have showed inconsistent results about T, °C) ) , the trenching treatment (NR) did not change soil temperature  , but inc% (v/v)) . The trev/v))    and incrrst year .R, T, and W of the CK, NR, and NRW treatments  also exhibited seasonal variation , clearly decreasing year by year (T (WET) was very similar to that of WERH, and WET was 2.6, 2.1, 2.0, and 1.8 °C (with an average of 2.1 °C) for 2011–2014, respectively , which meant WERH was affected by T, W, and WET. The factors T, W, and WET explained 13, 33, and 75% of WERH’s variation, respectively. A 1 °C increase in T and a 1% decrease in W would reduce 0.98 and 0.79% of WERH, respectively; however, a 1 °C increase in WET would cause a 21.78% increase in WERH (WET) control WERH.The factors  in WERH . This suRH acclimated to warmer temperature over the warming period822WERH in ecosystems with high soil organic carbon1830p > 0.05) decrease in NR and NRW comparing to CK during the 4-year measurement period, but they were larger in NRW than that in NR  analysis showed that warming increased Q10 in the later period of this study which showed opposite result to previous studies1117WERH remained stable during warming period, which also challenged “acclimatization” of microbial activity to warmer temperatureWET is likely to be the controlling factor in this study.Previous studies have shown that at in NR . In thisficially . Howevern period c and 4c.WERH (22.9%) was lower than that of Aguilos, et al.et al.et al.WET values in the experiments of Aguilos, et al.et al.et al.RH by an average of 21.0%WET of 3.9 and 6.7 °C  of the Chinese Ecological Research Network, which is located in Jingdong County, Yunnan Province, China. The annual mean air temperature was 11.3 °C, with a minimum monthly mean temperature of 5.7 °C in January and a maximum monthly mean temperature of 15.6 °C in July. Average annual rainfall was 1778 mm, with 86.0% of this falling in the rainy season (May–October)2 effluxes was established in October 2010. The system comprised 20 automatic chambers (90 × 90 × 50 cm) and a control box  at a 5-cm depth and air temperature  were measured inside each chamber using self-made thermocouplesW, % (v/v)) at a 10-cm depth was monitored using time domain reflectometry P, hPa) at a 30-cm height in the centre of the plot was measured by a pressure transducer Soil temperature (2 efflux (R) was calculated as follows:Soil C–COR is soil C–CO2 efflux (g C–CO2 m−2 d−1); M is the C molar mass; V0, P0 and T0 are constants ; Ta and P are air temperature (K) and pressure (hPa), respectively; H the is height of the respiration chamber (m); and dc/dt is the slope of CO2 concentration variation with time over the measurement period .where R (hourly), and 48 data points for T and W (twice per hour), except during periods of electrical failure. For each chamber, available data were used to calculate daily average values. Previous analysis showed that RH was unusual in the fifth group for both NR and NRW  and soil water content (W) with soil carbon efflux (R) as followsThe two-factor regression model was used to quantify the relationship of soil temperature   and on soil temperature  were calculated using the following equations:Warming effects on How to cite this article: Wu, C. et al. Heterotrophic respiration does not acclimate to continuous warming in a subtropical forest. Sci. Rep.6, 21561; doi: 10.1038/srep21561 (2016).


Oryza sativa L.) yield is a crucial challenge for modern agriculture. The ideal plant architecture is considered to be critical to enhance rice yield. Elite plant morphological traits should include compact plant type, short stature, few unproductive tillers, thick and sturdy stems and erect leaves. To reveal the genetic variations of important morphological traits, 523 germplasm accessions were genotyped using the Illumina custom-designed array containing 5,291 single nucleotide polymorphisms (SNPs) and phenotyped in two independent environments. Genome-wide association studies were performed to uncover the genotypic and phenotypic variations using a mixed linear model.Increasing rice (NAL1 and Rc) and new significant loci showed pleiotropy and gene linkage. In addition, a long genome region covering ~1.6 Mb on chromosome 11 was identified, which may be critical for rice leaf architecture because of a high association with flag leaf length and the ratio of flag leaf length and width. The pyramid effect of the elite alleles indicated that these significant loci could be beneficial for rice plant architecture improvements in the future. Finally, 37 elite varieties were chosen as breeding donors for further rice plant architectural modifications.In total, 126 and 172 significant loci were identified and these loci explained an average of 34.45 % and 39.09 % of the phenotypic variance in two environments, respectively, and 16 of 298 (~5.37 %) loci were detected across the two environments. For the 16 loci, 423 candidate genes were predicted in a 200-kb region (±100 kb of the peak SNP). Expression-level analyses identified four candidate genes as the most promising regulators of tiller angle. Known  contains supplementary material, which is available to authorized users. MOC1, a gene that is important in the control of rice tillering, was mapped on chromosome 6  . The array consisted of 5,291 single nucleotide polymorphisms (SNPs) that were chosen from the Rice Haplotype Map Project Database (http://202.127.18.221/RiceHap2/index.php) [Rice genomic DNA was extracted from the young leaf tissue of each of the 523 accessions, which had been grown in the experimental field of the China National Rice Research Institute in Hangzhou in 2014. All of the accessions were genotyped using Illumina custom-designed arrays by following the Infinium HD Assay Ultra Protocol [FST) among different subpopulations , were inferred using STRUCTURE version 2.2 [ad hoc statistic ΔK [Major allele frequency, gene diversity, polymorphic information content (PIC)  were calculated using Microsoft Office Excel 2007. The broad-sense heritability was computed through an analysis of variance using the following formula:A total of 12 morphological traits were evaluated in two environments. The statistical analyses of mean, standard error (SE) and broad-sense heritability (σg2 is genetic variance and σe2 is error variance. Correlation coefficients among the 12 agronomic traits were computed by R (http://www.r-project.org/). An ANOVA was carried out to evaluate the effects of genotype (G), environment (E) and genotype × environment (G × E) using SAS system 9.0 , and the percentage of phenotypic variation explained by population structure was also carried out in SAS 9.0.where An association analysis was performed using TASSEL version 4.0 , and they was the vector of phenotypic observation, α was the vector of SNP effects, β was the vector of population effects, μ was the vector of kinship effects, e was the vector of residual effects, Q was the matrix from STRUCTURE relating y to β, and X and K were incidence matrices of 1 s and 0 s relating y to α and μ, respectively [e × p × ln(p)) [P = a/n) [a = 1, 1/4,136 = 2.4 × 10−4) as the cutoff for all traits except for pericarp color .respectively, where ectively , 50. For× ln(p)) , 53 and P = a/n) , 55. In For the significant marker-trait associations, the elite and null alleles were used to determine the choice of elite accessions for further breeding by design. The formula for calculating phenotypic effect of alleles can be described asxi was the phenotypic value of accession carrying the i allele, yj was the phenotypic value accession carrying the j allele, and n and m represented the number of accessions. If a was positive, then the i allele was defined as an elite allele with an increasing effect and the j allele was defined as the null allele with a decreasing effect, and vice versa. In our study, we chose parental accessions with elite or null alleles based on the characteristics of different traits.where actin was used to standardize the data for each run. The expression experiment was performed at three replications for each sample. The qPCR primers and materials are summarized in Additional file The expression levels of 14 TA candidate genes were measured by quantitative real-time PCR (qPCR) using eight accessions with large TAs at the seedling, early and later tillering stages, respectively. Total RNA was extracted from the base of tiller tissue using a MiniBEST Plant RNA Extraction kit . Complementary DNA (cDNA) was synthesized by PrimeScript RT Master Mix . The reaction was performed on an ABI Step-One Plus Real Time PCR System . The expression level of O. sativa collected from 25 countries . The results showed that the LnP(D) value for each given K, from 1 to 12, increased but did not show a maximum value Fig. . The valups Fig. , and theups Fig. . In addiindica and japonica subspecies, only 469 indica accessions, most of which were distributed in southern China  and four groups under two subpopulations  + 0.9733 (R2 = 0.9422)   Fig. . TherefoHB2 was 71.43 %, ranging from 36.49 % for flag leaf length (FLL) to 99.61 % for pericarp color (PC). The average percentage of phenotypic variation explained by population structure was 21.76 %, indicating that phenotypic variation was influenced by population structure. Hence, the Q-matrix was necessary as a covariate for the GWAS analysis to control false positive associations.A wide range of phenotypic variations were observed for the 12 morphologic traits in LS and HZ (Table P < 0.001) except for the effect of E on panicle number  or significantly (P < 0.05) related to each other in LS  is shown in Fig. To control false positive trait-marker associations, four models, GLM, Q, K and Q + K, were compared with each other using the quantile-quantile (Q-Q) plot shown in Fig. P-values were maintained within a 200-kb window according to the average LD decay rate in the panel. Finally, in LS and HZ, 126 and 172 significant SNPs, including known and new candidate loci, respectively, were identified for the 12 traits and all of the significant loci explained ~34.45 % (10.29 to 72.19 %) and 39.09 % (20.10 to 71.76 %) of the phenotypic variation on average, respectively  was detected on chromosome 2, with the SNP explaining ~5.2 % of the total phenotypic variation, and 32 candidate genes were predicted within 200 kb (±100 kb of the peak SNP). For TA, a significant SNP (seq-rs3945) was identified on chromosome 8 both in LS (P = 4.52E-04) and HZ (P = 1.10E-06), explaining ~3.5 % and ~5.9 % of the total phenotypic variation, respectively. Moreover, 22 candidate genes were contained within 200 kb. For FLL in HZ, a significant locus of seq-rs5454 (P = 1.25E-05) was mapped on chromosome 11, explaining ~5.0 % of the total phenotypic variation and 34 candidate genes were obtained. For FLW in HZ, seq-rs4707 (P = 3.53E-06) was detected on chromosome 10, explaining ~5.6 % of the total phenotypic variation and 34 candidate genes were predicted. For panicle type (PT) in LS, a peak SNP of seq-rs1802 (P = 4.67E-06) was observed on chromosome 3, explaining ~5.4 % of the total phenotypic variation