AbstractAstatumen bartosi, Diphascon (Adropion) prorsirostre, D. (Diphascon) bullatum, D. (D.) pingue pingue, D. (D.) recamieri, D. (D.) rugosum, Hypsibius convergens, H. dujardini, H. cf. scabropygus, Isohypsibius ronsisvallei, I. sattleri, Macrobiotus harmsworthi harmsworthi, M. hufelandi hufelandi, Milnesium asiaticum, Milnesium tardigradum tardigradum, Minibiotus formosussp. n. and Paramacrobiotus richtersi. The new species is most similar to Minibiotus gumersindoi, but differs from it mainly by the presence of two types of cuticular pores, the absence of a triangular or pentagonal arrangement of pores above a single large pore on legs, the presence of granulation on all legs and a different macroplacoid length sequence. In this paper we also provide photographs and morphometrics of H. cf. scabropygus.In sixteen moss, lichen and mixed (moss/lichen) samples, collected from Estonia, Latvia and Lithuania, 291 specimens, 48 simplexes, including one exuvium with 6 eggs, and 8 free-laid eggs of eutardigrades were found. In total, 17 species, together with one new to science, were identified : PageBreak The temperate climate is intermediate between maritime and continental. Even though the phylum Tardigrada is cosmopolitan and currently comprises ca. 1,200 species  and Isohypsibius cf. marcellinoi , two from Latvia: Paramacrobiotus richtersi group and Macrobiotus hufelandi group and two from Lithuania: Macrobiotus sp. and Ramazzottius sp. . In 1988 Minibiotus maculartus, and within the last decade fourteen new Minibiotus species have been described. Several species have also been transferred to Minibiotus from the genus Macrobiotus based on characters defined by Minibiotus species amounts to as many as forty seven.In this study we report seventeen tardigrade species, which are all new records for the Baltic States. Moreover, one of these species is also new to science. The new species belongs to the genus Hypsibius cf. scabropygus, a rare species that belongs to a large group of hypsibiids with at least partially sculptured dorsal cuticle and pharynx with two macroplacoids and without the microplacoid.In addition to the description of the new species, we also provide morphometric data and photographs of PageBreakanimals were mounted on microscope slides in Hoyer’s medium. All specimens were examined measured and photographed using Phase Contrast Microscopy (PCM) or Scanning Electron Microscopy (SEM). In total 358 specimens (including 47 simplexes), one exuvium with 6 eggs, and 8 free-laid eggs were examined.Sixteen moss, lichen and mixed (moss/lichen) samples from trees, soil and stones were collected from 15 localities in Estonia, Latvia and Lithuania between the 29 April and the 5 May 2012 by the third author (more details below). Samples were collected and examined for tardigrades using standard methods see . After eMinibiotus and according to Hypsibius. The pt ratio is the ratio of the length of a given structure to the length of the buccal tube expressed as a percentage . In the species list Roman numbers indicate sample codes  and Arabic numbers indicate the number of specimens, exuvia/simplexes and eggs.For species identification and differentiation, keys in Minibiotus formosus sp. n. are deposited in the Tardigrada Register , date: 29.04.2012.55°25'59"N, 24°13'32"E, ca. 59 m asl: Lithuania, Kaunas county, Kėdainiai district municipality, Truskava city, near a church, lichens from tree and moss from soil (slide code: LT 2423), date: 29.04.2012.PageBreak55°17'12"N, 23°58'57"E, ca. 30 m asl: Lithuania, Kaunas county, Kėdainiai district municipality, Kėdainiai city, Kranto II street; moss from wall (slide code: LT 2424), date: 29.04.2012.55°17'13"N, 23°58'56"E, ca. 30 m asl: Lithuania, Kaunas county, Kėdainiai district municipality, Kėdainiai city, Paeismilgio street; moss from stone (slide code: LT 2425), date: 29.04.2012.55°43'35"N, 24°21'30"E, ca. 62 m asl: Lithuania, Panevėžys county, Panevėžys district municipality, Panevėžys city, Garden Street near Holy Trinity Rector; moss from tree (slide code: LT 2440), date: 05.05.2012.56°38'53"N, 23°43'18"E, ca. 7 m asl: Latvia, Zemgale region, Jelgava municipality, Jelgava city, City Park; moss from soil (slide code: ŁO 2426), date: 29.04.2012.57°10'33"N, 24°50'32"E, ca. 45 m asl: Latvia, Vidzeme region, Sigulda municipality, Gutmana Cave in the Gauja National Park; moss from rocks (slide code: ŁO 2427), date: 30.04.2012.56°23'55"N, 24°07'33"E, ca. 25 m asl: Latvia, Zemgale region, Bauska municipality, along Road No P103, 0.5 km from Saulaine; lichens from tree (slide code: ŁO 2428) date: 29.04.2012.57°09'55"N, 24°51'03"E, ca. 73 m asl: Latvia, Vidzeme region, Sigulda municipality, Turaida city, Turaida Castle; moss from stone (slide code: ŁO 2430), date: 30.04.2012.56°54'32"N, 24°08'45"E, ca. 10 m asl: Latvia, Riga Region, boundary of Ķekava municipality, along road no A2; moss from tree (slide code: ŁO 2431), date: 30.04.2012.57°09'59"N, 24°50'59"E, ca. 91 m asl: Latvia, Vidzeme region, Sigulda municipality, Sigulda city, Sigulda Castle; moss from stone (slide code: ŁO 2432), date: 30.04.2012.56°41'22"N, 23°47'43"E, ca. 4 m asl: Latvia, Zemgale region, Ozolnieki municipality, Ozolnieki city, about 100 m from the Ozolnieki Lake; moss from soil (slide code: ŁO 2433), date: 29.04.2012.57°45'43"N, 24°20'59"E, ca. 3 m asl: Latvia, Vidzeme region, Salacgriva municipality, Salacgriva city; moss from soil, near the beach (slide code: ŁO 2434), date: 01.05.2012.59°10'44"N, 24°30'06"E, ca. 59 m asl: Republic of Estonia, Harju county, Kernu Parish municipality, Road No 4, moss from tree (slide code: ES 2420), date: 04.05.2012.59°10'44"N, 24°30'06"E, ca. 59 m asl: Republic of Estonia, Harju county, Kernu Parish municipality, Road No 4, moss from tree (slide code: ES 2421), date: 04.05.2012.58°48'47"N, 24°24'46"E, ca. 32 m asl: Republic of Estonia, Rapla County, Märjamaa municipality, forest near Konuvere village, moss from tree (slide codes: ES 2487), date: 29.04.2012.PageBreakTumanov, 2006XV: 1 specimen.Milnesium asiaticum was originally described from Kirghizstan and subsequently found in the Svalbard archipelago  + 1 exuvium with 6 eggs.Specimens correspond perfectly with the redescription by Murray, 1905XIII: 1 specimen.Diphascon (Diphascon) patanei  and, as suggested by Diphascon (Diphascon) patanei, a re-description of Diphascon (Diphascon) bullatum is necessary based on material from locus typicus in Scotland   .PageBreakXV: 11 specimens.Diphascon (Diphascon) pingue because they corresponded perfectly to the partial redescriptions by pingue group and has been previously recorded from numerous localities throughout the World, however the majority of records should be verified based on the modern taxonomy II: 3 specimens.The species has previously been found in many localities in the Holarctic .SubgenusThulin, 1928XIV: 2 specimens, XV: 2 specimens.The species has been previously found in many localities, mostly in the Holarctic .IX: 6 specimens (including 2 simplexes).PageBreakconvergens-dujardini complex of species  species found throughout the world. The original Hypsibius convergens description no longer conforms to modern standards and therefore a redescription is required. Nevertheless, the examined specimens correspond perfectly with the original description and the Hypsibius convergens characteristics reviewed by Belonging to the cosmopolitan  species , HypsibiXIV: 2 specimens (including 1 simplex), XV: 5 specimens.Hypsibius dujardini belongs to the cosmopolitan convergens-dujardini complex of species , XV: 24 specimens (including 6 simplexes).(measurements in  Body transparent/white (after preparation), eyes present in 6 of 15 (40%) specimens mounted in Hoyer’s medium , and by Hypsibius scabropygus does not have the bar, then our specimens should probably be classified as a new species. Thus, until Hypsibius scabropygus is redescribed, our Latvian and Estonian records should be regarded as Hypsibius cf. scabropygus. As there is a possibility of our specimens belonging to a new species, we provide standard morphometrics XIV: 1 specimen.Astatumen bartosi proposed by Astatumen bartosi and Astatumen trinacriae being the absence/presence of cuticular bars on legs II–III. Due to the notorious difficulties in differentiating the two species, the actual distribution of Astatumen bartosi cannot currently be described with confidence. Our specimen corresponds perfectly with characters of Binda & Pilato, 1969III: 1 specimen.The species has previously been reported from several, mostly European, localities in the Holarctic .PageBreakPageBreakIX: 1 specimen, XI: 8 specimens (including 5 simplexes), XIV: 3 specimens, XV: 5 specimens, XVI: 1 specimen.The species has previously been reported from many localities throughout the World, thus it is considered cosmopolitan .Murray, 1907XI: 1 egg, XIV: 8 specimens, 1 egg.harmsworthi group which is widely distributed across a broad range of ecosystems throughout the world  and 3 unembryonated eggs).57°10'33"N, 24°50'32"E, ca. 45 m asl: Latvia, Vidzeme region, Sigulda municipality, Gutmana Cave in the Gauja National Park; moss from rock ..Given that we found the composition of small and large pores in the new species beautiful, we decided to name the animal after this impression .PageBreakPageBreakMacrobiotus gumersindoi Guil & Guidetti, 2005, but it differs from it by: the presence of two types of cuticular pores  in the new species vs pores of a uniform size in Macrobiotus gumersindoi, the absence of a triangular or a pentagonal arrangement of pores placed above a single large pore on legs, the presence of granulation on legs, a different macroplacoid length sequence (2<1<3 in the new species vs 1=2=3 in Macrobiotus gumersindoi), and by slightly larger macroplacoids .The new species is most similar to Minibiotus formosus sp. n. is similar by some characteristics of adult and/or egg morphology , include species listed below. The new species differs specifically from:Other species to which Macrobiotus bisoctus , the absence of star-shaped pores, the absence of four pores around the mouth opening, the presence of a single large pore on lateral sides of legs I-III, slightly shorter buccal tube (17.7–22.9 μm in the new species vs 24.7–34.2 μm in Macrobiotus eichhorni), stylet supports inserted in more anterior position (9.5–12.2 μm [pt=49.5–56.2] in the new species vs 16.2–23.8 μm [pt=65.4–70.6] in Macrobiotus eichhorni), a different macroplacoid sequence (2<1<3 in the new species vs 2<3<1 μm in Macrobiotus eichhorni), slightly shorter placoids, and by slightly smaller external claws I–IV  , the presence of a single large pore on each of legs I–III, the presence of granulation on legs, the absence of the oral cavity armature, stylet supports inserted in a more anterior position (pt=49.5–56.2 