Assessing the conservation impact of Chinese indigenous chicken populations between ex-situ and in-situ using genome-wide SNPs

Conservation programs require rigorous evaluation to ensure the preservation of genetic diversity and viability of conservation populations. In this study, we conducted a comparative analysis of two indigenous Chinese chicken breeds, Gushi and Xichuan black-bone, using whole-genome SNPs to understand their genetic diversity, track changes over time and population structure. The breeds were divided into five conservation populations(GS1, 2010, ex-situ;GS2, 2019, ex-situ;GS3, 2019, in-situ;XB1, 2010, in-situ;and XB2, 2019, in-situ) based on conservation methods and generations. The genetic diversity indices of three conservation populations of Gushi chicken showed consistent trends, with the GS3 population under in-situ strategy having the highest diversity and GS2 under ex-situ strategy having the lowest. The degree of inbreeding of GS2 was higher than that of GS1 and GS3. Conserved populations of Xichuan black-bone chicken showed no obvious changes in genetic diversity between XB1 and XB2. In terms of population structure, the GS3 population were stratified relative to GS1 and GS2. According to the conservation priority, GS3 had the highest contribution to the total gene and allelic diversity in GS breed, whereas the contribution of XB1 and XB2 were similar. We also observed that the genetic diversity of GS2 was lower than GS3, which were from the same generation but under different conservation programs(in-situ and ex-situ). While XB1 and XB2 had similar levels of genetic diversity. Overall, our findings suggested that the conservation programs performed in ex-situ could slow down the occurrence of inbreeding events, but could not entirely prevent the loss of genetic diversity when the conserved population size was small, while in-situ conservation populations with large population size could maintain a relative high level of genetic diversity.

S100 protein expression during induced Schwann cell-like cell differentiation of rat bone marrow mesenchymal cells in vitro

BACKGROUND： S100 protein can promote axonal growth. Therefore, transplantation of induced bone marrow-derived mesenchymal stem cells （MSCs） that can secrete S100 may provide a beneficial microenvironment for neural regeneration. OBJECTIVE： To explore the changes in S100 expression during rat MSCs differentiation into Schwann ceils in vitro. DESIGN, TIME AND SETTING： This cytology experiment was performed at the Jiangsu Key Laboratory of Neuroregeneration, Nantong University in China, from January 2006 to May 2007. MATERIALS： The rabbit anti-S100 polyclonal antibody was purchased from Dako, Denmark; the mouse anti-rat S100 monoclonal antibody was purchased from Sigma, USA. METHODS： MSCs were cultured from adult Sprague-Dawley rat femur and tibia. Cell proliferation was determined by the MTT method and CD markers, and cell cycle was measured by flow cytometry. MSCs were induced to differentiate into SC cells. SC cells were stained for S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor. S100 protein and mRNA levels were evaluated by flow cytometry, Western blot, and reverse transcription-polymerase chain reaction. MAIN OUTCOME MEASURES： S100 protein and mRNA expression. RESULTS： MSCs exhibited high amplification potential over eight passages. Prior to induction, the majority of MSCs were at the G0/G1 phase of the cell cycle. After induction, MSCs displayed morphology changes similar to Schwann cells. Moreover, induction increased S100 mRNA levels. Immunofluorescence showed that MSCs expressed S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor at 7 days of induction. Induction also increased S100 protein levels compared with untreated MSCs. CONCLUSION： MSCs are capable of differentiating into Schwann cells-like cells under conditional induction in vitro, with increasing S100 mRNA and protein expression.

Characterization of Xanthomonas citri pv.citri from China based on spoligotyping

Xanthomonas citri pv.citri(Xcc),a gram-negative bacterium,is the causal agent of citrus canker,one of the most devastating diseases threatening the citrus industry worldwide.Understanding the diversity and population structure of Xcc is a prerequisite for disease epidemiological monitoring and effective disease management.Recent characterization of the clustered regularly interspaced short palindromic repeats(CRISPR)/cas(CRISPR-associated proteins genes)system with a highly variable repeat number among species provides a new molecular typing method for bacterial genetic analysis.In this study,we performed systematic in silico analyses of 28 Xcc genomes and identified a credible CRISPR/cas in Xcc strains.Further analysis of CRISPR polymorphisms(repeat number and spacer types)in 129 Xcc A strains collected from six provinces in China identified 15 types of CRISPR arrays with 25 spacers.Phylogenetic analysis of Xcc strains based on the CRISPR locus produced a more reliable and accurate typing result compared to the commonly used loci.In addition,seven associated cas genes—cas1,cas2,cas3,cas4,cas5,cas7(csd2),and cas8(csd1)—were found located adjacent to the CRISPR array.BLAST results showed>99%similarity of seven cas genes among Xcc strains.Homology analysis of spacer sequences showed that six spacers had possible phage/prophage origin.The characterization of the CRISPR/cas system among Xcc strains provided an updated strain typing method for Xcc diversity analysis and yielded a panoramic view of CRISPR evolution for further studies of Xcc-phage interactions.

