The oilseed crop Camelina sativa exhibits salinity tolerance,but the effects on early growth stages across a range of different salts and in combination with salicylic acid(SA)have not been thoroughly evaluated.In thi...The oilseed crop Camelina sativa exhibits salinity tolerance,but the effects on early growth stages across a range of different salts and in combination with salicylic acid(SA)have not been thoroughly evaluated.In this study,seeds were germinated in varying concentrations of six salts(NaCl,CaCl_(2),ZnCl_(2),KCl,MgSO_(4),and Na2SO_(4))with or without 0.5 mM SA.Using the halotime model,we estimated salt thresholds for germination and parameters of seedling growth.Germination and seedling growth parameters of camelina significantly decreased with increasing salt concentration across all salt types.Salts containing Zn and SO_(4) were most detrimental to germination and seedling growth.Except for KCl,0.5 mM SA generally reduced the salinity tolerance threshold(Saltb(50))of camelina.Specifically,Saltb(50)was 21.5%higher for KCl and 16.1%,25.0%,54.9%,21.0%,and 5.6%lower for CaCl_(2),NaCl,MgSO_(4),Na2SO_(4),and ZnCl_(2),respectively,when 0.5 mM SA was compared to 0 mM SA.Furthermore,camelina seedling growth was consistently more sensitive than germination across all salt types.SA did not significantly enhance germination or seedling growth and was harmful when combined with certain salts or at the germination stage.It can be concluded that both the type of salt and the concentration of SA are as critical as the salt concentration in saline irrigation water.展开更多
Cryopreservation is a fundamental technology in biomedical research,regenerative medicine,and tissue engineering,enabling the long-term storage of cells,tissues,and organs.However,its effectiveness is limited by chall...Cryopreservation is a fundamental technology in biomedical research,regenerative medicine,and tissue engineering,enabling the long-term storage of cells,tissues,and organs.However,its effectiveness is limited by challenges such as intracellular ice formation,cryoprotectant toxicity,and reduced post-thaw viability.This review explores the crucial role of encapsulation in enhancing cryopreservation efficiency,with a focus on recent advances in materials science,bioengineering,and cryobiology.Emerging technologies,such as nanotechnology and stimuli-responsive polymers,are transforming encapsulation strategies.Innovations such as microfluidic systems offer precise control over cooling rates and cryoprotectant distribution,thereby mitigating conventional limitations.The review also addresses current obstacles related to scaling up encapsulation processes and ensuring the long-term biocompatibility and stability of preserved specimens.By synthesizing recent findings,this work provides a comprehensive resource for researchers and clinicians seeking to enhance biopreservation techniques and their applications in contemporary medicine and biotechnology.Finally,the review identifies critical knowledge gaps that must be addressed to improve the efficacy of cryopreservation strategies and advance their clinical translation.展开更多
Oocyte cryopreservation is an essential procedure in assisted reproductive technologies,aimed at preserving fertility,particularly for women undergoing IVF treatment or at risk of ovarian damage due to radiation,chemo...Oocyte cryopreservation is an essential procedure in assisted reproductive technologies,aimed at preserving fertility,particularly for women undergoing IVF treatment or at risk of ovarian damage due to radiation,chemotherapy,or surgery.Despite its growing use,the survival and fertilization rates of cryopreserved oocytes remain suboptimal,largely due to cryo-induced oxidative stress.The generation of Reactive Oxygen Species(ROS)during freezing and thawing causes considerable damage to key cellular components,including proteins,lipids,DNA,and mitochondria.This oxidative stress compromises oocyte quality and reduces developmental potential.To address these challenges,the use of additives-especially antioxidants-has shown significant promise in mitigating oxidative damage.Enzymatic antioxidants such as Superoxide Dismutase(SOD)and Catalase(CAT),along with non-enzymatic antioxidants like glutathione,melatonin,and resveratrol,have demonstrated the ability to neutralize ROS and improve oocyte viability and developmental outcomes.Recent studies highlight the potential of Mitoquinone(MitoQ),a mitochondria-targeted antioxidant,to effectively counteract mitochondrial ROS and enhance cellular defense mechanisms during cryopreservation.This review explores the cellular mechanisms of cryodamage,the role of oxidative stress in oocyte cryopreservation,and the potential of various antioxidant strategies to enhance oocyte survival and function.Developing effective antioxidant supplementation approaches may significantly improve the outcomes of cryopreservation in reproductive medicine.展开更多
