Identification of S-RNase genotype and analysis of its origin and evolutionary patterns in Malus plants

Identification of the S genotype of Malus plants will greatly promote the discovery of new genes,the cultivation and production of apple,the breeding of new varieties,and the origin and evolution of self-incompatibility in Malus plants.In this experiment,88 Malus germplasm resources,such as Aihuahong,Xishuhaitang,and Reguanzi,were used as materials.Seven gene-specific primer combinations were used in the genotype identification.PCR amplification using leaf DNA produced a single S-RNase gene fragment in all materials.The results revealed that 70 of the identified materials obtained a complete S-RNase genotype,while only one S-RNase gene was found in 18 of them.Through homology comparison and analysis,13 S-RNase genotypes were obtained:S_(1)S_(2)(Aihuahong,etc.),S_(1)S_(28)(Xixian Haitang,etc.),S_(1)S_(51)(Hebei Pingdinghaitang),S_(1)S_(3)(Xiangyangcun Daguo,etc.),S_(2)S_(3)(Zhaiyehaitang,etc.),S_(3)S_(51)(Xishan 1),S_(3)S_(28)(Huangselihaerde,etc.),S_(2)S_(28)(Honghaitang,etc.),S_(4)S_(28)(Bo 11),S_(7)S_(28)(Jiuquan Shaguo),S_(10)S_e(Dongchengguan 13),S_(10)S_(21)(Dongxiangjiao)and S_(3)S_(51)(Xiongyue Haitang).Simultaneously,the frequency of the S gene in the tested materials was analyzed.The findings revealed that different S genes had varying frequencies in Malus resources,as well as varying frequencies between intraspecific and interspecific.S_(3) had the highest frequency of 68.18%,followed by S_(1)(42.04%).In addition,the phylogenetic tree and origin evolution analysis revealed that the S gene differentiation was completed prior to the formation of various apple species,that cultivated species also evolved new S genes,and that the S_(50) gene is the oldest S allele in Malus plants.The S_(1),S_(29),and S_(33) genes in apple-cultivated species,on the other hand,may have originated in M.sieversii,M.hupehensis,and M.kansuensis,respectively.In addition to M.sieversii,M.kansuensis and M.sikkimensis may have also played a role in the origin and evolution of some Chinese apples.

An integrative database and its application for plant synthetic biology research

Plant synthetic biology research requires diverse bioparts that facilitate the redesign and construction of new-to-nature biological devices or systems in plants.Limited by few well-characterized bioparts for plant chassis,the development of plant synthetic biology lags behind that of its microbial counterpart.Here,we constructed a web-based Plant Synthetic BioDatabase(PSBD),which currently categorizes 1677 catalytic bioparts and 384 regulatory elements and provides information on 309 species and 850 chemicals.Online bioinformatics tools including local BLAST,chem similarity,phylogenetic analysis,and visual strength are provided to assist with the rational design of genetic circuits for manipulation of gene expression in planta.We demonstrated the utility of the PSBD by functionally characterizing taxadiene synthase 2 and its quan-titative regulation in tobacco leaves.More powerful synthetic devices were then assembled to amplify the transcriptional signals,enabling enhanced expression offlavivirus non-structure 1 proteins in plants.The PSBD is expected to be an integrative and user-centered platform that provides a one-stop service for diverse applications in plant synthetic biology research.

Associations of cholecystectomy with the risk of colorectal cancer: a Mendelian randomization study

Background:Cholecystectomy is a standard surgery for patients suffering from gallbladder diseases,while the causal effects of cholecystectomy on colorectal cancer(CRC)and other complications are still unknown.Methods:We obtained genetic variants associated with cholecystectomy at a genome-wide significant level(P value<5×10^(-8))as instrumental variables(IVs)and performed Mendelian randomization(MR)to identify the complications of cholecystectomy.Furthermore,the cholelithiasis was also treated as the exposure to compare its causal effects to those of cholecystectomy,and multivariable MR analysis was carried out to judge whether the effect of cholecystectomy was independent of cholelithiasis.The study was reported based on Strengthening the Reporting of Observational Studies in Epidemiology Using Mendelian Randomization guidelines.Results:The selected IVs explained 1.76%variance of cholecystectomy.Our MR analysis suggested that cholecystectomy cannot elevate the risk of CRC(odds ratio[OR]=1.543,95%confidence interval[CI]:0.607–3.924).Also,it was not significant in either colon or rectum cancer.Intriguingly,cholecystectomy might decrease the risk of Crohn’s disease(OR=0.078,95%CI:0.016-0.368)and coronary heart disease(OR=0.352,95%CI:0.164–0.756).However,it might increase the risk of irritable bowel syndrome(IBS)(OR=7.573,95%CI:1.096–52.318).Cholelithiasis could increase the risk of CRC in the largest population(OR=1.041,95%CI:1.010–1.073).The multivariable MR analysis suggested that genetic liability to cholelithiasis could increase the risk of CRC in the largest population(OR=1.061,95%CI:1.002–1.125)after adjustment of cholecystectomy.Conclusions:The study indicated that cholecystectomy might not increase the risk of CRC,but such a conclusion needs further proving by clinical equivalence.Additionally,it might increase the risk of IBS,which should be paid attention to in clinical practice.

