YOU CANNOT SUCCUMB TO CHOLERA,unless the sewage system breaks down-likewise deaths from violence vanish,as long as emotional support networks are working properly.Gut hygiene eliminates the one,societal hygiene the ot...YOU CANNOT SUCCUMB TO CHOLERA,unless the sewage system breaks down-likewise deaths from violence vanish,as long as emotional support networks are working properly.Gut hygiene eliminates the one,societal hygiene the other.Cholera was an inexorable plague-until Germ Theory was accepted and implemented.Violence too,will persist-until we agree and accept its equivalent in Societal Theory.The scientific method,especially Quantum Physics,sabotages philosophy-it cannot cope with the living fact that the elephant in the room is alive.Quantum Mechanics describes itself as being(1)more incomprehensible,(2)weirder,and(3)more uncertain than anything we’ve met before.But that’s chickenfeed,compared to what you find in clinical medicine,which though portrayed as being intrinsically woolly and subjective,wilfully aims to enhance life,whatever that might be.展开更多
Primordial germ cells(PGCs)are the stem-cell population of adult animal gametes,which develop into sperm or eggs.It can be propagated in vitro and injected into the host chicken for genome editing to obtain germline c...Primordial germ cells(PGCs)are the stem-cell population of adult animal gametes,which develop into sperm or eggs.It can be propagated in vitro and injected into the host chicken for genome editing to obtain germline chimeric chicken.However,it has the limitation that the host embryo contains endogenous PGCs,which raises complications,resultantly donor PGCs fail to compete,and transmission efficiency reduced.Therefore,to increase the transmission efficiency,we generated a novel sterile chicken with the inducible elimination of endogenous PGCs in the host.This is the first study that applied the herpes simplex virus thymidine kinase(HSV-TK)cell ablation system in avian.CRISPR/Cas9-mediated homology-directed repair was performed to localize the HSV-TK suicide gene to the last exon of the deleted in azoospermialike(DAZL)gene,and ganciclovir(GCV)was added to induce the apoptosis in the germ cells of the host embryo.The sterilized host embryo introduced genome-edited PGCs to produce chimeric chicken carrying exogenous germ cells only.It was observed that the germline transmission efficiency was 100%achieved,and the obtained chicks were purely from donor breeds.The technologies established in the current study have important applications in germplasm conservation and gene editing in chicken.展开更多
Peripheral nerve injuries result in the rapid degeneration of distal nerve segments and immediate loss of motor and sensory functions;behavioral recovery is typically poor.We used a plasmalemmal fusogen,polyethylene g...Peripheral nerve injuries result in the rapid degeneration of distal nerve segments and immediate loss of motor and sensory functions;behavioral recovery is typically poor.We used a plasmalemmal fusogen,polyethylene glycol(PEG),to immediately fuse closely apposed open ends of severed proximal and distal axons in rat sciatic nerves.We have previously reported that sciatic nerve axons repaired by PEG-fusion do not undergo Wallerian degeneration,and PEG-fused animals exhibit rapid(within 2–6 weeks)and extensive locomotor recovery.Furthermore,our previous report showed that PEG-fusion of severed sciatic motor axons was non-specific,i.e.,spinal motoneurons in PEG-fused animals were found to project to appropriate as well as inappropriate target muscles.In this study,we examined the consequences of PEG-fusion for sensory axons of the sciatic nerve.Young adult male and female rats(Sprague–Dawley)received either a unilateral single cut or ablation injury to the sciatic nerve and subsequent repair with or without(Negative Control)the application of PEG.Compound action potentials recorded immediately after PEG-fusion repair confirmed conduction across the injury site.The success of PEG-fusion was confirmed through Sciatic Functional Index testing with PEG-fused animals showing improvement in locomotor function beginning at 35 days postoperatively.At 2–42 days postoperatively,we anterogradely labeled sensory afferents from the dorsal aspect of the hindpaw following bilateral intradermal injection of wheat germ agglutinin conjugated horseradish peroxidase.PEG-fusion repair reestablished axonal continuity.Compared to unoperated animals,labeled sensory afferents ipsilateral to the injury in PEG-fused animals were found in the appropriate area of the dorsal horn,as well as inappropriate mediolateral and rostrocaudal areas.Unexpectedly,despite having intact peripheral nerves,similar reorganizations of labeled sensory afferents were also observed contralateral to the injury and repair.This central reorganization may contribute to the improved behavioral recovery seen after PEG-fusion repair,supporting the use of this novel repair methodology over currently available treatments.展开更多
Tissue interactions play a crucial role in tooth development.Notably,extracellular vesicle-mediated interactions between the mandible and tooth germ are considered essential.Here,we revealed that mandible extracellula...Tissue interactions play a crucial role in tooth development.Notably,extracellular vesicle-mediated interactions between the mandible and tooth germ are considered essential.Here,we revealed that mandible extracellular vesicles could modulate the proliferation and differentiation of dental mesenchymal cells by regulating the histone demethylase KDM2B.Further investigation showed that mandible derived extracellular vesicles could deliver miR-206 to KDM2B,thereby regulating tooth development.An animal study demonstrated that the miR-206/KDM2B pathway affected tooth morphogenesis and mineralization after eight weeks of subcutaneous transplantation in nude mice.In conclusion,this study suggested that the mandible played a critical role in tooth morphogenesis and mineralization,which could be a potential therapeutic target for abnormal tooth development and an alternative model for tooth regeneration.展开更多
The cell fate of primordial germ cell(PGC)in zebrafish is pre-determined by maternally deposited germ plasm,which is packaged into ribonucleoprotein complex in oocytes and inherited into PGC-fated cells in embryos.How...The cell fate of primordial germ cell(PGC)in zebrafish is pre-determined by maternally deposited germ plasm,which is packaged into ribonucleoprotein complex in oocytes and inherited into PGC-fated cells in embryos.However,the maternal factors regulating the assembly of germ plasm and PGC development remain poorly understood.In this study,we report that the maternal transcription factor Znf706 regulates the assembly of germ plasm factors into a granule-like structure localized perinuclearly in PGC during migration.Maternal and zygotic mutants of znf706 exhibit deficient germ plasm scattering at the early embryonic stage,decreased PGC numbers with some mislocation during PGC migration,and a lower female ratio in adulthood.Notably,the implementation of Znf706 CUT&Tag and RNA-seq on immature oocytes uncovers that Znf706 in stage I oocytes may promote transcription of several mitochondrial genes in addition to other functions.Hence,we propose that Znf706 is implicated in germ plasm assembly and PGC development in zebrafish.展开更多