Background Sperm cryopreservation is widely used in the cattle industry,as it allows for disassociating the localiza-tion of sires and the collection of semen from the timing of artificial insemination.While freeze-th...Background Sperm cryopreservation is widely used in the cattle industry,as it allows for disassociating the localiza-tion of sires and the collection of semen from the timing of artificial insemination.While freeze-thawing is known to impair sperm DNA integrity,whether the damage induced consists of single-(SSB)or double-strand breaks(DSB)has not been determined.In addition,no previous study has addressed if DNA breaks preferentially reside in specific genome regions such as those forming the toroid linker regions,or are rather spread throughout the regions linked to protamines.The main aim of the present work,therefore,was to elucidate the type and localization of the DNA damage generated by cryopreservation and to evaluate its impact on artificial insemination outcomes in cattle.Results The incidence of SSB and DSB was evaluated in 12 ejaculates before and after cryopreservation with the Comet assay,and the localization of the DNA breaks was assessed using pulsed-field gel electrophoresis(PFGE).Before cryopreservation,the incidence of SSB was 10.99%±4.62%and involved 20.56%±3.04%of sperm cells,whereas these figures significantly(P<0.0001)increased up to 34.11%±3.48%and 53.36%±11.00%in frozen-thawed sperm.In contrast,no significant differences in the incidence of DSB were observed(P>0.990)before and after cryopreservation(before:incidence of 13.91%±1.75%of sperm DNA affecting 56.04%±12.49%of sperm cells;after:incidence of 13.55%±1.55%of sperm DNA involving 53.36%±11.00%of sperm cells).Moreover,PFGE revealed that the percentage of sperm DNA fragments whose length was shorter than a toroid(<31.5 kb)was greater(P<0.0001)after(27.00%±4.26%)than before freeze-thawing(15.57%±4.53%).These differences indicated that the DNA breaks induced by cryopreservation affect the regions condensed in protamines,which are structured in toroids.On the other hand,in vivo fertility rates were associated to the incidence of SSB and DSB in frozen-thawed sperm(P=0.032 and P=0.005),but not with the size of the DNA fragments resulting from these breaks(P>0.05).Conclusion Cryopreservation of bovine sperm generates single-strand DNA breaks,which are mainly located in protamine-condensed toroidal regions.The incidence of DNA breaks in cryopreserved sperm has an impact on cat-tle fertility,regardless of the size of generated fragments.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: Cystinosis is a multisystemic autosomal recessive deficiency of the lysosomal membrane transporter protein (cystinosin) caused by mutations in CTNS gene. Objective: This study summarizes the Portuguese exp...Background: Cystinosis is a multisystemic autosomal recessive deficiency of the lysosomal membrane transporter protein (cystinosin) caused by mutations in CTNS gene. Objective: This study summarizes the Portuguese experience in the diagnosis and management of patients with this rare disease over the past few years and reports recurrent mutations in the CTNS gene. Methods: Unrelated patients from different pediatric and adult hospitals all over Portugal with non-nephrotic proteinuria, hypercalciuria, hypokalemia impaired proximal reabsorption of amino acids, glycosuria and hypophosphatemia, suggestive of a Fanconi syndrome and ocular problems, were studied. Intra-leukocyte cystine levels were determined and molecular analysis was performed, to determine the presence or absence of the 57-kb deletion in CTNS, followed by direct sequencing of the coding exons of CTNS. Results: From 1998 to 2017, twenty-one cystinotic patients were biochemically diagnosed. From the remaining seventeen (four deceased), eleven were studied for CTNS gene. Five out of eleven patients were homozygous for the 57-kb deletion (10/22;45.5%), and other five were compound heterozygous for this variant (15/22;68.2%). The other mutations found were p.Q128X (c.721 C>T;2/22), p.S139F (c.755 C>T;4/22) and c.18-21delGACT (p.T7FfsX7;1/22). All of these seventeen cystinotic patients are in treatment. Approximately 84% are adults, 16% are young children, and 54.5% are kidney transplant recipient. Conclusions: The authors would like to emphasize the importance of first screening for the 57-kb deletion since it is very common in our population. This genetic study is the first in our country and it could be the basis for future genetic counseling in Portuguese population.展开更多
