Deciphering the Plastomic Code of Chinese Hog-Peanut (Amphicarpaea edgeworthii Benth., Leguminosae): Comparative Genomics and Evolutionary Insights within the Phaseoleae Tribe.

The classification and phylogenetic relationships within the Phaseoleae tribe (Leguminosae) have consistently posed challenges to botanists. This study addresses these taxonomic intricacies, with a specific focus on the Glycininae subtribe, by conducting a comprehensive analysis of the highly conserved plastome in Amphicarpaea edgeworthii Benth., a critical species within this subtribe. Through meticulous genomic sequencing, we identified a plastome size of 148,650 bp, composed of 128 genes, including 84 protein-coding genes, 36 tRNA genes, and 8 rRNA genes. Comparative genomic analysis across seven Glycininae species illuminated a universally conserved circular and quadripartite structure, with nine genes exhibiting notable nucleotide diversity, signifying a remarkable genomic variability. Phylogenetic reconstruction of 35 Phaseoleae species underscores the affinity of Amphicarpaea with Glycine, placing Apios as a sister lineage to all other Phaseoleae species, excluding Clitorinae and Diocleinae subtribes. Intriguingly, Apios, Butea, Erythrina, and Spatholobus, traditionally clumped together in the Erythrininae subtribe, display paraphyletic divergence, thereby contesting their taxonomic coherence. The pronounced structural differences in the quadripartite boundary genes among taxa with unresolved subtribal affiliations demand a reevaluation of Erythrininae's taxonomic classification, potentially refining the phylogenetic contours of the tribe.

Evolutionary Comparison of the Complete Chloroplast Genomes in Convallaria Species and Phylogenetic Study of Asparagaceae.

The genus Convallaria (Asparagaceae) comprises three herbaceous perennial species that are widely distributed in the understory of temperate deciduous forests in the Northern Hemisphere. Although Convallaria species have high medicinal and horticultural values, studies related to the phylogenetic analysis of this genus are few. In the present study, we assembled and reported five complete chloroplast (cp) sequences of three Convallaria species (two of C. keiskei Miq., two of C. majalis L., and one of C. montana Raf.) using Illumina paired-end sequencing data. The cp genomes were highly similar in overall size (161,365-162,972 bp), and all consisted of a pair of inverted repeats (IR) regions (29,140-29,486 bp) separated by a large single-copy (LSC) (85,183-85,521 bp) and a small single-copy (SSC) region (17,877-18,502 bp). Each cp genome contained the same 113 unique genes, including 78 protein-coding genes, 30 transfer RNA genes, and 4 ribosomal RNA genes. Gene content, gene order, AT content and IR/SC boundary structure were nearly identical among all of the Convallaria cp genomes. However, their lengths varied due to contraction/expansion at the IR/LSC borders. Simple sequence repeat (SSR) analyses indicated that the richest SSRs are A/T mononucleotides. Three highly variable regions (petA-psbJ, psbI-trnS and ccsA-ndhD) were identified as valuable molecular markers. Phylogenetic analysis of the family Asparagaceae using 48 cp genome sequences supported the monophyly of Convallaria, which formed a sister clade to the genus Rohdea. Our study provides a robust phylogeny of the Asparagaceae family. The complete cp genome sequences will contribute to further studies in the molecular identification, genetic diversity, and phylogeny of Convallaria.

The complete chloroplast genome sequence of Elatostema stewardii Merr. (Urticaceae).

Elatostema stewardii is an important medicinal plant endemic to China. In this study, the complete chloroplast genome of E. stewardii was sequenced and assembled using next-generation sequencing technology. The complete chloroplast genome length of E. stewardii was 150,263 bp, including two inverted repeats (IRs) of 24,681 bp, which are separated by LSC and SSC of 83,791 bp and 17,110 bp, respectively. A total of 129 genes were included in the genome, consisting 85 protein-coding genes, eight rRNA genes, and 36 tRNA genes, the overall GC content of this genome was 36.3%. There are few studies on the genus Elatostema of Urticaceae, this chloroplast genome sequence will provide useful data for further research on solving the generic and familial relationships in Urticaceae.

Lipogenic stromal cells as members of the foam-cell population in human atherosclerosis: Immunocytochemical and ultrastructural assessment of 6 cases.