Germplasm resources are essential for the sustainable development of biodiversity and husbandry of local chickens, as well as for the breeding and industry of superior quality chickens. Unfortunately, many local and i...Germplasm resources are essential for the sustainable development of biodiversity and husbandry of local chickens, as well as for the breeding and industry of superior quality chickens. Unfortunately, many local and indigenous chicken breeds are at risk of declining numbers, emphasizing the need to conserve breed resources for endangered chickens. Primordial germ cells(PGCs) are crucial for preserving germplasm resources by inheriting genetic information from parents to offspring and ensuring stability of genetic material between germlines. In this study,PGCs were isolated from chicken embryos' gonads and cultured in FAcs medium without feeder cells. Over a period of approximately 40 d, the cells proliferated to a number of up to 10^(6), establishing various cell lines. Particularly, 18 PGC lines were created from Rugao Yellow chicken and Shouguang chicken, with an efficiency ranging from 39.1 to 45%. Furthermore, PGCs that had been cultured for 40 passages exhibited typical PGC characteristics, suchas glycogen staining reaction, and expression of pluripotency and reproductive markers. These results confirmthat PGCs maintain stem cell properties even after long-term in vitro culture. Additionally, PGCs cryopreserved for up to 120 d remained viable, maintained typical PGC morphologies, and possessed stable cell proliferation ability. Through intravascular injection into chicken embryos, green fluorescent protein(GFP)-PGCs were found in the recipient embryos' gonads and could develop into gametes to produce offspring, indicating that even after extended culture, PGCs retain their migratory and lineage-transmitting capabilities. This research offers valuable insights into the in vitro cultivation and preservation of PGCs of Chinese indigenous chickens. The findings of this study can be applied in transgenic chicken production and the preservation of genetic resources of indigenous chicken breeds.展开更多
Dear Editor,I would like to congratulate Mamsen et al.i on their extensive and scientifically valuable work.I analyzed their raw data presented in Table 1 of the original article from a different perspective and disco...Dear Editor,I would like to congratulate Mamsen et al.i on their extensive and scientifically valuable work.I analyzed their raw data presented in Table 1 of the original article from a different perspective and discovered an effect not mentioned in the article.My analysis showed that luteinizing hormone(LH)levels are significantly lower in patients at high infertility risk(HIR),whose testes lack A dark(Ad)spermatogonia and display an abnormal ratio of germ cells per crosssectional tubule(G/T).展开更多
Objective:Testicular germ cell tumors(TGCTs)represent the most common malignancy among young men aged 20–40 years.Transglutaminase 7(TG7),encoded by TGM7,is a poorly characterized enzyme whose function in TGCT remain...Objective:Testicular germ cell tumors(TGCTs)represent the most common malignancy among young men aged 20–40 years.Transglutaminase 7(TG7),encoded by TGM7,is a poorly characterized enzyme whose function in TGCT remains unknown.This study aimed to assess TG7 expression in clinical specimens and investigate its functional role in a testicular germ cell tumor cell line(NT2/D1).Methods:TG7 protein expression was evaluated in clinical testicular tissue samples via immunohistochemistry(IHC)and immunofluorescence(IF).Functional analysis was conducted in the NT2/D1 human testicular cancer cell line usingDicer-substrate small interferingRNAs(DsiRNAs)targeting TG7.Gene knockdown efficiency was confirmed by reverse transcription quantitative PCR(qRT-PCR),and protein suppression was validated by immunofluorescence.Cell viability was assessed using the MTT assay.The expression of inflammation and apoptosis-related genes was quantified via qRT-PCR.Results:TG7 expression was significantly elevated in testicular germ cell tumor tissues,showing approximately a 4.5-fold increase compared to normal testis,with strong localization in tumor nests and stromal compartments.In NT2/D1 cells,TG7 silencing using 20 nM DsiRNA3 led to a dose-dependent reduction in cell viability,with up to 48%inhibition observed at 200 nM(MTT assay,****p<0.0001).qRT-PCR analysis revealed significant upregulation of IL6(3.2-fold),TNFα(2.8-fold),and CASP3(2.5-fold)mRNA levels following TG7 knockdown(p<0.0001),while p53 expression remained unchanged.These findings support TG7’s role in modulating tumor cell survival,inflammation,and apoptosis via p53-independent pathways.Conclusion:Collectively,TG7 is significantly overexpressed in TGCT tissues and supports tumor cell viability in vitro.This study establishes TG7 as a novel biomarker and therapeutic target in testicular cancer,laying the groundwork for future studies on TG7-targeted interventions.展开更多
Dear Editor,We are much obliged that Hadziselimovic1 has used our data2 to highlight the substantial proportion of boys with cryptorchidism where gonadotropin insufficiency is an important factor related to the pathog...Dear Editor,We are much obliged that Hadziselimovic1 has used our data2 to highlight the substantial proportion of boys with cryptorchidism where gonadotropin insufficiency is an important factor related to the pathogenesis.We have recently presented a study on a series of 453 consecutive boys with bilateral nonsyndromic cryptorchidism,in which we conducted hormonal evaluations and assessed germ cell numbers in testicular biopsies.3 In this series,45%of the boys were classified as having gonadotropin insufficiency.3 Identifying these patients at the time of surgery is important.A recent follow-up study of 208 boys with nonsyndromic bilateral cryptorchidism from our department showed that the boys with gonadotropin insufficiency had an impaired fertility potential after surgery compared to boys with an intact hypothalamus–pituitary–gonadal feedback mechanism.4 In a review from 2022,Hadziselimovic5 suggested that infertility in patients diagnosed with cryptorchid testes is a consequence of a hormonal deficiency rather than temperature-induced cellular damage.展开更多
