High-quality haplotype-resolved genome assembly of cultivated octoploid strawberry

Cultivated strawberry(Fragaria×ananassa),a perennial herb belonging to the family Rosaceae,is a complex octoploid with high heterozygosity at most loci.However,there is no research on the haplotype of the octoploid strawberry genome.Here we aimed to obtain a high-quality genome of the cultivated strawberry cultivar,“Yanli”,using single molecule real-time sequencing and high-throughput chromosome conformation capture technology.The“Yanli”genome was 823 Mb in size,with a long terminal repeat assembly index of 14.99.The genome was phased into two haplotypes,Hap1(825 Mb with contig N50 of 26.70 Mb)and Hap2(808 Mb with contig N50 of 27.51 Mb).Using the combination of Hap1 and Hap2,we obtained for the first time a haplotype-resolved genome with 56 chromosomes for the cultivated octoploid strawberry.We identified a∼10 Mb inversion and translocation on chromosome 2-1.104957 and 102356 protein-coding genes were annotated in Hap1 and Hap2,respectively.Analysis of the genes related to the anthocyanin biosynthesis pathway revealed the structural diversity and complexity in the expression of the alleles in the octoploid F.×ananassa genome.In summary,we obtained a high-quality haplotype-resolved genome assembly of F.×ananassa,which will provide the foundation for investigating gene function and evolution of the genome of cultivated octoploid strawberry.

A splice site mutation in the FvePHP gene is associated with leaf development and f lowering time in woodland strawberry

Leaves and f lowers are crucial for the growth and development of higher plants.In this study we identified a mutant with narrow leaf lets and early f lowering(nlef)in an ethyl methanesulfonate-mutagenized population of woodland strawberry(Fragaria vesca)and aimed to identify the candidate gene.Genetic analysis revealed that a single recessive gene,nlef,controlled the mutant phenotype.We found that FvH4_1g25470,which encodes a putative DNA polymeraseαwith a polymerase and histidinol phosphatase domain(PHP),might be the candidate gene,using bulked segregant analysis with whole-genome sequencing,molecular markers,and cloning analyses.A splice donor site mutation(C to T)at the 5-end of the second intron led to an erroneous splice event that reduced the expression level of the full-length transcript of FvePHP in mutant plants.FvePHP was localized in the nucleus and was highly expressed in leaves.Silencing of FvePHP using the virus-induced gene silencing method resulted in partial developmental defects in strawberry leaves.Overexpression of the FvePHP gene can largely restore the mutant phenotype.The expression levels of FveSEP1,FveSEP3,FveAP1,FveFUL,and FveFT were higher in the mutants than those in‘Yellow Wonder’plants,probably contributing to the early f lowering phenotype in mutant plants.Our results indicate that mutation in FvePHP is associated with multiple developmental pathways.These results aid in understanding the role of DNA polymerase in strawberry development.

Three-dimensional structure of liver vessels and spatial distribution of hepatic immune cells

As the largest internal organ of the human body,the liver has an extremely complex vascularnetwork and multiple types of immune cells.It plays an important role in blood circulation,material metabolism,and immune response.Optical imaging is an effective tool for studying finevascular structure and immunocyte distribution of the liver.Here,we provide an overview of thestructure and composition of liver vessels,the threedimensional(3D)imaging of the liver,andthe spatial distribution and immune function of various cell components of the liver.Especially,we emphasize the 3D imaging methods for visualizing fine structure in the liver.Finally,wesummarize and prospect the development of 3D imaging of liver vesels and immune cells.

Successful surgical resection of large hepatocellular carcinoma with portal vein tumor thrombus after conversion therapy with mFOLFOX-HAIC combined with donafenib and sintilimab:two case reports and a literature review

The aim of our study was to evaluate the clinical efficacy of m FOLFOX-HAIC combined with donafenib and sintilimab conversion therapy followed by surgical resection of large hepatocellular carcinoma with portal vein tumor thrombus(PVTT).The clinical data of two patients with large hepatocellular carcinoma who were admitted to the Second Affiliated Hospital of Kunming Medical University were retrospectively collected.Both patients received m FOLFOX-HAIC combined with donafenib and sintilimab conversion therapy,followed by hepatectomy.Clinical data were reported,and clinical efficacy was evaluated.One patient had a 14.5×11.1 cm tumor with a tumor thrombus in the right portal vein.The other patient had a 12.1×8.3 cm tumor with portal and hepatic vein tumor thrombi.Both patients had CNLC stageⅢa prior to conversion therapy,which was reduced to stageⅠb after conversion therapy.Subsequently,the patient underwent open and laparoscopic right hemihepatectomies.Short-term high-intensity conversion therapy with m FOLFOXHAIC combined with donafenib and sintilimab is a feasible and effective treatment for patients with large hepatocellular carcinoma with PVTT.

