M3 Muscarinic Acetylcholine Receptor Antagonist Darifenacin Protects against Pulmonary Fibrosis through ERK/NF-κB/miR-21 Pathway

Idiopathic pulmonary fibrosis is an untreatable lethal lung disease, which is related to the aberrant proliferation of fibroblasts. M3 muscarinic acetylcholine receptor (M3-mAChR) activation exerts proliferative effect on various kinds of cells. However, whether M3-mAChR inhibition has a protective effect on pulmonary fibrosis remains unexplored. A rat model of pulmonary fibrosis was established by intratracheal instillation of bleomycin. Darifenacin was used to block M3-mAChR. Histological changes were observed using Masson’s Trichrome and hematoxylin and eosin (HE) staining. Hydroxyproline was measured by Hydroxyproline detection kit. Transforming growth factor β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). In vitro, pulmonary fibroblasts were isolated from lungs of neonatal rat. After treatment, the cell viability, Hydroxyproline level was measured by MTT and Hydroxyproline detection kit respectively. The expression level of extracellular signal-regulated kinase (ERK), nuclear factor kappa-B (N-NF-κB), and microRNA-21 (miR-21) was detected by western blot or quantitative real-time PCR (qRT-PCR). Darifenacin relieved the fibrotic effects provoked by bleomycin. The expression level of hydroxyproline, TGF-β1 and TNF-α level was all downregulated after darifenacin treatment. In lung fibroblasts, darifenacin decreased cell viability and hydroxyproline level induced by bleomycin. Besides, phosphorylation-ERK and nuclear N-NF-κB protein level was downregulated, as well as miR-21 level. M3-mAChR antagonist darifenacin attenuates bleomycin-induced pulmonary fibrosis in rats, which may relate to the ERK/NF-κB/miRNA-21 signaling pathway.

无筋豆新品种比较试验

针对西双版纳以种植无筋豆为主要冬季外销蔬菜生产中,存在种植品种单一,不能满足市场需求,以及多年种植的红豆王无筋豆品种抗病性降低等问题,从省内外引进12个新品种进行比较试验,筛选出适应不同地区市场需求的抗病性强、产量高,无筋,品质优的圆棍形品种3个,宽扁条形品种3个,特色品种2个。

Differential expression of microRNAs in dorsal root ganglia after sciatic nerve injury

This study investigated the possible involvement of microRNAs in the regulation of genes that participate in peripheral neural regeneration. A microRNA microarray analysis was conducted and 23 microRNAs were identiifed whose expression was signiifcantly changed in rat dorsal root ganglia after sciatic nerve transection. The expression of one of the downregulated microRNAs, microRNA-214, was validated using quantitative reverse transcriptase-PCR. MicroRNA-214 was predicted to target the 3′-untranslated region of Slit-Robo GTPase-activating protein 3. In situ hybridization veriifed that microRNA-214 was located in the cytoplasm of dorsal root ganglia primary neurons and was downregulated following sciatic nerve transection. Moreover, a com-bination of in situ hybridization and immunohistochemistry revealed that microRNA-214 and Slit-Robo GTPase-activating protein 3 were co-localized in dorsal root ganglion primary neu-rons. Western blot analysis suggested that Slit-Robo GTPase-activating protein 3 was upregulated in dorsal root ganglion neurons after sciatic nerve transection. These data demonstrate that mi-croRNA-214 is located and differentially expressed in dorsal root ganglion primary neurons and may participate in regulating the gene expression of Slit-Robo GTPase-activating protein 3 after sciatic nerve transection.