Primordial germ cells(PGCs)are the precursors of germline that are specified at the embryonic stage.Recent studies reveal that humans employ different mechanisms for PGC specification compared with model organisms suc...Primordial germ cells(PGCs)are the precursors of germline that are specified at the embryonic stage.Recent studies reveal that humans employ different mechanisms for PGC specification compared with model organisms such as mice.Moreover,the specific regulatory machinery remains largely unexplored,mainly due to the inaccessible nature of this complex biological process in humans.Here,we curate and integrate multi-omics data,including 581 RNA-seq,54 ATAC-seq,45 ChIP-seq,and 69 single-cell RNA-seq samples from different stages of human PGC development to recapitulate the precisely controlled and stepwise process,presenting an atlas in the human PGC database(hPGCdb).With these uniformly processed data and integrated analyses,we characterize the potential key transcription factors and regulatory networks governing human germ cell fate.We validate the important roles of some of the key factors in germ cell development by CRISPRi knockdown.We also identify the soma-germline interaction network and discover the involvement of SDC2 and LAMA4 for PGC development,as well as soma-derived NOTCH2 signaling for germ cell differentiation.Taken together,we have built a database for human PGCs(http://43.131.248.15:6882)and demonstrate that hPGCdb enables the identification of the missing pieces of mechanisms governing germline development,including both intrinsic and extrinsic regulatory programs.展开更多
BACKGROUND Urinary tract infections(UTIs)in kidney transplant patients are a challenge.AIM To evaluate epidemiology,clinical status,therapeutic management,and clinical outcome of kidney transplant patients in a univer...BACKGROUND Urinary tract infections(UTIs)in kidney transplant patients are a challenge.AIM To evaluate epidemiology,clinical status,therapeutic management,and clinical outcome of kidney transplant patients in a university hospital for UTI.METHODS We conducted a retrospective observational study,enrolling all kidney transplant patients hospitalized for UTI,with the objective to evaluate the epidemiology,clinical status,therapeutic management,and clinical outcome of kidney transplant patients.RESULTS From our real-life experience,infection with multidrug-resistant germs was confirmed as a risk factor for the severe evolution of the infection.At the same time,the re-evaluation of immunosuppressive therapy could be an important therapeutic strategy in the course of infection.CONCLUSION Prompt initiation of empiric antibiotic therapy upon initiation of microbiological investigations may reduce the risk of severe infection progression.展开更多
Dug well water in the working area of the Jongaya Community Health Center often fails to meet clean water quality standards due to low pH(6.1)and high organic matter content(14 mg/L),exceeding the limits set by Indone...Dug well water in the working area of the Jongaya Community Health Center often fails to meet clean water quality standards due to low pH(6.1)and high organic matter content(14 mg/L),exceeding the limits set by Indonesian Ministry of Health Regulation No.32 of 2017,which poses a risk of digestive disorders and skin diseases.Addressing this gap,this study evaluates the effectiveness of quicklime(CaO)and aluminum sulfate[Al_(2)(SO_(4))_(3)]in improving dug well water quality through a quasi-experimental pretest-posttest control group design.Water samples were purposively collected from contaminated wells and treated using quicklime,aluminum sulfate,and their combination at doses of 1 g,10 g,and 20 g,each replicated three times,with pH and organic matter content measured before and after treatment and analyzed using a paired t-test.Results showed that 1 g of CaO significantly increased pH from 6.1 to 7.88(p<0:05),meeting the clean water quality threshold(≥6:5),while 20 g of Al_(2)(_(S)O_(4))_(3)reduced organic matter content from 170.91 mg/L to 126.11 mg/L,and the combined treatment achieved the best outcome,with a 46.75%reduction in organic matter and a pH increase to 8.63,both meeting standards.All treatments were statistically significant(p<0:05),indicating that CaO and Al_(2)(SO_(4))_(3)are effective for improving dug well water quality in at-risk communities.The findings highlight the urgency of promoting proper application and optimal dosing,supported by long-term monitoring,and provide novel evidence on the combined use of these chemicals for addressing a local public health challenge.展开更多
Flight feathers represent a hallmark innovation of avian evolution.Recent comparative genomic analyses identified a 284 bp avian-specific highly conserved element(ASHCE)located within the eighth intron of the SIM bHLH...Flight feathers represent a hallmark innovation of avian evolution.Recent comparative genomic analyses identified a 284 bp avian-specific highly conserved element(ASHCE)located within the eighth intron of the SIM bHLH transcription factor 1(Sim1)gene,postulated to act as a cis-regulatory element governing flight feather morphogenesis.To investigate its functional significance,genome-edited(GE)primordial germ cell(PGC)lines carrying targeted ASHCE deletions were generated using CRISPR/Cas9-mediated editing,with germline chimeric males subsequently mated with wild-type(WT)hens to obtain GE progeny.The resulting GE chickens harbored 257-260 bp deletions,excising approximately half of the Sim1-ASHCE sequence.Reverse transcription-quantitative real-time polymerase chain reaction(RT-qPCR)analysis showed an average 0.32-fold reduction in Sim1 expression in the forelimbs of GE embryos at day 8(E8)compared to WT counterparts.Despite this,GE chickens developed structurally normal flight and tail feathers.In situ hybridization localized Sim1 expression to the posterior mesenchyme surrounding flight feather buds in E8 WT embryos,but not within the buds themselves.These results suggest that partial deletion of Sim1-ASHCE,despite diminishing Sim1 expression,does not disrupt flight feather formation.The excised region appears to possess enhancer activity toward Sim1 but is dispensable for flight feather development.Complete ablation of the ASHCE will be necessary to fully resolve the regulatory role of Sim1 in avian feather morphogenesis.展开更多