The sperm DNA fragmentation index(DFI)is a metric used to assess DNA fragmentation within sperm.During in vitro fertilizationembryo transfer(IVF-ET),high sperm DFI can lead to a low fertilization rate,poor embryo deve...The sperm DNA fragmentation index(DFI)is a metric used to assess DNA fragmentation within sperm.During in vitro fertilizationembryo transfer(IVF-ET),high sperm DFI can lead to a low fertilization rate,poor embryo development,early miscarriage,etc.A kinase anchoring protein(AKAP)is a scaffold protein that can bind protein kinase A(PKA)to subcellular sites of specific substrates and protects the biophosphorylation reaction.Sperm protein antigen 17(SPA17)can also bind to AKAP.This study intends to explore the reason for the decreased fertilization rate observed in high sperm DFI(H-DFI)patients during IVF-ET.In addition,the study investigates the expression of AKAP,protein kinase A regulatory subunit(PKARIl),and SPA17 between H-DFI and low sperm DFI(L-DFI)patients.SPA17 at the transcriptional level is abnormal,the translational level increases in H-DFI patients,and the expression of AKAP4/PKARIl protein decreases.H,O,has been used to simulate oxidative stress damage to spermatozoa during the formation of sperm DFI.It indicates that H,O,increases the expression of sperm SPA17 protein and suppresses AKAP4/PKARIl protein expression.These processes inhibit sperm capacitation and reduce acrosomal reactions.Embryo culture data and IVF outcomes have been documented.The H-DFI group has a lower fertilization rate.Therefore,the results indicate that the possible causes for the decreased fertilization rate in the H-DFI patients have included loss of sperm AKAP4/PKARIl proteins,blocked sperm capacitation,and reduced occurrence of acrosome reaction.展开更多
BACKGROUND Cholangiocarcinoma(CCA)is a highly malignant cancer,characterized by frequent mucin overexpression.MUC1 has been identified as a critical oncogene in the progression of CCA.However,the comprehensive underst...BACKGROUND Cholangiocarcinoma(CCA)is a highly malignant cancer,characterized by frequent mucin overexpression.MUC1 has been identified as a critical oncogene in the progression of CCA.However,the comprehensive understanding of how the mucin family influences CCA progression and prognosis is still incomplete.AIM To investigate the functions of mucins on the progression of CCA and to establish a risk evaluation formula for stratifying CCA patients.METHODS Single-cell RNA sequencing data from 14 CCA samples were employed for elucidating the roles of mucins,complemented by bioinformatic analyses.Subse-quent validations were conducted through spatial transcriptomics and immuno-histochemistry.The construction of a risk evaluation model utilized the least absolute shrinkage and selection operator regression algorithm,which was further confirmed by independent cohorts and diverse data types.RESULTS CCA tumor cells with elevated levels of MUC1 and MUC4 showed activated nucleotide metabolic pathways and increased invasiveness.MUC5AC-high cells were found to promote CCA progression through WNT signaling.MUC5B-high cells exhibited robust cellular oxidation activities,leading to resistance against antitumoral treatments.MUC13-high cells were observed to secret chemokines,recruiting and transforming macrophages into the M2-polarized state,thereby suppressing antitumor immunity.MUC16-high cells were found to promote tumor progression through interleukin-1/nuclear factor kappa-light-chain-enhancer of activated B cells signaling upon interaction with neutrophils.Utilizing the expression levels of these mucins,a risk factor evaluation formula for CCA was developed and validated across multiple cohorts.CCA samples with higher risk factors exhibited stronger metastatic potential,chemotherapy resistance,and poorer prognosis.CONCLUSION Our study elucidates the functional mechanisms through which mucins contribute to CCA development,and provides tools for risk stratification in CCA.展开更多