and 38 candidate genes were included. For PC, two significant SNPs, seq-rs1190 and gwseq-rs2, explained ~25.59 % of the phenotypic variation in LS and ~16.74 % in HZ were mapped on chromosomes 2 and 8, respectively. In addition, 29 and 27 candidate genes were predicted  containing 43 candidate genes  detected in two environments was chosen to test the association Fig. . FourteeNAL1 locus at ~31.8 Mb on chromosome 4 was closely related to FLW, FLLW and PN in LS. Moreover, the correlation coefficients of FLW-FLLW , FLW-PN  and FLLW-PN  were highly significant in LS  in LS, and the correlation coefficient was 0.41 (P < 0.001)  values except for markers seq-rs5397 and seq-rs5399  Fig. . In addi399 Fig. . The resThe effects of elite alleles for 10 traits that are directly related to rice plant architecture are summarized in Additional file Based on the characterization of the new plant type (NPT)  and the Rice is a staple food and provides energy for more than half of the world’s population. In recent decades, rice production has only increased slightly and improving rice yields has become a pressing problem. Plant type modification is a potential way to solve these challenges. The NPT was characterized by Khush et al. in the 1990s to change rice plant architecture to enhance rice production , 28. Theindica subspecies to control false positive associations. In addition, the program STRUCTURE was used to uncover population structures in the indica panel for a further GWAS analysis [Although GWAS is a feasible tool to reveal complicated genetic variations in rice, it may be highly affected by population structure, resulting in false positive associations . Structuanalysis , 41. Fouanalysis   and relative kinship (K matrix) play key roles in controlling type I error. Therefore, combining the two factors has been suggested to reduce the false positive loci in association mapping , 62. In Arabidopsis [LD plays an extremely important role in association mapping and determines the resolution of an association study . In our bidopsis , than inbidopsis . Marker bidopsis , 28, 31,sd1 [MOC1 [D53 [LC1 [indica panel. The third possible reason was false negative associations because of the strict threshold value.To date, many morphological genes (or QTLs) have been identified in rice using the bi-parental mapping strategy , 15–22. sd1 , MOC1 [2OC1 [D53  and LC1 Rc gene at ~6.1 Mb on chromosome 7, which was highly associated with both PC and HC traits in LS   Fig. . Previouin yield –14. Our  LS Fig. , and the LS Fig. . In addi LS Fig. . This reThe results of the allelic analysis demonstrated that with the continual pyramid of elite alleles, the average phenotypic value increases significantly (Additional file indica accessions. Moreover, 16 significant loci, including 423 candidate genes, were obtained across two environments. Relative expression analyses identified four candidate genes as the most promising regulators of tiller angle. In addition, many significant loci showed pleiotropy and gene linkage. A long genomic region covering ~1.6 Mb may be critical for rice leaf architecture. The genetic variants and elite alleles will be useful for rice improvement. Furthermore, the biological validation of the candidate genes will be focused on in future research. Genome selection and breeding by design using the elite varieties as parents will also be of interest in future.In this work, we identified a number of significant marker-trait associations and candidate genes, and also uncovered the genetic variations of rice morphological traits using the GWAS strategy in 469 The 5291 SNPs genotype data for the 523 accessions was deposited in the Dryad digital repository: http://dx.doi.org/10.5061/dryad.cp25h.


AhHMA4 from Arabidopsis thaliana encodes Zn/Cd export protein that controls Zn/Cd translocation to shoots. The focus of this manuscript is the evaluation of AhHMA4 expression in tomato for mineral biofortification (more Zn and less Cd in shoots and fruits). Hydroponic and soil-based experiments were performed. Transgenic and wild-type plants were grown on two dilution levels of Knop’s medium  with or without Cd, to determine if mineral composition affects the pattern of root/shoot partitioning of both metals due to AhHMA4 expression. Facilitation of Zn translocation to shoots of 19-day-old transgenic tomato was noted only when plants were grown in the more diluted medium. Moreover, the expression pattern of Zn-Cd-Fe cross-homeostasis genes  was changed in transgenics in a medium composition-dependent fashion. In plants grown in soil (with/without Cd) up to maturity, expression of AhHMA4 resulted in more efficient translocation of Zn to shoots and restriction of Cd. These results indicate the usefulness of AhHMA4 expression to improve the growth of tomato on low-Zn soil, also contaminated with Cd.The online version of this article (doi:10.1007/s11270-016-2877-0) contains supplementary material, which is available to authorized users. Zinc (Zn) is an essential microelement for living organisms. Its deficiency in plants retards growth and reduces crop yields. Consequently, plant-derived foods are low in Zn, thus contributing to Zn malnutrition in humans, and Zn deficiency is a major risk factor in human health . A transgenic approach involves, among others, ectopic expression of transporters responsible for loading metals into xylem vessels in roots, which determines the efficiency of the translocation process  encoding a plasma membrane-localized efflux protein that participates in Zn/Cd xylem loading  to transfer high amounts of Zn to shoots is conferred by tenfold higher expression of AhHMA4 than in the non-accumulating A. thaliana  not only in AtHMA4 under the 35SCaMV promoter was used to transform tobacco to engineer high metal contents in the shoots for phytoremediation purposes  and, unexpectedly, in a decreased Cd level. Similarly, expression of AhHMA4p1::AhHMA4 from A. halleri in tomato facilitated root-to-shoot Zn translocation and induced Zn uptake in a Zn-supply dependent manner   which was triggered by exposure to excess Zn . Tomato expressing AhHMA4p1::AhHMA4 : (i) wild type and (ii) two independent transgenic homozygous lines (no. 6 and no. 8) with the expression of the AhHMA4p1::AhHMA4 at the similar level 2, 1.5 mM KNO3, 1.2 mM MgSO4, 1.0 mM KH2PO4, microelements: 40 μM NaFeEDTA, 25 μM H3BO3, 2.0 μM MnCl2, 2.0 μM ZnSO4, 0.1 μM CuSO4, 0.5 μM Na2MoO4, 5.0 μM KJ, and 0.1 μM CoCl2  pre-culture in 1/4 Knop’s medium for 2 days (to adjust to hydroponic conditions), (ii) growth on 1/10 or 1/2 Knop’s medium for 6 days (nutrient solution refreshed every 3 days), followed by 4-day growth under the same conditions (1/10 or 1/2 Knop’s medium) with or without the presence of 0.25 μM Cd added as CdClAt the end of each experiment, the plants’ appearance was assessed visually. Roots and shoots were separated, leaves were cut off, and all plant parts were dried in an oven at 56 °C for 3 days. The dry biomass was determined. Collected samples were used for evaluation of Cd, Zn, and Fe concentrations in roots and leaves.Roots and leaves (2nd and 3rd counting upward) were also collected for determination of the expression level of selected metal-homeostasis genes see “”. For th2 solution. Afterwards, the soil was kept for 2 weeks before planting with daily mixing and dampening with equal amounts of distilled water according to Wojas et al.  and in soil to which Cd was added (10 mg Cd/kg dry matter of soil) as a CdCls et al. .One-week-old seedlings were transferred from glass jars to liquid 1/4 Knop’s medium and grown for 2 weeks. Three-week-old plants were transferred for further growth to pots containing soil with or without the addition of Cd, one plant/pot containing 1.35 kg of soil. Plants were grown on soil for 101 days. Five plants were used for each experimental regimen. The plants were watered regularly with distilled water. The side shoots were removed on a regular basis. Only two inflorescences were left on the main shoot. The growth and development of plants were monitored throughout the experiments, and their appearance was assessed visually. At the end of the experiments, the roots and shoots were separated. Leaves were cut off the stem and collected in two groups: (i) lower leaves (leaves 1 to 7 counting upward) and (ii) upper leaves (remaining leaves above the “lower leaves”). Roots were thoroughly washed under tap water, then in distilled water. Fruits were collected upon maturity throughout the whole growth period. Seeds were removed from ripe fruits, washed, gently dried with filter paper, and counted. Collected plant parts were dried in an oven at 56 °C, and their dry biomass was determined. Dried plant material was used for the determination assay of Cd, Zn, and Fe concentrations.Dried plant parts  gene was used as the reference gene.The levels of gene expression were evaluated in the roots and leaves collected from hydroponically grown plants “”. The RNt test. Analysis was performed with Excel 2007 for Windows.Statistical significance was calculated at the 0.05 probability level using Student’s AhHMA4p1::AhHMA4 to a better performance of transgenic plants under low mineral medium composition with more focus on root-borne processes. In soil experiments, it is not possible to examine in roots precisely metal accumulation and expression of chosen genes (the majority of fine roots are lost when soil is removed from the root system). Therefore, the experimental setup used in this study involved simulation of low and rich medium with the use of hydroponic experiments. In plants grown hydroponically, metals are available for uptake (as from soil solution), and applied concentrations are comparable to those identified in the soil solution of agricultural soils. For example, in soil solutions extracted from variety of soils, Zn concentrations varied between 0.14 and 33 μM  versus rich medium (1/2 Knop’s) were used to simulate soil poor and rich in nutrients and to evaluate a possible contribution of the expression of AhHMA4 varied over a range of Zn supply . In all experimental trials, the plants had green healthy looking leaves, and exposure to 0.25 μM Cd did not induce any visual symptoms of toxicity (data not shown). Roots and leaves biomass production remained at a similar level in transgenic and wild-type plants. A difference was detected, however, for leaves between all lines grown on 1/10 and on 1/2 Knop’s medium. Their biomass was lower for those grown on more diluted medium  encoding the Fe/Zn/Cd uptake protein  involved in the regulation of metal cross-homeostasis , encoding nicotianamine synthase (NAS) mediating the biosynthesis of nicotianamine (NA), a chelator of several metals , not yet characterized in tomato, however, from the studies on Arabidopsis, maize and rice, it is known that Zn, Fe, and Cd could be involved in ZIP4 regulation  as compared with 1/2 Knop’s medium, both with and without the presence of Cd . It is worth noting, however, that the expression of this gene was twofold higher in the leaves than in the roots  in the presence and absence of Cd, with a view towards