Primordial germ cells(PGCs),the precursors of oocytes and spermatozoa,are highly pluripotent.In recent years,the in vitro induction of human primordial germ cell-like cells(h PGCLCs)has advanced significantly.However,...Primordial germ cells(PGCs),the precursors of oocytes and spermatozoa,are highly pluripotent.In recent years,the in vitro induction of human primordial germ cell-like cells(h PGCLCs)has advanced significantly.However,the stability and efficacy of obtaining h PGCLCs in vitro still require improvement.In the current study,we identified a novel induction system using Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12(DMEM/F-12)as the basal medium,supplemented with B27 and N2(referred to as N2B27)in combination with four cytokines:bone morphogenetic protein 4,stem cell factor,epidermal growth factor,and leukemia inhibitory factor.The h PGCLCs induced under these conditions closely resembled PGCs from 4-to 5-week-old embryos at the transcriptomic level.Compared with traditional GK15(GMEM supplemented with 15%Knockout?Serum Replacement)-based induction conditions,the N2B27 system significantly increased the speed and efficacy of h PGCLC induction.RNA sequencing analysis revealed that this improvement was due to an increased cellular capacity to cope with hypoxic stress and avoid apoptosis.The N2B27 medium promoted enhanced mitochondrial activity,enabling cells to better manage hypoxic stress while reducing the production of reactive oxygen species.Moreover,through gradient concentration experiments,we demonstrated that the addition of the common antioxidant N-acetyl-L-cysteine at an optimized concentration further enhanced the efficiency of PGCLC induction under GK15 conditions.In summary,we have established an optimized induction system that enhances the efficiency of h PGCLC differentiation by improving cellular resilience to hypoxic stress and apoptosis.展开更多
[Objective] The aim of this study was to breed new strains which have higher inhibitory effects on the pathogens of watermelon fusarium wilt.[Method] The endophytic Bacillus subtilis B47 strain was obtained from tomat...[Objective] The aim of this study was to breed new strains which have higher inhibitory effects on the pathogens of watermelon fusarium wilt.[Method] The endophytic Bacillus subtilis B47 strain was obtained from tomato stems by UV mutagenesis for two consecutive times,then genetic stability as well as physiological and biochemical properties of mutant strains were studied.[Result] The antibacterial activity of all the three mutant strains F303,F304 and F305 was higher than that of B74 strain.After subculture of 10 successive generations,the antibacterial activity of all the three mutant strains for the pathogens of watermelon fusarium wilt decreased,but the antibacterial activity of F305 strain decreased the least,indicating its best genetic stability among the tested strains.The antibacterial circle diameter of F305 strain was 5 mm larger than that of wild strain B47 under the same condition.The mutant strain F305 was in logarithmic growth phase within 36 h and in stationary phase within 36-96 h,while its optimum growth temperature was 35 ℃.F305 strain could grow in sodium salt with the concentration of 1%-10%,but it grew best at the concentration of 1%.Physiological and biochemical responses of F305 strain were in accordance with those of wild strain B47.[Conclusion] This study lays the foundation for the factorial production of antagonistic substance by B47 strain and new methods of preventing from the pathogens watermelon fusarium wilt.展开更多
[Objective] The paper was to establish embryonic stem cell system of goats. [Method] Numerous primordial germ cell colonies were derived from gonadal ridge and the surrounding tissues in 20 millimeter fetuses of down ...[Objective] The paper was to establish embryonic stem cell system of goats. [Method] Numerous primordial germ cell colonies were derived from gonadal ridge and the surrounding tissues in 20 millimeter fetuses of down producing goat. Primordial germ cells and goats embryonic fibroblasts obtained from conceptus of equivaient gestational age were co-cultured. [Result] The colonies showed some characteristics of embryonic stem cells, such as the morphology of nest-like, they continued to be AKP positive and the ability to be continuously passed [Conclusion] These cells were pluripotent and ES-like cells.展开更多
[Objective] This research aimed to study the FTIR spectra of corn germs and endosperms so as to provide a scientific way for identifying corn of different types. [Method] The corn germs and endosperms of three types w...[Objective] This research aimed to study the FTIR spectra of corn germs and endosperms so as to provide a scientific way for identifying corn of different types. [Method] The corn germs and endosperms of three types were studied by using Fourier transform infrared spectroscopy（FTIR） technology, combined with cluster analysis. [Result] The overall characteristics of original FTIR spectra were basically similar within the range of 700-1 800 cm^-1. The FTIR spectra were mainly composed by the absorption peaks of polysaccharides, proteins and lipids. Within the wavelength range of 700-1 800 cm^-1, there were only tiny differences in original FTIR spectra among the corn germs and endosperms of three different types. The spectra were then processed by using first derivative and second derivative. The second derivative spectra were used for hierarchical cluster analysis（HCA）. The results showed that with the wavelength range of 700-1 800 cm^-1, the second derivative spectra of the 52 samples could be better clustered according to the tree types and corn germ and corn endosperm. The clustering correct rate reached 96.1%.[Conclusion] FTIR technology, combined with cluster analysis, can be used to identify different types of corn germs and endosperms, and it is characterized by convenience and rapidness.展开更多
Wheat germ meal is the by production of oil extracting, and a great quantity of it has been wasted, thus the quantity of lost protein is great. In order to use wheat germ meal proteins adequately, wheat germ proteins ...Wheat germ meal is the by production of oil extracting, and a great quantity of it has been wasted, thus the quantity of lost protein is great. In order to use wheat germ meal proteins adequately, wheat germ proteins were hydrolyzed to anti-oxidation peptides by using alkaline protease. Through the single factor analysis and regression analysis, the optimized experiment conditions of hydrolysising wheat germ meal to wheat germ peptides were enzymatic quantity 0.8%（w/w）, material to liquid ratio 1∶12.3, enzymolysis time 2.1 h. Under these conditions, the scavenging effect was 49.78%,the DH was 22% and peptides content in enzymatic hydrolysate was 1.9%（w/w）.By SDS-PAGE electrophoresis,the molecular weight range of wheat germ peptides were below 10 ku and most were between 4.54 and 5.63 ku.The wheat germ proteins could be used ful y and grain resources would be saved.展开更多
文摘YOU CANNOT SUCCUMB TO CHOLERA,unless the sewage system breaks down-likewise deaths from violence vanish,as long as emotional support networks are working properly.Gut hygiene eliminates the one,societal hygiene the other.Cholera was an inexorable plague-until Germ Theory was accepted and implemented.Violence too,will persist-until we agree and accept its equivalent in Societal Theory.The scientific method,especially Quantum Physics,sabotages philosophy-it cannot cope with the living fact that the elephant in the room is alive.Quantum Mechanics describes itself as being(1)more incomprehensible,(2)weirder,and(3)more uncertain than anything we’ve met before.But that’s chickenfeed,compared to what you find in clinical medicine,which though portrayed as being intrinsically woolly and subjective,wilfully aims to enhance life,whatever that might be.