Egyptian species of the leafhopper genus Exitianus Ball,1929,E.capicola(St?l,1855),E.nanus(Distant,1908),and E.pondus Ross,1968 are reviewed.Illustrations,morphological descriptions,and a key for their identification ...Egyptian species of the leafhopper genus Exitianus Ball,1929,E.capicola(St?l,1855),E.nanus(Distant,1908),and E.pondus Ross,1968 are reviewed.Illustrations,morphological descriptions,and a key for their identification are provided.In this study,we used molecular techniques to confirm morphological identification and detect phylogeny among the three Exitianus species.The partial nucleotide sequences of the amplified products obtained were determined by Macrogen Korea.The nucleotide sequences of 28S rDNA and COX genes of the EGY-ARC-9,EGY-ARC-4,and EGY-ARC-5 were determined by Macrogen Korea,blasted into BLAST at the National Center for Biotechnology Information website(NCBI)and compared with those deposited in the GenBank DNA database.The results represented the homology percentage between the partial sequences of the 28S rRNA and COX genes from each species and related species obtained from GenBank DNA database.The morphological identifications of the three Exitianus species were confirmed by molecular characterization and sequencing of 28S rDNA and COX genes and identified as E.capicola,E.nanus,and E.pondus.Also,their sequences of 28srDNA and COX genes were deposited in GenBank with an accession number(LC670610,LC670607,and OQ196105)for 28srDNA gene and(LC775357,LC775358,and LC775359)for the COX gene.展开更多
AIM:To find out the association of secreted protein acidic and rich in cysteine(SPARC)-related modular calcium binding 2(SMOC2)gene variants rs2255680 and rs13208776 with genotypic and phenotypic characteristics in bo...AIM:To find out the association of secreted protein acidic and rich in cysteine(SPARC)-related modular calcium binding 2(SMOC2)gene variants rs2255680 and rs13208776 with genotypic and phenotypic characteristics in both familial and non-familial primary open angle glaucoma(POAG)patients.METHODS:A total of 212 POAG patients,comprising 124 familial and 88 non-familial,were enrolled.For genotyping the SMOC2 variant rs2255680,amplification refractory mutation system(ARMS)-polymerase chain reaction(PCR)method and PCR-restriction fragment length polymorphism(PCR-RFLP)were utilized for analyzing rs13208776 variant.RESULTS:The mean age of familial POAG patients was 50.92±9.12y,with 78 males and 46 females.The mean age of non-familial POAG patients was 53.14±13.44y,with 52 males and 36 females.The SMOC2 gene variant rs13208776 showed the significant association with POAG between familial and non-familial groups.The homozygous G/G variant was frequent among non-familial(60.2%)whereas the heterozygous G/A variant was more frequent in familial POAG patients(46%).There were significant differences in G/A variant between familial and non-familial glaucoma patients,and the risk was decreased to 0.53-fold in non-familial glaucoma patients[odds ratio(OR):0.53;95%confidence interval(CI):0.29-0.94;P=0.033]in codominant model.The risk was further reduced to 0.49-fold(95%CI:0.28-0.86;P=0.012)in dominant model for non-familial patients.No significant association of SMOC2 gene variant rs2255680 between familial and non-familial glaucoma patients was found in our population.The haplotype analysis showed the decreased risk for TA[OR:0.48(95%CI:0.29-0.79);P=0.004]and an increased risk for TG[OR=2.28(95%CI:1.22-4.25);P=0.01]haplotypes.CONCLUSION:Current findings show significant association of SMOC2 gene variant rs13208776 with POAG between familial and non-familial Pakistani patients.展开更多
Although climate changes are predicted to be an increasingly dominant threat to plant biodiversity, the degradation of ecosystems witnessed to date has been largely driven by factors such as human-induced habitat loss...Although climate changes are predicted to be an increasingly dominant threat to plant biodiversity, the degradation of ecosystems witnessed to date has been largely driven by factors such as human-induced habitat loss and fragmentation, overexploitation, pollution and the introduction of invasive species. Given the evidence that climate changes and anthropogenic pressures have greatly increased the extinction of natural populations of species, we can recognize that human-induced land use and climate changes are perhaps the greatest threats to terrestrial biodiversity. In this context, effective prioritization of conservation efforts is critical for the sustainability of biodiversity, as current environmental changes are likely to continue in the future. Countries with limited financial resources for conservation projects may be at greater risk from habitat loss, direct harvesting and invasive species, and may also lead to unsustainable exploitation of resources, further accelerating species loss through direct harvesting and causing rapid loss of biodiversity. In this context, the protection of biodiversity is an important issue that concerns the entire world population. Causes such as anthropogenic pressures, great fires, introduction of new species from different regions, invasion of cultivars and dominant species cause a dramatic impact on plant biodiversity as well as an