错配修复基因蛋白在结直肠癌诊治中的临床应用

目的:探讨错配修复基因(mismatch repair gene,MMR)蛋白MLH1、MSH2、MSH6、PMS2在结直肠癌中的表达及在临床中的应用。方法:选取四川省人民医院2015年1月至2016年9月收治的607例结直肠癌患者,采用免疫组织化学法检测手术标本中MMR蛋白的表达情况,研究其与临床病理学的关系,并评价其在Lynch综合征和散发性结直肠癌筛查中的价值。结果:607例患者中MMR表达缺失率为35.58%。MMR蛋白表达缺失的阴性组与表达正常的阳性组,在年龄、性别、肿瘤大小、P53、CD34、D2-40的比较,差异均无统计学意义(P>0.05);两组患者在肿瘤位置、分化程度、TNM分期、淋巴结转移、VEGF、Ki-67的比较,差异均有统计学意义(P<0.05)。联合检测MLH1、MSH2、PSM2、MSH6蛋白可以作为初步筛选Lynch综合征患者的方法。结论:对结直肠癌患者的手术标本进行MMR检测,筛查Lynch综合征患者和家族成员,进行管理及干预,可降低部分人群患结直肠癌的风险。

产权性质、大股东持股比例与债务异质性

以信用债企业为样本，讨论了我国企业的债务异质性特征，并探究了大股东持股比例对企业债务异质性程度的影响，研究发现：我国信用债企业整体债务异质性程度高，但对银行借款的依赖程度较强，债券融资比率呈逐年上升趋势，由于大股东持股比例的增加有助于提高公司信息披露质量，从而拓宽了债务融资渠道，故大股东持股比例与债务异质性程度正相关，但民营企业的大股东更倾向于采用信息欺诈手段来侵占小股东利益.从而削弱了大股东激励效应对信息披露质量的正向影响，故民营企业弱化了大股东持股比例与债务异质性程度的正相关关系.

Activation of paternally expressed imprinted genes in newly derived germline-competent mouse parthenogenetic embryonic stem cell lines

单性生殖的胚胎的茎(足) 细胞为学习位于印的 genomic 下面的分子的机制提供一个珍贵试管内模型系统。然而，足房间的 thepluripotency 和父亲一般地表示的印的基因的表示侧面一直不是充分探索了。在这研究，三根老鼠足房间线被建立，在扩大文化的这些房间的区别潜力被评估。无差别的房间有正常核型和同型结合的染色体，并且表示了 ES-cell-specific 分子的标记。房间在超过 50 个段落以后仍然保持无差别并且展出了 pluripotentdifferentiation 能力。确定的 ES 房间的所有三根线生产了 teratomas；ES 房间的二根线生产了怪物和细菌线传播。而且，在这些房间的父亲一般地表示的印的基因 Snrpn， U2af1-rs1， Peg3，影响， Zfp127， Dlk1 和 Mest 的激活被检测。父亲一般地表示的印的基因被发现在单性生殖的联盟者的胚囊阶段被表示的一些激活胚胎试管内，他们的表示水平与扩大的足细胞培养增加了。而且，我们在足房间的这些父亲一般地表示的印的基因的激活在 ther 的甲基化与一个变化被联系的数据表演鼓舞了差别 methylated 区域。这些调查结果为足房间的 thepluripotency 提供直接证据并且在足房间表明在脱氧核糖核酸甲基化模式和父亲一般地表示的印的基因的激活之间的协会。因此，确定的 ES 房间线为在哺乳动物的开发学习 epigenetic 规定提供一个珍贵模型。

A 14 nucleotide deletion mutation in the coding region of the PpBBX24 gene is associated with the red skin of“Zaosu Red”pear(Pyrus pyrifolia White Pear Group):a deletion in the PpBBX24 gene is associated with the red skin of pear