Overexpression of the Watermelon Ethylene Response Factor ClERF069 in Transgenic Tomato Resulted in Delayed Fruit Ripening

Watermelon fruit undergoes distinct development stages with dramatic changes during fruit ripening.To date,the molecular mechanics of watermelon ripening remain unclear.Genetic and transcriptome evidences suggested that the ethylene response factor(ERF)gene ClERF069 may be an important candidate factor affecting watermelon fruit ripening.To dissect the roles of ClERF069 in fruit ripening,structure and phylogenetic analysis were performed using the amplified full-length sequence.Normal-ripening watermelon 97103,non-ripening watermelon PI296341-FR and the RIL population were used to analyze ClERF069 expression dynamics and the correlation with fruit ripening indexs.The results indicated that ClERF069 belongs to ERF family group VI and show high homology(83%identity)to melon ERF069-like protein.ClERF069 expression in watermelon flesh was negatively correlated with fruit lycopene content and sugar content during fruit ripening progress.Further transgenic evidences indicated that overexpression of 35S:ClERF069 in tomato noticeably delayed the ripening process up to 5.2 days.Lycopene,β-carotenoid accumulation patterns were altered and ethylene production patterns in transgenic fruits was significantly delayed during fruit ripening.Taken together,watermelon ethylene response factor ClERF069 was concluded to be a negative regulator of fruit ripening.

Evaluation of the mold-filling ability of alloy melt in squeeze casting

The mold-filling ability of alloy mclt in squceze casting process was evaluated by means of the maximum length of Archimedes spiral line. A theoretical evaluating model to predict the maximum filling length was built based on the flowing theory of the incompressible viscous fluid. It was proved by experiments and calculations that the mold-tilling pressure and velocity are prominent influencing factors on the mold-filling ability of alloy melt. The mold-filling ability increases with the increase of the mold-filling pressure and the decrease of the proper mold-filling velocity. Moreover, the pouring temperature relatively has less effect on the mold-filling ability under the experimental conditions. The maximum deviation of theoretical calculating values with experimental results is less than 15%. The model can quantitatively estimate the effect of every factor on the mold-filling ability.

Genetic relationship and pedigree of Chinese watermelon varieties based on diversity of perfect SNPs

Watermelon(Citrullus lanatus)is one of the world’s most important fruit crops,and China produces the most watermelons in the world.Recently,a watermelon variome consisting of 414 key resequenced accessions was reported.However,the genetic relationships and pedigree of Chinese watermelon varieties in the seed market remain unclear.In this study,241 evenly distributed perfect single nucleotide polymorphisms(SNPs)derived from the watermelon variome were selected for variety identification.The diversity of 247 Chinese watermelon varieties was identified based on their SNP genotypes.The 247 watermelon varieties were clustered into five subpopulations:the East Asian ecotype,intermediate ecotype,small fruit with red flesh ecotype,small fruit with yellow flesh ecotype,and American ecotype.We further established the pedigree of four subpopulations,of which JingXinNo.1,ZaoChunHongYu,HuangXiaoYu and XiaoLan,and Sugarlee were the main doner of the East Asian ecotype,small fruit with red flesh ecotype,small fruit with yellow flesh ecotype,and American ecotype,respectively.Thirty-two core SNPs were selected and applied in watermelon variety identification.They were also validated by the Kompetitive allele-specific PCR(KASPar)platform.The present study furthered our understanding of the genetic relationships and pedigree of watermelon varieties in China,and will help to manage the plant variety protection in watermelon.

Immunological rejection of acellular heterogeneous nerve transplant for bridging the sciatic nerve in rats

BACKGROUND：Artificial materials composed of acellular heterogeneous nerves can resolve donor shortage problems for the repair of peripheral nerve defects.However,it remains unclear whether artificial materials can overcome immunological rejection of heterogeneous nerve grafts and obtain similar effects as allogeneic nerve grafts.OBJECTIVE：To analyze regeneration and immunological rejection of defective sciatic nerves in rats through the use of acellular heterogeneous nerve grafts.DESIGN,TIME AND SETTING：A randomized,controlled study was performed at the Department of Anatomy,China Medical University and the Experimental Center,First Affiliated Hospital,China Medical University between January and December 2008.MATERIALS：TritonX-100 （Sigma,USA） and deoxycholate （Pierce,USA） were used.METHODS：Bilateral sciatic nerves were collected from adult rabbits and treated with TritonX-100 and sodium deoxycholate to prepare acellular sciatic nerves,which were used to bridge 1 -cm defective sciatic nerves in adult rats.MAIN OUTCOME MEASURES：The lymphocyte percentage in leukocytes was quantified following hemocyte staining.Neural regeneration and the recovery of motor end plates in the gastrocnemius muscle were observed under optical and electronic microscopy following toluidine blue staining,as well as acetylcholinesterase and succinate dehydrogenase histochemical staining.RESULTS：There was no significant difference in the lymphocyte percentage in leucocytes between transplanted and normal rats （P 〉 0.05）.At 3 months after surgery,the rat toes on the operated side were separated and the rats could walk.In addition,the footplates exhibited an escape response when acupunctured.A large number of regenerated nerve fibers were observed in the transplant group,and acetylcholinesterase-positive motor end plates were visible in fibers of the gastrocnemius muscle.CONCLUSION：Acellular heterogeneous nerve transplants for the repair of defective sciatic nerves in rats promote neural regeneration without significant immunological rejection.