in the new species vs pt≈68.4 in Macrobiotus furcatus), a different macroplacoid length sequence (2<1<3 in the new species vs 2<3<1 in Macrobiotus furcatus), and by egg processes without an obvious flexible portion (and never bifurcated).PageBreakMacrobiotus harrylewisi Meyer & Hinton, 2009 by: the absence of tri- and quadrilobed cuticular pores, the presence of two types of pores  over the entire cuticle in the new species vs small pores present only in the anterior part of the body and large pores present only in the posterior part of the body in Macrobiotus harrylewisi, the presence of a single large pore on each of legs I–III, stylet supports inserted in a more anterior position (pt=49.5–56.2 in the new species vs pt=61.4–67.6 in Macrobiotus harrylewisi), a different macroplacoid length sequence (2<1<3 in the new species vs 2≤3<1 μm in Macrobiotus harrylewisi), a different shape of egg processes , a smaller diameter of eggs without and with processes (44.1–45.7 μm and 55.1–55.6 μm in the new species vs 66.1–80.0 μm and 78.2–101.9 μm in Macrobiotus harrylewisi), a slightly lower number of processes on egg circumference (29–30 in the new species vs 32–41 in Macrobiotus harrylewisi), and by smaller egg processes (4.5–5.3 μm in the new species vs 7.6–12.8 μm in Macrobiotus harrylewisi).Macrobiotus jonesorum Meyer et al., 2011 by: the absence of trilobed and polygonal pores, the presence of two types of cuticular pores  in the new species vs small pores present only in the anterior part of the body, intermediate in size in the middle of the body and large pores in the posterior part of the body in Macrobiotus jonesorum), the presence of a single large pore on each of legs I-III, the presence of granulation on all legs, a slightly shorter buccal tube (17.7–22.9 μm in the new species vs 24.4–29.6 μm in Macrobiotus jonesorum), stylet supports inserted in a more anterior position (pt=49.5–56.2 in the new species vs pt=63.0–65.6 in Macrobiotus jonesorum), a slightly smaller external diameter of the buccal tube (1.3–2.0 μm [pt=7.0-9.9] in the new species vs 2.1–2.6 μm [pt=7.4–10.7] in Macrobiotus jonesorum), a different macroplacoid length sequence (2<1<3 in the new species vs 1<2<3 in Macrobiotus jonesorum), slightly shorter macroplacoids II and III , a slightly shorter macroplacoid row (4.5–6.8 μm [24.3–33.7] in the new species vs 7.0–8.4 μm [pt=27.0–34.4] in Macrobiotus jonesorum), the presence of a microplacoid, and by slightly shorter primary and secondary branches of external claws I–IV  in the new species vs pores uniform in size (ca. 1.0 μm) in Macrobiotus keppelensis), the presence of a single large pore on each of legs I-III, a slightly shorter buccal tube (17.7–22.9 μm in the new species vs 24.9–28.4 μm in Macrobiotus keppelensis), stylet supports inserted in a more anterior position (pt=49.5–56.2 in the new species vs pt≈60.6 in Macrobiotus keppelensis), a different macroplacoid length sequence (2<1<3 in the new species vs 2=3<1 in Macrobiotus keppelensis), a slightly shorter macroplacoid row (4.5–6.8 μm in the new species vs 7.0–7.6 μm in Macrobiotus keppelensis), the lack of a membrane around egg processes, a smaller diameter of eggs with processes (55.1–55.6 μm in the new species vs 65.0–85.0 μm in Macrobiotus keppelensis), a larger number of processes on egg circumference (29–30 in the new species vs ca. 11 in Macrobiotus keppelensis), smaller egg processes (4.5–5.3 μm in the new species vs 11.0–16.0 μm in Macrobiotus keppelensis), narrower egg processes bases (2.4–3.4 μm in the new species vs 9.0–12.0 μm in Macrobiotus keppelensis), and by slightly smaller distances between egg processes (1.8–3.9 μm in the new species vs 4.0–6.0 μm in Macrobiotus keppelensis).PageBreakMacrobiotus orthofasciatus Fontoura et al., 2009 by: cuticular pores arranged in 9–10 transverse bands (11 transverse bands present in Macrobiotus orthofasciatus), the absence of tri- and quadrilobed cuticular pores, the presence of two types of pores  in the new species vs all pores of similar size in Macrobiotus orthofasciatus, the presence of a single large pore on each of legs I–III, the presence of granulation on all legs, stylet supports inserted in a more anterior position (pt=49.5–56.2 in the new species vs pt=66.5–67.8 in Macrobiotus orthofasciatus), a different shape of egg processes , a slightly larger number of processes on egg circumference (29–30 in the new species vs ca. 24 in Macrobiotus orthofasciatus), and by smaller distances between egg processes (1.8–3.9 μm in the new species vs 6.4–6.9 μm in Macrobiotus orthofasciatus).Macrobiotus poricinctus Claxton, 1998 by: cuticular pores arranged in 9–10 transverse bands (8 transverse bands in Macrobiotus poricinctus), the presence of two types of pores  in the new species vs uniform pore size in Macrobiotus poricinctus), the presence of a single large pore on each of legs I–III, stylet supports inserted in a more anterior position (pt=49.5–56.2 in the new species vs pt≈59.5 in Macrobiotus poricinctus), a different macroplacoid length sequence (2<1<3 in the new species vs 2=3<1 in Macrobiotus poricinctus), a different shape of egg processes , the absence of granulation on egg shell, a larger number of processes on egg circumference (29–30 in the new species vs 18–20 in Macrobiotus poricinctus), slightly smaller egg processes (4.5–5.3 μm in the new species vs 6.5–7.0 μm in Macrobiotus poricinctus), and by smaller distances between egg processes (1.8–3.9 μm in the new species vs 6.0–8.0 μm in Macrobiotus poricinctus).Macrobiotus pustulatus  by: the absence of triangular and polygonal pores, the presence of two types of cuticular pores  in the new species vs small pores present only in the anterior part of the body, intermediate in size in the middle of the body and the large pores in the posterior part of the body in Macrobiotus pustulatus), the presence of a single large pore on each of legs I–III and, egg processes without a filiform bristle.Macrobiotus ramazzottii Binda & Pilato, 1992 by: pores arranged in bands, the presence of two types of pores  in the new species vs universal pores size in Macrobiotus ramazzottii), the presence of a single large pore on each of legs I–III, the absence of the oral cavity armature, stylet supports inserted in a more anterior position (pt=49.5–56.2 in the new species vs pt=68.2–68.3 in Macrobiotus ramazzottii), a different macroplacoid length sequence (2<1<3 in the new species vs 3<2<1 in Macrobiotus ramazzottii), and by a lower number of processes on egg circumference (29–30 in the new species vs ca. 34–41 in Macrobiotus ramazzottii).Macrobiotus subintermedius  by the presence of cuticular pores, the presence of granulation on all legs, and by fully developed lunules .PageBreakMacrobiotus vinciguerrae Binda & Pilato, 1992 by: pores arranged in bands, the absence of tri- and quadrilobed pores, the presence of two types of pores  in the new species vs uniform pore size in Macrobiotus vinciguerrae), the presence of a single large pore on each of legs I–III, the absence of the oral cavity armature, a larger mean body size (184 μm in the new species vs 380 μm in Macrobiotus vinciguerrae), stylet supports inserted in a more anterior position (pt=49.5–56.2 in the new species vs pt=66.1–68.7 in Macrobiotus vinciguerrae), a different macroplacoid length sequence (2<1<3 in the new species vs 2<3<1 μm in Macrobiotus vinciguerrae), a smaller diameter of eggs without and with processes (44.1–45.7 μm and 55.1–55.6 μm in the new species vs ca. 76.4 μm and ca. 88.0 μm in Macrobiotus vinciguerrae), a slightly larger number of processes on egg circumference (29–30 in the new species vs ca. 26 in Macrobiotus vinciguerrae), egg processes without flexible filaments, smaller egg processes (4.5–5.3 μm in the new species vs ca. 8.2 μm in Macrobiotus vinciguerrae), and by narrower bases of egg processes (2.4–3.4 μm in the new species vs ca. 5.0 μm in Macrobiotus vinciguerrae).Macrobiotus weglarskae Michalczyk et al., 2005 by: the absence of bi-, trilobed and star-shaped pores, the presence of two types of pores  in the new species vs uniform pore size in Macrobiotus weglarskae), the absence of 3–5 large triangular or irregularly shaped pores on the caudo-dorsal cuticle above hind legs, the presence of a single large pore on each of legs I-III, a different shape of egg processes , a slightly larger number of processes on egg circumference (29–30 in the new species vs ca. 24 in Macrobiotus weglarskae), and by slightly wider bases of egg processes (2.4–3.4 μm in the new species vs 1.6–2.0 μm in Macrobiotus weglarskae).Macrobiotus xavieri Fontoura et al., 2009 by: the absence of trilobed pores, the presence of two types of pores  in the new species vs all pores of similar size in Macrobiotus xavieri), the presence of a single large pore on each of legs I–III, the presence of granulation on all legs, a smaller body size (113–236 μm in the new species vs 275–410 μm in Macrobiotus xavieri), stylet supports inserted in a more anterior position (pt=49.5–56.2 in the new species vs pt=66.1–67.9 in Macrobiotus xavieri), a different macroplacoid length sequence (2<1<3 in the new species vs 2<3<1 in Macrobiotus xavieri). shorter macroplacoids , a shorter microplacoid (0.5–0.9 μm [pt=2.4–4.7] in the new species vs 1.5–2.0 [5.0–6.2] in Macrobiotus xavieri), a shorter macroplacoid row (4.5–6.8 μm [pt=24.3–33.7] in the new species vs 9.8–12.6 μm [pt=35.6–38.5] in Macrobiotus xavieri), a shorter placoid row (5.2–7.9 μm [pt=27.1–39.1] in the new species vs 10.9–13.9 μm [39.6–43.3] in Macrobiotus xavieri), a different shape of egg processes , egg shell and processes without granulation, a smaller diameter of eggs without and with processes (44.1–45.7 μm and 55.1–55.6 μm in the new species vs 56.0–79.0 μm and 80.0–99.2 μm in Macrobiotus xavieri), a larger number of processes on egg circumference (29–30 in the new species vs 20–23 in Macrobiotus xavieri), smaller egg processes (4.5–5.3 μm in the new species vs 10.6–19.0 μm in Macrobiotus xavieri), and by slightly narrower bases of egg processes (2.4–3.4 μm in the new species vs 3.7–6.6 μm in Macrobiotus xavieri).PageBreakXI: 2 specimens (including 1 simplex) and 1 egg.Paramacrobiotus species (until recently a collection of species within Macrobiotus) can be divided into three groups: areolatus, huziori and richtersi, with respect to the combination of two traits: the presence/absence of the microplacoid in the pharynx and the type of egg areolation. Paramacrobiotus richtersi, considered cosmopolitan, is recognised as the nominal species for a group of very similar taxa that require careful taxonomic examination of adults and egg morphology for correct identification. In the last decade many new species of this group have been described from various localities (e.g. ies e.g. , with moies e.g. .