Penpulimab,an anti-PD-1 antibody,for heavily pretreated metastatic nasopharyngeal carcinoma:a single-arm phase II study

Penpulimab is an anti-programmed cell death-1(PD-1)IgG1 antibody with no Fc gamma receptor(FcγR)binding activity,and thus theoretically reduced immune-related adverse events(irAEs)while maintaining efficacy.This single-arm,phase II trial conducted across 20 tertiary care centers in China enrolled adult patients with metastatic nasopharyngeal carcinoma(NPC)who had failed two or more lines of previous systemic chemotherapy.Patients received 200-mg penpulimab intravenously every 2 weeks(4 weeks per cycle)until disease progression or intolerable toxicities.The primary endpoint was objective response rate(ORR)per RECIST(version 1.1),as assessed by an independent radiological review committee.The secondary endpoints included progression-free survival(PFS)and overall survival(OS).One hundred thirty patients were enrolled and 125 were efficacy evaluable.At the data cutoff date(September 28,2022),1 patient achieved complete response and 34 patients attained partial response.The ORR was 28.0%(95%CI 20.3–36.7%).The response was durable,with 66.8%still in response at 9 months.Thirty-three patients(26.4%)were still on treatment.The median PFS and OS were 3.6 months(95%CI=1.9–7.3 months)and 22.8 months(95%CI=17.1 months to not reached),respectively.Ten(7.6%)patients experienced grade 3 or higher irAEs.Penpulimab has promising anti-tumor activities and acceptable toxicities in heavily pretreated metastatic NPC patients,supporting further clinical development as third-line treatment of metastatic NPC.

罗丹明6G酰肼水杨醛席夫碱锌离子配合物的合成及光致变色性能表征综合实验

设计了一个6学时的综合化学实验,结合光致变色这一化学研究热点,向学生传授有机合成、波谱分析等相关知识。在本实验中,学生通过两步反应合成罗丹明6G酰肼水杨醛席夫碱分子,进而与锌离子结合,得到具有可逆光致变色性能的罗丹明6G酰肼水杨醛席夫碱锌离子配合物。学生通过观察溶液和固体基质中该配合物在光照前后的颜色和紫外光谱变化,了解光致变色现象。结合课堂讲解、文献阅读,了解光致变色现象产生的机理。本实验所需的试剂仪器简单、成本低廉,适于在化学或相关专业的本科实验教学中推广。此外,本实验现象明显,具有较强的趣味性,通过本实验不但可以提高学生的知识水平和专业素养,也可以培养其对化学的兴趣和热爱。

Effects of pituitary-specific overexpression of FSHα/β on reproductive traits in transgenic boars

Background： Follicle-stimulating hormone（FSH） is a gonadotropin synthesized and secreted by the pituitary gland.FSH stimulates follicle development and maturation in females. It also plays an important role in spermatogenesis in males, including humans and mice. However, the effects of FSH on male pigs are largely unknown. In this study,we generated transgenic pigs to investigate the effects of FSHα/β overexpression on reproductive traits in boars.Results： After five transgenic F0 founders were crossed with wide-type pigs, 193 F_1 animals were obtained. Of these,96 were confirmed as transgenic. FSHα and FSHβ m RNAs were detected only in pituitary tissue. Transgenic boars exhibited significantly higher levels of FSHα and FSHβ m RNA, serum FSH, and serum testosterone, compared to ful-sib non-transgenic boars. Significant increases in testis weight, vas deferens diameter, seminiferous tubule diameter,and the number of Leydig cel s were observed, suggesting that the exogenous FSHα/β affects reproductive traits. Final y,transgenic and non-transgenic boars had similar growth performance and biochemical profiles.Conclusions： Pituitary-specific overexpression of FSHα/β genes is likely to impact reproductive traits positively, as indicated by enhancements in serum testosterone level, testis weight, the development of vas deferens, seminiferous tubules, and Leydig cells in transgenic boars. A high level of serum FSH induces secretion of serum testosterone,possibly by boosting the number of Leydig cells, which presumably increases the libido and the frequency of sexual activity in transgenic boars. Our study provides a preliminary foundation for the genetic improvement of reproductive traits in male pigs.