To identify the nature of foam cells in atherosclerosis, carotid atherosclerotic plaques (CAPs) from six patients were studied. Hematoxylin-and-eosin, Congo Red and Oil Red O staining were used to study histopathologic alterations in CAPs. CD31, α-smooth-muscle actin (α-SMA), CD68, desmin and S100 were stained immunohistochemically. The ultrastructure of foam cells was analyzed by transmission electron microscopy (TEM). CAPs were shown to be composed of a fibrous cap covering a dome-shaped mass with a peripheral, circumferential fringe merging with a basal band which itself met the tunica media, the latter consisting of smooth-muscle cells (SMCs). The interior of the dome-shaped mass exhibited fibrosis, neovascularization, hemorrhage, necrosis and calcification. Lipid droplets identified by histological stains and TEM were found in the rounded epithelioid foam cells regarded as macrophages, as well as in spindled cells interpreted here as lipoleiomyocytes (lipid-containing SMCs), lipofibroblasts and lipomyofibroblasts; and all these cells were located in different regions of the CAPs. All of these lipid-laden cells were strongly positive for CD68 but negative for desmin. Foam cells were weakly positive for α-SMA, CD31 and S100. The results indicate that the light microscopically identifiable population of foam/lipid-laden cells hide a spectrum of diverse differentiation ranging from the expected macrophage phenotype to non-macrophage phenotypes. The origin of these diverse cell phenotypes in terms of multipotential mesenchymal precursors and the origin of the intracellular lipid are discussed.

Activation of UCP2 by anethole trithione suppresses neuroinflammation after intracerebral hemorrhage.

Intracerebral hemorrhage (ICH) is a devastating disease, in which neuroinflammation substantially contributes to brain injury. Uncoupling protein 2 (UCP2) is a member of the mitochondrial anion carrier family, which uncouples oxidative phosphorylation from ATP synthesis by facilitating proton leak across the mitochondrial inner membrane. UCP2 has been reported to modulate inflammation. In this study we investigated whether and how UCP2 modulated neuroinflammation through microglia/macrophages following ICH in vitro and in vivo. We used an in vitro neuroinflammation model in murine BV2 microglia to mimic microglial activation following ICH. ICH in vivo model was established in mice through collagenase infusion into the left striatum. ICH mice were treated with anetholetrithione (ADT, 50 mg· kg-1 ·d-1, ip) or the classical protonophoric uncoupler FCCP (injected into hemorrhagic striatum). We showed that the expression and mitochondrial location of microglial UCP2 were not changed in both in vitro and in vivo ICH models. Knockdown of UCP2 exacerbated neuroinflammation in BV2 microglia and mouse ICH models, suggesting that endogenous UCP2 inhibited neuroinflammation and therefore played a protective role following ICH. ADT enhanced mitochondrial ROS production thus inducing mitochondrial uncoupling and activating UCP2 in microglia. ADT robustly suppressed neuroinflammation, attenuated brain edema and improved neurological deficits following ICH, and these effects were countered by striatal knockdown of UCP2. ADT enhanced AMP-activated protein kinase (AMPK) activation in the hemorrhagic brain, which was abrogated by striatal knockdown of UCP2. Moreover, striatal knockdown of AMPK abolished the suppression of neuroinflammation by ADT following ICH. On the other hand, FCCP-induced mitochondrial uncoupling was independent of UCP2 in microglia; and striatal knockdown of UCP2 did not abrogate the suppression of neuroinflammation by FCCP in ICH mice. In conclusion, the uncoupling activity is essential for suppression of neuroinflammation by UCP2. We prove for the first time the concept that activators of endogenous UCP2 such as anetholetrithione are a new class of uncouplers with translational significance.

The complete chloroplast genome sequence of Japanese buttercup Ranunculus japonicus Thunb.

Ranunculus japonicus is an important medicinal herb widely used in East Asia. In this study, we report the first complete chloroplast genome sequence of Ranunculus japonicus using next-generation sequencing technology. The chloroplast genome size of R. japonicus was 156,981 bp. A total of 129 genes were included, consisting 84 protein-coding genes, eight rRNA genes, and 37 tRNA genes. Thirteen protein-coding genes had intron (ycf3 gene, rps12 gene, rps12 gene, clpP gene contained two introns). A further phylogenomic analysis of Ranunculaceae, including 10 taxa, was conducted for assessing the placement of R. japonicus. It will provide valuable genetic information for this medicinally important species.