The Liver X receptors (LXRs), Liver X receptor A (LXRA) and Liver X receptor B (LXRB), regulate lipid metabolism and antimicrobial response. LXRs have a crucial role in the control of Mycobacterium tuberculosis (M.tb). Lacking LXRs mice is more susceptibility to infection M.tb, developing higher bacterial burdens and an increase in the size and number of granulomatous lesions. We aimed to assess the associations between single nucleotide polymorphisms (SNPs) in LXRs and risk of tuberculosis.We sequenced the LXRs genes to detect SNPs and to examine genotypic frequencies in 600 patients and 620 healthy controls to investigate for associations with tuberculosis (TB) in the Chinese Han population. DNA re-sequencing revealed eight common variants in the LXRs genes.The G allele of rs1449627 and the T allele of rs1405655 demonstrated an increased risk of developing TB , and the T allele of rs3758673, the T allele of rs2279238, and the C allele of rs1449626 in LXRA and the C allele of rs17373080, the G allele of rs2248949, and the C allele of rs1052677 in LXRB were protective against TB patients compared to healthy controls . All SNP genotypes were significantly associated with TB. An estimation of the frequencies of haplotypes revealed two potential risk haplotypes,GGCG in LXRB  and TTCG in LXRA . Moreover, three protective haplotypes, TTAT and CCAT in LXRA and CATC in LXRB, were significantly “protective”  for TB. Furthermore, we determined that the LXRs SNPs were nominally associated with the clinical pattern of disease.Our study data supported that LXRs play a fundamental role in the genetic susceptibility to TB and to different clinical patterns of disease. Thus, further investigation is required in larger populations and in additional areas. Approximately one-third of the population is infected with Mycobacterium tuberculosis (M.tb) worldwide, which is the main etiological agent of tuberculosis (TB). M.tb is a prototypic intracellular macrophage pathogen, and resides for a long period of time to avoid recognition of the human immune system during clinical dormancy. The host protective immunity plays a pivotal role in the primary immune response and active immune response, which is central in inflammation processes.Liver X receptors (LXRs), Liver X receptor α  and Liver X receptor β , are a subset of nuclear receptor transcription factors that have emerged as master regulators of lipid In a mice study, Hannelie et al. reported that mice deficient in LXRs were more susceptible to M.tb infection, and demonstrated a decrease clearance of bacterial load in the lungs. These data support the crucial role of LXRs in the control of mycobacterial infection As a control population, we included 620 individuals  who were older than 18 years of age and did not demonstrated inflammatory, autoimmune disease or a history of chronic infectious disease. These patients had no history of tuberculosis and no evidence of prior tuberculosis noted using chest radiography. Individuals of the control group were matched with the case group according to age, sex and ethnic.A total of 600 patients  were recruited from Shanghai Pulmonary Hospital in China. All patients were diagnosed on the basis of radiographic and clinical presentation, and demonstrated a positive smear and culture for M.tb. According to the different location of infection, the case subjects were then further divided into two subgroups: (1) pulmonary tuberculosis patients  and (2) extra-pulmonary patients , as indicated in All controls and patients were Chinese, and all subjects were HIV-negative. Written consent was obtained from all subjects, and the Ethics Committee of the Shanghai Pulmonary Hospital Affiliated to Tongji University School of Medicine, China, provided authorization for the study.http://frodo.wi.mit.edu/cgi-bin/primer3/primer3_www.cgi). The primers are shown in PCR primers were designed using Primer 3 software (Genomic DNA was extracted from 0.2 ml EDTA-anticoagulated blood samples using the QIAamp DNA Blood Mini Kit (Qiagen) according to the manufacturer’s protocol. Genotyping of single nucleotide polymorphisms (SNPs) was performed using the SNaPshot kit purchased from Applied Biosystems. This is a rapid and robust assay used to simultaneously genotype SNPs using single nucleotide primer extension (mini-sequencing) and can obtain population genetic data.2+, 0.3 mM dNTP, 0.1 µM of each primer, 1U HotStarTaq polymerase (Qiagen Inc.) and 1 µl template DNA. The cycling program was the following: 95°C for 15 min; 11 cycles of 94°C for 20 s, (62°C −0.5°C/cycle) for 40 s, 72°C for 1.5 min; 26 cycles of 94°C for 20 s, 56°C for 30 s, 72°C for 1.5 min; and 72°C for 5 min.A 20 µl mixture was prepared to amplify all fragments in a Multiple PCR reaction and included 1x HotStarTaq buffer, 2.8 mM MgTTTTCAGGGGCTTTGGGAAGAGG, rs3758673SF: TTTTTTTTCCCTATCCCAGGCCTGCCA, rs2279238SR: TTTAGGCCTTGTCCCCACACAC, rs1405655SF: TTTTTTTTTTTTCTGAGAAAATTGAGGATCAGGC, rs1449626SF: TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTGATCCGCCTCCAGGCCCAG, rs1449627SR: TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCAGATGCCAAAGGAAAA, rs17373080SF: TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCGCATGAATGAGCTAAAGCCA, rs1052677SF: TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTATGGCTCTCCCCCCTAGCC. The SNaPshot Extension Reaction mixture included 5 µl SNaPshot mix, 2 µl purified PCR product, 2 µl ultrapure water and 2 µl extension primer mix (0.8 µM for each extension primer). The cycling protocol was the following: 96°C for 1 min; 28 cycles of 96°C for 10 s, 52°C for 5 s, and 60°C for 30 s. To purify the extension products, we added 1 U SAP to the extension products, incubated at 37°C for 60 min and then incubated at 75°C for 15 min. One microlitre of purified extension product was then mixed with 9 µl HiDi Formamide and 0.5 µl Liz120 (Applied Biosystems) size standard and denatured at 95°C for 5 min. Next, we loaded the mixture on the ABI 3730×l DNA Analyser (Applied Biosystems).After the completion of multiple amplification, we used shrimp alkaline phosphatase (SAP) and ExonucleaseI (ExoI) to purify the PCR product. Next, 2U SAP and 1U ExoI were added to 5 µl of PCR product. The mixture was incubated at 37°C for 80 min, followed by incubation at 75°C for 15 min. Next, the purified PCR products were used as templates for the mini-sequencing reaction using the commercially available SNaPshot Kit. To detect polymorphisms, we used the following mini-sequencing extension primers rs2248949SF: http://analysis.Bio-x.cn/SHEsisMain.htm) was used to analyze the haplotypes and determine linkage disequilibrium (LD) in the patient cases and controls. This system used a Full-Precise-Iteration (FPI) algorithm in haplotype reconstruction and frequency estimation between the patient and control The allele and genotypic frequencies were determined by direct quantification. The distribution of the LXR SNPs was compared between TB patients and healthy controls using the X2 or Fisher’s exact test. The data were analyzed for appropriateness between the observed and expected genotypic values, and then evaluated for fit in Hardy-Weinberg equilibrium (HWE). HWE was calculated to ensure that all of the data were within population equilibrium. The SHEsis system  and re- sequencing results, using reasonable inclusion criteria. These SNPs occurred at allele frequencies of ≥0.1, according to the international Hap Map project data for Chinese populations. Three SNPS were chosen using the binning algorithm of LDSelect (http://droog.gs.washington.edu/IdSelect.pI) at γ2 thresholds of 0.5 and minor allele frequencies of ≥0.1.The tagging SNPs were selected on the basis of information provided by public data-bases   = 1.62(1.38–1.92); p = 0.002, OR (95%CI)  = 1.46 (1.14–1.87), respectively); the T allele of rs3758673, T allele of rs2279238, and C allele of rs1449626 in LXRA and the C allele of rs17373080, G allele of rs2248949, C allele of rs1052677 in LXRB were less in TB patients compared to healthy controls  = 0.71(0.60–0.85), p = 0.006, OR (95%CI)  = 0.79(0.67–0.94), p<0.001, OR (95%CI)  = 0.71(0.60–0.83), p = 0.004, OR (95%CI)  = 0.70(0.55–0.89), p = 0.008, OR (95%CI)  = 0.74(0.59–0.92), p = 0.003, OR (95%CI)  = 0.69(0.53–0.88), respectively).Analysis of LXRs genotype frequencies demonstrated that the rs3758673 T/T, rs2279238 T/T, rs1449627 G/G, and rs1449626 C/C in LXRA were significantly more common in TB patients than healthy controls . In the case of LXRB gene, rs17373080 C/C, rs1052677 C/C, rs2248949 G/G and rs1405655 T/T genotype variants were associated with TB .Statistical significant differences in the allele , and genotypes , however, were lost after Bonferroni correction of the p values (Pc).2, and the LD structure (LXRA and LXRB) was shown in The LD pattern across SNPs was assessed using both D’ and rThe most common haplotypes in healthy controls and TB patients were TTCG in LXRA and GGCG in LXRB. Haplotypes analysis revealed two haplotypes in LXRB (CATC and GGCG) and three haplotypes in LXRA , which were significantly associated with TB. The GGCG haplotype and TTCG haplotype was a significant “at-risk” haplotype   = 1.440 (1.120–1.852); p<0.001, OR (95%CI)  = 1.437 (1.222–1.691)). The TTAT and CCAT in LXRA and CATC in LXRB was a significant “protective” haplotype   = 0.654 (0.478–0.895), P<0.001, OR (95%CI)  = 0.689 (0.576–0.825); P  = 0.031, OR (95%CI)  = 0.779 (0.621–0.978)). Only the relationship between TTCG and CCAT in LXRA, and GGCG in LXRB and TB were still significant after Bonferroni correction.To further understand the association between LXR SNPs and the risk of different clinical patterns, we evaluated the SNP contribution on clinical variables related to tuberculosis. No significant association was detected between the allele of LXRs and the risk of different clinical forms of tuberculosis (date not shown). However, the genotype at the LXRA SNP rs1449627 and LXRB SNPs rs17373080, rs1405655 and rs1052677 was associated nominally associated to PTB, p = 0.006, p = 0.013, p = 0.004 and p = 0.0007, respectively. We also performed haplotype analyses using the LXR SNPs in two clinical groups. Seven haplotypes were observed in samples of two subgroups, but only the CATC haplotype in LXRB demonstrated a significant association with an increase in the risk of PTB (p = 0.045 OR (95%CI)  = 1.511 (1.007–2.269)). After Bonferroni correction, CATC haplotype and LXRB SNPs rs17373080 were lost significant associated with the different clinical forms of the disease.Listeria monocytogenesLeishmaniaMycobacterium tuberculosisTuberculosis, a chronic inflammatory disease that occurs within the entire body, is a complex process, and can cause multi-system complications, such as pulmonary, osteoarticular, and renal, central nerve systems (CNS). The initiation and progression of tuberculosis involves a dynamic interplay between numerous signaling pathways and cell types, where macrophages, dendritic cells (DCs), T cells, and other cellular elements present in the infection sites contribute to its pathogenesis. Many studies have investigated the potential role of LXRs in various bacterial infections, such as We have analyzed eight common genetic variations in LXRs, LXRA and LXRB, for an association with tuberculosis in a Chinese Han population and for an association with different disease clinical patterns in two subgroups of tuberculosis patients. We assessed the hypothesis that sequence variations in the LXR gene may contribute to TB susceptibility. We screened LXR genes for sequence variations in 24 healthy controls and 24 TB patients, resulting in eight representative SNPs across genes, four SNPs for each respective gene.In this study, we observed an association of LXR SNPs with TB. Frequencies of the allele and genotype in the LXR genes were different between the healthy controls and TB patients. The G allele of rs1449627 in LXRA and the T allele of rs1405655 in LXRB were more common in TB patients compare to healthy controls. The T allele of rs3758673, the T allele of rs2279238, and the C allele of rs1449626 in LXRA and the C allele of rs17373080, the G allele of rs2248949, the C allele of rs1052677 in LXRB exhibited a significant protective effect. Moreover, all genotypes of LXRs were significantly associated with TB. Statistical significant differences in the allele , and genotypes , however, were lost after Bonferroni correction of the p values (Pc).To investigate a potential role of LXR SNPs in two subgroups of TB patients, we evaluated the SNP contribution on the clinical variables related to tuberculosis. No LXR allele associated to the investigated phenotypes in two patient subgroups was observed. In addition, the genotype at the LXRA SNP rs1449627 and LXRB SNPs s17373080, rs1405655, and rs1052677 was nominally associated to pulmonary TB, and the haplotype CATC in LXRB was significantly associated with an increased risk of PTB. After Bonferroni correction, CATC haplotype and LXRB SNPs rs17373080 were lost significant associated with disease risk.The LXRB SNP rs17373080 in the promoter region was nominally associated with TB, particularly with PTB. A previously study revealed that the LXRB promoter SNP rs17373080, resides proximally to the transcription start site and could potentially contribute to the transcription potential of the LXRB gene promoter The LXRs, LXRA and LXRB, are ligand-activated transcription factors that belong to the nuclear hormone receptor super-family, and have emerged as important regulators of lipid metabolism and inflammation. LXRA is highly expressed in liver, and at lower levels in the intestine, macrophages, adipose tissue, lung, adrenal glands and kidney, all of which play important roles in lipid homeostasis. In contract, LXRB is ubiquitously expressed In conclusion, our results suggested associations between common genetic variants of LXRs and the risks of pulmonary tuberculosis, confirming the recent study by Korf H et al. This investigation suggested that gene polymorphisms in LXRs are likely to affect the risk of developing tuberculosis, and demonstrated the potential of LXR modulators as therapeutic targets for tuberculosis. However, these findings must be interpreted in other large population samples and performing meta-analyses are necessary before a link between LXRs gene variability and tuberculosis can definitely be established. Moreover, understanding the molecular mechanisms behind the LXRs SNP-disorder association will require further experimental research.