biofortification. The mineral composition of the soil and the mineral concentrations in the water extract are shown in Table The aim was to determine how the expression of The development of plants was monitored throughout the experiment. No differences were detected between transgenic and wild-type plants in their appearance (data not shown). On control soil, the growth rate was comparable between the transgenic and wild-type plants. However, transgenics grew faster on Cd-contaminated soil. Their shoots were significantly higher until the 77th day of growth ; howeverThe transgenic plants exhibited a number of features indicating better than wild-type performance in the generative phase, both under the control conditions and in the presence of Cd. The first two, upper and lower, bunches of fruit were analyzed in each plant. The total number of fruits and their biomass were higher in the transgenic than in the wild-type plants Fig. . SimilarAhHMA4p1::AhHMA4 on the development  of the reproductive organs and, to a lesser extent, on growth in the vegetative phase. This suggests that expression of AhHMA4 provides a protective effect against Cd when plants are exposed to its low level.In summary, these data point to the positive impact of the expression of AhHMA4p1::AhHMA4 expression in tomato plants modifies Zn, Cd, and Fe accumulation, their concentrations in the roots, leaves, fruits, and seeds were measured.To determine how AhHMA4 contributed to moderately enhanced Zn level in the leaves, but only in the presence of Cd   might suggest that Fe deficiency-inducible IRT1, as a broad substrate range transporter mediating distribution of not only Fe, but also of Zn, Mn, Cd, and Ni, primarily at low Fe  and LeNRAMP1 was also higher than in wild-type plants , chelator of metals . Recent study implicated a role of AhNAS2 in Zn and Cd translocation to shoots of A. halleri. A decrease in NA concentration in roots through RNAi targeting NAS2 resulted in reduction in Zn and Cd level in leaves  compared with 1/2 Knop’s medium, and the presence of Cd did not affect the transcript level , in uptake and/or redistribution , and in the regulation of metal cross-homeostasis (LeChln), likely as a response to alteration of a plant metal status due to export activity of HMA4 , the level of expression of the transgene was lower in roots than in leaves  accumulated more Cd when grown on the more diluted medium (1/10)  Fig. . This is10) Fig. .The advantage of using liquid media for plant growth is the ability to control the mineral composition, which is necessary to learn about specific plant responses at the transcriptome and metabolome levels to the conditions that have been set up. However, it is known that plants grown in nature have to cope with complex, variable soil conditions. These include interactions with microorganisms and the composition of the soil solution, which depends on the content of minerals, presence of organic matter, as well as water availability. For example, the reduction of soil salinity decreased distribution of heavy metals by tomato and amaranth  bunch of transgenic plants and, not the second, upper one , suggest the usefulness of this transformation for improving the growth of tomato on low-Zn soil, also contaminated with Cd, with a chance of reducing the concentration of this toxic metal in the aerial parts.In summary, expression of ESM 1a) and leaves (b) of 19-day-old tomato plants expressing AhHMA4 , and wild-type (WT) grown hydroponically in 1/2 and 1/10 Knop’s medium without and with 0.25 μM Cd for 4 days. Values correspond to arithmetic means + SD (n = 6). (PDF 51.5 kb)Dry biomass of roots , and wild-type (WT), grown fopr 101 days in control soil (a), spiked with 10 mg Cd/kg d.m. (b). Values correspond to means ± SD (n = 3); those significantly different from the WT (Student’s t test) are indicated by arrows (P ≤ 0.05). (PDF 60.7 kb)Height of tomato plants expressing ESM 3a), lower leaves (b), roots (c) of tomato plants expressing AhHMA4 , and wild-type (WT), grown for 101 days in soil spiked with 10 mg Cd/kg d.m. and without Cd (control soil). Values correspond to means ± SD (n = 3); Different letters represent significantly different values at P < 0.05 for wild-type plants grown upon different medium composition; . Fe concentration from fruits collected from upper and lower bunches, and from seeds collected from these fruits, were not significantly different between transgenic and WT plants. Fe concentrations [mg/kg d.m..] are as follow: fruits form upper bunches: (-Cd) Line 6: 50,89 ± 9,54; Line 8: 42,22 ± 3,17; WT 55,85 ± 15,69; (+Cd) Line 6: 47,78 ± 9,67; Line 8: 49,85 ± 8,04; WT 50,30 ± 4,13;fruits from lower bunches: (-Cd) Line 6: 44,84 ± 7,02; Line 8: 38,83 ± 3,51; WT 49,60 ± 7,16; (+Cd) Line 6: 42,76 ± 12,64; Line 8: 40,67 ± 5,25; WT 49,57 ± 0,42, seeds form upper bunches: ( -Cd) Line 6: 76,03 ± 37,96; Line 8: 68,93 ± 7,27; WT 72,67 ± 24,22; (+Cd) Line 6: 40,21 ± 4,47; Line 8: 88,84 ± 14,97; WT 37,55 ± 4,92, seeds from lower bunches: (-Cd) Line 6: 48,14 ± 4,48; Line 8: 67,34 ± 18,49; WT 41,65 ± 6,86, (+Cd) Line 6: 59,65 ± 9,43; Line 8: 52,95 ± 3,67; WT 42,76 ± 14,06. (PDF 53.1 kb)Fe concentration in upper leaves (Below is the link to the electronic supplementary material.


Over 20 % of ovarian cancer patients have preoperative thrombocytosis or hyperfibrinogenemia. We aimed to demonstrate the clinical and prognostic significance of thrombocytosis and hyperfibrinogenemia in high-grade serous ovarian cancer (HGSC).9/L, and hyperfibrinogenemia was defined as a fibrinogen level higher than 4.00 g/L. Progression-free survival (PFS) and overall survival (OS) were analyzed with the Kaplan-Meier method and log-rank tests for univariate analyses. For the multivariate analyses, Cox regression analysis was used to evaluate the effects of the prognostic factors, which are expressed as hazard ratios (HRs).We retrospectively investigated HGSC patients who underwent primary staging or debulking surgery between April 2005 and June 2013 in our institution. None of these patients had received neoadjuvant chemotherapy. Data, including age, performance status, FIGO stage, serum CA125, platelet count, fibrinogen level, and surgical residual disease, were collected. Thrombocytosis was defined as a platelet count greater than 450 × 109/L, and 121 (13.8 %) women had thrombocytosis. The median (IQR) preoperative fibrinogen level was 3.85 (3.19–4.45) g/L, and 332 (45.9 %) of the patients had hyperfibrinogenemia. Both preoperative thrombocytosis and hyperfibrinogenemia were associated with an advanced FIGO stage , an increased CA125 level , more extensive ascites , more extensive residual disease  and chemosensitivity . In the univariate analyses, hyperfibrinogenemia was associated with reduced PFS (p < 0.001) and OS (p < 0.001). However, thrombocytosis was not found to be a potential predictor of PFS (P = 0.098) or OS (p = 0.894). In the multivariate analyses, hyperfibrinogenemia was an independent predictor of OS (p = 0.014) but not PFS (p = 0.062).A total of 875 consecutive HGSC patients were identified. The median follow-up time was 29 (1–115) months. The median  preoperative platelet count was 301 (235–383) × 10Preoperative thrombocytosis and hyperfibrinogenemia reflected tumor burden to some extent and thus influenced treatment outcomes, and the fibrinogen level was found to be useful as a prognostic predictor in the HGSC patients. Ovarian cancer is the seventh most commonly diagnosed cancer and the eighth leading cause of cancer death among females worldwide. There were 238,700 estimated new cases and 151,900 deaths in 2012 , and appSerologic coagulation parameters, including platelet count and fibrinogen level, have emerged as potential preoperative predictors of clinical outcomes , 5, 7. SFurthermore, previous studies have shown that epithelial ovarian cancer is not a single disease but a group of heterogeneous tumors based on distinctive morphologic and molecular genetic features . PreviouThe clinical data were collected retrospectively from women who underwent primary staging or debulking surgery for HGSC between April 2005 and June 2013 at Fudan University Shanghai Cancer Center. This study was approved by the ethics committee of Fudan University Shanghai Cancer Center. All participants had provided written informed consent form. Patients were excluded if they had received neoadjuvant therapy, had been treated for recurrent disease, or were found to have other histological diagnoses on pathological review.vs R2), date of surgery, date of progression or recurrence, date of last follow-up, and the patient’s disease status at last contact, were collected.Clinical and pathological data were obtained from the medical records, cancer registries, and pathology reports. Patient characteristics including age, FIGO stage, preoperative laboratory data , presence of ascites, surgical outcomes  after surgery. R1 was defined as a maximal diameter of the macroscopic residual disease after cytoreduction of <1 cm. R2 was defined as a maximal diameter of residual disease of ≥1 cm. The optimal surgery was defined as the combination of R0 and R1. Chemosensitivity was defined as a time interval of 6 months or longer between the completion of platinum-based chemotherapy and the detection of relapse. Chemoresistance was defined as disease progression during adjuvant chemotherapy or within the 6-month interval between the completion of platinum-based chemotherapy and the detection of relapse.Thrombocytosis was defined as platelet count greater than 450 × 10Progression-free survival (PFS) was defined as the time interval from the date of primary surgery to the date of disease progression or recurrence. Overall survival (OS) was defined as the time interval from the date of the primary surgery to the date of death or the last follow-up.P < 0.05 was considered statistically significant, and all reported P values were 2-sided.SPSS statistical software  was used for the statistical analyses. Descriptive statistics were used for the demographic data and are summarized as the means with the standard deviations (SD), the medians with the interquartile ranges (IQRs) or ranges, or the frequencies with the percentages. The categorical data were compared with chi-square or Fisher’s exact tests as appropriate. The PFS and OS were analyzed with the Kaplan-Meier method, and log-rank tests were used in the univariate analyses. For the multivariate analyses, Cox regression analysis was used to evaluate the effects of the prognostic factors, which are expressed as hazard ratios (HRs). 