Red skin is an important quality trait for pear fruits and is determined by the concentration and composition of anthocyanins.The regulatory mechanism underlying anthocyanin accumulation is a popular topic in fruit research.Red mutants are ideal materials for studying the molecular mechanism of color diversity in pear.Although several red pear mutants have been cultivated and are in production,no exact locus containing the responsible genetic mutation has been identified.In this study,by combining the bulked segregant analysis with whole-genome sequencing,we identified a 14 nucleotide deletion mutation in the coding region of the PpBBX24 gene from the red pear mutant“Zaosu Red”.We further verified that the deletion was present only in the red mutant of“Zaosu”and in its red offspring,which was different from that which occurred in other red pear fruits.This deletion results in a coding frame shift such that there is an early termination of the PpBBX24 gene and loss of key NLS and VP domains from PpBBX24.The lost domains may reduce or alter the normal function of PpBBX24.In addition,we found that the transcript levels of the PpMYB10 and PpHY5 genes in red samples were significantly higher than those in green samples,whereas the results for the normal-type PpBBX24 gene were the opposite.We ultimately revealed that the 14 nucleotide deletion mutation in the coding region of the PpBBX24 gene is associated with the red skin of the“Zaosu Red”pear.This finding of somatic mutational events will be helpful for breeding new red pear cultivars and for understanding the regulatory mechanisms involved in pear skin pigmentation.

Integrated DNA-based/biochemical screening for early diagnosis of multiple endocrine neoplasia type 2A (MEN2A)

Multiple endocrine neoplasia type 2A （MEN2A）, a subtype of MEN2, is characterized by medullary thyroid cancer, pheochromocytoma, and primary hyperparathyroidism. A Han Chinese pedigree with MEN2A was investigated following confirmation of the proband＇s diagnosis by pathological findings and DNA/biochemical screening. DNA samples from 4 other family members were collected and exon 5, 8, 10, 11, 13, 16 and 18 of the RET proto-oncogene were sequenced and then analyzed. A missense mutation of TGG （Trp） to TGC （Cys） at codon 634 （the classic MEN2A mutation） in exon 11 of the RET gene was detected in 3 family members, including the proband. Sequencing data were compared with the human gene mutation database. Elevated serum calcitonin level was detected initially; medullary thyroid carcinoma was revealed in the 3 cases and adrenal pheochromocytoma was also found in the proband. Elective operations were successfully performed on the adrenal and thyroid glands because of pheochromocytoma and medullary thyroid carcinoma. Our case study confirms that integrated DNA-based/biochemical screening is crucial for early diagnosis of MEN2A and is helpful in the screening of their relatives. In addition, DNA-based screening may occasionally uncover a previously unknown RET sequence.

Auxin-induced AUXIN RESPONSE FACTOR4 activates APETALA1 and FRUITFULL to promote flowering in woodland strawberry

Flowering time is known to be regulated by numerous pathways,such as the autonomous,gibberellin,aging,photoperiod-mediated,and vernalization pathways.These regulatory mechanisms involve both environmental triggers and endogenous hormonal cues.Additional flowering control mechanisms mediated by other phytohormones,such as auxin,are less well understood.We found that in cultivated strawberry(Fragaria×ananassa),the expression of auxin response factor4(FaARF4)was higher in the flowering stage than in the vegetative stage.Overexpression of FaARF4 in Arabidopsis thaliana and woodland strawberry(Fragaria vesca)resulted in transgenic plants flowering earlier than control plants.In addition,FveARF4-silenced strawberry plants showed delayed flowering compared to control plants,indicating that FaARF4 and FveARF4 function similarly in regulating flowering.Further studies showed that ARF4 can bind to the promoters of the floral meristem identity genes APETALA1(AP1)and FRUITFULL(FUL),inducing their expression and,consequently,flowering in woodland strawberry.Our studies reveal an auxin-mediated flowering pathway in strawberry involving the induction of ARF4 expression.

Woodland strawberry WRKY71 acts as a promoter of flowering via a transcriptional regulatory cascade

The WRKY proteins are a large family of transcription factors that play important roles in stress responses and plant development.However,the roles of most WRKYs in strawberry are not well known.In this study,FvWRKY71 was isolated from the woodland strawberry‘Ruegen’.FvWRKY71 was highly expressed in the shoot apex and red fruit.Subcellular localization analysis showed that FvWRKY71 was located in the nucleus.Transactivation analysis showed that FvWRKY71 presented transcriptional activation activity in yeast.Overexpression of FvWRKY71 in Arabidopsis and woodland strawberry revealed early flowering in the transgenic plants compared with the wild-type control.Gene expression analysis indicated that the transcript levels of the flowering time and development integrator genes AP1,LFY,FT,AGL42,FUL,FPF1,SEP1,SEP2,and SEP3 were increased in FvWRKY71-overexpressing Arabidopsis and strawberry plants compared with the wild-type controls,which may result in accelerated flowering in transgenic plants.Furthermore,FvWRKY71 was proven to directly bind to the W-boxes(TTGACT/C)of the FvFUL,FvSEP1,FvAGL42,FvLFY,and FvFPF1 promoters in vitro and in vivo.Taken together,our results reveal a transcriptional regulatory cascade of FvWRKY71 involved in promoting flowering in woodland strawberry.