Microalbuminuria has been shown to be a risk factor for cardiovascular and renal disease in patients with hypertension and diabetes as well as in the general population. Urinary albumin excretion over 24 h is considered a ‘gold standard’ to detect microalbuminuria. Few studies have used 24-h urinary albumin excretion to analyze the prevalence of and related factors for microalbuminuira in a general Chinese population.This study included 1980 adults aged 18–69 years from the Shandong-Ministry of Health Action on Salt and Hypertension (SMASH) Project 2011 survey. Blood pressure, height, weight and waist circumference were measured, and a venous blood and timed 24-h urine samples were collected from each participant. Linear and logistic regression analyses were used to test associations between established cardiovascular risk factors and microalbuminuria.The median (25th–75th percentile) of 24-h urinary albumin excretion was 6.1 mg/d (4.5–8.7 mg/d) for all adults, 6.0 mg/d (4.4–8.5 mg/d) for men and 6.2 mg/d (4.6–8.9 mg/d) for women. The overall prevalence of microalbuminuria was 4.1% , 3.7% (95% CI: 2.9–4.5%) for men and 4.6% (95% CI: 3.7–5.5%) for women. Microalbuminuria was present in 8.1% (95% CI: 6.9–9.3%) of individuals with hypertension, 11.4% (95% CI: 10.0–12.8%) of those with diabetes and 15.6% (95% CI: 14.0–17.2%) of those with both. Multiple logistic regression analysis indicated that systolic blood pressure  and fasting blood glucose  were the independent risk factors for microalbuminuria.Adults in the general population of Shandong Province have a moderate prevalence of microalbuminuria. Those with hypertension and diabetes are at high risk of having microalbuminuria, suggesting the need for screening and early intervention for microalbuminuria among these individuals. Microalbuminuria (MAU), an abnormal increase in the urinary excretion of albumin, is a risk factor for cardiovascular and renal disease in patients with hypertension and diabetes –4. EarlyMAU has also been shown to be predictive of cardiovascular events and all cause mortality in the general population –11. In mThe amount of urinary albumin excreted over a 24-h period is considered the ‘gold standard’ for defining MAU . HoweverThe study subjects had participated in the Shandong-Ministry of Health Action on Salt and Hypertension (SMASH) project 2011 survey; the methods used for this survey have been described in detail . Using a2, ≥18.5 but <24 kg/m2, ≥24 but <28 kg/m2 and ≥28 kg/m2 respectively according to Chinese guidelines  of 1.19 (95% CI: 1.05–1.35)  and diabetes (11.4%). Disease duration, blood pressure and serum glucose level were associated with MAU; therefore, both blood pressure and glucose should be controlled as much as possible to prevent the progression of MAU.The prevalence of MAU in Shandong adults was lower in other regions in China (4.5–15%) , 24–26. In agreement with previous studies, we found that hypertensive individuals were at high risk for MAU, with blood pressure linearly associated with MAU , 28, 29.MAU is the first sign of diabetic nephropathy. The percentage of our diabetic patients with MAU was much lower than the previously reported prevalence in China and other countries –33. The We also found that MAU was associated with obesity and dyslipidemia , 36, witThe utility of albuminuira screening in the general population is unclear , 39–42. A major strength of our study was our use of the gold standard 24-h UAE to classify individuals with microalbuminuria, unlike previous studies in Chinese populations. Furthermore, this study included a representative sample of the general population, using rigorously standardized methods for data collection and strict quality control. However, our study has several limitations. First, we collected only one 24-h urine sample per participant, thus preventing a determination of day-to-day variability for each individual . Second,This study showed that the prevalence of MAU in the general adult population of Shandong Province, China, is moderate. Rates of MAU were higher in subjects with than without hypertension and diabetes, with these two factors together associated with MAU. Screening of subjects with hypertension and/or diabetes for MAU may result in earlier diagnosis and treatment.


Anaerobic digestion is a promising alternative to disposal organic waste and co-digestion of mixed organic wastes has recently attracted more interest. This study investigated the effects of temperature and carbon-nitrogen (C/N) ratio on the performance of anaerobic co-digestion of dairy manure (DM), chicken manure (CM) and rice straw (RS). We found that increased temperature improved the methane potential, but the rate was reduced from mesophilic (30∼40°C) to thermophilic conditions (50∼60°C), due to the accumulation of ammonium nitrogen and free ammonia and the occurrence of ammonia inhibition. Significant ammonia inhibition was observed with a C/N ratio of 15 at 35°C and at a C/N ratio of 20 at 55°C. The increase of C/N ratios reduced the negative effects of ammonia and maximum methane potentials were achieved with C/N ratios of 25 and 30 at 35°C and 55°C, respectively. When temperature increased, an increase was required in the feed C/N ratio, in order to reduce the risk of ammonia inhibition. Our results revealed an interactive effect between temperature and C/N on digestion performance. Anaerobic digestion is an effective way of converting agricultural waste into biogas that can be used to generate energy, which is especially efficient in rural western China. In the past decade, this technology has received great attention in both scientific research and practice. However, the efficiency of anaerobic digestion may be limited by inadequate amount and diversity of waste from a single resource, which is insufficient for large-scale digesters, as well as the drawbacks of using single substrates, such as improper carbon-nitrogen (C/N) ratios, low pH of the substrate itself, poor buffering capacity, and high concentrations of ammonia et al. revealed that swine manure co-digested with corn stalks at a C/N ratio of 20 obtained increased cumulative biogas production up to 11-fold and increased cumulative net methane volume up to 16-fold, when compared to swine manure digested alone et al. also suggested that co-digestion of dairy manure, chicken manure and wheat straw, had better digestion performance with stable pH and low concentrations of total ammonium nitrogen (TAN) and free ammonia (FA) at adjusted C/N ratios of 25 and 30 Co-digestion of various biosolid wastes, a process that utilizes the nutrients and bacterial diversity in those wastes to optimize the digestion process, is an attractive approach for improving the efficiency of biotransformation −1 at a pH of 7.5 −1 in the ammonia toxicity test, a gradual decrease of 21% was observed in biogas Although many studies indicated that the optimal C/N ratios in methane fermentation were 25∼30 Base on previous studies mentioned above, there are interactive effects between temperature and ammonia in the digestion process and the digestion efficiency is dramatically affected by the temperature and C/N ratio. Thus, to investigate this interaction, we first examined the effect of a series of temperatures on the mixtures of certain ratios of C/N (25), and secondly, compared the digestion performance of mixtures with a series of C/N ratios by adjusting the proportions of each substrate, dairy manure (DM), chicken manure (CM) and rice straw (RS) under mesophilic and thermophilic conditions.DM and CM were collected from a livestock farm located in Yangling, China. RS was obtained from a local villager. Before being put into the reactor, the air-dried RS was cut into pieces (2∼3 cm). The substrates were individually homogenized and subsequently stored at 4°C for further use. The chemical characterization of each substrate tested in this study is shown in The collections of DM and CM were permitted by livestock farms belonging to ‘Besun’ group in Yangling, China. The RS was provided voluntarily by a local villager in Qishan, Baoji, China. The inoculum was obtained from a household biogas digester in a biogas demonstration village named Cuixigou in Yangling and the collection was permitted by the hosts. The all experimental procedures conformed to the regulations established by the Ethics Committee of the Research Center of Recycle Agricultural Engineering and Technology of Shaanxi Province, China.Experiment 1: Three mixture sets were investigated in this experiment: set A (DM+ RS), set B (CM+RS), and set C (DM+CM+RS). For set A and set B, the C/N ratio was 25, achieved by adjusting the DM/RS or CM/RS ratio. For set C, based on a DM/CM ratio of 1∶1, multi-component substrates were prepared by adding RS to the DM-CM mixtures in order to adjust the C/N ratio to 25. The proportions of all substrates in each mixture were in a volatile solid (VS) state. The operation temperatures were 20, 30, 40 (mesophilic), 50, and 60°C (thermophilic), respectively.Experiment 2: For all mixture sets, RS was added into the DM-CM mixtures with a VS ratio of 1∶1, in order to adjust the C/N ratio to selected levels. C/N ratios of 15, 20, 25, 30 and 35 were selected in tests at a temperature of 35°C, but ratios of 20, 25, 30, 35 and 40 were selected in tests at a temperature of 55°C.The initial VS ratio of substrate to inoculum was kept at 1∶2 for all experimental setups. Each reactor had a 1 L capacity and contained 600 mL of total liquid, including 200 mL of inoculum and mixed substrate of 15gVS/L. The inoculum used for digestion at 20, 30, 35 and 40°C was digested cattle manure, taken from a lab-scale reactor operated at 35°C with a hydraulic retention time (HRT) of 15 days. Additionally, digestion at 50, 55 and 60°C was inoculated with digested cattle manure from the lab-scale reactor operated at 55°C with a HRT