9/L, and 121 (13.8 %) women had preoperative thrombocytosis. The median (IQR) fibrinogen level was 3.85 (3.19–4.45) g/L, and 332 (45.9 %) had preoperative hyperfibrinogenemia. Seventy-seven patients had coexisting thrombocytosis and hyperfibrinogenemia. Seventy-five (8.6 %) patients were early stage (I-II) and 800 (91.4 %) were advanced stage (III-IV).The patient characteristics are shown in Table p = 0.008 and <0.001, respectively), an increased CA125 level  and more extensive ascites . There were no significant correlations of age with thrombocytosis or hyperfibrinogenemia .The correlations of the clinical characteristics with preoperative thrombocytosis and hyperfibrinogenemia are summarized in Tables p < 0.001) and hyperfibrinogenemia  increased. Pairwise comparisons revealed significant differences in the prevalences of hyperfibrinogenemia between the R0, R1 and R2 groups . Regarding the platelet counts, a smaller proportion of the patients in the R0 group had thrombocytosis compared with the R1 and R2 groups (R0 vs R1: p = 0.001 and R0 vs R2: p < 0.001). However, the difference in the prevalence of thrombocytosis between the R1 and R2 groups was not significant (p =0.092).After primary surgery, 272 (31.1 %) of the patients were debulked to R0, and 434 (49.6 %) were debulked to <1 cm with macroscopic disease (R1). As the extent of residual disease increased, the numbers of patients who presented with thrombocytosis  and hyperfibrinogenemia  compared with the chemosensitive patients. Next, we investigated the correlations of chemosensitivity with thrombocytosis and hyperfibrinogenemia after stratification according to surgical outcome. The prevalence of thrombocytosis was no longer associated with chemosensitivity in the R0, R1 or R2 subgroups . Regarding the fibrinogen levels, greater proportions of chemosensitive patients in the R0 and R1 groups had hyperfibrinogenemia compared with the corresponding chemoresistant patients (pR0 = 0.001 and pR1 = 0.004). However, the difference in hyperfibrinogenemia between the chemosensitive and chemoresistant patients was not significant in the R2 group (p =1.000).Following primary surgery, 849 (97.0 %) patients had received platinum-based adjuvant chemotherapy. Among these patients, 568 (66.9 %) were chemosensitive Table . GreaterThe median(range) follow-up time was 29 (1–115) months. One hundred four (11.9 %) women experienced disease progression during adjuvant chemotherapy, 499 (57.0 %) patients exhibited documented recurrence, and 345 (39.4 %) deaths were documented. The median (95 % CI) PFS was 18 (16.8–19.2) months, and the 2-year and 5-year PFSs were 38.1 and 19.4 %, respectively. The median (95 % CI) OS was 58 (51.4–64.6) months, and the 2-year and 5-year OSs were 79.3 and 48.8 %, respectively.p < 0.001 and <0.001, respectively), the presence of residual disease , and chemoresistance  were confirmed in the univariate analyses.Regarding PFS and OS, the known negative influences of advanced FIGO stage (vs 21.0 (17.8–24.2) months, p < 0.001, Fig. vs 19.0 (17.5–20.5) months, respectively, p=0.098; Fig. p = 0.062; Table In the univariate analysis, preoperative hyperfibrinogenemia was associated with impaired PFS (15.0 (13.8–16.2) .0 17.8–2.2 monthsvs 72 (58.5–85.5) months, p < 0.001) in the univariate analysis, but thrombocytosis was not correlated with OS (54.0 (41.0–67.0) vs 58.0 (50.6–65.4) months, p = 0.894; Fig. p = 0.001; Table Hyperfibrinogenemia was associated with a shorter OS (42 35.2–48.8) .2–48.8 vIn this large mono-institutional study, we demonstrated that both thrombocytosis and hyperfibrinogenemia were associated with clinicopathological characteristics and treatment outcomes. Additionally, hyperfibrinogenemia was correlated with poorer overall survival in the Chinese patients with HGSC.We found that both thrombocytosis and hyperfibrinogenemia were associated with increased tumor stage, elevated CA125 level, and more extensive ascites. These finding are in accordance with the previous published data from patients with epithelial ovarian cancer regardless of histologic type , indicatp = 0.035). Another multicenter study reported that an elevated fibrinogen level is associated with the presence of increased postoperative residual tumor mass (p < 0.001), which might be due to the increased operative complexity that results from increased tumor burden. Our study confirmed that both thrombocytosis and hyperfibrinogenemia were correlated with more extensive post-surgical residual disease.Because they were correlated with increased disease burden, platelet count and plasma fibrinogen level could be considered to be predictors of surgical outcomes –6, 16. MAdditionally, we investigated the relationship between hypercoagulability and chemosensitivity in HGSC patients for the first time. We observed that more patients with chemoresistant disease had documented thrombocytosis and hyperfibrinogenemia compared with the chemosensitive patients. Previous studies have demonstrated that coagulation system components can promote tumor growth, angiogenesis, and metastasis and thus interact with cytotoxic drugs , 18–21. Several previous studies have also shown that platelet count and fibrinogen level are potential prognostic indicators in ovarian cancer patients –6, 11. OThe limitation of our study is that it was a retrospective study that depended on accurate documentation, and thus the potential for recall bias existed. Additionally, our center is one of the tertiary referral hospitals in China, and over 90 % of patients in such hospitals have advanced disease. Previous studies have shown that other coagulation biomarkers, including D-dimer and von Willebrand factor, can also reflect disease burden and serve as prognostic predictors , 8. HoweNotwithstanding its limitation, this was a mono-institutional retrospective study that included a large number of HGSC patients. This study involved a group of homogenous patients with the same histology who were treated within 9 years and underwent similar surgical procedures and consistent platinum-based chemotherapy regimens. And this could increase the generalizability of our study findings.In conclusion, our study suggests that both preoperative thrombocytosis and hyperfibrinogenemia reflect tumor burden to some extent and thus are predictive factors of treatment outcomes and that fibrinogen level could be regarded as a prognostic predictor in HGSC patients.


Polygonum hydropiper L decoctions are traditionally used in the treatment of various ailments including inflammation, dyspepsia, diarrhea, menorrhagia, hemorrhoids, helminthiasis and CNS disorders. Present study was undertaken to investigate P. hydropiper L. for heavy metals content, phytoconstituents, Phytotoxic and anthelmintic activities to explore its toxicological and pharmacological potentials and rationalize its ethnomedicinal uses.Pheretima posthuma and Ascaridia galli were used to study anthelmintic potential of the plant using albendazole and levamisole HCl as standard drugs.Plant crude powder, methanolic extract, fractions and soil samples were analyzed for heavy metals using atomic absorption spectrophotometer. Qualitative phytochemical analysis of the plant extracts was carried out for the existence of alkaloids, flavonoids, glycosides, anthraquinones, saponins, terpenoids, sterols and tannins. Radish seeds phytotoxicity assay was used to study phytotoxic action of plant extracts. n-hexane (Ph.Hex), chloroform (Ph.Chf), ethyl acetate (Ph.EtAc), n-Butanol (Ph.Bt), aqueous (Ph.Aq), saponins (Ph.Sp) and soil samples were found to contain copper (Cu), iron (Fe), chromium (Cr), zinc (Zn), lead (Pb), nickel (Ni), cadmium (Cd) and lead (Pb) in different concentrations. In crude powder of the plant, heavy metals concentrations were within WHO specified limits, whereas different fractions and soil samples exhibited high metals content. Ph.Cr was tested positive for the presence of alkaloids, flavonoids, saponins, tannins, triterpenoids and anthraquinone glycosides. Among different fractions Ph.EtAc, Ph.Sp, Ph.Chf and Ph.Bt were most effective causing 89.32, 89.25, 86.68 and 85.32% inhibition of seeds in phytotoxicity assay, with IC50 values of 50, 60, 35 and 100 μg/ml respectively. In anthelmintic study, Ph.Sp, Ph.Chf, Ph.EtAc and Ph.Cr were most effective against P. posthuma at 10 mg/ml concentration with an average death time of 50, 64.67, 68.67 and 71 minutes respectively. Ph.EtAc, Ph.Chf and Ph.Aq were most effective against A. galli with average death time of 7, 9 and 10 min respectively at 1 mg/ml concentration.Plant crude powder, methanolic extract (Ph.Cr), its subsequent fractions; P. hydropiper contains different heavy metals and secondary metabolites. Different fractions exhibited phytotoxic and anthelmintic activites comparable to control drugs, thus provide pharmacological basis for ethnomedicinal uses of this plant.Our findings indicate that  Pakistan is naturally gifted with a wide variety of medicinal plants due to its diverse climate and edaphic conditions. At least six thousand flowering plants have been reported currently from Pakistan among which 400–600 are of high medicinal value. Even thPolygonum hydropiper, also known as "smartweed" belong to Polygonaceae, a family comprising of about 50 genera and 1200 species[P. hydropiper and other species of Polygonaceae are used to treat fever, chill, joint pain, oedema and infectious diseases for more than 300 years in Chinese folk medicine[P. hydropiper for antioxidant, anticholinestrase activities and its potential use in the treatment of neurodegenerative diseases[Polygonaceae including Polygonum persicaria and Polygonum plebejum are reported for cytotoxic and phytotoxic activities[ species. Domesti species. Ethnomemedicine. Moreovemedicine, Prostatmedicine, insomnimedicine, hypertemedicine, 12. We diseases. Other stivities.Though plant based therapies are known to be free from side effects in comparison to allopathic drugs, still the presence of toxic heavy metals in such products can cause serious health consequences, 16. RecPhytochemicals are broadly grouped into two categories namely, primary constituents and secondary metabolites. Primary constituents include amino acids, proteins, common sugars and chlorophyll whereas secondary constituents include alkaloids, phenolic compounds, flavonoids, saponins, essential oils, tannins and terpenoids. MajoritP.hydropiper extracts and crude saponins.Globally there are two billion people of different ages which are parasitic worms carriers. These hAlbendazole and Levamisole HCl used as standard drugs in anthelmintic activity were purchased from GlaxoSmithKline and ICI Pakistan respectively. Paraquat (Sigma-Aldrich EC 231-791-2) was used as positive control in phytotoxic assay. Solvents used in extraction, metals analysis and reagents for phytochemical tests were of pure analytical grade.P. hydropiper whole plant was collected from Talash Valley, Khyber Pakhtoonkhwa Pakistan in July, 2013 and was identified by Dr. Gul Rahim, Arid Agriculture University, Rawalpindi, Pakistan. A voucher (H.UOM.BG.107) of the specimen was deposited at the herbarium of University of Malakand, Chakdara (Dir), Pakistan. Plant material was cleansed with distilled water and was shade dried for 30 days. Thereafter, it was coarsely crushed using cutter mill. The powder material (4.5 kg) was soaked in 80% methanol (22 L) in large container for 15 days with frequent shaking. Extraction with methanol was repeated three times, added to original extract and filtered through muslin cloth and then through filter paper[P. hydropiper (Ph-Cr) was suspended in 500 ml of distilled water