In vitro and in vivo studies on pure Mg, Mg-1Ca and Mg-2Sr alloys processed by equal channel angular pressing

In the present work,the biomedical as-cast pure Mg,Mg–1 Ca and Mg–2 Sr alloys were processed with equal channel angular pressing(ECAP)technique to develop ultrafine microstructure within the materials,and their microstructures,mechanical properties,degradation behavior,cytocompatibility in vitro and biocompatibility in vivo were studied comprehensively.Finer-gained microstructures and improved mechanical properties of these three materials after ECAP were confirmed compared to their as-cast counterparts.Moreover,after ECAP the degradation rate of pure Mg was increased while that of Mg–1 Ca or Mg–2 Sr alloys decreased compared to the ascast counterparts.Additionally,good in vitro cytocompatibility and in vivo biocompatibility of these three materials were revealed by cell cultural tests using osteoblastic MC3 T3-E1 and human mesenchymal stem cells(h MSC)and in vivo animal tests at the lateral epicondyle of SD-rats’femur.This study offers an alternative powerful avenue to achieve good comprehensive properties of magnesium-based biodegradable metals.It might also help to extend the applied range of magnesium-based biodegradable metals in orthopedic field.

Efficient production of pronuclear embryos in breeding and nonbreeding season for generating transgenic sheep overexpressing TLR4

Background： Brucella is a zoonotic Gram-negative pathogen that causes abortion and infertility in ruminants and humans. TLR4 is the receptor for LPS which can recognize Brucella and initiate antigen-presenting cell activities that affect both innate and adaptive immunity. Consequently, transgenic sheep over-expressing TLR4 are an suitable model to investigate the effects of TLR4 on preventing Brucellosis. In this study, we generated transgenic sheep overexpressing TLR4 and aimed to evaluate the effects of different seasons（breeding and non-breeding season） on superovulation and the imported exogenous gene on growth.Results： In total of 43 donor ewes and 166 recipient ewes in breeding season, 37 donor ewes and 144 recipient ewes in non-breeding season were selected for super-ovulation and injected embryo transfer to generate transgenic sheep.Our results indicated the no. of embryos recovered of donors and the rate of pronuclear embryos did not show any significant difference between breeding and non-breeding seasons（P 〉 0.05）. The positive rate of exogenous TLR4 tested were 21.21 % and 22.58 % in breeding and non-breeding season by Southern blot. The expression level of TLR4 in the transgenic sheep was 1.5 times higher than in the non-transgenic group（P 〈 0.05）. The lambs overexpressing TLR4 had similar growth performance with non-transgenic lambs, and the blood physiological parameters of transgenic and non-transgenic were both in the normal range and did not show any difference.Conclusions： Here we establish an efficient platform for the production of transgenic sheep by the microinjection of pronuclear embryos during the whole year. The over-expression of TLR4 had no adverse effect on the growth of the sheep.

Research progress on immune checkpoint inhibitors in neoadjuvant therapy for gastric cancer

In recent years,immune checkpoint inhibitors(ICIs)have become an important treatment strategy for advanced gastric cancer.Immunotherapy has gradually transitioned from a later-line to a first-line treatment for advanced gastric cancer.Simultaneously,more and more researchers have begun to pay attention to whether immunotherapy can be used for resectable gastric cancer.The current use of ICIs in the neoadjuvant treatment of gastric cancer is still in its exploratory stage,with a number of clinical trials currently underway.However,the available data show good application prospects.This article reviews the research progress on ICIs in the neoadjuvant therapy for gastric cancer and evokes some unresolved problems.