of 15 days. A control with only inoculum was used to determine biogas production due to endogenous respiration. Each treatment was performed in triplicate. All reactors were tightly closed with rubber septa and screw caps. The headspace of each reactor was flushed with nitrogen gas for about 3 min to assure anaerobic conditions prior to starting the digestion tests. To provide mixing of the reactor contents, all reactors were shaken manually for about 1 min, once a day prior to measurement of biogas volume.et al.et al. Total solids, VS, pH, total Kjeldahl nitrogen (TKN), and total ammonium nitrogen (TAN) analysis were performed according to APHA Standard Methods −1VS) in each substrate and N1, N2 and N3 were the nitrogen content (g kg−1VS) in each substrate.Where W1, W2 and W3 were the VS weight in a single substrate in the mixture, C1, C2 and C3 were the organic carbon content  and temperature  was evaluated by the following equations: Y = 0.0302e1.82T in the mixture of DM+RS, Y = 0.0216e2.0T in the mixture of CM+RS and Y = 0.101e1.65T in the mixture of DM+CM+RS. On average, the mixture of DM+CM+RS had significantly higher TAN and FA concentrations than the mixture of DM+RS, but was lower than the mixture of CM+RS (A linear correlation between TAN and temperature (20 – 60°C) was observed and the highest TAN value was 1,261 mg L at 50°C . The relof CM+RS  and 4.With the increase of temperature, methane potential continuously increased, but the increasing rate was lower under thermophilic than under mesophilic conditions . The mix−1 and 314, 108 mg L−1, respectively. Treatments with C/N ratios of 25, 30 and 35 resulted in low and stable TAN and FA during the anaerobic process. The average concentrations of TAN were 985, 739 and 568 mg L−1 when C/N ratios were of 25, 30, and 35, respectively and the average concentrations of FA were 9.1, 7.5 and 2.2 mg L−1 when C/N ratios were of 25, 30, and 35, respectively.The pH value and the concentrations of TAN and FA were significantly influenced by C/N ratios at 35°C. For digesters with C/N ratios of 15 and 20, the pH values were higher than 7.0 during the whole digestion process, and the final pH values reached to 8.09 and 7.68, respectively .The aver−1 by day 10 and reached peaks as high as 2415 and 1932 mg L−1, respectively (−1 when C/N ratios were of 20 and 25. For C/N ratios between 30 and 40, TAN and FA concentrations were in the range of 430∼1426 mg L−1 and 2∼131 mg L−1, respectively (Under 55°C, pH values were between 7.0 and 7.92 in treatments with C/N ratios of 20 and 25. Stable pH values around 7.0 were observed when C/N ratios were of 30 and 35. When the C/N ratio was increased to 35, the pH value was lower, at around 6.2 . The conectively . FA increctively .−1VS and with a C/N ratio of 30 at 50°C with 286 mL g−1VS, respectively (Methane potential increased first and then decreased with increases of C/N ratios. The highest methane potential was observed with a C/N ratio of 25 at 35°C with 272 mL gectively . The qua−1 VS.Where Y was methane potential and X was the C/N ratio. The optimum conditions for maximum methane potential were calculated as a C/N of 26.76 at 35°C and a C/N ratio of 30.67 at 55°C, respectively. Accordingly, the highest methane potential was estimated as 265.7 and 309.9 mL get al., ammonia inhibition occurs in the range of 1500∼3000 mg L−1 TAN when the pH value is over 7.4 −1 at temperatures between 20 and 60°C (−1 FA was inhibitory for methanogens −1 at 60°C (−1 FA at 37°C and by 690 mg L−1 FA at 55°C According to the study by Calli and 60°C . Compare at 60°C , indicatet al. Due to the potential role of the C/N ratio in regulating the inhibitory effects of ammonia, digestions with different C/N ratios were tested in experiment 2 under mesophilic and thermophilic conditions to further obtain optimal C/N ratios with less ammonia inhibition. We found that the mixture of DM+CM+RS had better digestion performance in methane potential than the mixtures of DM+RS and CM+RS , which m−1 and FA increased up to final concentrations of 314 and 461 mg L−1, when the C/N ratio was of 15 at 35°C and was 20 at 55°C, respectively (−1VS at 35°C and 169 mLg−1VS at 55°C, accounting for just 53.0% and 52.5%, compared with their maximum values (Substrates that have low C/N ratios contain relatively high concentrations of ammonia, exceeding concentrations necessary for microbial growth, and probably inhibiting anaerobic digestion ectively  and 3. Tm values . Under bAmmonia inhibition under mesophilic and thermophilic conditions has been compared in previous studies. It has been observed that an increase in temperature resulted in a reduction of the biogas yield, due to the increased inhibition of FA under higher temperature This study demonstrated an interactive effect between C/N ratio and temperature on the performance of anaerobic co-digestion of dairy manure, chicken manure and rice straw. Our results suggest that increased temperature from mesophilic to thermophilic conditions resulted in ammonia inhibition, however, this kind of inhibition could be reduced or avoided by increasing the C/N ratio of mixed feedstock to an appropriate level. In anaerobic co-digestion of DM, CM and RS, the optimal C/N level was 26.76 at 35°C and 30.67 at 55°C. Adjusting the proportions of mixture substrates in anaerobic co-digestion to obtain suitable feed characteristics, such as the C/N ratio, pH and nutrients, is an effective way to achieve desired digestion performance.


Obtained results revealed that radiation of millet grains at a dose level higher than 0.5 kGy caused significant (P < 0.05) reduction on the percentage of fungal incidence and the free fatty acid of the seeds, while, no significant change in the germination capacity was observed of the grains after radiation. Additionally, the radiation process caused significant (P < 0.05) reduction on both tannins and phytic acid content and gradual increment on in vitro protein digestibility of the grains. On the other hand, the treatments significantly (P < 0.05) increased the protein solubility of the grains. Obtained results indicate that gamma irradiation might improve the quality characteristics of millet grains, and can be used as a postharvest method for disinfestations and decontamination of millet grains.The aim of this study was to evaluate the effect of gamma radiation processing of millet grains on fungal incidence, germination, free fatty acids content, protein solubility, digestible protein, and antinutritional factors (tannin and phytic acid). The grains were exposed to gamma radiation at doses 0.25, 0.5, 0.75, 1.0, and 2.0  Pennisetum gluucum L.) is considered to be the staple food for most people in Asia and Africa. It is considered as a good source of needed elements  served as control.Radiation processing was done at Kaila irradiation processing unit, Sudanese Atomic Energy Corporation (SAEC). About 250 g of millet grains packed in polyethylene bags were irradiated, using a The fungal incidence and the colony formation unit per gram (cfu/g) of treated and untreated samples was determined after platted on double strength Sabaroud Dextrose Agar and incubated at 25°C for 5 days according to standard methods AOAC .The germination of grains was determined according to the international Seed Testing Association ISTA . Twenty Free fatty acid of millet was determined according to Aibara et al.  cited byTannins content of grains was estimated according to Price et al. . Phytic The crude protein was determined following the Kjeldahl method described by AOAC . The digSoluble protein solubility was determined in millet grains after extracted by water, using the method described by Hagenmaier .P < 0.05), using least significant difference (LSD).All data were the average of triplicates. Data were analyzed using one‐way analysis of variance (ANOVA). Significant differences were calculated  reduced to 190.9, 190.6, and 190.5 mg/100 g, respectively. Significant reduction on FFA content might be due to lipase activity reduction in treated grains, which result in dropping the FFA formation. Pankaj et al.  reduced tannin content of millet grains. The reduction in tannin content was found to be 38.9, 44.4, 50, and 52.8, and 74.9% when millet grains were irradiated at dose 0.25, 0.50, 0.75, 1.0, and 2.0 kGy, respectively, compared to control one. These findings are in agreement with those stated by several researchers. Hassan et al.  decreased. The level of reduction increased with an increase in radiation dose. Decreases in phytic acid were 3.9, 29.9, 42.5, 43.3, and 52.8%, at dose 0.25, 0.50, 0.75, 1.0, and 2.0 kGy, respectively. This reduction might be the result of the action of free radicals, since they are able to cleave to the phytate ring  decreased the level of phytic acid of legumes and cereal grains.On the other hand, before radiation treatment, the phytic acid of millet grains was found to be 1.27 mg/g Fig. . After rFigure P < 0.05) to 11.32, 11.82, 13.04, 12.74, and 13.44% after radiation at 0.25, 0.50, 0.75, 1.0, and 2.0 kGy, respectively. Increase in protein solubility after radiation is likely due to the high proteolytic activity during radiation, which may lead to hydrolysis of the stored proteins.Protein solubility is doubtless the most important function among the functional properties of proteins. Data in Figure The obtained results revealed that gamma irradiation processing of millet grains up to 2 kGy significantly reduced the fungal incidence and free fatty acids content of the grains. On the other hand, it caused a decrease in the amount of antinutrients namely, tannin and phytic acid and gradually increase the in vitro protein digestibility and protein solubility of the grains. According to these results, therefore, gamma radiation can be applied as a safe postharvest method in order to extend the shelf life of millet grains.None declared.