and consequently partitioned with n-hexane (3 × 500 ml), chloroform (3 × 500 ml), ethyl acetate (3 × 500 ml), n-butanol (3 × 500 ml) and finally aqueous fraction was left. Extraction yield were 68 g (27.2%) for Ph.Hex, 27 g (10.8%) for Ph.Chf, 13 g (5.2%) for Ph.EtAc, 11 g (4.4%) for Ph.Bt & 37 g (14.8%) for Ph.Aq fractions.er paper. The filn-butanol was added to aqueous fraction. The combined aqueous - butanol mixture was washed two times with 5% NaCl solution. Finally solvents were evaporated using water bath to get saponins (9 g) with a percent yield of 15%[Plant material (powdered) weighing 60 g was transferred to a conical flask and was soaked with 100 ml of 20% ethanol. The mixture was heated for 4 h at 55°C using water bath and constant shaking. Thereafter, it was filtered and was again extracted with 200 ml of 20% ethanol. Volume of the liquid extracted was reduced to 40 ml using water bath and transferred it to a separating funnel. Diethyl ether (20 ml) was added to it with vigorous shaking to separate the two phases (Diethyl ether and water). Organic layer was discarded, whereas 60 ml of d of 15%.For metal analysis chemicals of analytical grade were used to prepare sample of the crude drug and different fractions. From each fraction a sample weighing 2 g was taken in a crucible and was ignited in a muffle furnace at 550°C for 6 h. The ash produced was digested in 5 ml of concentrated nitric acid followed by evaporation on hot plate. Small amount of distilled water was added to the digested residue, filtered and volume was made to 30 ml using distilled water. The solutions formed were quantitatively analyzed using atomic absorption spectrophotometer  for the heavy metals including, iron (Fe), lead (Pb), copper (Cu), cadmium (Cd), zinc (Zn), nickel (Ni) and chromium (Cr).2SO4 was then added from the sides of the test tube and was observed for the formation of a brown ring at the junction of two layers. Formation of green color at the upper layer and deep red color at the lower layer indicate presence of steroids and triterpenoids respectively. For the detection of glycosides "Keller Killiani" Test was used. Test solution was added a few drops of glacial acetic acid and Ferric chloride solution and were properly mixed. A few drops of concentrated H2SO4 were added to this mixture and observed for the formation of two layers. Lower reddish brown layer and upper acetic acid layer which turns bluish green indicates positive test for glycosides. Presence of saponins was detected based on the formation of froth upon vigorous shaking using diluted samples. Anthraquinones were detected by boiling the test sample with 1 ml of H2SO4 in a test tube followed by filtration. After cooling the filtrate was shacked with equal volume of chloroform and lower layer was separated and shacked with dilute ammonia. Formation of rose pink to red color ammonical layer indicate presence of anthraquinone glycosides. Ph.Cr was treated with gelatin solution and the appearance of white precipitate was observed which indicate the presence of tannins.Qualitative phytochemical analysis of the plant extracts was carried out for the existence of alkaloids, flavonoids, glycosides, anthraquinones, saponins, terpenoids, sterols and tannins using the method reported previously. The pre(HgCl2) 0.1% solution was prepared in distilled water and radish seeds were put in it to be surface sterilized for 2–3 minutes. The seeds were rinsed with autoclaved distilled water and were dried using sterilized blotting paper. 0.5 ml of each sample solution was transferred to sterilized Petri dish containing Whatman paper and methanol was vacuum evaporated. Methanol (5 ml per Petri dish) was used as negative control (blank). To each test group Petri dish 5 ml of distilled water was added, and 25 radish seeds were placed in each plate at sufficient distance using sterilized forceps. All plates were incubated at 25°C in dim light. After 3–5 days of incubation, number of seeds germination and percent inhibition of root length was calculated using formula:Radish seeds were used to study phytotoxic potential of plant extracts employing method previously described. MercuriThe test was performed in triplicate and data were analyzed by ANOVA.Pheretima posthuma L. Vaill) and roundworms  were used to investigate anthelmintic potential of Ph.Cr, Ph.Hex, Ph.Chf, Ph.EtAc, Ph.Bt, Ph.Aq and Ph.Sp using method previously described[in-vitro investigation due to their high physiological and anatomical resemblance with human intestinal roundworm Ascaris lumbricoides[. The earth worms were collected from marshy soil having small pellets on the surface in the locality of University of Malakand, Chakdara Khyber Pakhtoonkhwa, Pakistan, with an average length of 8-9 cm. For collection of round worms intestines of freshly slaughtered fowls were dissected and were maintained in normal saline solution. average length of roundworms was 5-7 cm. Different dilutions  of plant extracts, Ph.Sp and standard drugs  were prepared. From these solutions, 25 ml each were transferred to sterile Petri dishes and six worms were added to each Petri dish with the help of spatula. Time for complete paralysis of the worms, when no movement (except with vigorous shaking) was observed. Similarly, death time was noted when no movement was observed even with vigorous shaking and exposure to hot water at 50°C temperature.Adult earthworms  were calculated at 95% confidence intervals.One-way ANOVA followed by Dunnett’s multiple comparison test were applied for the comparison of positive control with the test groups. P. hydropiper are summarized in Table P. hydropiper, concentrations of all heavy metals including Cu, Fe, Cr, Zn, Ni and Cd were 7.21 ± 0.23, 95.00 ± 0.55, 1.51 ± 0.45, 7 ± 0.06, 0.35 ± 0.154 and 0.199 ± 0.001 ppm while the concentration of Pb in crude powder was below the detectable range. On the other hand, concentrations of these metals were high in soil samples as compared with the Ph.Cr and different fractions extracted from crude extract. Cr concentration was below detection limit in almost all fractions except Ph.Cr.The concentrations of various metals in dry powder, Ph.Cr and different fractions i.e. Ph.Hex, Ph.Chf, Ph.EtAc, Ph.Bt, Ph.Aq, Ph.Sp of Ph.Cr was tested positive for the presence of alkaloids, flavonoids, saponins, tannins, triterpenoids anthraquinones, glycosides, while tested negative for the presence of steroids Table .Table 250 values of 35, 50 and 60 μg/ml, respectively. This was followed by Ph.Cr, Ph.Hex, Ph.Bt and Ph.Aq fractions which showed 68.00 ± 2.8% (IC50 310 μg/ml), 77.32 ± 1.3% (IC50 215 μg/ml), 85.32 ± 3.5% (IC50 100 μg/ml), 73.32 ± 3.5% (IC50 485 μg/ml) inhibition of seed germination respectively at 1 mg/ml concentration. Ph.Bt, Ph.Sp, Ph.EtAc and Ph.Cr showed highest inhibitory effect on root length causing 86.26 ± 1.25% (IC50 8 μg/ml), 91.90 ± 0.75% (IC50 10 μg/ml), 92.34 ± 1.30% (IC50 11 μg/ml) and 82.38 ± 0.68% (IC50 11 μg/ml) respectively, at 1 mg/ml concentration , Ph.Hex (21.00 ± 3.00), Ph.Chf (9.67 ± 2.51), Ph.EtAc (12.67 ± 0.57), Ph.Bt (8.33 ± 1.53), Ph.Aq (22.67 ± 2.51) and Ph.Sp (11.00 ± 0.58) minutes. While for albendazole and levamisole HCl the paralysis times were 7.67 ± 0.57 and 8.33 ± 0.58 minutes respectively at 10 mg/ml concentration. In anthelmintic activity against A. galli, Ph.EtAc, Ph.Chf and Ph.Aq were most effective with average death time of 7, 9 and 10 minutes respectively at 1 mg/ml concentration. Activity of these fractions were comparable to control drugs albendazole and levamisole HCl which killed all tested roundworms in 10 and 9 minutes respectively at the same concentration. All other fractions were effective in concentration dependent manner  of crude powder so high concentrations of these metals are concentrated in these fractions. High concentrations of these metals can be attributed to their concentration in the soil and marshy place from where the plant was collected.High levels of heavy metals, like lead, cadmium, chromium and manganese which are present in soil and waste water utilized for agricultural irrigation, can be accumulated into these systems. Because of their non-biodegradability features, toxicity effects onto several crops and consequently their bioavailability in human beings may be hazardous. Presence of heavy metals and their interactions with essential trace elements can cause serious health consequences. WHO recommends that plant materials, which are used to make finished products for use, may be checked for the occurrence of heavy metals contamination. Pb and Cd are non-essential trace elements having no useful function in the body. Pb poisoning causes convulsions, chronic nephritis, central nervous system disorders and colic whereas Cd after chronic accumulation in the body causes liver and kidney damages. The consequences of chronic Cr intake include skin rash, nasal itch, kidney and liver damage, bleeds, stomach upsets and lungs cancer. AccordiP.hydropiper could be a significant source of natural herbicides for weeds control in a sustainable manner to increase per acre yield.Different weeds are amongst the most key factors responsible for significant reduction in crops yield. In Sindh and Punjab provinces of Pakistan almost forty weed species have been reported which account for about 40% loss in wheat crop yield on the average. To counteract these unwanted weeds different chemicals are used. However, such chemicals are frequently associated with environmental pollution, residual toxicity, carcinogenesis, high cost and hence their use is restricted, 44. ConP. posthuma and A. galli are mentioned in Figures P. posthuma at 10 mg/ml concentration. Whereas Ph.EtAc, Ph.Chf and Ph.Aq were most effective against A. galli. Anthelmintic activities of these fractions were comparable to positive control drugs  at the same concentration.Anthelmintic drugs are losing their efficacy due to resistance of different nematodes to these drugs especially as a single dose regimen. WidesprP. hydropiper contain Fe, Pb, Cu, Cd, Zn, Cr, Co, secondary metabolites like; alkaloids alkaloids, flavonoids, saponins, tannins, terpenoids, anthraquinones and glycosides. The plant crude extract, its various fractions and crude saponins exhibited concentration dependent phytotoxic and anthelmintic actions, and therefore may be excellent sources of phytotoxic and anthelmintic constituents that warrant their isolation and purification. Further investigations including activity guided isolation is in progress in our laboratory.This study reveals that