Genetic architecture of maize yield traits dissected by QTL mapping and GWAS in maize

The study of yield traits can reveal the genetic architecture of grain yield for improving maize production.In this study, an association panel comprising 362 inbred lines and a recombinant inbred line population derived from X178 × 9782 were used to identify candidate genes for nine yield traits. High-priority overlap(HPO) genes, which are genes prioritized in a genome-wide association study(GWAS), were investigated using coexpression networks. The GWAS identified 51 environmentally stable SNPs in two environments and 36 pleiotropic SNPs, including three SNPs with both attributes. Seven hotspots containing 41 trait-associated SNPs were identified on six chromosomes by permutation. Pyramiding of superior alleles showed a highly positive effect on all traits, and the phenotypic values of ear diameter and ear weight consistently corresponded with the number of superior alleles in tropical and temperate germplasm. A total of 61 HPO genes were detected after trait-associated SNPs were combined with the coexpression networks. Linkage mapping identified 16 environmentally stable and 16 pleiotropic QTL.Seven SNPs that were located in QTL intervals were assigned as consensus SNPs for the yield traits.Among the candidate genes predicted by our study, some genes were confirmed to function in seed development. The gene Zm00001 d016656 encoding a serine/threonine protein kinase was associated with five different traits across multiple environments. Some genes were uniquely expressed in specific tissues and at certain stages of seed development. These findings will provide genetic information and resources for molecular breeding of maize grain yield.

Correction: The NAC transcription factor ClNAC68 positively regulates sugar content and seed development in watermelon by repressing ClINV and ClGH3.6

In this article 1,author name Yanping Wang was incorrectly tagged as corresponding author.Prof.Wang should be the co-first author,instead of the corresponding author.The original article has been corrected.

Nutrient Composition of Different Foxtail Millet Seeds

Ten millet cultivars from China and abroad were analyzed for their crude fat, amylose and amylopectin, crude protein and amino acid content. A wide variation was observed in amylopectin contain, ranging from 54.47% to 69.26%, while amylose content ranged from 11.26% to 23.22%. The average crude fat in all cultivates was 3.46%, and most cultivates ranged from 3.1% to 3.7%. Protein contents ranged from 8.61% to 15.54% with a mean value of 11.94%. For amino acid composition, the ten cultivates were particularly rich in leucine, and the mean was 34.57 mg/g. The millet cultivars were also high in glycine, glutamic acid and cysteine. Millet was known to be limiting in the essential amino lysine, and the mean was 5.18 mg/g. Other amino acids, tyrosine, histidine and arginine were also very low in the ten cultivars. In general, significant nutrient composition differences were observed between the different millet cultivars, and A2 （a kind of mother line of the hybrid millet） and hybrid millet registered a higher level of protein and amino acid composition which would be useful in millet breeding.