increase in the number of threatened species. Plant biodiversity constitutes the natural source of products used in the food and pharmaceutical industries and also provides basic different raw materials. On the other hand, plant biodiversity is important in the development of species and more productive species that are more resistant to biological and environmental stresses, and in providing new genetic information for feeding programs. Advances in plant biotechnology, particularly in vitro cultures and molecular biology, have been a powerful tool in the control and conservation of plant biodiversity. Today, biotechnological methods include the most suitable methods for the pathogen-free short-, medium- and long-term preservation of ornamental plants, medicinal and aromatic plants and woody species that are in danger of extinction. In vitro conservation strategies are especially important in the protection of plant species that are vegetatively propagated and have seeds that are intolerant to desiccation. In addition, in vitro techniques provide a reliable platform for the international exchange of plant material, enable the creation of large collections using minimal space, enable the acquisition of valuable materials for wild species recovery, and facilitate molecular research and ecological studies.展开更多
Developing high-yield maize hybrids is critical for sustaining maize production,especially in the face of rapid climate changes and the growing global population.Exploring the genetic diversity and combining ability i...Developing high-yield maize hybrids is critical for sustaining maize production,especially in the face of rapid climate changes and the growing global population.Exploring the genetic diversity and combining ability in parental inbreds is needed for developing such high-yielding hybrids.Consequently,this study aimed at evaluating parental genetic diversity employing simple sequence repeats(SSR)markers,estimating effects of general(GCA)and specific(SCA)combining abilities for grain yield and yield contributing characters,identifying high yielding hybrids,and evaluating the association of SCA effects and performance of hybrids with genetic distance.Half-diallel mating scheme was utilized to develop 21 F_(1) hybrids from seven diverse maize inbred lines.The F_(1) hybrids along with check hybrid(SC-10),were investigated in a field trial over two growing seasons under arid conditions.The assessed F_(1) hybrids displayed significant genetic variations across all recorded traits.The inbreds P_(1) and P_(3) were detected as effective combiners to develop early maturing hybrids.Additionally,P_(3) and P_(4) were recognized as better combiners for improving grain yield and yield attributed characters.The hybrids P_(1)×P_(5) and P_(4)×P_(7) displayed significant SCA effects coupled with favorable agronomic performance.These hybrids are recommended for further evaluation and release as variety for arid environments to increase total maize production and contribute to food security.The alleles per locus differed between 2 and 5,with average of 3.5 alleles/locus.The polymorphic information content(PIC)altered between 0.21 to 0.74,with a mean of 0.56.Unweighted neighbor-joining tree grouped the inbred lines into three clusters,providing a valuable tool to decrease the crosses needed to be assessed in the trial field.Parental genetic distance varied from 0.63 to 0.90,averaging 0.79.The relationship between genetic diversity assessed through SSR markers and SCA effects was insignificant for all considered traits.Otherwise,SCA demonstrated a significant correlation with hybrid performance,suggesting that SCA serves as a reliable predictor for hybrid performance.The assessed maize inbred lines and developed hybrids revealed substantial genetic variability,offering valuable resources for enhancing maize productivity under arid conditions.The identified promising inbred lines(P_(1),P_(3),and P_(4))might be regarded as effective combiners for developing early-maturing genotypes and excellent combiners for enhancing yield attributes.Notably,the developed hybrids P_(1)×P_(5) and P_(4)×P_(7) possessed significant SCA alongside superior yield traits.SCA demonstrated a significant correlation with hybrid performance,suggesting its potential as a reliable predictor for the performance of developed hybrids.展开更多