Mycobacterium species, and misdiagnosis can result in inappropriate treatment and increased mortality of patients. Differential diagnosis among Mycobacterium species has been made by conventional phenotypic and biochemical tests after a long culture period. Specialized molecular diagnostics of mycobacteria allows rapid detection and species identification; however, such tests are not available in public health programs. Our aim was to demonstrate the clinical implications of erroneous diagnosis by performing molecular genotyping of mycobacterial infections in patients that were diagnosed based on symptoms, culture and bacilloscopy.The establishment of therapeutic regimens for mycobacteriosis depends on the accurate identification of Culture samples of mycobacterial infections from 55 patients clinically diagnosed as tuberculosis in 2013 and 2014, based on conventional methods, were identified by PCR -RFLP and results are discussed.M. tuberculosis, but 18 (32.7 %) were identified as non-tuberculous mycobacteria  and two were negative. Regarding clinical diagnosis, 61.8 % (34/55) was classified as pulmonary tuberculosis. It is important to emphasize that 36.4 % (20/55) of samples were misdiagnosed by conventional methods, and 11 (61.1 %) of the HIV positive patients (18/55) were NTM-coinfected.We have confirmed 35 (63.6 %) positive samples as The identification of species in mycobacterial infections is essential for correct diagnosis and choice of treatment regimen, and misdiagnosis by conventional tools can lead to chronic disease, increased resistance and death.The online version of this article (doi:10.1186/s12879-016-1474-6) contains supplementary material, which is available to authorized users. Mycobacterium species , and only 11 % (6/55) had a substitute treatment for atypical mycobacteria, which included auxiliary drugs to the standard treatment, due to the chronic infection. Among those 6 chronic patients, 5 (7.3 %) were NTM-infected and had their treatment replaced by the appropriate treatment, and one patient was negative  of samples is consistent with other studies that found a higher prevalence of MTB; 67 % [The identification of TB; 67 % , 57.5 % TB; 67 % , 78.6 % TB; 67 %  and 77 %TB; 67 % . DespiteTB; 67 % . The incM. avium, M. fortuitum, M.kansasii, M. chelonae, M. gordonae and M. abscessus [M.avium type 1, 11.1 % M. avium type 2, 5.5 % M. intracellulare, M. kansasii type 1 (11.1 %) and M. chelonae (5.5 %). We also found two rarely pathogenic species, such as 1 M. mucogenicum (5.5 %) and 1 M. terrae type 3 (5.5 %), although they are not commonly associated with diseases they are found in the environment and in potable water distribution systems due to its resistance potential in disinfection procedures [In most studies similar to this, the NTM complex most commonly associated with the clinical disease were bscessus . In thisocedures . We alsoM. tuberculosis (2-8 weeks), 5 MTB cultures presented fast growth (<15 days)  the mycobacterium is the etiological agent, in which diagnosis is supported by clinical signs and by repeated isolation of this bacillus in the same patient, or by a single isolation in the case of samples taken aseptically; (b) the mycobacterium may have colonized the sample, but has no clinical significance, which may have occurred due to the use of contaminated equipment, a common phenomenon called “pseudo-infection”; or (c) the detection of mycobacteria in clinical samples comes from laboratory contamination (contaminated solutions), which can be easily verified with proper controls or by observing if the same mycobacteria was isolated from other samples analyzed in the same day .The diagnosis performed at the Clinics’ Hospital of Uberlândia, takes into account only bacilloscopy, culture time and symptoms for clinical diagnosis, and among 55 patients diagnosed as tuberculosis, 35 were correctly identified as MTB (63.6 %) when verified by PCR-RFLP, but 36.4 % (20/55) of these samples were either NTM-infected (18) or negative (2), configuring misdiagnosis. The disease caused by NTM in most cases is pulmonary or disseminated , but ourM. avium and M. kansasii complex, which constitute most of the cases described in the literature [The recommended treatment for MTB applied to NTM-infected patients, or for NTM applied to MTB patients, or for NTM with non-specific schemes may lead to major complications, since different therapeutic regimens should be adopted for each group. The antimicrobial and the treatment time varies according to the species, and some NTM are resistant to the drugs used for tuberculosis treatment, and not all NTM are sensitive to the same treatments, which may result in chronic infections . Availabterature , 35.Briefly, we have demonstrated that conventional methods used to detect mycobacterial infections, AFB and culture, have led to significant uncertain diagnoses, and considering that species identification is essential to choose the correct therapeutic regimen, it is expected then that mycobacteria resistance, chronic infections, and poorer outcomes may be direct consequences of such misdiagnosis. We suggest that urgent molecular identification of mycobacteria species and resistance tests are incorporated into public health systems in regions with resource-poor settings in order to reduce morbidity by adopting appropriate therapeutic regimens, and to prevent the increased burden of antibiotic-resistant mycobacteria.All the data supporting our findings are contained within this work.


Trisiniotus Jeannel of the pselaphine tribe Batrisini comprises two species distributed in North India and southern Myanmar. Here, a third species, T.taoismus Feng & Yin sp. nov., is described from Mao-shan, Jiangsu Province, East China. The new species can be readily distinguished from both congeners by the unmodified male antennae.The genus  Trisiniotus Jeannel certainly falls into this category. No specific comments were provided on generic characteristics, nor was the genus compared to potentially related genera. In the key to genera  of a median sulcus on the pronotum appears to be homoplastic in a number of Asian batrisine genera, as pointed out by Trisiniotus may indicate, to a certain degree, close relationships with the Japanese species of Batriscenaulax Jeannel and Physomerinus Jeannel, which have the same type of the aedeagus and are separated from each other mainly by different locations of the male sexual characters  among material from southern Myanmar, and transferred the species to Trisiniotus. A third, distinct but unnamed species that has an enlarged antennomere 9 and a medially sulcate pronotum was reported by Trisiniotus, both named species are easily recognizable by the swollen antennomere 10 in the male.In his review of the pselaphine fauna of North India, o genera : 409, Traracters . The typArchiclavigergaofani Yin, Hlaváč & Cuccodoro in Batrisini, which is evidently related to the two Trisiniotus species from India and Myanmar based on the shape of the pronotum and aedeagus. As no attempt is made here to clarify the relationships of Trisiniotus with potentially related groups, we simply describe the new species and place it as a member of Trisiniotus.In August 2020, we organized a collecting trip to Mao-shan Scenic Area, Jiangsu Province in the hope of finding additional material of a distinctive clavigerite beetle (described as SNUC). The label data of the material are quoted verbatim.The type material of the new species described in this paper is deposited in the Insect Collection of Shanghai Normal University, Shanghai  was used for image stacking. All images were modified and grouped into plates using Adobe Photoshop CC 2020.Measurements were taken as follows: total body length was measured from the anterior margin of the clypeus to the apex of the abdomen; head length was measured from the anterior margin of the clypeus to the head base, excluding the occipital constriction; head width was measured across the eyes; the length of the pronotum was measured along the midline; the width of the pronotum is its maximum width; the length of the elytra was measured along the suture; the width of the elytra was measured as the maximum width across both elytra; the length of the abdomen is the length of the dorsally exposed part of the abdomen along its midline; the width of the abdomen is its maximum width. Abdominal tergites and sternites are numbered following Taxon classificationAnimaliaColeopteraStaphylinidaeFeng & Yinsp. nov.D8E7A2BB-8914-5299-BF9E-B1CFDF2440D8http://zoobank.org/C2995259-9A1F-42FF-B22D-A0169BB25162Holotype(15 exx.). : China: ♂, ‘China: Jiangsu, Jurong City, Mao Shan, 31°47'41.99"N, 119°18'43.38"E, leaf litter, sifted, 140 m, 24.viii.2020, Ting Feng leg., 江苏句容市茅山风景区’ (SNUC). Paratypes: China: 6 ♂♂, 3 ♀♀, same data as that of holotype; 1 ♂♂, 2 ♀♀, ‘China: Jiangsu, Jurong City, Yaji-shan, 31°39'24.06"N, 119°17'253.71"E, leaf litter, sifted, 100 m, 23.viii.2020, Zi-Wei Yin leg., 江苏句容市丫髻山脚; 2 ♀♀, China: Jiangsu, Jurong City, nr. Wawu-shan, 31°39'6.28"N, 119°16'20.99"E, leaf litter, sifted, 100 m, 22.viii.2020, Ting Feng leg., 江苏句容市瓦屋山上杆湖农庄 .Male. Body length approximately 1.9 mm. Head sub-rectangular at base; vertex with large and setose foveae, with transverse sulcus at anterior portion; antenna elongate; antennomeres more or less elongate, lacking modifications. Pronotum lacking a median longitudinal sulcus. Discal stria of elytron long, extending posteriorly to approximately apical 3/4 of elytral length. Mesotibia with small apical spine. Metaventrite with setose admesal longitudinal ridges. Tergite 1 (IV) predominantly large, dorsally longer than 2–4 (V–VII) combined, lacking modifications. Aedeagus strongly asymmetrical; median lobe with restricted basal capsule and triangular foramen, ventral stalk erect, narrowing towards apex in lateral view; dorsal lobe narrowed at base, broadened towards apex. Female. Body length approximately 1.8 mm, legs and metaventrite lacking modifications, genitalia as in Fig. Male. Body , length 0.47–0.50 mm, width 0.64–0.68 mm. Tergite 1 (IV) in dorsal view longer than 2–4 (V–VII) combined, lacking basal sulcus, with one pair of basolateral foveae and a short discal carina; tergites 2–4 each with one pair of small basolateral foveae, 4 as long as 2 and 3 combined along middle, 5 (VIII) semicircular, posterior margin roundly emarginate at middle. Sternite 2 (IV) with mediobasal and two basolateral foveae, with a pair of short lateral carinae; midlength of sternite 2 as long as 3–5 (V–VII) combined, 3–5 each short at middle, lacking fovea, 6 (VIII) transverse, posterior margin emarginate at middle, 7 (IX) membranous or absent.Aedeagus Fig.  0.30 mm Female. Similar to male in external morphology; antenna shorter; each compound eye composed of approximately 30 ommatidia; elytra constricted at bases, humerus not prominent; metathoracic wings absent; mesotibia lacking apical spine, metaventrite lacking admesal ridges. Measurements : body length 1.79–1.83 mm; length/width of head 0.37/0.42–0.43 mm, pronotum 0.41–0.43/0.45 mm, elytra 0.56–0.57/0.69 mm; abdomen 0.52–0.53/0.65 mm; length of antenna 0.85–0.87 mm; maximum width of genitalia (Fig. lia Fig.  0.23 mm.Trisiniotustaoismus sp. nov. shares with its two congeners the lack of a median longitudinal sulcus on the pronotum, as well as a similar aedeagus. This species can be readily separated by the simple male antennomere 10, which is swollen in both species from India and Myanmar.East China: Jiangsu Fig. .All individuals were collected by sifting the leaf litter layer in broad-leaved forests Fig. .The specific epithet reflects that Mao-shan is a Taoist sacred mountain in eastern China.