基金supported by the National Key R&D Program of China(2021YFD1300100)Guangxi Key R&D Program,China(AB21220005)+1 种基金Reproductive Medicine,Guangxi Medical and Health Key Discipline Construction Project of the Affiliated Hospitalthe National Natural Science Foundation of China(32360180)。
文摘Primordial germ cells(PGCs)are the stem-cell population of adult animal gametes,which develop into sperm or eggs.It can be propagated in vitro and injected into the host chicken for genome editing to obtain germline chimeric chicken.However,it has the limitation that the host embryo contains endogenous PGCs,which raises complications,resultantly donor PGCs fail to compete,and transmission efficiency reduced.Therefore,to increase the transmission efficiency,we generated a novel sterile chicken with the inducible elimination of endogenous PGCs in the host.This is the first study that applied the herpes simplex virus thymidine kinase(HSV-TK)cell ablation system in avian.CRISPR/Cas9-mediated homology-directed repair was performed to localize the HSV-TK suicide gene to the last exon of the deleted in azoospermialike(DAZL)gene,and ganciclovir(GCV)was added to induce the apoptosis in the germ cells of the host embryo.The sterilized host embryo introduced genome-edited PGCs to produce chimeric chicken carrying exogenous germ cells only.It was observed that the germline transmission efficiency was 100%achieved,and the obtained chicks were purely from donor breeds.The technologies established in the current study have important applications in germplasm conservation and gene editing in chicken.
基金supported by the Department of Defense AFIRMⅢW81XWH-20-2-0029 grant subcontractLone Star Paralysis gift,UT POC19-1774-13 grant+1 种基金Neuraptive Therapeutics Inc.26-7724-56 grantNational Institutes of Health R01-NS128086(all to GDB)。
文摘Peripheral nerve injuries result in the rapid degeneration of distal nerve segments and immediate loss of motor and sensory functions;behavioral recovery is typically poor.We used a plasmalemmal fusogen,polyethylene glycol(PEG),to immediately fuse closely apposed open ends of severed proximal and distal axons in rat sciatic nerves.We have previously reported that sciatic nerve axons repaired by PEG-fusion do not undergo Wallerian degeneration,and PEG-fused animals exhibit rapid(within 2–6 weeks)and extensive locomotor recovery.Furthermore,our previous report showed that PEG-fusion of severed sciatic motor axons was non-specific,i.e.,spinal motoneurons in PEG-fused animals were found to project to appropriate as well as inappropriate target muscles.In this study,we examined the consequences of PEG-fusion for sensory axons of the sciatic nerve.Young adult male and female rats(Sprague–Dawley)received either a unilateral single cut or ablation injury to the sciatic nerve and subsequent repair with or without(Negative Control)the application of PEG.Compound action potentials recorded immediately after PEG-fusion repair confirmed conduction across the injury site.The success of PEG-fusion was confirmed through Sciatic Functional Index testing with PEG-fused animals showing improvement in locomotor function beginning at 35 days postoperatively.At 2–42 days postoperatively,we anterogradely labeled sensory afferents from the dorsal aspect of the hindpaw following bilateral intradermal injection of wheat germ agglutinin conjugated horseradish peroxidase.PEG-fusion repair reestablished axonal continuity.Compared to unoperated animals,labeled sensory afferents ipsilateral to the injury in PEG-fused animals were found in the appropriate area of the dorsal horn,as well as inappropriate mediolateral and rostrocaudal areas.Unexpectedly,despite having intact peripheral nerves,similar reorganizations of labeled sensory afferents were also observed contralateral to the injury and repair.This central reorganization may contribute to the improved behavioral recovery seen after PEG-fusion repair,supporting the use of this novel repair methodology over currently available treatments.
基金supported by the National Natural Science Foundation of China(No.82071078,82370939)the Shaanxi Provincial High-level Talent Program and Young Talent Support Plan of Xi’an Jiaotong University.
文摘Tissue interactions play a crucial role in tooth development.Notably,extracellular vesicle-mediated interactions between the mandible and tooth germ are considered essential.Here,we revealed that mandible extracellular vesicles could modulate the proliferation and differentiation of dental mesenchymal cells by regulating the histone demethylase KDM2B.Further investigation showed that mandible derived extracellular vesicles could deliver miR-206 to KDM2B,thereby regulating tooth development.An animal study demonstrated that the miR-206/KDM2B pathway affected tooth morphogenesis and mineralization after eight weeks of subcutaneous transplantation in nude mice.In conclusion,this study suggested that the mandible played a critical role in tooth morphogenesis and mineralization,which could be a potential therapeutic target for abnormal tooth development and an alternative model for tooth regeneration.