Design of single-phased magnesium alloys with typically high solubility rare earth elements for biomedical applications:Concept and proof

Rare earth elements(REEs)have been long applied in magnesium alloys,among which the mischmetal-containing WE43 alloy has already got the CE mark approval for clinical application.A considerable amount of REEs(7 wt%)is needed in that multi-phased alloy to achieve a good combination of mechanical strength and corrosion resistance.However,the high complex RE addition accompanied with multiple second phases may bring the concern of biological hazards.Single-phased Mg-RE alloys with simpler compositions were proposed to improve the overall performance,i.e.,“Simpler alloy,better performance”.The single-phased microstructure can be successfully obtained with typical high-solubility REEs(Ho,Er or Lu)through traditional smelting,casting and extrusion in a wide compositional range.A good corrosion resistance with a macroscopically uniform corrosion mode was guaranteed by the homogeneously single-phased microstructure.The bimodal-grained structure with plenty of sub-grain microstructures allow us to minimize the RE addition to<1 wt%,without losing mechanical properties.The single-phased Mg-RE alloys show comparable mechanical properties to the clinically-proven Mg-based implants.They exhibited similar in-vitro and in-vivo performances(without local or systematic toxicity in SD-rats)compared to a high purity magnesium.In addition,metal elements in our single-phased alloys can be gradually excreted through the urinary system and digestive system,showing no consistent accumulation of RE in main organs,i.e.,less burden on organs.The novel concept in this study focuses on the simplification of Mg-RE based alloys for biomedical purpose,and other biodegradable metals with single-phased microstructures are expected to be explored.

Perceiving the connection between the bone healing process and biodegradation of biodegradable metal implants through precise bioadaptability principle

The implants made of metallic biomaterials help healing the bone fracture but also affect the bone repair process.As proposed in Matter 4(2021)2548–2650 by Wang et al.,a precisely adaptable biomaterial ought to recapitulate the targeted tissue with spatiotemporal precision and hierarchical accuracy,ranging from atoms and molecules(genes,proteins,etc.)to cells(including organelles)and to tissues and organs.In comparison to traditional bio-inert metallic bone implants such as Co-based alloys and Ti alloys,biodegradable metal(Mg and Zn alloys)bone implants had been developed and might arise many unexpected variables in the bone repair,due to their bioactive nature.In this paper,the bone repair without and with the presence of metallic implants is compared.Thereafter,the perspectives concerning the interactions between the bone tissues and biodegradable metal implants are put forward,and how to better mimic in vivo biodegradation by in vitro experiments is proposed for further research and development of biodegradable metals.

Rational design of Prussian blue analogue-derived manganese-iron oxides-based hybrids as high-performance Li-ion-battery anodes

The unique components and architecture of Prussian blue analogous(PBAs) offer great potential for the construction of various functional nanostructures. Herein, we reported the preparation of a series of Mn–Fe oxides-based hybrids using Mn–Fe PBA as a template and an organic carbon source by calcination.The study focuses on revealing the interaction between the microstructure and electrochemical performance of the products obtained at different calcination temperatures. Notably, the as-derived porous Fe–Fe0.33Mn0.67O/C nanocubes(i.e., M600) exhibited the best rate capability and cycle life compared with other samples(~890 m Ah/g at 0.1 A/g, 626.8 m Ah/g after 1000 cycles at 1.0 A/g with a 99% capacity retention). These can be attributed to the fact that the porous structure provides shorter Li+diffusion path and promotes the penetration of electrolyte. Besides, the N-doped C formed by the carbonization of organic ligands can buffer the volume change and prevent the aggregation of Fe_(0.33)Mn_(0.67)O nanoparticles during the discharge/charge cycles. Moreover, the presence of metallic Fe enhances the conductivity and the electrochemical activity, which accelerates the electrochemical reactions. Therefore, reasonable design of microstructure and compositions of functional nanocomposites is the key to obtain ideal electrochemical properties.

Rational design of self-sacrificial template derived quasi-Cu-MOF composite as anodes for high-performance lithium-ion batteries

MOF-based composites have aroused widespread concern due to their controllable morphology and pore characteristics.Nevertheless,the poor conductivity and volume expansion hinder its practical application in LIBs.Herein a classical structure HKUST-1,as the precursor,was used to fabricate quasi-Cu-MOF composite through a facile thermal decomposition strategy.The results showed that quasi-Cu-MOF composite had superior reversible specific capacity(627.5 m Ah/g at 100 m A/g)and outstanding cycle stability(514.6 m Ah/g at 500 m A/g after 400 cycles)as anodes for LIBs.The results demonstrated that the lowtemperature calcination strategy played a significant role in morphology retaining during cycling and the derived copper framework play a crucial part in conductivity improvement.This work is helpful to the design of high-performance electrodes with advanced three-dimensional hierarchical structures.