Dear Editor,Genome-wide association studies(GWASs)have reported a genetic association between certain populations and the severity of COVID-19.In severe patients,changes in the locus 3p21.31(including SLC6A20,LZTFL1,C...Dear Editor,Genome-wide association studies(GWASs)have reported a genetic association between certain populations and the severity of COVID-19.In severe patients,changes in the locus 3p21.31(including SLC6A20,LZTFL1,CCR9,FYCO1,CXCR6,and XCR1)are associated with COVID-19 hospitalization(Ellinghaus et al.,2020).Moreover,variations in 9p34.2 region(including genes related to the ABO blood grouping)can affect both susceptibility and severity of COVID-19;in particular,individuals with blood group A may have an increased risk of COVID-19 infection,while those with blood group O may have a protective effect(Ellinghaus et al.,2020;Shelton et al.,2021).展开更多
BACKGROUND Thus far,genetic analysis of patients clinically diagnosed with glycogen storage diseases(GSDs)in Thailand has not been reported.AIM To evaluate the clinical and biochemical profiles,molecular analysis and ...BACKGROUND Thus far,genetic analysis of patients clinically diagnosed with glycogen storage diseases(GSDs)in Thailand has not been reported.AIM To evaluate the clinical and biochemical profiles,molecular analysis and long-term outcomes of Thai children diagnosed with hepatic GSD.METHODS Children aged<18 years diagnosed with hepatic GSD and followed up at King Chulalongkorn Memorial Hospital were recruited.Whole-exome sequencing(WES)was performed to identify the causative gene variants.Medical records were assessed.RESULTS All eight children with histopathologically confirmed diagnosis were classified by WES into subtypes Ia(n=1),III(n=3),VI(n=3),and IX(n=1).A total number of 10 variants were identified including G6PC(n=1),AGL(n=4),PYGL(n=5),and PHKA2(n=1).AGL had two novel variants.The clinical manifestations were hepatomegaly(n=8),doll-like facies(n=3),wasting(n=2),and stunting(n=5).All patients showed hypoglycemia,transaminitis,and dyslipidemia.The mainstay of treatment was cornstarch supplementation and high-protein and low-lactosefructose diet.After a median follow-up time of 9.59 years,height turned to normal for age in 3/5 patients and none had malnutrition.Liver enzymes,blood sugar,and lipid profiles improved in all.CONCLUSION Hepatomegaly,transaminitis,and hypoglycemia are the hallmarks of GSD confirmed by liver histopathology.Molecular analysis can confirm the diagnosis or classify the subtype that might benefit from personalized treatment,prognosis,and long-term care.展开更多
Aim： To detect the expression of hepatitis B virus （HBV） genes （HB S and C genes） in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fer...Aim： To detect the expression of hepatitis B virus （HBV） genes （HB S and C genes） in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fertilization （IVF） technique. Methods： Human sperm-mediated HBV genes were delivered into zona-free hamster oocytes by the IVF method. Polymerase chain reaction （PCR） was used to detect HB S and pre-Core/Core （pre-C/C） coding genes both in one- and two-cell embryos. Reverse transcription-PCR （RT-PCR） analysis was used to study the expression of the two genes. Fluorescence in situ hybridization （FISH） analysis using the full-length HBV DNA as the hybridization probe was performed to confirm the integration of viral DNA in the host embryonic genome. Results： Both HB S and pre-C/C coding genes are present and transcribed in one- and two-cell embryos originated from hamster ova IVF with human spermatozoa carrying HBV DNA sequences. Conclusion： Sperm-mediated HBV genes are able to replicate and express themselves in early embryonic cells. These results provide direct evidence that HBV DNA could transmit vertically to the next generation via the male germ line.展开更多
The combination of PCR amplification of 16S rRNA genes with denaturing gradient gel electrophoresis （DGGE） analysis was used to reveal the compositions and dynamics of bacterial communities in a sewage treatment pla...The combination of PCR amplification of 16S rRNA genes with denaturing gradient gel electrophoresis （DGGE） analysis was used to reveal the compositions and dynamics of bacterial communities in a sewage treatment plant with two systems, i.e., an anoxic- anaerobic-aerobic system （inverted A2O） and an anaerobic-anoxic-aerobic one （conventional A2O） over a period from February to July 2009, during which both systems experienced serious sludge bulking problems. The DGGE patterns showed that there were many common bands in both systems, suggesting the high similarity of bacterial communities of the two systems. Meanwhile, the moving window correlation analysis showed that the two systems experienced different microbial community structure changes during the period, which might be related with the different situations of the occurrence and disappearance of sludge bulking, as being reflected by sludge volume index （SVI） values. Major bands of DGGE patterns of sludge samples were further sequenced. Phylogenetic affiliation indicated that the majority of the sequences obtained were affiliated with Actinobacteria, Firmicutes, Bacteroidetes/Chlorobi group and α- and β-Proteobacteria. Two sequences showed high similarities to typical filamentous bacteria Microthrix parvicella and Nostocoida limicola I, indicating that these bacterial species have been involved in the sludge bulking problems.展开更多