基金supported by the National Natural Science Foundation of China(31988101 to A.M.)the National Key Research and Development Program of China(2023YFA1800300 to X.W.and 2018YFC1003304 to A.M.)the Yunnan Provincial Science and Technology Project at Southwest United Graduate School(202302A0370011 to A.M.).
文摘The cell fate of primordial germ cell(PGC)in zebrafish is pre-determined by maternally deposited germ plasm,which is packaged into ribonucleoprotein complex in oocytes and inherited into PGC-fated cells in embryos.However,the maternal factors regulating the assembly of germ plasm and PGC development remain poorly understood.In this study,we report that the maternal transcription factor Znf706 regulates the assembly of germ plasm factors into a granule-like structure localized perinuclearly in PGC during migration.Maternal and zygotic mutants of znf706 exhibit deficient germ plasm scattering at the early embryonic stage,decreased PGC numbers with some mislocation during PGC migration,and a lower female ratio in adulthood.Notably,the implementation of Znf706 CUT&Tag and RNA-seq on immature oocytes uncovers that Znf706 in stage I oocytes may promote transcription of several mitochondrial genes in addition to other functions.Hence,we propose that Znf706 is implicated in germ plasm assembly and PGC development in zebrafish.
基金supported by the National Key Research and Development Program of China (2021YFD1200301 and 2021YFD1200302)the Natural Science Foundation of Jiangsu Province, China (BK20210813)+1 种基金the National Natural Science Foundation of China (32102534)the Yangzhou International Science and Technology Cooperation Projects, China (YZ2021175)。
文摘Germplasm resources are essential for the sustainable development of biodiversity and husbandry of local chickens, as well as for the breeding and industry of superior quality chickens. Unfortunately, many local and indigenous chicken breeds are at risk of declining numbers, emphasizing the need to conserve breed resources for endangered chickens. Primordial germ cells(PGCs) are crucial for preserving germplasm resources by inheriting genetic information from parents to offspring and ensuring stability of genetic material between germlines. In this study,PGCs were isolated from chicken embryos' gonads and cultured in FAcs medium without feeder cells. Over a period of approximately 40 d, the cells proliferated to a number of up to 10^(6), establishing various cell lines. Particularly, 18 PGC lines were created from Rugao Yellow chicken and Shouguang chicken, with an efficiency ranging from 39.1 to 45%. Furthermore, PGCs that had been cultured for 40 passages exhibited typical PGC characteristics, suchas glycogen staining reaction, and expression of pluripotency and reproductive markers. These results confirmthat PGCs maintain stem cell properties even after long-term in vitro culture. Additionally, PGCs cryopreserved for up to 120 d remained viable, maintained typical PGC morphologies, and possessed stable cell proliferation ability. Through intravascular injection into chicken embryos, green fluorescent protein(GFP)-PGCs were found in the recipient embryos' gonads and could develop into gametes to produce offspring, indicating that even after extended culture, PGCs retain their migratory and lineage-transmitting capabilities. This research offers valuable insights into the in vitro cultivation and preservation of PGCs of Chinese indigenous chickens. The findings of this study can be applied in transgenic chicken production and the preservation of genetic resources of indigenous chicken breeds.
文摘Dear Editor,I would like to congratulate Mamsen et al.i on their extensive and scientifically valuable work.I analyzed their raw data presented in Table 1 of the original article from a different perspective and discovered an effect not mentioned in the article.My analysis showed that luteinizing hormone(LH)levels are significantly lower in patients at high infertility risk(HIR),whose testes lack A dark(Ad)spermatogonia and display an abnormal ratio of germ cells per crosssectional tubule(G/T).
文摘Objective:Testicular germ cell tumors(TGCTs)represent the most common malignancy among young men aged 20–40 years.Transglutaminase 7(TG7),encoded by TGM7,is a poorly characterized enzyme whose function in TGCT remains unknown.This study aimed to assess TG7 expression in clinical specimens and investigate its functional role in a testicular germ cell tumor cell line(NT2/D1).Methods:TG7 protein expression was evaluated in clinical testicular tissue samples via immunohistochemistry(IHC)and immunofluorescence(IF).Functional analysis was conducted in the NT2/D1 human testicular cancer cell line usingDicer-substrate small interferingRNAs(DsiRNAs)targeting TG7.Gene knockdown efficiency was confirmed by reverse transcription quantitative PCR(qRT-PCR),and protein suppression was validated by immunofluorescence.Cell viability was assessed using the MTT assay.The expression of inflammation and apoptosis-related genes was quantified via qRT-PCR.Results:TG7 expression was significantly elevated in testicular germ cell tumor tissues,showing approximately a 4.5-fold increase compared to normal testis,with strong localization in tumor nests and stromal compartments.In NT2/D1 cells,TG7 silencing using 20 nM DsiRNA3 led to a dose-dependent reduction in cell viability,with up to 48%inhibition observed at 200 nM(MTT assay,****p<0.0001).qRT-PCR analysis revealed significant upregulation of IL6(3.2-fold),TNFα(2.8-fold),and CASP3(2.5-fold)mRNA levels following TG7 knockdown(p<0.0001),while p53 expression remained unchanged.These findings support TG7’s role in modulating tumor cell survival,inflammation,and apoptosis via p53-independent pathways.Conclusion:Collectively,TG7 is significantly overexpressed in TGCT tissues and supports tumor cell viability in vitro.This study establishes TG7 as a novel biomarker and therapeutic target in testicular cancer,laying the groundwork for future studies on TG7-targeted interventions.