A chromosome-level genome assembly reveals that tandem-duplicated CYP706V oxidase genes control oridonin biosynthesis in the shoot apex of Isodon rubescens

The ent-kaurenoids(e.g.,oridonin and enmein)from the Isodon genus(Lamiaceae)are one class of diterpenoids with rich structural diversity and intriguing pharmaceutical activity.In contrast to the well-established gibberellin pathway,oxidative modifications diversifying the ent-kaurene skeleton in Isodon have remained undetermined for half a century.Here we report a chromosome-level genome assembly of I.rubescens,a well-recognized oridonin producer long favored by Asian people as a traditional herb with antitumor effects.The shoot apex was confirmed to be the actual region actively producing ent-kaurene diterpenoids.Through comparative genomics and phylogenetic analyses,we discovered a cluster of tandem-duplicated CYP706V oxygenase-encoding genes located on an ancient genomic block widely distributed in eudicots,whereas almost exclusively emerged in Isodon plants.In the shoot apex,IrCYP706V2 and IrCYP706V7 oxidized the ent-kaurene core in the initial stage of oridonin biosynthesis.Loss of CYP706Vs in other Lamiaceae plants offered an explanation for the specific kaurenoid production in Isodon plants.Moreover,we found that the Isodon genomes encode multiple diterpenoid synthases that are potentially involved in generating diterpenoid diversity.These findings provided new insights into the evolution of the lineage-specific diterpenoid pathway and laid a foundation for improving production of bioactive ent-kaurene-type diterpenoids by molecular breeding and synthetic biology approaches.

Consensus on the application of negative pressure wound therapy of diabetic foot wounds

Because China is becoming an aging society,the incidence of diabetes and diabetic foot have been increasing.Diabetic foot has become one of the main health-related killers due to its high disability and mortality rates.Negative pressure wound therapy(NPWT)is one of the most effective techniques for the treatment of diabetic foot wounds and great progress,both in terms of research and its clinical application,has been made in the last 20 years of its development.However,due to the complex pathogenesis and management of diabetic foot,irregular application of NPWT often leads to complications,such as infection,bleeding and necrosis,that seriously affect its treatment outcomes.In 2020,under the leadership of Burns,Trauma and Tissue Repair Committee of the Cross-Straits Medicine Exchange Association,the writing group for‘Consensus on the application of negative pressure wound therapy of diabetic foot wounds’was established with the participation of scholars from the specialized areas of burns,endocrinology,vascular surgery,orthopedics and wound repair.Drawing on evidence-based practice suggested by the latest clinical research,this consensus proposes the best clinical practice guidelines for the application and prognostic evaluation of NPWT for diabetic foot.The consensus aims to support the formation of standardized treatment schemes that clinicians can refer to when treating cases of diabetic foot.

Chinese expert consensus on conversion therapy for hepatocellular carcinoma(2021 edition)

Recent advances in systemic and locoregional treatments for patients with unresectable or advanced hepatocellular carcinoma(HCC)have resulted in improved response rates.This has provided an opportunity for selected patients with initially unresectable HCC to achieve adequate tumor downstaging to undergo surgical resection,a‘conversion therapy’strategy.However,conversion therapy is a new approach to the treatment of HCC and its practice and treatment protocols are still being developed.Review the evidence for conversion therapy in HCC and develop consensus statements to guide clinical practice.Evidence review:Many research centers in China have accumulated significant experience implementing HCC conversion therapy.Preliminary findings and data have shown that conversion therapy represents an important strategy to maximize the survival of selected patients with intermediate stage to advanced HCC;however,there are still many urgent clinical and scientific challenges for this therapeutic strategy and its related fields.In order to summarize and learn from past experience and review current challenges,the Chinese Expert Consensus on Conversion Therapy for Hepatocellular Carcinoma(2021 Edition)was developed based on a review of preliminary experience and clinical data from Chinese and non-Chinese studies in this field and combined with recommendations for clinical practice.Sixteen consensus statements on the implementation of conversion therapy for HCC were developed.The statements generated in this review are based on a review of clinical evidence and real clinical experience and will help guide future progress in conversion therapy for patients with HCC.