The complete chloroplast genome sequence of spleen amaranth (Amaranthus dubius Mart. ex Thell., Amaranthaceae).

Amaranthus dubius is a leafy vegetable widely cultivated in Asia and Africa. The complete chloroplast genome of Amaranthus dubius was sequenced and assembled in this study. The complete chloroplast genome is 150,520 bp. A total of 130 genes were identified, including 85 protein-coding genes, eight rRNA genes, and 37 tRNA genes. The overall GC content of this genome was 36.6%. The phylogenetic tree based on 10 chloroplast genomes in Amaranthaceae supports that A. dubius is sister to A. hypochondriacus and A. caudatus.

The complete chloroplast genome sequence of wild Japanese pepper Tubocapsicum anomalum Makino (Solanaceae).

As an important medicinal herb, no complete organelle molecular data has been reported for Tubocapsicum anomalum. In this study, the first complete chloroplast genome of Tubocapsicum anomalum Makino was sequenced and assembled. The genome is 155,802 bp in length and contained 124 encoded genes in total, including 75 protein-coding genes, 10 ribosomal RNA genes, and 39 transfer RNA genes. The phylogenomic analysis showed that Tubocapsicum anomalum was closely related to Withania somnifera according the current sampling extent.

The complete chloroplast genome of balsam aster (Aster ageratoides Turcz. var. scaberulus (Miq.) Ling., Asteraceae).

The first complete chloroplast genome of Aster ageratoides Turcz. var. scaberulus (Miq.) Ling. is reported in this study. The total chloroplast genome size of A. ageratoides var. scaberulus was 153,071 bp and comprised of a large single-copy region (LSC with 84,896 bp), a small single-copy region (SSC with 18,269 bp), and two inverted repeat regions (IR with 24,953 bp). A total of 122 genes were included in the genome, including 83 protein-coding genes, 8 rRNA genes, and 37 tRNA genes. Eleven protein-coding genes had intron (ycf3, clpP and rps12 gene contained two introns. Further phylogenomic analysis of Asteraceae, including 13 taxa, was conducted for the placement of A. ageratoides var. scaberulus.

Medical expenditure for liver cancer in urban China: A 10-year multicenter retrospective survey (2002-2011).

OBJECTIVE: This study aims to understand the medical expenditure for liver cancer during 2002-2011 in urban areas of China. MATERIALS AND METHODS: This is a retrospective study. Based on a stratified cluster sampling method, a medical expenditure survey collected basic personal information from related medical records. Two-tailed independent sample t-test, variance analysis, and Student-Newman-Keuls Tests were used in cost analysis for the corresponding data types. RESULTS: A total of 12,342 liver cancer patients were included in the analysis. Overall average medical expenditure per case for liver cancer diagnosis and treatment in China has increased from ¥21, 950 to ¥40, 386 over the study period. For each liver cancer patient diagnosed between 2009 and 2011, the average expenditures were 29,332 CNY for stage I, 35,754 CNY for stage II, 34,288 CNY for stage III, and 30,275 CNY for stage IV diseases (P < 0.001). Pharmaceuticals accounted for the biggest part of the medical expenditure and it rose from 48.01% to 52.96% during these ten years, and the share of nursing fee expenses was the lowest (around 1%). Over the entire 10-year data period, the per capita expenditure of the east region (32,983 CNY) was higher than that of the west region (26,219 CNY) and slightly higher than the central region (31,018 CNY, P < 0.001). DISCUSSION: As a major cancer in China, liver cancer accounts for a large portion of health economic burden and its medical expenditure is heavy for families. Early diagnosis and treatment for liver cancer will save medical expenditure. CONCLUSION: The economic burden of liver cancer is high in China and related medical expenditure has increased.

Progress and Challenges of Ultrasonic Testing for Stress in Remanufacturing Laser Cladding Coating.