CRISPR/Cas9-mediated mutagenesis of CiBG1 decreased seed size and promoted seed germination in watermelon

Abscisic acid(ABA)is a critical regulator of seed development and germination.β-glucosidases(BGs)have been suggested to be contributors to increased ABA content because they catalyze the hydrolysis of ABA-glucose ester to release free ABA.However,whether BGs are involved in seed development is unclear.In this study,a candidate gene,CiBG1,in watermelon was selected for targeted mutagenesis via the CRISPR/Cas9 system.Seed size and weight were significantly reduced in the Clbg1-mutant watermelon lines,which was mainly attributed to decreased cell number resulting from decreased ABA levels.A transcriptome analysis showed that the expression of 1015 and 1429 unique genes was changed 10 and 18 days after pollination(DAP),respectively.Cytoskeleton-and cell cycle-related genes were enriched in the differentially expressed genes of wild type and Clbg1-mutant lines during seed development.Moreover,the expression of genes in the major signaling pathways of seed size control was also changed.In addition,seed germination was promoted in the Cibg1-mutant lines due to decreased ABA content.These results indicate that ClBG1 may be critical for watermelon seed size regulation and germination mainly through the modulation of ABA content and thereby the transcriptional regulation of cytoskeleton-,cell cycle-and signaling-related genes.Our results lay a foundation for dissecting the molecular mechanisms of controlling watermelon seed size,a key agricultural trait of significant economic importance.

The NAC transcription factor CINAC68 positively regulates sugar content and seed development in watermelon by repressing CIINV and CIGH3.6

NAC(NAM,ATAF1/2,and CUC2)transcription factors play important roles in fruit ripening and quality.The watermelon genome encodes 80 NAC genes,and 21 of these NAC genes are highly expressed in both the flesh and vascular tissues.Among these genes,CINAC68 expression was signi fi cantly higher in flesh than in rind.However,the intrinsic regulatory mechanism of CINAC68 in fruit ripening and quality is still unknown.In this study,we found that ClNAC68 is a transcriptional repressor and that the repression domain is located in the C-terminus.Knockout of CINAC68 by the CRISPR-Cas9 system decreased the soluble solid content and sucrose accumulation in mutant flesh.Development was delayed,germination was inhibited,and the IAA content was signi ficantly decreased in mutant seeds.Transcriptome analysis showed that the invertase gene CUNV was the only gene involved in sucrose metabolism that was upregulated in mutant flesh,and expression of the indole-3-acetic acid-amido synthetase gene CIGH3.6 in the IAA signaling pathway was also induced in mutant seeds.EMSA and dual-luciferase assays showed that CINAC68 directly bound to the promoters of CUNV and CIGH3.6 to repress their expression.These results indicated that CINAC68 positively regulated sugar and IAA accumulation by repressing CUNV and CIGH3.6.Our findings provide new insights into the regulatory mechanisms by which NAC transcription factors affect fruit quality and seed development.

Genome-wide Association Analysis of Maize Flowering Traits

Flowering regulation is important for maize to adapt to a variety of environments as well as associated with high yield.In this study,the genetic mechanism of three flowering traits of 310 maize inbred lines with rich genetic background was investigated in three years at three different environments such as days to tasseling(DTT),days to silking(DTS)and days to pollen shedding(DTP).Based on mean performance,the longest flowering time was observed in Zhanyi(2018),whereas the shortest in Shizong(2019).The coefficient of variance depicted the range from 3.62%to 9.06%for three flowering traits under all environments.Therefore,we have integrated these flowering traits corresponding to SNP molecular markers for genome-wide association study(GWAS).Results showed that 22 SNPs markers were significantly associated with DTT according to physical position and average linkage disequilibrium(LD)decay distance,and a total of 234 candidate genes were identified near these significantly associated SNP markers.Moreover,KEGG and GO analysis showed that these genes were enriched in the regulation of the physiological pathways for flowering.In more details,16 genes involved in development of floral organs are more worthy of our attention in future studies.