文摘Dear Editor,We are much obliged that Hadziselimovic1 has used our data2 to highlight the substantial proportion of boys with cryptorchidism where gonadotropin insufficiency is an important factor related to the pathogenesis.We have recently presented a study on a series of 453 consecutive boys with bilateral nonsyndromic cryptorchidism,in which we conducted hormonal evaluations and assessed germ cell numbers in testicular biopsies.3 In this series,45%of the boys were classified as having gonadotropin insufficiency.3 Identifying these patients at the time of surgery is important.A recent follow-up study of 208 boys with nonsyndromic bilateral cryptorchidism from our department showed that the boys with gonadotropin insufficiency had an impaired fertility potential after surgery compared to boys with an intact hypothalamus–pituitary–gonadal feedback mechanism.4 In a review from 2022,Hadziselimovic5 suggested that infertility in patients diagnosed with cryptorchid testes is a consequence of a hormonal deficiency rather than temperature-induced cellular damage.
基金supported by the National Natural Science Foundation of China awarded to D.C.(32270835)Zhejiang Natural Science Foundation awarded to D.C.(Z22C129553)Dr.Li Dak Sum&Yip Yio Chin Development Fund for Regenerative Medicine,Zhejiang University,awarded to D.C.
文摘Primordial germ cells(PGCs)are the precursors of germline that are specified at the embryonic stage.Recent studies reveal that humans employ different mechanisms for PGC specification compared with model organisms such as mice.Moreover,the specific regulatory machinery remains largely unexplored,mainly due to the inaccessible nature of this complex biological process in humans.Here,we curate and integrate multi-omics data,including 581 RNA-seq,54 ATAC-seq,45 ChIP-seq,and 69 single-cell RNA-seq samples from different stages of human PGC development to recapitulate the precisely controlled and stepwise process,presenting an atlas in the human PGC database(hPGCdb).With these uniformly processed data and integrated analyses,we characterize the potential key transcription factors and regulatory networks governing human germ cell fate.We validate the important roles of some of the key factors in germ cell development by CRISPRi knockdown.We also identify the soma-germline interaction network and discover the involvement of SDC2 and LAMA4 for PGC development,as well as soma-derived NOTCH2 signaling for germ cell differentiation.Taken together,we have built a database for human PGCs(http://43.131.248.15:6882)and demonstrate that hPGCdb enables the identification of the missing pieces of mechanisms governing germline development,including both intrinsic and extrinsic regulatory programs.
文摘BACKGROUND Urinary tract infections(UTIs)in kidney transplant patients are a challenge.AIM To evaluate epidemiology,clinical status,therapeutic management,and clinical outcome of kidney transplant patients in a university hospital for UTI.METHODS We conducted a retrospective observational study,enrolling all kidney transplant patients hospitalized for UTI,with the objective to evaluate the epidemiology,clinical status,therapeutic management,and clinical outcome of kidney transplant patients.RESULTS From our real-life experience,infection with multidrug-resistant germs was confirmed as a risk factor for the severe evolution of the infection.At the same time,the re-evaluation of immunosuppressive therapy could be an important therapeutic strategy in the course of infection.CONCLUSION Prompt initiation of empiric antibiotic therapy upon initiation of microbiological investigations may reduce the risk of severe infection progression.
文摘Dug well water in the working area of the Jongaya Community Health Center often fails to meet clean water quality standards due to low pH(6.1)and high organic matter content(14 mg/L),exceeding the limits set by Indonesian Ministry of Health Regulation No.32 of 2017,which poses a risk of digestive disorders and skin diseases.Addressing this gap,this study evaluates the effectiveness of quicklime(CaO)and aluminum sulfate[Al_(2)(SO_(4))_(3)]in improving dug well water quality through a quasi-experimental pretest-posttest control group design.Water samples were purposively collected from contaminated wells and treated using quicklime,aluminum sulfate,and their combination at doses of 1 g,10 g,and 20 g,each replicated three times,with pH and organic matter content measured before and after treatment and analyzed using a paired t-test.Results showed that 1 g of CaO significantly increased pH from 6.1 to 7.88(p<0:05),meeting the clean water quality threshold(≥6:5),while 20 g of Al_(2)(_(S)O_(4))_(3)reduced organic matter content from 170.91 mg/L to 126.11 mg/L,and the combined treatment achieved the best outcome,with a 46.75%reduction in organic matter and a pH increase to 8.63,both meeting standards.All treatments were statistically significant(p<0:05),indicating that CaO and Al_(2)(SO_(4))_(3)are effective for improving dug well water quality in at-risk communities.The findings highlight the urgency of promoting proper application and optimal dosing,supported by long-term monitoring,and provide novel evidence on the combined use of these chemicals for addressing a local public health challenge.
基金supported by the Ministry of Agriculture and Rural Affairs of the People's Republic of China(125A0607)Department of Science and Technology of Yunnan Province(XDYC-KJLJ-2022-0004)。
文摘Flight feathers represent a hallmark innovation of avian evolution.Recent comparative genomic analyses identified a 284 bp avian-specific highly conserved element(ASHCE)located within the eighth intron of the SIM bHLH transcription factor 1(Sim1)gene,postulated to act as a cis-regulatory element governing flight feather morphogenesis.To investigate its functional significance,genome-edited(GE)primordial germ cell(PGC)lines carrying targeted ASHCE deletions were generated using CRISPR/Cas9-mediated editing,with germline chimeric males subsequently mated with wild-type(WT)hens to obtain GE progeny.The resulting GE chickens harbored 257-260 bp deletions,excising approximately half of the Sim1-ASHCE sequence.Reverse transcription-quantitative real-time polymerase chain reaction(RT-qPCR)analysis showed an average 0.32-fold reduction in Sim1 expression in the forelimbs of GE embryos at day 8(E8)compared to WT counterparts.Despite this,GE chickens developed structurally normal flight and tail feathers.In situ hybridization localized Sim1 expression to the posterior mesenchyme surrounding flight feather buds in E8 WT embryos,but not within the buds themselves.These results suggest that partial deletion of Sim1-ASHCE,despite diminishing Sim1 expression,does not disrupt flight feather formation.The excised region appears to possess enhancer activity toward Sim1 but is dispensable for flight feather development.Complete ablation of the ASHCE will be necessary to fully resolve the regulatory role of Sim1 in avian feather morphogenesis.