Long-term exposure to genistein inhibits the proliferation of gallbladder cancer by downregulating the MCM complex

Soy isoflavones are natural tyrosine kinase inhibitors closely associated with decreased morbidity and mortality of various tumors.The activation of tyrosine kinases such as ERBB2 is the mechanism by which cholecystitis transforms into gallbladder cancer(GBC),therefore,it is important to investigate the relationship between long-term exposure to soy isoflavones and the occurrence and progression of GBC.This case-control study(n=85 pairs)found that the high level of plasma soy isoflavoneDgenistein(GEN)was associated with a lower risk of gallbladder cancer(≥326.00 ng/m L compared to≤19.30ng/m L,crude odds ratio 0.15,95%CI 0.04–0.59;P for trend=0.016),and that the level of GEN exposure negatively correlated with Ki67 expression in GBC tissue(n=85).Consistent with these results,the proliferation of GBC cells was inhibited in the long-term exposure models of GEN in vitro and in vivo.The long-term exposure to GEN reduced the tyrosine kinase activity of ERBB2 and impaired the function of the PTK6-AKT-GSK3βaxis,leading to downregulation of the MCM complex in GBC cells.In summary,long-term exposure to GEN associated with soy products intake might play a certain role in preventing GBC and even inhibiting the proliferation of GBC cells.

Improved production of docosahexaenoic acid in batch fermentation by newly-isolated strains of Schizochytrium sp.and Thraustochytriidae sp.through bioprocess optimization

Thraustochytrids,rich in docosahexaenoic acid(DHA,C22:6u3),represent a potential source of dietary fatty acids.Yet,the effect of culture conditions on growth and fatty acid composition vary widely among different thraustochytrid strains.Two different thraustochytrid strains,Schizochytrium sp.PKU#Mn4 and Thraustochytriidae sp.PKU#Mn16 were studied for their growth and DHA production characteristics under various culture conditions.Although they exhibited similar fatty acid profiles,PKU#Mn4 seemed a good candidate for industrial DHA fermentation while PKU#Mn16 displayed growth tolerance to a wide range of process conditions.Relative DHA content of 48.5%and 49.2%(relative to total fatty acids),respectively,were achieved on glycerol under their optimal flask culture conditions.Maximum DHA yield(Yp/x)of 21.0%and 18.9%and productivity of 27.6 mg/L-h and 31.9 mg/L-h were obtained,respectively,in 5-L bioreactor fermentation operated with optimal conditions and dual oxygen control strategy.A 3.4-and 2.8-fold improvement of DHA production(g/L),respectively,was achieved in this study.Overall,our study provides the potential of two thraustochytrid strains and their culture conditions for efficient production of DHA-rich oil.

Two-stage Hydrolysis of Invasive Algal Feedstock for Ethanol Fermentation

The overall goal of this work was to develop a saccharification method for the production of third generation biofuel （i.e. bioethanol） using feedstock of the invasive marine macroalga Gracilaria salicornia. Under optimum conditions （120 ℃ and 2% sulfuric acid for 30 min）, dilute acid hydrolysis of the homogenized invasive plants yielded a low concentration of glucose （4.1 mM or 4.3 g glucose/kg fresh algal biomass）. However, two-stage hydrolysis of the homogenates （combination of dilute acid hydrolysis with enzymatic hydrolysis） produced 13.8 g of glucose from one kilogram of fresh algal feedstock. Batch fermentation analysis produced 79.1 g EtOH from one kilogram of dried invasive algal feedstock using the ethanologenic strain Escherichia coil KOll. Furthermore, ethanol production kinetics indicated that the invasive algal feedstock contained different types of sugar, including C5-sugar. This study represents the first report on third generation biofuel production from invasive macroalgae, suggesting that there is great potential for the production of renewable energy using marine invasive biomass.

Cultivation and diversity analysis of novel marine thraustochytrids

Thraustochytrids are a group of unicellular marine heterotrophic protists,and have long been known for their biotechnological potentials in producing squalene,polyunsaturated fatty acids(PUFAs)and other bioactive products.There are less than a hundred known strains from diverse marine habitats.Therefore,the discovery of new strains from natural environments is still one of the major limitations for fully exploring this interesting group of marine protists.At present,numerous attempts have been made to study thraustochytrids,mainly focusing on isolating new strains,analyzing the diversity in specific marine habitats,and increasing the yield of bioactive substances.There is a lack of a systematic study of the culturable diversity,and cultivation strategies.This paper reviews the distribution and diversity of culturable thraustochytrids from a range of marine environments,and describes in detail the most commonly used isolation methods and the control of culture parameters.Furthermore,the perspective approaches of isolation and cultivation for the discovery of new strains are discussed.Finally,the future directions of novel marine thraustochytrid research are proposed.The ultimate goal is to promote the awareness of biotechnological potentials of culturable thraustochytrid strains in industrial and biomedical applications.