Treatment of femur supracondylar fracture with retrograde interlocking intramedullary nails in elderly patients

Objective： To report 32 cases of femur supracondylar fracture treated with retrograde interlocking intramedullary nails in elderly patients. Methods： According to the AO classification, all of 32 cases were classified as extra-articular type A. 32 cases were treated with interlocking intramedullary nail by closed insertion from intercondylar fossa of the knee. All cases accepted CPM exercise as early as possible after operation. Results： Following up 5 to 15 months, all fractures united within an average duration of 5.3 months （4-7 months）. According to the Shelbourne scale, the excellent rate of the knee function was 86.3%. Conclusion： Retrograde interlocking intramedullary nail is useful alternative implant for the treatment of osteoporotic supracondylar fracture of femur, particularly of the type A fracture in the elderly population. Its merits include stable fixation, high rate of fracture union and few complications.

太阳极轨天文台

太阳极轨天文台利用高轨道倾角(≥80°)和小椭偏率轨道,将首次实现正面对太阳极区磁场和速度场高精度成像观测,结合多波段遥感和原位测量,为解决太阳磁活动周起源的世纪难题提供决定性观测,为破解“原始”高速太阳风的起源效应之谜提供直接观测支撑,为理解日球层整体结构突破性创建数据驱动的日球数值模型.

ClSnRK2.3 negatively regulates watermelon fruit ripening and sugar accumulation

Watermelon(Citrullus lanatus) as non-climacteric fruit is domesticated from the ancestors with inedible fruits. We previously revealed that the abscisic acid(ABA) signaling pathway gene ClSnRK2.3 might infuence watermelon fruit ripening. However,the molecular mechanisms are unclear. Here,we found that the selective variation of ClSnRK2.3 resulted in lower promoter activity and gene expression level in cultivated watermelons than ancestors, which indicated ClSnRK2.3 might be a negative regulator in fruit ripening. Overexpression (OE) of ClSnRK2.3 significantly delayed watermelon fruit ripening and suppressed the accumulation of sucrose, ABA and gibberellin GA4. Furthermore,we determined that the pyrophosphate-dependent phosphofructokinase(ClPFP1) in sugar metabolism pathway and GA biosynthesis enzyme GA20 oxidase(Cl GA20ox) could be phosphorylated by ClSnRK2.3 and thereby resulting in accelerated protein degradation in OE lines and finally led to low levels of sucrose and GA4. Besides that, ClSnRK2.3 phosphorylated homeodomain-leucine zipper protein(ClHAT1) and protected it from degradation to suppress the expression of the ABA biosynthesis gene 9’-cis-epoxycarotenoid dioxygenase 3(Cl NCED3). These results indicated that ClSnRK2.3 negatively regulated watermelon fruit ripening by manipulating the biosynthesis of sucrose, ABA and GA4. Altogether, these findings revealed a novel regulatory mechanism in non-climacteric fruit development and ripening.

Karyotype Stability and Unbiased Fractionation in the Paleo-Allotetraploid Cucurbita Genomes

The Cucurbita genus contains several economically important species in the Cucurbitaceae family. Here, we report high-quality genome sequences of C. maxima and C. moschata and provide evidence supporting an allotetraploidization event in Cucurbita. We are able to partition the genome into two homoeologous subgenomes based on different genetic distances to melon, cucumber, and watermelon in the Benincaseae tribe. We estimate that the two diploid progenitors successively diverged from Benincaseae around 31 and 26 million years ago （Mya）, respectively, and the allotetraploidization happened at some point between 26 Mya and 3 Mya, the estimated date when C. maxima and C. moschata diverged. The subgenomes have largely maintained the chromosome structures of their diploid progenitors. Such long-term karyotype stability after polyploidization has not been commonly observed in plant polyploids. The two subgenomes have retained similar numbers of genes, and neither subgenome is globally dominant in gene expression. Allele-specific expression analysis in the C. maxima ×C. moschata interspecific F1 hybrid and their two parents indicates the predominance of trans-regulatory effects underlying expression divergence of the parents, and detects transgressive gene expression changes in the hybrid correlated with heterosis in important agronomic traits. Our study provides insights into polyploid genome evolution and valuable resources for genetic improvement of cucurbit crops.