超声评估远端指间关节指伸肌腱附着点炎对炎性关节病的临床诊断价值

目的探讨超声识别远端指间关节(DIP)指伸肌腱附着点炎对炎性关节病(IA)的诊断价值。方法纳入2021年6月至2022年6月于福建医科大学附属第一医院确诊的IA患者140例(IA组),其中类风湿关节炎(RA)患者102例(RA组),脊柱关节炎(SpA)患者38例(SpA组);以及骨性关节炎(OA)患者104例(OA组)。对比IA组与OA组DIP指伸肌腱附着点的灰阶与能量多普勒超声特征,采用单因素及多因素Logistic回归分析IA的独立预测指标。并对RA组与SpA组的DIP指伸肌腱附着点炎超声检出率进行比较分析。结果多因素Logistic分析显示,DIP指伸肌腱附着点炎灰阶超声特征(肌腱厚度、骨质增生、骨侵蚀)不是IA的独立预测指标(P均>0.05);DIP指伸肌腱附着点炎的能量多普勒超声信号阳性是IA的独立预测指标(OR值:23.20,95%CI:10.37~51.92,P<0.01),可作为鉴别IA与OA的超声特征。RA组与SpA组的DIP附着点炎超声检出率分别为67.64%(69/102)、84.21%(32/38),二者差异无统计学意义(χ^(2)=3.13,P>0.05)。结论超声检测DIP指伸肌腱附着点炎,尤其是能量多普勒超声对于IA与OA的鉴别诊断,具有一定的临床价值。

Hydrogen bond-bridged phosphorene flexible film for photodynamic inhibiting Staphylococcus aureus

Antibiotics are a widely used and effective treatment for bacterial infections.However,bacteria can gradually evolve during infection,leading to developing resistance to antibiotics,which renders previously effective treatments ineffective.Finding a useful and convenient manner to treat bacterial infections is a great challenge.Here,we report a flexible hydrogen-bond-bridged phosphorene film with photodynamic antibacterial properties and excellent mechanical properties,fabricated from electrochemical exfoliation of black phosphorus(BP).When illuminated under 700 nm light,the hydrogen bond-bridged phosphorene flexible film is capable of converting ground-state triplet oxygen(O_(2))into excited-state singlet oxygen(^(1)O_(2)),destroying the structure of the membrane of Staphylococcus aureus,and eventually leading to bacterial death,via breaking the C=C of unsaturated fatty acids within the bacterial cell membrane after the reaction between^(1)O_(2)and unsaturated fatty acids,thus realizing a highly efficient antibacterial approach,which is supported by gas chromatography-mass spectrometry(GC-MS)technique.This work establishes an effective phototherapy platform for treating bacterial traumatic infections.

Dietary supplementation with Clostridium butyricum improves growth performance of broilers by regulating intestinal microbiota and mucosal epithelial cells

Clostridium butyricum has been widely considered an antibiotic substitute in recent years.It can promote growth performance,improve the immune response and enhance the intestinal barrier function of the host.In the present study,1-d-old Arbor Acres(AA)broilers were fed C.butyricum(1×109 cfu/kg)for 28 d.The transcriptomic characteristics of epithelial cells of the cecal mucosa were determined by RNA-sequence,and the cecal microbiota composition was explored by 16 S ribosomal RNA gene sequencing.The changes in the intestinal mucosa of broilers were then analyzed by tissue staining.Gene Ontology(GO)annotations identified substance transport and processes and pathways that might participate in intestinal development and cell viability.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis revealed that the differentially expressed genes are involved in numerous pathways related to amino acid and vitamin metabolism and antioxidant and defensive functions,among others.The relative expression of some genes associated with intestinal barrier function(claudins 2,15,19,and 23,tight junction proteins 1,2,and 3 and mucin 1)was significantly increased in the treatment group(P<0.05 or P<0.01).Moreover,the proportion of Firmicutes was higher in the C.butyricum-treated group,whereas the proportion of Proteobacteria was higher in the control group.At the genus level,the relative abundances of Butyricicoccus and Lactobacillus,among other bacteria,were increased after C.butyricum supplementation.The tissue staining analysis showed that the cecal mucosa of broilers was significantly ameliorated after the addition of C.butyricum(P<0.05 or P<0.01).These results showed that dietary supplementation with C.butyricum can enhance the antioxidant capacity,mucosal barrier function,and stabilize the cecal microbiota,resulting in improving the growth performance.