Ecosystem C storage was composed of 0.23 ± 0.01 (0.7%) above-ground biomass, 1.38 ± 0.14 (4.5%) below-ground biomass, and 29.37 ± 1.2 (94.8%) Pg C in the 0–100 cm soil layer. Carbon storage calculated by the grassland classification methods (18 grassland types) was closer to the mean value than those calculated by the spatial interpolation methods. Differences in integrative methods may partially explain the high uncertainty in C storage estimates in different studies. This first evaluation demonstrates the importance of multi-methodological approaches to accurately estimate C storage in large-scale terrestrial ecosystems.The accurate estimate of grassland carbon (C) is affected by many factors at the large scale. Here, we used six methods  to estimate C storage of Chinese grasslands based on published data from 2004 to 2014, and assessed the uncertainty resulting from different integrative methods. The uncertainty  of grassland C storage was approximately 4.8% for the six methods tested, which was mainly determined by soil C storage. C density and C storage to the soil layer depth of 100 cm were estimated to be 8.46 ± 0.41 kg C m Similarly, estimates of soil organic C (SOC) storage range from 16.70 Pg C16 to 53.72 Pg C2. The large difference among these estimates may be resulted from the difference in data used for the estimation. In addition, the application of the different methods would be another factor affecting the estimates.Some studies have estimated C storage of different components in Chinese grasslands using various data source. Fang Different methods have been used to estimate C storage in the different components of Chinese grasslands, including remote sensingIn this study, we collected field-measured data from papers published from 2004 to 2014, including AGB, BGB, and SOC, and used six integrative methods  to calculate the C storage of different components in Chinese grasslands. We have addressed two fundamental questions in this paper: (1) To investigate the uncertainty of the estimates of C density and storage in grasslands resulting from the selected integrative methods, and (2) To evaluate C storage in all components of Chinese grasslands  on the basis of different integrative methods at the national scale.−2, and the highest and lowest CVs for AGBC were in R1 (13.1%) and R5 (1.3%). The range of BGBC was 0.061 ~ 1.314 kg C m−2, and the highest and the lowest CVs for BGBC were in R6 (22.9%) and R1 (4.1%). For SOC, R1 and R3 had higher CVs, which were 10.5% and 7.3%, respectively. The range of SOC was 1.892 ~ 24.856 kg C m−2, and the CV value was about 5.8%. Overall, the CV values of C density were highest in BGBC, followed by SOC and AGBC  of C density estimated by three spatial interpolation methods was different among six grassland regions  and diffand AGBC .Furthermore, the CVs of grassland C density differed among regions, with a mean CV of approximately 5.5% . The CV P < 0.01 for AGBC; F = 11.46, P < 0.01 for BGBC; F = 13.25, P < 0.01 for SOC; F = 11.99, P < 0.01 for grassland C density; −2 using three spatial interpolation methods, which estimated AGBC, BGBC, and SOC as 0.08 ± 0.01, 0.42 ± 0.03, and 8.16 ± 0.48 kg C m−2, respectively  and in desert (28.2%), and the range of BGBC was 0.041 ~ 1.844 kg C m−2. The CV value of SOC in all grassland types was 5.2%, with the highest CV in shrub-tussock (13.9%), and the range of SOC was 2.197 ~ 21.850 kg C m−2. The CV values of grassland C density ranged from 5.2% to 13.0% across all grassland types, with a mean CV of 2.2%.The CV values resulting from different grassland classification methods were different for the different components in Chinese grasslands . The CV P < 0.01 for AGBC; F = 60.49, P < 0.01 for BGBC, F = 46.32, P < 0.01 for SOC, and F = 50.55, P < 0.01 for grassland; −2, respectively. Correspondingly, total C storage was estimated as 30.47 ± 0.28 Pg C for Chinese grasslands (down to 100 cm soil layer), with AGB, BGB, and SOC containing 0.24 ± 0.02, 1.39 ± 0.21, and 28.84 ± 0.14 Pg C, respectively.The C density differed significantly in the different components of various grassland types  among all components. However, the CV value of grassland C density was close to that of SOC, where they were 4.8% and 4.9%, respectively. The C density of Chinese grasslands was 8.46 ± 0.41 kg C m−2, most of which stored as SOC.When combining all grassland areas, the C storage of grassland , estimated by the six integrative methods, ranged from 29.49 to 32.53 Pg C, with a mean of 30.98 Pg C . The C s−2 to 3.44 kg C m−2. Although expanding the soil depth to 100 cm may result in overestimation of BGBC to some extent, the influence on the ecosystem C estimates was not noticeable because of the small contribution of BGBC in deeper soil layers to total BGBC  of grassland C density than the grassland classification methods (8.25 ± 0.18 kg C m−2). One possible explanation is that the spatial interpolation methods may be affected more by the number of sampled sites and nearby sites, particularly those with higher C density. The grassland C estimate was mainly decided by SOC. The SOC estimate in the 0–100 cm soil layer obtained using the six integrative methods was 7.96 kg C m−2 in this study, which was similar to the previous results of (7.80 kg C m−2) Yang et al.−2) Fang et al.−2 in the 0–20 cm soil layer, which was about 41.8% of that in the 0–100 cm and was similar to the average value of global grasslands (42%)et al.The CV values of grassland C density calculated from the three spatial interpolation methods (5.5%) were higher than those calculated from the three grassland classification methods (2.2%). Our results demonstrated that the spatial interpolation methods have higher uncertainty of estimates, because they were more easily affected by the data quantity and spatial distribution of sampling sites8.25 ± 0. kg C m−2et al.16−2, respectively. In addition, Yang et al.The estimation of grassland C storage at the national scale was limited and was apparently different among the previous studies .One poss1013141322−2 with 47.4% variation, whereas the C density in BGB was 0.41 ± 0.27 kg C m−2 with 66.4% variation. It was estimated that the C density of BGB was approximately 7 times higher than that of AGB, which was consistent with previous results suggesting that 86–88% of vegetation C density is stored in BGB21in situ measurements of BGB, despite such measurement being important for the estimation of vegetation C density. The R/S ratio has been verified as a good parameter for inferring BGB at the regional scale; however, this measure also generates uncertainty due to various disturbances . Furthermore, previous studies using global mean soil C density2A second possible explanation for these differences is the lack of data for key components in grasslands, especially synchronous measurements. Vegetation C density could be estimated by many methods, such as remote sensing, modelling, and field-investigation. However, SOC is difficult to be estimated by remote sensing and modelling, although SOC is the most important component in grassland ecosystems. As shown in et al.−2 and 0.04 kg C m−2. Therefore, multiple approaches should be used to estimate the C storage in terrestrial ecosystems and improve the accuracy of C storage estimates at large scales. The uncertainty of C storage in Chinese grasslands using the six integrative methods was approximately 4.8%. The level of uncertainty differed among different components, with the highest values observed in BGBC. Our findings emphasize the underlying influence of integrative methods for estimating C storage in terrestrial ecosystems at a large scale. Based on the six integrative methods, C storage in Chinese grasslands was estimated as 0.23 ± 0.01, 1.38 ± 0.14, 29.37 ± 1.2, and 30.98 ± 1.25 Pg C in AGBC, BGBC, SOC, and grasslands, respectively. This first assessment of C storage in all main components of Chinese grasslands at the national scale will help to determine the potential roles of Chinese grasslands in response to global climate change.Additionally, this study provided the first estimate of three components by field-measurement data and demonstrated that integrative methods have an important influence on the estimation of grassland C storage at the national scale. However, previous studies only used one method to estimate one or two components. Nihttp://epub.cnki.net) (C density data for different components of Chinese grasslands were derived from: (1) field-measured data of 210 papers publicly published from 2004 to 2014 in the Web of Science (www. Webofknowledge.com) and in the China National Knowledge Infrastructure (nki.net) , in addinki.net) .8 ha.Furthermore, we obtained the Chinese grassland areas and the spatial distribution of different grassland types from the map of grassland resources in China at 1:4M−2) records were used directly. If the AGB data were measured monthly in the published papers, we chose the values measured in August, which is commonly considered to be the peak period for AGB in Chinese grasslands−2) in grasslands was estimated to comprise 45% of the plant dry matter−2) from AGB and the coefficient of C content (0.45).Field-measured above-ground biomass  was calculated from BGB and the coefficient of C content (0.45). In practice, the BGB data were directly used if the sampling depth was 100 cm. For data obtained at a soil depth of less than 100 cm, we estimated the BGBC as a 0–100 cm soil layer based on the 0–20 cm data using the following procedure. We first established the relationships of BGBC between 0–20 cm and 0–100 cm using 99 sampling sites where data were simultaneously collected at 20 cm and 100 cm depths. We found that BGBC showed a significantly positive correlation between 0–20 cm and 0–100 cm  was calculated using equation 2, unless SOC data were reported directly in the published papers. In practice, we commonly used the field-measured bulk density. For soil samples without bulk density records, we used the mean values of bulk density from the second soil survey in China (1.3 g cm−3) as a substitute 2 = 0.95)  .−2), Hj is soil thickness (cm), cj is the concentration of SOC (%), bj is bulk density (g cm−3).where SOC is SOC density , Inverse Distance Weighted interpolation (M2), and Empirical Bayesian Kriging interpolation (M3). Among these methods, M2 only considers the distance, whereas M1 and M3 consider both the spatial orientation and the distance.The geo-statistical principle assumes that grasslands gradually change with climate. Theoretically, the biomass at one sampling site may have the highest similarity to that at the nearest site. Based on this assumption, spatial interpolation methods may be used to estimate the C storage of grasslands in China. In practice, we selected three spatial interpolation methods, For statistical and comparative analyses, the grasslands were divided into six regions according to the nationwide grassland resource survey of DAHV & GSAHVGrasslands exhibit different ecological characteristics based on climate (temperature and precipitation), topography, and soil. Therefore, grasslands may be divided into different artificial types. Based on this assumption, C storage in Chinese grasslands may be calculated from grassland type and the corresponding area at different scales . The majFor the statistical and comparative analyses, the Chinese grasslands were classified at three different scales. First, 5 grassland types of China’s vegetation were determined at 1:1000000 resolutionP = 0.05.The coefficient of variation  was used to assess the uncertainty of the six methods for estimating C storage33How to cite this article: Ma, A. et al. Carbon storage in Chinese grassland ecosystems: Influence of different integrative methods. Sci. Rep.6, 21378; doi: 10.1038/srep21378 (2016).