The maximum elongation to failure of 287.12% was achieved in this alloy at 573 K and 5 × 10−4 s−1. It was found that strain-induced grain coarsening at 523 K is much weaker than the strain-induced grain coarsening at 573 K. Thus, the ductility of 228.05% is suitable for application in high strain rate superplastic forming. The stress exponent of 3 and the average activation energy for deformation of 50.06 kJ/mol indicate that the rate-controlling deformation mechanism is dislocation-glide controlled by pipe diffusion.To explore new approaches to severe plastic deformation and the ductility of a multicomponent magnesium–lithium alloy, an ultralight microduplex Mg-9.55Li-2.92Al-0.027Y-0.026Mn alloy was made by novel multidirectional forging and asymmetrical rolling, and the superplasticity behavior was investigated by optical microscope, hot tensile test, and modeling. The average grain size is 1.9 μm in this alloy after multidirectional forging and asymmetrical rolling. Remarkable grain refinement caused by such a forming, which turns the as-cast grain size of 144.68 μm into the as-rolled grain size of 1.9 μm, is achieved. The elongation to failure of 228.05% is obtained at 523 K and 1 × 10 The Mg-Li alloy, the lightest nontoxic metallic alloy, has been investigated extensively in recent years  and has Tm, where Tm is the absolute melting temperature, and a certain strain rate [Superplasticity is the capability of materials to exhibit large ductility and requires (i) fine and ultra-fine grain sizes of less than 10 μm; (ii) temperature of more than 0.5ain rate . Superplain rate . Accordiain rate ,13,14, hain rate ,16, fricain rate ,18, multain rate , and difain rate  have beeain rate . HoweverIn this work, the research contents include several aspects: (i) to fabricate a multicomponent Mg-Li-Al-Y-Mn alloy via MDF and asymmetric rolling; (ii) to investigate its microstructural phenomenon and superplasticity behavior; (iii) to establish a constitutive model and investigate the deformation mechanism at elevated temperature. It is expected that this first report on MDF and asymmetric rolling will stimulate the interests of Mg-Li alloy researchers.3. Our previous reports on multidirectional forging are available elsewhere [The melting and casting process of alloy ingot adopts Jackson’s flux-argon atmosphere protection method. The analyzed composition of the ingot was Mg-9.55Li-2.92Al-0.027Y-0.026Mn. After milling of the ingot surface, the milled ingot was homogenized at 473 K for 16 h. The ingot was cut into billets with dimensions of 40 × 32 × 22 mmlsewhere ,22. The 3. After being annealed at 448 K for 60 min and held at designated testing temperatures for 15 min, the tensile tests were performed on Shimidazu- AG-Xplus 100 kN tester in the temperature range of 423~573 K and strain rate range of 1 × 10−2~5 × 10−4 s−1.Dog-boned specimens for tensile deformation and microstructural observation were taken along the longitudinal rolling direction and made by spark discharge processing. The specimen dimensions were 13 × 3 × 2 mm4 + 90% ethyl alcohol. The operating voltage of twin-jet was 12 V. The current was 30 mA. The temperature of electrolytic polishing was 233 K (−40 °C). The operating parameters of ion thinning were 4 kV ± 3 for 5 min, 3 kV ± 3 for 10 min, and 2 kV ± 2 for 10 min. The temperature of ion thinning ranged from 143 K (−130 °C) to 173 K (−100 °C). FEI Tecnai F30 field emission transmission electron microscope was used for dislocation observation.Optical examination specimens were cut, ground, and polished by the conventional metallographic method. The etched solution was a solution of 10 vol.% hydrochloric acid + 90% vol.% ethyl alcohol or a solution of 5 g picric acid + 5 g acetic acid + 10 mL deionized water + 100 mL absolute alcohol. Optical microstructures were observed on Olympus-DSX500 optical microscope. Image-pro-plus (IPP) software was used to measure the grain size. The high temperature tensiled specimens were mechanically ground and polished to 80 μm. Then, discs with 3 mm diameter were punched. Chemical twin-jet and liquid nitrogen cooled ion thinning were conducted to prepare the samples for transmission electron microscopy (TEM) observation. The electrolyte was a solution of 10% HClO−2 to 5 × 10−4 s−1. The engineering stress decreases with the increase in tensile temperatures from 423 to 573 K. This is because, with the decrease in strain rate and the increase in temperature, the tensile time prolongs and thermal activation accelerates, dislocation density decreases, and stress decreases. As shown in −2 s−1, which demonstrates the low temperature and high strain rate quasi-superplasticity. As shown in −2 s−1, which demonstrates the high strain rate quasi-superplasticity. As shown in −4 s−1.−2 s−1 and 573 K and 5 × 10−4 s−1. Average grain sizes under both conditions are 6.42 and 22.32 µm, respectively. Compared to the grain size of 4.14 µm in −2 s−1. In addition, compared to −4 s−1. This means that 287.12% elongation is obtained in this coarse grained microstructure. Furthermore, the average grain sizes of α-Mg and β-Li phases are 2.03 and 6.91 μm, respectively, at 523 K and 1 × 10−2 s−1. The average grain sizes of α-Mg and β-Li phases are 9.64 and 26.27 μm, respectively, at 573 K and 5 × 10−4 s−1. As the temperature increases and the strain rate decreases, the average grain sizes of the dual phase increase. This indicates the occurrence of phase coarsening.−2 s−1 and 573 K and 5 × 10−4 s−1. Some stacking faults exist in the alloy. Since the samples for TEM examination have been exposed to the ambient environment for 365 days, with dislocations existing in high energy and non-equilibrium state, when the high temperature tensile test was conducted, they reacted and dissociated to become the current low-energy and equilibrium states as shown in m value) under different conditions. m = σ is the true stress and m values range from 0.169 to 0.423, most of which lie between 0.2 and 0.3, indicating that the dominant deformation mechanism is dislocation creep. The m value of 0.375 (stress exponent n = 1/m = 2.66 ≈ 3) corresponds to the maximum elongation to failure of 287.12%, indicative of the occurrence of quasi-superplasticity or superplasticity-like behavior. The stress exponent n = 2.66 ≈ 3 reveals that dislocation viscous glide governs the rate-controlling process under this condition.G is the shear modulus, a function of temperature, b is the magnitude of Burgers vector of dislocation, k is Boltzmann’s constant, T is the absolute temperature, d is the grain size, p is the grain size exponent, σ is the applied stress, n is the stress exponent , D0 is the frequency factor for diffusion, Q is the activation energy for deformation, and R is the universal gas constant. Here, power-law constitutive modeling is performed to elucidate the deformation mechanism at elevated temperature and is suitable for the application in superplastic forming process control. In order to determine the threshold stress, n-value, and Q-value, true stress and true strain formulae are used on the basis of The power-law constitutive equation at elevated temperature is generally expressed as (1)ε˙=ADn1/ relation in this alloy to determine the threshold stress and stress exponent. At the true strain of 0.2, the values of threshold stress n1/ relation. When n = 4 and 5, the threshold stresses become negative. Hence, n = 4 and n = 5 are excluded. When n is 3, the determination coefficient, R2, is 0.9858 with the best correlation, which is higher than the determination coefficient, R2, of 0.9759 when n = 2. Thus, the true stress exponent is determined to be 3.As per Equation (1), the deformation activation energy is given byp is equal to zero [As dislocation creep is predominant in this alloy,  to zero .E = 48,700 − 8.59T − 0.0195T2 [E, Poisson’s ratio υ, and shear modulus G is given byυ of Mg is 0.28 [Young’s modulus of Mg is given by 0.0195T2 . The rel is 0.28 .R2, is 0.946. Thus, the power-law constitutive equation is obtained as the following:The method of MDF + asymmetrical rolling is proposed in this manuscript and put into effect via experimental forming. The total imposed strain during the MDF + asymmetrical rolling processing is 6.2. The principle behind the combined forming is that the accumulated strain, 6.2, through MDF + asymmetrical rolling is much larger than the accumulative strain of MDF, 3.6, and the strain of asymmetrical rolling, 2.6. Here, the strain of asymmetrical rolling, 2.6, is calculated using the true strain formula in reference . Thus, tCompared to the average grain size of 5.5 μm in our previous work  in Mg-6.In recent years, the issue of grain coarsening including static grain coarsening ,30,31 an−2 s−1 and 523 K, the elongation to failure of 228.05% is obtained at 523 K and 1 × 10−2 s−1. It is found that strain-induced grain coarsening at 523 K is much weaker than the strain-induced grain coarsening at 573 K. Thus, the ductility of 228.05% is especially suitable for application in high strain rate quasi-superplastic forming. Furthermore, due to pronounced grain coarsening (grain size 22.32 µm) at 573 K, the elongation of 287.12% is obtained in this alloy at 573 K and 5 × 10−4 s−1. The causes of grain coarsening are analyzed as follows.As shown in b), b is the magnitude of Burgers vector of dislocation, 2.86 × 10−10 m (Al), Dgb is the grain boundary diffusivity of Al, R is the gas constant, and T is the absolute temperature. Thus, the grain boundary diffusivity of Al at 573 K is calculated to be 1.92 × 10−12 m2 s−1. The grain boundary diffusivity of Mg at 573 K is 2.88 × 10−11 m2 s−1 [Mgb is equal to Dgb/kT based on the Einstein