Stress in laser cladding coating is an important factor affecting the safe operation of remanufacturing components. Ultrasonic testing has become a popular approach in the nondestructive evaluation of stress, because it has the advantages of safety, nondestructiveness, and online detection. This paper provides a review of ultrasonic testing for stress in remanufacturing laser cladding coating. It summarizes the recent research outcomes on ultrasonic testing for stress, and analyzes the mechanism of ultrasonic testing for stress. Remanufacturing laser cladding coating shows typical anisotropic behaviors. The ultrasonic testing signal in laser cladding coating is influenced by many complex factors, such as microstructure, defect, temperature, and surface roughness, among others. At present, ultrasonic testing for stress in laser cladding coating can only be done roughly. This paper discusses the active mechanism of micro/macro factors in the reliability of stress measurement, as well as the impact of stress measurement on the quality and safety of remanufacturing components. Based on the discussion, this paper proposes strategies to nondestructively, rapidly, and accurately measure stress in remanufacturing laser cladding coating.

Enhanced Performance of E-Bike Motive Power Lead-Acid Batteries with Graphene as an Additive to the Active Mass.

The effects of both graphene nanoplatelets and reduced graphene oxide as additives to the negative active material in valve-regulated lead-acid batteries for electric bikes were investigated. Low-temperature performance, charge acceptance, cycle performance, and water loss were investigated. The test results show that the low-temperature performance, charge acceptance, and large-current discharge performance of the batteries with graphene additives were significantly improved compared to the control battery, and the cycle life under 100% depth of discharge condition was extended by more than 52% from 250 to 380 cycles. Meanwhile, the amount of water loss from the batteries with graphene changed only slightly compared with the control cells. The excellent performance of the batteries can be ascribed to the graphene promoting the negative-plate charge and discharge processes and suppressing the growth of lead sulfate crystals.

Melamine as a single source for fabrication of mesoscopic 3D composites of N-doped carbon nanotubes on graphene.

Integration of two-dimensional graphene and one-dimensional carbon nanotubes (CNTs) to create potentially useful 3D mesoscopic carbon structures with enhanced properties relative to the original materials is very desirable. Here, we report a novel and simple route using chemical vapor deposition (CVD) methods to fabricate bead-like nitrogen-doped CNT/graphene composites (NCNT/G) via a simple pyrolysis of the N-rich melamine in the presence of graphene oxide (GO) as a substrate using a Mn-Ni-Co ternary catalyst. We have characterized these structures by field-emission scanning electron microscopy, transmission electron microscopy, X-ray diffraction, Raman spectra, isothermal analyses, and X-ray photoelectron spectroscopy. The three dimensional NCNT/G hybrids have unique network structures, moderate graphitization, high specific surface area, good mesoporosity, and N doping, which makes them promising materials for applications in energy storage and conversion.

Effects of shRNA-Mediated Silencing of PKM2 Gene on Aerobic Glycolysis, Cell Migration, Cell Invasion, and Apoptosis in Colorectal Cancer Cells.

This study aims to explore the effects of shRNA-mediated silencing on Pyruvate kinase type M2 (PKM2) gene during aerobic glycolysis in colorectal cancer (CRC) cells. CRC tissues and adjacent normal tissues were obtained from 136 patients diagnosed with qRT-PCR, Western blotting, and immunohistochemistry (IHC) were performed to detect mRNA and protein expressions of PKM2. CRC cells were divided into a blank, vector, and PKM2-shRNA groups. Hexokinase (HK) and PKM2 activity were both determined by glucose-6-phosphate dehydrogenase (G-6-PD) coupled colorimetric assay and enzyme coupling rate method. The extracellular lactate concentration was measured by ultraviolet spectrophotometer and caspase activity was measured using spectrophotometry. The proliferation, cell cycle, apoptosis, invasion, and migration of CRC cells were detected by cell counting kit-8 (CCK-8) assay, flow cytometry, transwell assay, and scratch test. Three groups of nude mice were injected with 0.2 mL single-cell suspension from the blank, vector, and PKM2-shRNA groups, respectively. PKM2 protein content in CRC tissues was higher than that in adjacent normal tissues. Results showed that the PKM2-shRNA group exhibited significantly lower mRNA and protein expressions of PKM2, decreased PKM2 activity, reduced lactate metabolism level, increased cell apoptosis rate, elevated caspase-3 and caspase-9 activity, weakened proliferation, and a reduction in cell invasion and migration ability compared to the vector and blank groups. The optical density (OD) value was lower in the PKM2-shRNA group than in the blank and vector groups. These findings indicate that shRNA-mediated silencing of PKM2 gene promotes apoptosis and inhibits aerobic glycolysis, proliferation, migration, and invasion in CRC cells. J. Cell. Biochem. 118: 4792-4803, 2017. © 2017 Wiley Periodicals, Inc.