Acacia tortilis polysaccharide (AEATP) from gum exudates and its role in comorbidities associated with diabetes in STZ-nicotinamide induced diabetic rats. Male albino Wistar rats were divided into control, diabetic control, glimepiride treated (10 mg/kg), and diabetic rats treated with 250, 500, and 1000 mg/kg dose of AEATP groups and fasting blood glucose, glycated hemoglobin, total cholesterol, triglyceride, LDL, VLDL, HDL, SGOT, and SGPT levels were measured. STZ significantly increased fasting blood glucose level, glycated hemoglobin, total cholesterol, triglyceride, LDL, VLDL, SGOT, and SGPT levels, whereas HDL level was reduced as compared to control group. After 7 days of administration, 500 and 1000 mg/kg dose of AEATP showed significant reduction (P < 0.05) in fasting blood glucose level compared to diabetic control. AEATP has also reduced total cholesterol, triglyceride, LDL, VLDL, SGOT, and SGPT levels and improved HDL level as compared to diabetic control group. Our study is the first to report the normalization of fasting blood glucose level, lipid profile, and liver enzyme in AEATP treated diabetic rats. Thus, it can be concluded that AEATP may have potentials for the treatment of T2DM and its comorbidities.The present study was designed to investigate the antidiabetic activity of aqueous extract of  Diabetes mellitus (DM) is characterized by hyperglycemia and is associated with a group of metabolic disorders, that is, abnormalities in carbohydrate, fat and protein metabolism which further result in chronic complications including microvascular, macrovascular, and neuropathic disorder . It is dThe currently available oral hypoglycemic and antihyperglycemic drugs for type-II diabetes have their own limitations, adverse effects, and secondary failures. Therefore, to reduce their cost, limitation, and adverse effects, focus has been shifted towards the medicinal herbs for safe and effective use. Recently a lot of medicinal herbs are being investigated for their role in pharmacotherapy of diabetes . Acacia tortilis and are widespread distributed around the globe . In India, this tree was introduced in 1958 from Israel [α2-adrenoceptor related diseases [ Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans [Israeli babool and umbrella thorn are the other names ofm Israel . Variousdiseases , antimicalbicans , in-vitralbicans , and hypalbicans . Acacia arabica, Acacia catechu, Acacia mollissima, Acacia polyacantha, and so forth [ Phellinus linteus [ Acacia tortilis has been also found to have an antihyperglycemic activity [Moreover, various species of Acacia are reported to have antidiabetic activity likeso forth –15. Prev linteus , Ascophy linteus , Taxus [ linteus , Acantho linteus , and And linteus  demonstractivity . Acacia tortilis (Forsk.) Hayne ssp. raddiana polysaccharide from gum exudates.Thus, with the same line of research, the present study was designed to explore the antidiabetic activity of aqueous extract ofStreptozotocin, Glimepiride, and Nicotinamide were procured form Sigma-Aldrich, Milwaukee, USA, and all the other chemicals were of analytical grade. Acacia tortilis was collected from Central Arid Zone Research Institute Campus, Jodhpur, India.Gum exudates from the stem and branches of ad libitum under controlled environmental condition of temperature (24–28°C), relative humidity 60–70%, and natural light/dark cycle (12 : 12) and maintained on standard food pellets and tap water ad libitum.Male albino Wistar rats (150–200 gm) were used in this study and experimental protocol was approved by Institutional Animal Ethics Committee. Animals were kept as per the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Ministry of Environment and Forest, Government of India . Animals were fed normal chow diet andAcute oral toxicity studies were performed according to OECD (Organization for Economic Cooperation and Development) 423 guidelines at the dose of 300, 2,000, and 5,000 mg/kg. The general behavior was observed continuously for 48 hr, 3 days and mortality was observed for 14 days .Gum exudate was crushed into fine particles using laboratory grinder. Fine powder of gum exudates (100 gm) was stirred vigorously in distilled water (200 mL) for 6 hours at room temperature and centrifuged to remove water-insoluble part. The supernatant solution was decanted off. The concentrated aqueous solution was poured into 3 times its volume of ethanol with constant stirring. The polysaccharide was precipitated out in the form of a fluffy precipitate. The precipitate was again dissolved in water and added to ethanol. Precipitate was treated successively with dry solvent ether and acetone. It was filtered under vacuum and dried in vacuum desiccators at room temperature .The pure polysaccharide was subjected to hydrolysis with sulfuric acid (2N) for 18 hr on steam bath. The hydrolyzate was cooled, neutralized with saturated solution of barium carbonate by dropwise addition till the pH of the solution reached at 7, filtered, and the residue washed with water. The combined filtrate was concentrated at or below 40°C in rotary evaporator under reduced pressure. This hydrolyzed mass was used for paper chromatography.In experimental animals, diabetes was induced by intraperitoneal injection of Nicotinamide (230 mg/kg) 15 min before streptozotocin  administration . The STZ Group 1: control: no intervention. Group 2: diabetic control + vehicle distilled water. Group 3: diabetic rats + glimepiride (10 mg/kg body weight). Group 4: diabetic rats + 250 mg/kg AEATP. Group 5: diabetic rats + 500 mg/kg AEATP. Group 6: diabetic rats + 1000 mg/kg AEATP.After 29 days of continuous treatment, blood sample was collected from retroorbital plexus under anesthesia for biochemical estimation of total cholesterol, triglycerides, LDL, HDL, VLDL, HbA1c, SGOT, and SGPT by commercially available kits of Reckon Diagnostics Pvt., Ltd. Body weight was measured before induction of diabetes and during treatment period. Fasting blood glucose level of overnight fasted rats was measured using glucometer on the 1st, 7th, 14th, 21st, and 28th days of pharmacological interventions. Serum insulin was measured by using ELISA kits (EMD Millipore-EZRMI-13 K). The assay is based on Sandwich ELISA technique and enzyme activity was measured spectrophotometrically by the increased absorbency at 450 nm, corrected from the absorbency at 590 nm, after acidification of formed products. Pancreatic insulin content was extracted by taking 0.2 g pancreas portion with 5.0 mL of ice-cold acid-alcohol in a centrifuge tube, homogenized, followed by sonication, stored at −20°C overnight, and finally centrifuged at 3000 rpm at 4°C for 15 minutes. The supernatant was transferred into a new centrifuge tube and stored at −20°C, while the pellet was subjected to extraction again. After extraction insulin level was measured at room temperature by ELISA assay kit [Further, animals with blood glucose level above 250 mg/dL were selected and divided into six groups comprising ten animals in each group.ssay kit . post hoc multiple comparison test.Statistical analysis was performed using Graph pad Prism 6. Values are expressed as mean ± SEM and statistical analysis was carried out by using by ANOVA followed by Tukey's asThe pure polysaccharide was extracted as amorphous white powder with a percentage yield of 24.5% from the gum exudates. Complete hydrolysis of the polysaccharide followed by paper chromatography revealed the presence of four spots, corresponding to D-galactose, D-glucose, L-rhamnose, and D-glucuronic acids, respectively. Acacia tortilis polysaccharide did not exhibit any sign of toxicity, behavioral changes, and mortality. Thus Acacia tortilis polysaccharide was found to be nontoxic and therefore can be safely used.In the present study, oral toxicity was carried out according to OECD guidelines, up to an elevated concentration of 5,000 mg/kg. However, at this doseAnimals of the same weight range were used in experimental protocol. During the study, the body weight of control group was increased naturally, whereas body weight was found to be significantly attenuated in STZ-induced diabetic group as compared to control group . After 2The basal values of fasting blood glucose level were almost the same and statistically no significant difference was observed while including the animals for experimentation. Fasting blood glucose level of control group ranged from 106.5 ± 6.64 to 110.16 ± 3.33 mg/dL in 28 days of study, while there was a significant increase in fasting blood glucose level in STZ + nicotinamide treated rat 400.5 ± 7.8 mg/dL as compared to control group . FastingGlycated hemoglobin was significantly increased in STZ-induced diabetic group 9.79 ± 0.2% as compared to control group 3.92 ± 0.19% . After 2The present result showed that total cholesterol level in serum was significantly elevated in diabetic control group 206.1 ± 3.7 mg/dL as compared to control group 115.1 ± 3.1 mg/dL . DiffereThere was significant increase in total triglyceride (TG), low density lipoprotein (LDL), and very low density lipoprotein (VLDL) levels in diabetic control group when compared to control group. The administration of AEATP and glimepiride for 28 days significantly attenuated diabetes induced high level of TG, LDL, and VLDL. 250, 500, and 1000 mg/kg AEATP and glimepiride significantly reduced the TG level by 106.73 ± 2.6, 104.87 ± 2.0, 102.3 ± 3.7, and 123.14 ± 4.8 mg/dL, respectively, as compared to diabetic control group 194.67 ± 3.41 mg/dL . SimilarHDL level was found to be significantly reduced in diabetic control group 17.75 ± 2.73 mg/dL compared to control group 51.43 ± 2.95 mg/dL. After 28 days of intervention with 250, 500, and 1000 mg/kg of AEATP, a significant increase was observed in HDL level 28.35 ± 1.28 mg/dL, 31.55 ± 2.79 mg/dL, and 32.87 ± 3.3 mg/dL, respectively, as compared to diabetic control group 17.75 ± 2.73 mg/dL. Administration of glimepiride also increased HDL level of diabetic rats 24.44 ± 1.3 .A significant elevation in SGOT and SGPT was observed in serum of STZ-induced diabetic rats 284.5 ± 10.21 and 161.9 ± 5.21 unit/lit. as compared to control group 177.0 ± 2.4 and 55.76 ± 3.09 unit/lit., respectively. A statistically significant attenuation was observed in SGOT enzyme level after 28 days of administration of 250, 500, and 1000 mg/kg of AEATP  and glimepiride (88.50 ± 5.22 unit/lit.) as compared to diabetic control 284.5 ± 10.21 unit/lit. . SimilarA significant decrease was observed in fasting insulin level (0.373 ± 0.026 ng/mL) and pancreatic insulin content (56.0 ± 2.81 ng/mg pancreas) in STZ-induced diabetic rats as compared to control group 0.820 ± 0.024 ng/mL and 103.9 ± 5.24 ng/mg pancreas, respectively. Fasting insulin level was statistically increased after 28 days administration of 250, 500, and 1000 mg/kg of AEATP  and glimepiride (0.723 ± 0.024 ng/mL) as compared to diabetic control (0.373 ± 0.026 ng/mL) . Similarβ-cells by necrosis [β-cell mass producing type 2 diabetes [The present study was designed to investigate antidiabetic activity of AEATP along with its effect on lipid profile and liver enzymes. STZ (N-nitro derivative of glucosamine) has potent alkylating property  and is snecrosis . Recentldiabetes . Like prdiabetes , 30. AdmInduction of diabetes is associated with the characteristic loss of body weight , 32, whi Acacia tortilis was also reported to have flavonoids content [Glycated hemoglobin (HbA1c) is the standard biochemical marker in assessment of diabetes. In our study, diabetic rats showed higher level of glycated hemoglobin indicating their poor glycemic control which is also supported by other previously reported studies , 34. Ora content  and mighLipid plays an important role in the pathogenesis of complications associated with diabetes mellitus. The elevated level of serum cholesterol and reduced level of serum HDL cholesterol in diabetic condition, poses to be a risk factor for developing microvascular complication leading to atherosclerosis and cardiovascular diseases like coronary heart disease. The abnormal high concentration of serum lipid in diabetic mainly due to increased mobilization of free fatty acids from peripheral fat depots, since insulin inhibits the hormone sensitive lipase, insulin deficiency, or insulin resistance may be responsible for dislipidemia . The preThere was increase in liver enzymes in serum of diabetic rats like earlier reports  which mi+-ATP channels, membrane depolarization, and stimulation of Ca2+ influx, an initial key step in insulin secretion. As studies have shown that the gut incretin content is altered in animal models of type 2 diabetes [The possible mechanism of action of AEATP could be correlated with promoting insulin secretion by closure of Kdiabetes , therefodiabetes . AEATP e Acacia tortilis polysaccharide in prevention and treatment of diabetes, hyperlipidemia, and its associated complications. However, the precise mechanism by which AEATP reduced fasting blood glucose level in diabetic rats will require further detailed study. Therefore, future research and clinical trials in this area may lead to the use of Acacia tortilis polysaccharide as a new type of therapeutic agent in treatment of type 2 diabetes.This study revealed first time the antidiabetic activity of AEATP and its effective role in maintaining lipid profile and liver toxicity in diabetic rats. This new insight allows an understanding of the use of