equation, where k is Boltzmann’s constant, Mgb (Mg)/Mgb (Al) = Dgb (Mg)/Dgb (Al). Hence, at 573 K, Mgb (Mg)/Mgb (Al) = Dgb (Mg)/Dgb (Al) = 15:1. Our quantitative calculation of Al and Mg diffusivity reveals that the grain boundary diffusivity or mobility of Mg is 15 times as much as that of Al. Higher mobility leads to higher boundary migration. Higher boundary migration results in strain-induced grain coarsening. This means that strain-induced grain coarsening in Mg alloys is higher than that in Al alloys. It is not surprising that strain-induced grain coarsening is a common phenomenon and feature in Mg alloys at a certain temperature, regardless of the strain-induced grain coarsening in the present Mg-9.55Li-2.92Al-0.027Y-0.026Mn alloy. The aforementioned viewpoint is supported by our result in this Mg-Li-Al-Y-Mn alloy, Kim et al.’s thought [Firstly, the experimental phase proportion of α-Mg phase to β-Li phase, which is 15.59:84.41, indicates that the present alloy is a β-Li phase-dominated alloy with a small volume fraction of α-Mg phase. As such, the capability of hard α-Mg phase in restricting the coarsening of soft β-Li phase is weaker at 573 than at 523 K. Secondly, based on the report on the diffusivities of α-Mg and β-Li phases , the dif1 m2 s−1 . As the  thought , and Fig thought  in ZK60 In addition, the superplasticity of Mg-Li-Al system alloy and binary Mg-Li alloy are analyzed. Dutkiewicz et al.  have recDgb is the grain boundary diffusivity, Dl is the lattice diffusivity, and T is the absolute temperature in Kelvin. Due to Dp = Dgb [Dp is the coefficient of pipe diffusion or pipe diffusivity, in consideration of D = D0 exp(−Q/RT), where D0 = 1 × 10−4 m2 s−1 [−11 m2 s−1 and the pipe diffusivity of 5.79 × 10−11 m2 s−1 at 573 K in The activation energy for deformation and stress exponents were determined so as to judge the deformation mechanism at elevated temperature. The phase proportion of the α-Mg phase to β-Li phase is calculated to be 16.30:83.70 as per the binary Mg-Li phase diagram , and theDp = Dgb , where D4 m2 s−1 , and EquQp, is 68.4 kJ/mol, and the theoretical lattice diffusion activation energy, Ql, is 107 kJ/mol in the temperature range of 473–573 K. As shown in It is noted in TM is Taylor factor, =4.5 [k is Boltzmann’s constant, T is the absolute temperature, σ is the true stress, and b3, where b is the magnitude of Burgers vector of dislocation, is considered as a volume unit. Here, the b value of Mg is 3.21 × 10−10 m. The V*/b3 is taken as the normalized activation volume. V* = 100 − 300b3, in this case, trans-granular dislocation slip occurs, but when V* < 1 − 10b3, grain boundary sliding occurs in nanometer material. The experimental grain sizes of 6.42 and 22.32 μm fall into the range of 6–40 μm. Since the ratio of V*/b3 is in the range of 25–366 b3, as shown in Furthermore, the activation volume is estimated to judge the deformation mechanism at elevated temperature. The activation volume, formula (8)V*≡MTor, =4.5  for the −2 s−1. The number of dislocations inside a grain is given by the following relation [N is the number of dislocations, d is the grain size, d = 6.42 μm ), and b is the magnitude of Burgers vector, 3.21 × 10−10 m for Mg. Thus, N = 28.14 ≈ 29. There are 29 pieces of dislocations inside a grain under this condition. In consideration of experimental evidence of stacking faults dissociated from the dislocation reaction in To further validate the dislocation activity, an estimation was made to calculate the number of dislocations inside a grain at 523 K and 1 × 10 6.42 μm a, σ is t(1) An ultralight microduplex Mg-9.55Li-2.92Al-0.027Y-0.026Mn alloy was made by novel multidirectional forging and asymmetrical rolling. The average grain size was 1.9 μm in the present alloy fabricated by multidirectional forging + asymmetrical rolling. Remarkable grain refinement caused by such a forming, which turns the as-cast grain size of 144.68 μm into the as-rolled grain size of 1.9 μm, was achieved.−2 s−1, which demonstrates the high strain rate quasi-superplasticity. The maximum elongation to failure of 287.12% was demonstrated in this alloy at 573 K and 5 × 10−4 s−1. It was found that strain-induced grain coarsening at 523 K is much weaker than the strain-induced grain coarsening at 573 K. Thus, the ductility of 228.05% is suitable for application in high strain rate superplastic forming. Theoretical analysis of atomic diffusion shows that the grain-coarsening velocity of the present alloy was two times as much as that of Mg at 573 K. This indicates that the appropriate quasi-superplastic deformation temperatures for the present Mg-9.55Li-2.92Al-0.027Y-0.026Mn alloy is from 473 to 523 K in which strain-induced grain coarsening is not obvious.(2) The elongation to failure of 228.05% was obtained at 523 K and 1 × 10(3) The power-law constitutive equation was established. The stress exponent was determined to be 3. The average activation energy for deformation was 50.06 kJ/mol, which is close to the theoretical activation energy of pipe diffusion, 68.4 kJ/mol. The results of estimation of stress exponent, activation energy, and activation volume indicate that the rate-controlling deformation mechanism is dislocation glide controlled by pipe diffusion.


The X-ray photoelectron spectroscopy result implies partial oxidation of Sn film on the diamond surface. The leakage current and capacitance–voltage properties of Al/SnOx/H-diamond metal-oxide-semiconductor diodes were investigated. The maximum leakage current density value at −8.0 V is 1.6 × 10−4 A/cm2, and the maximum capacitance value is measured to be 0.207 μF/cm2. According to the C–V results, trapped charge density and fixed charge density are determined to be 2.39 × 1012 and 4.5 × 1011 cm−2, respectively. Finally, an enhancement-mode H-diamond field effect transistor was obtained with a VTH of −0.5 V. Its IDMAX is −21.9 mA/mm when VGS is −5, VDS is −10 V. The effective mobility and transconductance are 92.5 cm2V−1 s−1 and 5.6 mS/mm, respectively. We suspect that the normally-off characteristic is caused by unoxidized Sn, whose outermost electron could deplete the hole in the channel.SnO T. Tx/H-diuations: (1)VFB=ϕ4.90 eV) ; q is th12 cm−2) , and TiO12 cm−2) . The orix film, CV curves were measured from different directions. As shown in x film was believed to be the incompletely oxidized Sn. The trapped charge density (tQ) could be obtained by Equation (4)In order to investigate trapped charge density in SnOtQ was calculated to be 2.39 × 1012 cm−2. CV curves measured at 10 kHz, 100 kHz, 500 kHz, 1 MHz, and 5 MHz were shown in The TH) is extracted from the transfer characteristics  keeps increasing when VGS shifts from VTH to the negative direction, reaches the maximum value of about 5.6 mS/mm at VGS of −4.2 V, and then decreases. G) and on/off ratio of the H-diamond FET, and the gate voltages (VGS) were measured from 2 to −8 V. IG was measured when the drain voltage (VDS) was −10 V, which was 6.8 × 10−12 A at VGS = −8 V. Such a low large leakage current indicates that a dense, insulating oxide film was formed by thermal oxidation. The small leakage current makes the thermal oxidation SnOx film suitable for H-diamond MOSFET with low power consumption. The on/off ratio was evaluated to be ~108, which is high enough for practical applications. In combination with the XPS results, the normally-off characteristic may be caused by partial oxidation of Sn.Next, the I-V characteristic of the MOSFET was investigated by an Agilent B1505A  power device analyzer. The gate length and gate width of this device are 8 μm and 100 μm, respectively. The threshold voltage (Veristics a at VDS D) versus drain-source voltage (VDS) output characteristic curves at different gate voltage (VGS) varying from 2 to −5 with a step of −0.5V. With the increase in |VGS|, |ID| increased, indicating that it was a p-type channel. Additionally, the distinct pinch-off and saturation characteristics were observed in Dmax) is −21.9 mA/mm at VGS = −5 V and VDS = −10 V. Compared with the reported device in ONR for the FET channel.ONR is on state resistance, CHR is the resistance of the channel, and SDR is the resistance between source/drain and H-diamond. GL is channel length, GW is channel width, and OXC is the capacitance of the MOS structure. Thus, the effμ was calculated to be 92.5 cm2V−1 s−1, as shown in effμ of H-diamond MOSFETs are 38.7, 34.2, 39.0, and 88 cm2V−1 s−1 for the gate dielectrics of HfO2 [2/Al2O3 bilayer [x/ZrO2 bilayer [x [x film can form a good interface with the H-diamond surface, which provided a high electrical transport channel.Equation (5) provided the relationship between effective mobility and  of HfO2 , ZrO2/Al bilayer , SiNx/Zr bilayer , and TiOlayer [x , respectx dielectric layer deposited by thermal oxidation of Sn. The SnOx/H-diamond MOS structure showed a low leakage current density, and the capacitance value is 0.207 μF/cm−2. A normally-off H-diamond FET was obtained with a VTH of −0.5 V. Its IDMAX is −21.9 mA/mm at VGS = −5 V and VDS = −10 V. The effective mobility and transconductance are 92.5 cm2V−1 s−1 and 5.6 mS/mm, respectively. According to the C–V results, the fixed charge density and trapped density are 4.5 × 1011 and 2.39 × 1012 cm−2, respectively. In combination with the XPS results, the normally-off characteristic may be caused by partial oxidation of Sn. This is a simple and effective strategy for the fabrication of normally-off H-diamond FET. However, it still has the problem of low current density, which may be improved by reducing contact resistance and scattering in subsequent studies.In summary, we report herein an investigation of the H-diamond FET with SnO