Genome mining combined metabolic shunting and OSMAC strategy of an endophytic fungus leads to the production of diverse natural products

Endophytic fungi are promising producers of bioactive small molecules.Bioinformatic analysis of the genome of an endophytic fungus Penicillium dangeardii revealed 43 biosynthetic gene clusters,exhibited its strong ability to produce numbers of secondary metabolites.However,this strain mainly produce rubratoxins alone with high yield in varied culture conditions,suggested most gene clusters are silent.Efforts for mining the cryptic gene clusters in P.dangeardii,including epigenetic regulation and one-strain-many-compounds(OSMAC)approach were failed probably due to the high yield of rubratoxins.A metabolic shunting strategy by deleting the key gene for rubratoxins biosynthesis combining with optimization of culture condition successfully activated multiple silent genes encoding for other polyketide synthases(PKSs),and led to the trace compounds detectable.As a result,a total of 23 new compounds including azaphilone monomers,dimers,turimers with unprecedented polycyclic bridged heterocycle and spiral structures,as well as siderophores were identified.Some compounds showed significant cytotoxicities,anti-inflammatory or antioxidant activities.The attractive dual PKS s gene clusters for azaphilones biosynthesis were mined by bioinformatic analysis and overexpression of a pathway specific transcription factor.Our work therefor provides an efficient approach to mine the chemical diversity of endophytic fungi.

A Turn-On Fluorescence Chemosensor for Cyanide in Aqueous Media Based on a Nucleophilic Addition Reaction

We synthesized a new cyanide （CN-） chemosensor CX based on a nucleophilic addition reaction prompted by cyanide ion, which could be used for highly selective and sensitive fluorescence turn-on detection of cyanide in aqueous media. The CX showed selective fluorescence recognition for CN-, the miscellaneous competitive anions （F-, Cl-, Br-, I-, AcO-, H2PO4, HSO4, ClO4, S2-, PO4^3-, CO3^2- and SCN- ） did not lead to any significant interference. The detection limit of the sensor towards CN is 1.15 × 10^-7 mol.L- 1. The sensor has been successfully applied to estimate the cyanide ion in seeds of cherries. Test strips based on CX were fabricated, which could be used as a convenient and efficient CN test kit to detect CN in aqueous solution for ＂in-the-field＂ measurement.

民族认同构建视角下雅利安主义在伊朗的散布与影响

雅利安主义在欧洲的起源及其在伊朗的散布是世界民族国家史上的一个奇特现象。脱胎于欧洲东方语言学的雅利安种族主义思想自19世纪后期传入伊朗后,经历本土化和纳粹德国的影响,而成为伊朗界定自我、区别他者的重要认同工具。作为自我东方化的错位认同,伊朗雅利安主义以所谓共同的雅利安身份而将西方视为“我们”,以非雅利安的阿拉伯人等族群为“他者”,渲染雅利安种族优越与伊朗文明的辉煌。它迎合了伊朗巴列维王朝民族国家构建与西方化改革的需要,成为伊朗的官方意识形态并得以广泛散布传播。1979年伊斯兰革命后,伊朗雅利安主义经过一段时期的边缘化后再次公开出现,在官方伊斯兰主义主导下继续参与伊朗民族认同的构建,但雅利安主义和伊斯兰主义二者内在的排他性使得伊朗并未走出民族认同困境。雅利安主义对伊朗的民族认同、族群关系和对外交往都产生了重要的影响。

E3 ligase Herc4 regulates Hedgehog signalling through promoting Smoothened degradation

Hedgehog(Hh)signalling plays conserved roles in controlling embryonic development;its dysregulation causes many diseases including cancers.The G protein-coupled receptor Smoothened(Smo)is the key signal transducer of the Hh pathway,whose posttranslational regulation has been shown to be critical for its accumulation and activation.Ubiquitination has been reported an essential posttranslational regulation of Smo.Here,we identify a novel E3 ligase of Smo,Herc4,which binds to Smo,and regulates Hh signalling by controlling Smo ubiquitination and degradation.Interestingly,our data suggest that Herc4-mediated Smo degradation is regulated by Hh in PKA-primed phosphorylation-dependent and independent manners.