Identification of citrullinated peptides in the synovial fluid of patients with rheumatoid arthritis using LC-MALDI-TOF/TOF.

The objective of the study is to investigate potential citrullinated autoantigens as targets of anti-citrullinated protein antibodies (ACPAs) response in synovial fluids (SFs) of patients with rheumatoid arthritis (RA). SFs from six RA patients and six osteoarthritis (OA) patients as controls were collected. The citrullinated proteins in SFs were extracted by immunoprecipitation with rabbit anti-citrulline antibodies. Matrix-assisted laser desorption/ionization time of flight mass spectrometry/time of flight mass spectrometry (MALDI-TOF/TOF) mass spectrometry was subsequently performed to discover a characteristic neutral loss to finally determine citrullinated autoantigens. A total of 182 citrullinated peptides and 200 citrullinated sites were identified in RA SFs, while 3 citrullinated peptides and 4 citrullinated sites were identified in OA SFs. The 182 citrullinated peptides from RA SFs and the 3 citrullinated peptides from OA SFs were derived from 83 and 3 autoantigens, respectively. Eighty-three autoantigens except protein-arginine deiminase type-2 (PADI2) and protein-arginine deiminase type-2 (PADI4) were over-citrullinated compared with controls, and the citrullinated sites of PADI2 and PADI4 were different in two groups. Interestingly, citrullinated histone H3.3 (H3F3A) was found in OA controls, but not in RA groups. The differential citrullinated proteins identified in RA SFs suggested potential autoantigens were targeted for ACPAs response and might contribute to the induction and perpetuation of complement activation and joint inflammation in RA.

Incongruent range dynamics between co-occurring Asian temperate tree species facilitated by life history traits.

Postglacial expansion to former range limits varies substantially among species of temperate deciduous forests in eastern Asia. Isolation hypotheses (with or without gene flow) have been proposed to explain this variance, but they ignore detailed population dynamics spanning geological time and neglect the role of life history traits. Using population genetics to uncover these dynamics across their Asian range, we infer processes that formed the disjunct distributions of Ginkgo biloba and the co-occurring Cercidiphyllum japonicum (published data). Phylogenetic, coalescent, and comparative data suggest that Ginkgo population structure is regional, dichotomous (to west-east refugia), and formed ˜51 kya, resulting from random genetic drift during the last glaciation. This split is far younger than the north-south population structure of Cercidiphyllum (~1.89 Mya). Significant (recent) unidirectional gene flow has not homogenized the two Ginkgo refugia, despite 2Nm > 1. Prior to this split, gene flow was potentially higher, resulting in conflicting support for a priori hypotheses that view isolation as an explanation for the variation in postglacial range limits. Isolation hypotheses (with or without gene flow) are thus not necessarily mutually exclusive due to temporal variation of gene flow and genetic drift. In comparison with Cercidiphyllum, the restricted range of Ginkgo has been facilitated by uncompetitive life history traits associated with seed ecology, highlighting the importance of both demography and lifetime reproductive success when interpreting range shifts.

miR-10a controls glioma migration and invasion through regulating epithelial-mesenchymal transition via EphA8.

MicroRNAs (miRNAs) play a critical role in the development of cancers. However, the role of miRNAs in glioma is still poorly understood. In this study, we demonstrate that microRNA-10a (miR-10a) promotes cell migration and invasion by negatively regulating the expression of Eph tyrosine kinase receptor A8 (EphA8). Ectopic expression of EphA8 counteracts the promotion of migration and invasion induced by miR-10a. We further demonstrate that miR-10a and EphA8 regulate epithelial-mesenchymal transition (EMT) to affect cell migration and invasion. Collectively, we unveil a branch of the miR-10a/EphA8 pathway that regulates the progression of glioma.

[Influence of serum storage on the laboratory results of prostate-specific antigen].