The first all-season sample set for mapping global land cover with Landsat-8 data

We report the world's first all-season training and validation sample sets for global land cover classification with Landsat-8 data.Prior to this,such samples were only available at a single date primarily from the growing season.It is unknown how much limitation such a single-date sample has to mapping global land cover in other seasons of the year.To answer this question,we selected available Landsat-8 images from four seasons and collected training and validation samples from them.We compared the performances of training samples in different seasons using Random Forest algorithm.We found that the use of training samples from any individual season would result in the best overall classification accuracy when validated by samples in the same season.The global overall accuracy from combined best seasonal results was 67.2% when classifying the 11 Level-1 classes in the Finer Resolution Observation and Monitoring of Global Land Cover(FROM-GLC) classification system.The use of training samples from all seasons(named all-season training sample set hereafter) produced an overall accuracy of 67.0%.We also tested classification within 10° latitude 60° longitude zones using all-season training subsample within each zone and obtained an overall accuracy of 70.2%.This indicates that properly grouped subsamples in space can help improve classification accuracies.All the results in this study seem to suggest that it is possible to use an all-season training sample set to reach global optimality with universal applicability in classifying images acquired at any time of a year for global land cover mapping.

Experimental study of effect of post processing on fracture toughness and fatigue crack growth performance of selective laser melting Ti-6Al-4V

For Ti-6Al-4V,a titanium alloy increasingly used in aerospace structure,selective laser melting(SLM)is an attractive additive manufacturing technology,which is attributed to its complex construction capability with high accuracy and good surface quality.In order to obtain qualified mechanical properties,SLM parameters and post processing should be tailored for diverse service conditions.Fracture toughness and fatigue crack growth(FCG)behavior are critical characteristics for damage tolerance evaluation of such metallic structures,and they are affected by post processing technologies significantly.The objective of this study is to obtain the fracture toughness and fatigue crack growth behavior of Ti-6Al-4V manufactured by SLM,and to evaluate the influence of post-SLM thermomechanical treatment and surface machining.Fracture toughness and FCG tests were performed for SLM Ti-6Al-4V in three types of post processing status:as-built,heat treated and hot isostatically pressed(HIPed),respectively.Specimens with as-built and machined surface were tested.The microstructure and fractography were analyzed as well in order to investigate the relevance among manufacture process,microstructure and mechanical properties.The results demonstrate that as-built SLM Ti-6Al-4V presents poor ductility and FCG behavior due to martensitic microstructure and residual stresses.Both heat treatment and hot isostatic pressing improve the plane-stress fracture toughness and FCG performance considerably,while surface machining shows slight effect.

Natural variation in the NAC transcription factor NONRIPENING contributes to melon fruit ripening

ThThe NAC transcription factor NONRIPENING(NOR)is a master regulator of climacteric fruit ripening.Melon(Cucumis melo L.)has climacteric and nonclimacteric fruit ripening varieties and is an ideal model to study fruit ripening.Two natural CmNAC-NOR variants,the climacteric haplotype CmNAC-NOR^(S,N) and the non-climacteric haplotype CmNAC-NOR^(A,S),have effects on fruit ripening;however,their regulatory mechanisms have not been elucidated.Here,we report that a natural mutation in the transcriptional activation domain of CmNAC-NORS,Ncontributes to climacteric melon fruit ripening.CmNAC-NOR knockout in the climacteric-type melon cultivar“BYJH”completely inhibited fruit ripening,while ripening was delayed by 5-8 d in heterozygous cmnac-nor mutant fruits.CmN AC-NOR directly activated carotenoid,ethylene,and abscisic acid biosynthetic genes to promote fruit coloration and ripening.Furthermore,CmNAC-NOR mediated the transcription of the“CmNAC-NOR-CmNAC73-CmCWINV2”module to enhance flesh sweetness.The transcriptional activation activity of the climacteric haplotype CmNAC-NORS,Non these target genes was significantly higher than that of the nonclimacteric haplotype CmNAC-NOR^(A,S).Moreover,CmNAC-NORS,Ncomplementation fully rescued the non-ripening phenotype of the tomato(Solanum lycopersicum)cr-nor mutant,while CmNAC-NOR^(A,S) did not.Our results provide insight into the molecular mechanism of climacteric and non-climacteric fruit ripening in melon.