Dual role of MdSND1 in the biosynthesis of lignin and in signal transduction in response to salt and osmotic stress in apple

Clarifying the stress signal transduction pathway would be helpful for understanding the abiotic stress resistance mechanism in apple(Malus×domestica Borkh.)and could assist in the development of new varieties with high stress tolerance by genetic engineering.The key NAC transcription factor SND1,which is involved in the lignin biosynthesis process in apple,was functionally analyzed.The results of the stress treatments indicated that MdSND1 could be induced by salt,mannitol and ABA.Compared with wild-type GL-3 plants,MdSND1-overexpressing apple plants with greater antioxidant capacity and lignin were more resistant to salt and simulated osmotic stress,while RNAi plants were more vulnerable.Additionally,molecular experiments confirmed that MdSND1 could regulate the biosynthesis of lignin by activating the transcription of MdMYB46/83.Moreover,genes known to be involved in the stress signal transduction pathway(MdAREB1A,MdAREB1B,MdDREB2A,MdRD29A,and MdRD22)were screened for their close correlations with the expression of MdSND1 and the response to salt and osmotic stress.Multiple verification tests further demonstrated that MdSND1 could directly bind to these gene promoters and activate their transcription.The above results revealed that MdSND1 is directly involved in the regulation of lignin biosynthesis and the signal transduction pathway involved in the response to both salt and osmotic stress in apple.

基于Ansoft的汽车车窗电子开关PCB电磁兼容性的仿真分析

在汽车电子电路硬件设计中,最突出的问题是由于高频高速所造成的电磁兼容性问题。传统的PCB设计由于缺乏高速分析和仿真指导,信号的质量无法得到保证,本文针对汽车车窗电子开关PCB设计中高频干扰所带来的电磁兼容性问题,提出了一种新的“自上而下”的设计方法,采用软件Protel 99SE和Ansoft Designer相结合的方法对PCB板图进行电磁场仿真分析,根据仿真分析曲线对PCB进行电磁兼容性能的优化改进,通过优化改进以提高产品的电磁兼容性能。

The process and mechanism for cesium and rubidium extraction with saponified 4-tert-butyl-2-(a-methylbenzyl) phenol

Cesium(Cs)and rubidium(Rb)separation from brine is an important and application-oriented topic.4-tert-butyl-2-(a-methylbenzyl)phenol(t-BAMBP)has been used for Cs and Rb extraction.However,the traditional extraction technology is base and acid consumed.In the present work,an innovative process for Cs and Rb extraction with t-BAMBP is developed,which consists of saponification,extraction,scrubbing and stripping.Both infrared spectrum and electrostatic potential analysis indicate the hydrogen of phenolic hydroxyl is dissociated from t-BAMBP during saponification and the oxygen of phenolic hydroxyl is the binding site for alkali metal ions.Saponified organic phase shows an excellent extraction effect for Cs^(+)and Rb^(+).The extraction reaches equilibrium in 5 min,with 99.5%Cs^(+)and 46.7%Rb^(+)are loaded into the organic phase in the single-stage extraction.Slope method indicates the structure of the extraction complex is MOR·3ROH(M=Cs^(+),Rb^(+),K^(+)),where the electrostatic attraction between M^(+)and the oxygen of phenolic hydroxyl is dominant,and the cation–p interaction has a significant effect also.The extraction complex of MOR·3ROH dissociates in the acid environment while scrubbing and stripping is completed.The Cs^(+)and Rb^(+)are separated from the mixture phase,the proton H bonds to the phenolic hydroxyl group,and the extractant is regenerated.

Paclitaxel-induced stress granules increase LINE-1 mRNA stability to promote drug resistance in breast cancer cells

Abnormal expression of long interspersed element-1(LINE-1)has been implicated in drug resistance,while our previous study showed that chemotherapy drug paclitaxel(PTX)increased LINE-1 level with unknown mechanism.Bioinformatics analysis suggested the regulation of LINE-1 mRNA by drug-induced stress granules(SGs).This study aimed to explore whether and how SGs are involved in drug-induced LINE-1 increase and thereby promotes drug resistance of triple negative breast cancer(TNBC)cells.We demonstrated that SGs increased LINE-1 expression by recruiting and stabilizing LINE-1 mRNA under drug stress,thereby adapting TNBC cells to chemotherapy drugs.Moreover,LINE-1 inhibitor efavirenz(EFV)could inhibit drug-induced SG to destabilize LINE-1.Our study provides the first evidence of the regulation of LINE-1 by SGs that could be an important survival mechanism for cancer cells exposed to chemotherapy drugs.The findings provide a useful clue for developing new chemotherapeutic strategies against TNBCs.