AIM:To improve hepatic differentiation of human mesenchymal stem cell(MSC)using insulin growth factor 1(IGF-Ⅰ),which has important role in liver development,hepatocyte differentiation and function.METHODS:Bone marrow...AIM:To improve hepatic differentiation of human mesenchymal stem cell(MSC)using insulin growth factor 1(IGF-Ⅰ),which has important role in liver development,hepatocyte differentiation and function.METHODS:Bone marrow of healthy donors was aspirated from the iliac crest.The adherent cells expanded rapidly and were maintained with periodic passages until a relatively homogeneous population was established.The identification of these cells was carried out by immunophenotype analysis and differentiation potential into osteocytes and adipocytes.To effectively induce hepatic differentiation,we designed a protocol based on a combination of IGF-Ⅰ and liver specificfactors(hepatocyte growth factor,oncostatin M and dexamethasone).Morphological features,hepatic functions and cytological staining were assessed to evaluate transdifferentiation of human marrow-derived MSCs.RESULTS:Flow cytometric analysis and the differentiation potential into osteoblasts and adipocytes showed that more than 90% of human MSCs which were isolated and expanded were positive by specif ic markers and functional tests.Morphological assessment and evaluation of glycogen storage,albumin and α-feto protein expression,as well as albumin and urea secretion revealed a statistically signif icant difference between the experimental groups and control.CONCLUSION:In vitro differentiated MSCs using IGF-Ⅰwere able to display advanced liver metabolic functions,supporting the possibility of developing them as potential alternatives to primary hepatocytes.展开更多
We have yet to develop a fundamental understanding of the molecular complexities of human spermatozoa.This encompasses the unique packaging and structure of the sperm genome along with their paternally derived RNAs in...We have yet to develop a fundamental understanding of the molecular complexities of human spermatozoa.This encompasses the unique packaging and structure of the sperm genome along with their paternally derived RNAs in preparation for their delivery to the egg.The diversity of these transcripts is vast,including several anti-sense mol- ecules resembling known regulatory micro-RNAs.The field is still grasping with its delivery to the oocyte at fertiliza- tion and possible significance.It remains tempting to analogize them to maternally-derived transcripts active in early embryo patterning.Irrespective of their role in the embryo,their use as a means to assess male factor infertility is promising.展开更多
基金the Genetics and Agricultural Biotechnology Institute of Tabarestan (GABIT) Sari Agricultural Sciences and Natural Resources University (SANRU) for the use of the services and financial supports of this research
文摘The oilseed crop Camelina sativa exhibits salinity tolerance,but the effects on early growth stages across a range of different salts and in combination with salicylic acid(SA)have not been thoroughly evaluated.In this study,seeds were germinated in varying concentrations of six salts(NaCl,CaCl_(2),ZnCl_(2),KCl,MgSO_(4),and Na2SO_(4))with or without 0.5 mM SA.Using the halotime model,we estimated salt thresholds for germination and parameters of seedling growth.Germination and seedling growth parameters of camelina significantly decreased with increasing salt concentration across all salt types.Salts containing Zn and SO_(4) were most detrimental to germination and seedling growth.Except for KCl,0.5 mM SA generally reduced the salinity tolerance threshold(Saltb(50))of camelina.Specifically,Saltb(50)was 21.5%higher for KCl and 16.1%,25.0%,54.9%,21.0%,and 5.6%lower for CaCl_(2),NaCl,MgSO_(4),Na2SO_(4),and ZnCl_(2),respectively,when 0.5 mM SA was compared to 0 mM SA.Furthermore,camelina seedling growth was consistently more sensitive than germination across all salt types.SA did not significantly enhance germination or seedling growth and was harmful when combined with certain salts or at the germination stage.It can be concluded that both the type of salt and the concentration of SA are as critical as the salt concentration in saline irrigation water.