基金funded by the National Key R&D Program(Grant Nos.2022YFC2702800 and 2021YFC2700302 to Y.Y.)the National Natural Science Foundation of China(Grant Nos.82122025 to Y.Y.,82221005 to J.S.,and 82201763 to L.L.)。
文摘Primordial germ cells(PGCs),the precursors of oocytes and spermatozoa,are highly pluripotent.In recent years,the in vitro induction of human primordial germ cell-like cells(h PGCLCs)has advanced significantly.However,the stability and efficacy of obtaining h PGCLCs in vitro still require improvement.In the current study,we identified a novel induction system using Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12(DMEM/F-12)as the basal medium,supplemented with B27 and N2(referred to as N2B27)in combination with four cytokines:bone morphogenetic protein 4,stem cell factor,epidermal growth factor,and leukemia inhibitory factor.The h PGCLCs induced under these conditions closely resembled PGCs from 4-to 5-week-old embryos at the transcriptomic level.Compared with traditional GK15(GMEM supplemented with 15%Knockout?Serum Replacement)-based induction conditions,the N2B27 system significantly increased the speed and efficacy of h PGCLC induction.RNA sequencing analysis revealed that this improvement was due to an increased cellular capacity to cope with hypoxic stress and avoid apoptosis.The N2B27 medium promoted enhanced mitochondrial activity,enabling cells to better manage hypoxic stress while reducing the production of reactive oxygen species.Moreover,through gradient concentration experiments,we demonstrated that the addition of the common antioxidant N-acetyl-L-cysteine at an optimized concentration further enhanced the efficiency of PGCLC induction under GK15 conditions.In summary,we have established an optimized induction system that enhances the efficiency of h PGCLC differentiation by improving cellular resilience to hypoxic stress and apoptosis.
基金Supported by the Fund of Science and Technology in GuangXi Zhuang Autonomous Region(0009018)~~
文摘[Objective] The aim of this study was to breed new strains which have higher inhibitory effects on the pathogens of watermelon fusarium wilt.[Method] The endophytic Bacillus subtilis B47 strain was obtained from tomato stems by UV mutagenesis for two consecutive times,then genetic stability as well as physiological and biochemical properties of mutant strains were studied.[Result] The antibacterial activity of all the three mutant strains F303,F304 and F305 was higher than that of B74 strain.After subculture of 10 successive generations,the antibacterial activity of all the three mutant strains for the pathogens of watermelon fusarium wilt decreased,but the antibacterial activity of F305 strain decreased the least,indicating its best genetic stability among the tested strains.The antibacterial circle diameter of F305 strain was 5 mm larger than that of wild strain B47 under the same condition.The mutant strain F305 was in logarithmic growth phase within 36 h and in stationary phase within 36-96 h,while its optimum growth temperature was 35 ℃.F305 strain could grow in sodium salt with the concentration of 1%-10%,but it grew best at the concentration of 1%.Physiological and biochemical responses of F305 strain were in accordance with those of wild strain B47.[Conclusion] This study lays the foundation for the factorial production of antagonistic substance by B47 strain and new methods of preventing from the pathogens watermelon fusarium wilt.
基金Supported by Project of Baotou University(BSY2010-23)~~
文摘[Objective] The paper was to establish embryonic stem cell system of goats. [Method] Numerous primordial germ cell colonies were derived from gonadal ridge and the surrounding tissues in 20 millimeter fetuses of down producing goat. Primordial germ cells and goats embryonic fibroblasts obtained from conceptus of equivaient gestational age were co-cultured. [Result] The colonies showed some characteristics of embryonic stem cells, such as the morphology of nest-like, they continued to be AKP positive and the ability to be continuously passed [Conclusion] These cells were pluripotent and ES-like cells.
基金Supported by National Natural Science Foundation of China(30960179)Natural Science Foundation of Yunnan Province(2007A048M)~~
文摘[Objective] This research aimed to study the FTIR spectra of corn germs and endosperms so as to provide a scientific way for identifying corn of different types. [Method] The corn germs and endosperms of three types were studied by using Fourier transform infrared spectroscopy（FTIR） technology, combined with cluster analysis. [Result] The overall characteristics of original FTIR spectra were basically similar within the range of 700-1 800 cm^-1. The FTIR spectra were mainly composed by the absorption peaks of polysaccharides, proteins and lipids. Within the wavelength range of 700-1 800 cm^-1, there were only tiny differences in original FTIR spectra among the corn germs and endosperms of three different types. The spectra were then processed by using first derivative and second derivative. The second derivative spectra were used for hierarchical cluster analysis（HCA）. The results showed that with the wavelength range of 700-1 800 cm^-1, the second derivative spectra of the 52 samples could be better clustered according to the tree types and corn germ and corn endosperm. The clustering correct rate reached 96.1%.[Conclusion] FTIR technology, combined with cluster analysis, can be used to identify different types of corn germs and endosperms, and it is characterized by convenience and rapidness.
文摘Wheat germ meal is the by production of oil extracting, and a great quantity of it has been wasted, thus the quantity of lost protein is great. In order to use wheat germ meal proteins adequately, wheat germ proteins were hydrolyzed to anti-oxidation peptides by using alkaline protease. Through the single factor analysis and regression analysis, the optimized experiment conditions of hydrolysising wheat germ meal to wheat germ peptides were enzymatic quantity 0.8%（w/w）, material to liquid ratio 1∶12.3, enzymolysis time 2.1 h. Under these conditions, the scavenging effect was 49.78%,the DH was 22% and peptides content in enzymatic hydrolysate was 1.9%（w/w）.By SDS-PAGE electrophoresis,the molecular weight range of wheat germ peptides were below 10 ku and most were between 4.54 and 5.63 ku.The wheat germ proteins could be used ful y and grain resources would be saved.