Early identification of acute hypoxia based on brain NADH fluores cence and cerebral blood flow

Hypoxia is closely related to many diseases and often leads to death.Early detection andidentification of the hypoxia causes may help to promptly determine the right rescue plan andreduce the mortality.We proposed a new multiparametric monitoring method employingmitochondrial reduced nicotinamide adenine dinucleotide(NA DH)fluorescence,regional reflectance,regional cerebral blood flow(CBF),electrocardiography(ECG),and respiration under sixkinds of acute hypoxia in four categories to investigate a correlation bet ween the parametervariances and the hypoxia causes.The variation patterns of the paramet ers were discussed,andthe combination of NADH and CBF may contribute to the identification of the causes of hypoxia.

Scrolled Production of Large-Scale Continuous Graphene on Copper Foils

We report an efficient and economical way for mass production of large-scale graphene films with high quality and uniformity.By using the designed scrolled copper-graphite structure,a continuous graphene film with typical area of 200×39 cm^2 could be obtained in 15 min,and the production rate of the graphene film and space utilization rate of the CVD reactor can reach 520 cm 2⋅min−1 and 0.38 cm−1⋅min−1,respectively.Our method provides a guidance for the industrial production of graphene films,and may also accelerate its large-scale applications.

Cerebrospinal fluid and blood biomarkers in the diagnostic assays of Alzheimer's disease

The anti-amyloid-j(anti-Aβ)fibrils and soluble oligomers antibody aducanumab were approved to effectively slow down the progression of A lzheimer's disease(AD)at higher doses in 2019,reaffirming the therapeutic effects of targeting the core pathology of AD.A timely and accurate diagnosis in the prodromal or pre-dementia stage of AD is essential for patient recruitment,stratifcation,and monitoring of treat ment effects.A D core biomarkers amyloid-B(Aβ1-42),total tau(t-tau),and phosphorylated tau(p-tau)have been clinically validated to reflect AD-type pathological changes through cerebrospinal fuid(CSF)measurement or positron-emission to-mography(PET)and found to have high diagnostic performance for AD identification in the stage of mild cognitive impairment.The development of ultrasensitive immunoassay technology enables AD pathological proteins such as tau and neurofilament light(NFL)to be measured in blood samples.However,combined biomarker detection or targeting multiple biomarkers in immunoassays will increase detection sensitivity and specifcity and improve diagnostic accuracy..This review summarizes and analyzes the performance of current detection methods for early diagnosis of AD,and provides a concept of detection method based on multiple biomarkers instead of a single target,which may become a potential tool for early diagnosis of AD in the future.

VISUALIZATION OF HEAD AND NECK CANCER MODELS WITH A TRIPLE FUSION REPORTER GENE

The development of experimental animal models for head and neck tumors generally rely on the biol uminescence imaging to achieve the dynamic monitoring of the tumor growth and metastasis due to the complicated anatomical structures.Since the bioluminescence imaging is largely affected by the intracellular luciferase expression level and external D-luciferin concentrations,its imaging accuracy requires further confirmation.Here,a new triple fusion reportelr gene,which consists of a herpes simplex virus type 1 thymidine kinase(TK)gene for radioactive imaging,a far-red fuorescent protein(mLumin)gene for fuorescent imaging,and a firefly luciferase gene for bioluminescence imaging,was introduced for in vrivo observation of the head and neck tumors through multi-modality imaging.Results show that fuorescence and bioluminescence signals from mLumin and luciferase,respectively,were clearly observed in tumor cells,and TK could activate suicide pathway of the cells in the presence of nucleotide analog-ganciclovir(GCV),demonstrating the effecti veness of individual functions of each gene.Moreover,subcutaneous and metastasis animal models for head and neck tumors using the fusion reporter gene-expressing cell lines were established,allowing multi-modality imaging in vio.Together,the established tumor models of head and neck cancer based on the newly developed triple fusion reporter gene are ideal for monitoring tumor growth,assessing the drug therapeutic efficacy and verifying the effec-tiveness of new treatments.

Morphological and molecular basis of ovarian serous carcinoma

Serous carcinoma is the most common type of epithelial ovarian cancer. In this review, we provide a com- prehensive picture of ovarian serous cancers from multiple aspects： the first part of this review summarizes the morphological, histological, and immunological signatures of ovarian serous carcinoma; subsequently, we review the history of the evolvement of different grading systems used in ovarian serous cancer; in the end, we focus on characterizing the genetics that underlie the 2-tiered pathways through which ovarian serous cancers are believed to arise： the low-grade and the high-grade pathways.