OBJECTIVE: To investigate the influence of serum storage on the laboratory results of serum T-PSA, F-PSA and FPSA%. METHODS: Using automated chemiluminescence, we detected and compared the values of serum T-PSA, F-PSA and F-PSA% in the serum stored in different conditions. RESULTS: When the serum was stored at 4 degrees C or at the room temperature (22 - 26 degrees C), FPSA was unstable as compared with T-PSA. Compared with the initial value, after 4 hours at the room temperature, F-PSA was decreased to (0.392 +/- 0.246) microg/L (P < 0.01), while T-PSA and F-PSA% to (1.522 +/- 1.085) microg/L and (25.03 +/- 5.94)%, respectively, with no significant difference; after 8 hours at the room temperature, T-PSA and F-PSA were reduced to (1.513 +/- 1.083) and (0.389 +/- 0.247) microg/L (P < 0.05 and P < 0.01). At 4 degrees C, T-PSA, F-PSA and F-PSA% were decreased to (9.418 +/- 7.965) microg/L, (2.168 +/- 1.558) micro/L and (26.6 +/- 6.63)%, respectively, after 2 days (P < 0.05), and to (9.203 +/- 7.736) microg/L, (2.047 +/- 1.478) microg/L and (25.64 +/- 6.56)% after 1 week (P < 0.01). At -40 degrees C, T-PSA, F-PSA and F-PSA% were (4.532 +/- 4.393) microg/L, (1.178 +/- 1.034) microg/L and (24.45 +/- 8.81)% after 4 weeks. When the serum was stored at -40 degrees C and after 3 freeze-thaws, F-PSA and T-PSA were (5.982 +/- 5.314) and (1.341 +/- 1.029) microg/L, respectively, with no significant difference from the initial values. CONCLUSION: Different conditions of serum storage have different influences on the laboratory results of serum TPSA, F-PSA and F-PSA%, more on F-PSA than on T-PSA, while F-PSA% is relatively stable. At -40 degrees C, T-PSA and F-PSA may remain stable for a month at least. Repeated freeze-thaws of the serum do not affect the laboratory results of F-PSA and T-PSA.

Isolation of compound microsatellite markers for the endangered plant Neolitsea sericea (Lauraceae).

PREMISE OF THE STUDY: The development of compound microsatellite markers was conducted in Neolitsea sericea to investigate genetic diversity and population genetic structure of this endangered insular species. • METHODS AND RESULTS: Using the compound microsatellite marker technique, 10 compound microsatellite markers that were successfully amplified showed polymorphism when assessed in 55 individuals from two populations in East China and Japan. Overall, the number of alleles ranged from 3 to 17, with an average of 7.9 alleles per locus. In addition, these primers could be easily amplified in Neolitsea aurata var. paraciculata and N. aurata var. chekiangensis. • CONCLUSIONS: The highly polymorphic markers developed and characterized in this study will be useful for population genetic studies of N. sericea.

[Preparation of ASODN-protamine-HSA-PLGA nanoparticles and initial evaluation of their nucleus targeting property in vitro].

OBJECTIVES: To develop a novel non-viral gene delivery system-SODN-Protamine-HSA-PLGA (ASODN-P/H-PLGA-NP) and investigate its nucleus targeting potential in vitro. METHODS: ASODN-P/H-PLGA-NP was prepared by mixing the protamine sulfate and HSA. Then the PLGA nanoparticles were prepared using double-emulsion evaporation technique, followed by addition of ASODN to the prepared P/H complex. The morphology of ASODN-P/H-PLGA-NP was observed by transmission electron microscopy. The diameter, PDI, and surface charge of ASODN-P/H-PLGA-NP were measured by photo correlation spectroscopy (PCS). The encapsulation efficiency of ASODN was determined by double step method. The cytotoxicity of ASODN-P/H-PLGA-NP was investigated by MTT assays. The ability to enter the squamouse carcinoma: Hep-2 cell line and its nucleus targeting property were observed by confocal laser scanning microscope. RESULTS: The average diameter, PDI, zeta potential, and encapsulation efficiency of ASODN-P/H-PLGA-NP were 128 nm, 0.234, -23.3 mV, and 78.45%, respectively. ASODN-P/H-PLGA-NP could protect the ASODN from the shear force in the ultrasound process during preparation. ASODN-P/H-PLGA-NP couldenter Hep-2 cells and have certain level of nucleus targeting property. CONCLUSION: ASODN-P/H-PLGA-NP can be prepared easily with small particle sizes and low cytotoxicity, which might be employed as a good non-viral vector for applications in ASODN delivery to nucleus.