Rheum tanguticum resource in Baihe Pasture, Zoige County, Sichuan Province. The amount of R. tanguticum from images is M = S*ρ and S is vegetation coverage obtained by satellite imaging, whereas ρ is R. tanguticum density obtained by low-altitude imaging. Only the R. tanguticum which coverages exceeded 1 m2 could be recognized from the remote sensing image because of the 0.1 m resolution of the remote sensing image , and the results of ground investigation represented the amounts of R. tanguticum resource in all sizes . The data in paper showed that the present available amount of R. tanguticum accounted for 4% to 5% of the total quantity. The quantity information and the population structure of R. tanguticum in the Baihe Pasture were initially confirmed by this system. It is feasible to monitor the quantitative distribution for natural medicinal plants with scattered distribution.Remote sensing has been extensively applied in agriculture for its objectiveness and promptness. However, few applications are available for monitoring natural medicinal plants. In the paper, a multilevel monitoring system, which includes satellite and aerial remote sensing, as well as ground investigation, was initially proposed to monitor natural  Rheum tanguticum in the Baihe Pasture, Zoige County, Sichuan Province, to determine the quantitative distribution and population structure of R. tanguticum in this region.Remote sensing has been extensively applied in agriculture in recent years. This approach can provide objective, accurate, and timely information on the ecological environment of crops. Many remote sensing applications for medicinal plants are focused on cultivated medicinal plants. By contrast, natural medicinal plants are largely ignored because of their scattering distribution characteristic and small coverage area. A multilevel monitoring system, which includes satellites, aerial remote sensing, and ground investigation was proposed in this paper to monitor natural R. tanguticum are mainly distributed in temperate Asian zones, including the Greater Khingan Mountains, Taihang Mountains, Qinling Mountain, Daba Mountains, and Yunnan-Guizhou Plateau [ R. tanguticum is a peculiar Chinese perennial herb that belongs to the Polygonaceae family. This herb mainly thrives in Southern Gansu Province, Northwest Sichuan Province, and Northeast Tibet. These areas are characterized by high altitudes and short frost-free periods. R. tanguticum grows slowly in these regions and has rich substances with good qualities. The popular “Xining Rheum” and “Quanshui Rheum” originated from these areas. Recent studies on R. tanguticum are mainly focused on ecological suitability, chemical analysis, and pharmacology [ R. tanguticum as an ingredient for Tibetan medicine, health products, and specialty food has gradually increased [ R. tanguticum in the Qinghai-Tibet Plateau proliferates slowly because of the cold climate and fragile ecological environment. Moreover, the unreasonable excavation of R. tanguticum leads to a significant reduction in the quantity of this resource and may even lead to desertification. Determining the updated status of the quantitative distribution and population structure of R. tanguticum is necessary for ecological protection and resource development [ R. tanguticum [ R. tanguticum in the Baihe Pasture. The viability and effectiveness of the multilevel remote system were evaluated by ground-sampled surveys. This paper presented a new technical method and platform for monitoring wild and scattered medicinal plant resources.A number of studies indicate that domestic Plateau , 2. R. tmacology –7. The dncreased . R. tangelopment . Thus, tnguticum –12. Sate2 with an altitude of 3,400 m to 3,900 m. Zoige County is located on a plateau with a cold, temperate, and humid monsoon climate with a relative humidity of 68%. The average temperatures in January and July are −9.4 and 11.5°C, respectively. The average annual temperature is 1.7°C, and the highest and lowest temperatures are 25.4 and −29.5°C, respectively. The annual accumulated temperature higher than 10 is 718.4°C. Annual sunshine duration, rainfall, evaporation, gale day, and dust day are 2,506.7 h, 543.2 mm to 761.6 mm, 1,188.24 mm, 11.2 d, and 0.6 d, respectively. Zoige County has abundant water resources and contains several main branches of the Yellow River upstream, including the Heihe, Baihe, and Jiasong Rivers. Zoige County receives significant sunshine and heat energy, thus contributing to its rich 8,084 km2 natural grasslands that contain various medicinal plants, including Fritillaria, Cordyceps, Gentiana, Rhubarb, and Saussurea. These plants amount to 121 families and 1,094 species. Rheum palmatum, R. tangulicum and Rheum officinale all have distribution in Sichuan Province, but only wild R. tangulicum scatters in the studied region, and the other two species are mainly cultivated in farmlands or around farmhouses. The studied region is shown in 2. Most areas of the Baihe Pasture are covered with natural grasslands, except for the 1,000 ha of crops.Zoige County , which is located in the northeastern margin of the Tibetan Plateau, belongs to the northernmost area of Aba, Sichuan Province. This area is bordered by 4 counties of Gansu Province  and Sichuan Province . Zoige County is an important region of the Northwest Sichuan Pasture and covers an area of 10,620 kmSatellite Image. Landsat Thematic Mapper (TM) 5 image with a 0.45 μm to 2.35 μm spectrum range was used to determine the vegetation area in the Baihe Pasture. The image parameters were as follows: N33°25′ E102°14′ image center, blue B1 band, 0.45 μm to 0.52 μm spectrum range, and 30 m resolution; green B2 band, 0.52 μm to 0.60 μm spectrum range, and 30 m resolution; red B3 band, 0.63 μm to 0.69 μm spectrum range, and 30 m resolution; near-infrared B4 band, 0.76 μm to 0.90 μm spectrum range, and 30 m resolution; short-wave infrared B5 band, 1.55 μm to 1.75 μm spectrum range, and 30 m resolution; thermal infrared B6 band, 10.4 μm to 12.5 μm spectrum range, and 120 m resolution; middle-infrared B7 band, 2.08 μm to 3.35 μm spectrum range, and 30 m resolution. TM imaging is suitable for large-scale object monitoring because of its moderate resolution and low cost. The TM5 image shot on 18 July 2008 was chosen in the monitoring of the Baihe Pasture vegetation based on the biological features of natural R. tanguticum. R. tanguticum was initially conducted in the Baihe Pasture of Zoige County by the low-altitude remote sensing system.The low-altitude remote sensing system consisted of a flight platform, digital camera, single-axis stabilized platform, and equipment control system . The fliGround Survey. A total of 10 fenced zones with an area of 200 m × 200 m were selected for the R. tanguticum field survey. Four 10 m × 10 m plots were established in the 10 fenced zones based on the plot-setting principle. The ground survey for R. tanguticum was conducted separately in the fenced zones in July 2008, 2010, and 2011. R. tanguticum in vegetation was determined by using low-altitude images with 0.1 m resolutions. Finally, the amount of natural R. tanguticum resource was calculated based on the vegetation area from the satellite image and the R. tanguticum distribution density from low-altitude remote sensing. The results were compared and assessed with those of the ground survey.The complete technical road map is summarized in  R)/(NIR + R), where NIR denotes the near-infrared band reflectivity and R stands for red band reflectivity. An NDVI value in the range of zero to one generally refers to vegetation regions. The vegetation region of the Baihe Pasture was determined based on the NDVI analysis in Bands 4, 3, and 2 in the TM image were fused to distinguish the vegetation and water. The Baihe Pasture boundary in the TM image was determined via the registration function by the feature points. Thus, the Baihe Pasture satellite image was detached from the original TM image. NDVI is an internationally accepted criterion for vegetation coverage level. Consider that NDVI = , area (M), and vegetation area (M1) of the Baihe Pasture were P = 305676/(305676 + 189658) = 62%, M = (305676 + 189658) × 30 m × 30 m = 445 km2 , and M1 = 445 × 62% = 276 km2 , respectively. The results were consistent with the 43,333 ha provided by the Baihe Pasture. Therefore, remote sensing technology was feasible in monitoring the fields.Based on NDVI anlysis, the pixel numbers of 0.1 < NDVI < 1 and NDVI < 0.1 were 305,676 and 189,658, with each representing the vegetation and nonvegetation regions, respectively. Therefore, the vegetation area ratio  could be interpreted from the image. Thus, the average density of available R. tanguticum could be calculated by the interpretation results of many low-altitude images. The effective resources of R. tanguticum in the entire Baihe Pasture were that the Baihe Pasture vegetation area multiplied by the average R. tanguticum density, which was equal to 350,000. Therefore, the amount of available natural R. tanguticum resources in the Baihe Pasture in 2008 was 350,000.As a matter of experience, onlyTen vegetation fences, each with an area of 200 m × 200 m, except for the third fence with 150 m × 150 m, were established in the Baihe Pasture, and there were four 10 m × 10 m quadrats in every fence. Ground survey work was conducted in July 2008, 2010, and 2011 in the above fences and quadrats . R. tanguticum per 100 m²; thus, the total amount of R. tanguticum in the Baihe Pasture was M1(276 km2) × 2.6/100 m² = 718 × 104.The ground survey results in 2008 indicated the existence of 2.6 R. tanguticum in each fence was approximately 1,520, except for approximately 855 in the third fence. Thus, approximately 3.8 R. tanguticum per 100 m² existed, and the total amount of R. tanguticum was 1,049 × 104.The ground survey results in 2010 indicated that the number of R. tanguticum in each fence was approximately 1,220; the minimum amount was approximately 300 in the second fence, and the amount was approximately 1600 in the first, fourth, and sixth fences. The calculation result was 3.1 R. tanguticum per 100 m². Thus, the total amount of R. tanguticum in 2011 was 856 × 104.The results in 2011 indicated that the average number of R. tanguticum based on the results of different years because of operation errors and other factors. However, the total quantity of R. tanguticum was approximately 750 × 104 to 1,000 × 104. The current available amount and the future total amount of natural R. tanguticum resources in 2008 were, respectively, 35 × 104 and 718 × 104 based on the results of the multilevel remote sensing and ground survey. The population structure of natural R. tanguticum, that is, the available amount of R. tanguticum, comprised 4% to 5% of the total amount. Therefore, the safe digging quantity of R. tanguticum in the Baihe Pasture was 4% to 5% for the protection of natural resources.Quantity variance existed in the total amount ofRemote sensing technology has become an important means of obtaining geographic environment information. This technology has remarkably extended and improved the ability of humans to recognize their surrounding environment. Technology innovation in remote sensing provides a new method and platform for surveying natural Chinese medicine resources R. tanguticum in the Baihe Pasture. Based on the satellite image results, the Baihe Pasture area was 445 km2 , which was consistent with the 43,333 ha provided by the administrative department of the Baihe Pasture. The vegetation coverage in the Baihe Pasture was 276 km2. R. tanguticum density was 2.6 to 3.8 per 100 m² based on the result of the multilevel remote sensing. Compared with the ground investigation, the multilevel remote sensing technology system was fast and convenient. Results indicated that the available amount of R. tanguticum comprised 4% to 5% of the total amount. The quantity information and population structure of R. tanguticum in the Baihe Pasture of Zoige County were initially confirmed by this multilevel monitoring system. This system was also successfully applied in monitoring Ferula sinkiangensis in the fourth survey of the Chinese medicine resources of Xinjiang.(1) Remote sensing has been extensively applied in agriculture in recent years –25. Remo R. tanguticum in low-altitude aerial images. Only R. tanguticum with coverage greater than 1 m2 could be extracted from the low-altitude aerial image because of the spatial resolution limitations. Therefore, the amount of R. tanguticum obtained from multilevel remote sensing was the current available resources, whereas the results from the ground investigation were the total resources because all R. tanguticum sizes could be observed through ground surveys. The amount of R. tanguticum obtained from the ground survey represented the total available resource in three years to five years. The results from the two methods  represented the present and the future amounts of R. tanguticum. Thus, the corresponding method should be selected to satisfy different demands and purposes.(2) An object-oriented image-recognition method was initially adopted to extract R. tanguticum through remote sensing was early July because R. tanguticum was in bloom and taller than the surrounding vegetation during this period. The ground survey results in three years indicated that the total quantity of R. tanguticum was approximately 750 × 104 to 1,000 × 104. The available amount of natural R. tanguticum resources was 35 × 104 in 2008 based on the results of the multilevel remote sensing. However, the total amount in 2008 was 718 × 104 based on the ground survey results. So, the safety digging quantity of R. tanguticum in the Baihe Pasture was 4% to 5% for the protection of natural resources.(3) The optimum time for monitoringWe conclude that the use of low-altitude remote sensing technology to monitor the scattered distribution of natural medicinal plants was feasible. This study provided a new technical system for monitoring objects with scattered distributions.