基金supported by the National Natural Science Foundation of China(82172114)the"Challenge and Response"project for key and common technology research of Hefei(GJ2022SH08).
文摘Cryopreservation is a fundamental technology in biomedical research,regenerative medicine,and tissue engineering,enabling the long-term storage of cells,tissues,and organs.However,its effectiveness is limited by challenges such as intracellular ice formation,cryoprotectant toxicity,and reduced post-thaw viability.This review explores the crucial role of encapsulation in enhancing cryopreservation efficiency,with a focus on recent advances in materials science,bioengineering,and cryobiology.Emerging technologies,such as nanotechnology and stimuli-responsive polymers,are transforming encapsulation strategies.Innovations such as microfluidic systems offer precise control over cooling rates and cryoprotectant distribution,thereby mitigating conventional limitations.The review also addresses current obstacles related to scaling up encapsulation processes and ensuring the long-term biocompatibility and stability of preserved specimens.By synthesizing recent findings,this work provides a comprehensive resource for researchers and clinicians seeking to enhance biopreservation techniques and their applications in contemporary medicine and biotechnology.Finally,the review identifies critical knowledge gaps that must be addressed to improve the efficacy of cryopreservation strategies and advance their clinical translation.
基金Anhui Province Clinical Medical Research Translation Special Program(No.2204295107020002).
文摘Oocyte cryopreservation is an essential procedure in assisted reproductive technologies,aimed at preserving fertility,particularly for women undergoing IVF treatment or at risk of ovarian damage due to radiation,chemotherapy,or surgery.Despite its growing use,the survival and fertilization rates of cryopreserved oocytes remain suboptimal,largely due to cryo-induced oxidative stress.The generation of Reactive Oxygen Species(ROS)during freezing and thawing causes considerable damage to key cellular components,including proteins,lipids,DNA,and mitochondria.This oxidative stress compromises oocyte quality and reduces developmental potential.To address these challenges,the use of additives-especially antioxidants-has shown significant promise in mitigating oxidative damage.Enzymatic antioxidants such as Superoxide Dismutase(SOD)and Catalase(CAT),along with non-enzymatic antioxidants like glutathione,melatonin,and resveratrol,have demonstrated the ability to neutralize ROS and improve oocyte viability and developmental outcomes.Recent studies highlight the potential of Mitoquinone(MitoQ),a mitochondria-targeted antioxidant,to effectively counteract mitochondrial ROS and enhance cellular defense mechanisms during cryopreservation.This review explores the cellular mechanisms of cryodamage,the role of oxidative stress in oocyte cryopreservation,and the potential of various antioxidant strategies to enhance oocyte survival and function.Developing effective antioxidant supplementation approaches may significantly improve the outcomes of cryopreservation in reproductive medicine.
基金Ministry of Science,Innovation and Universities,Spain(NextGeneration EU fundsMaría Zambrano Program 124/MTAI/22+2 种基金and PID2020-113320RB-I00)Agency for Management of University and Research Grants,Regional Government of Catalonia,Spain(2021-SGR-00900)Catalan Institution for Research and Advanced Studies(ICREA).
文摘Background Sperm cryopreservation is widely used in the cattle industry,as it allows for disassociating the localiza-tion of sires and the collection of semen from the timing of artificial insemination.While freeze-thawing is known to impair sperm DNA integrity,whether the damage induced consists of single-(SSB)or double-strand breaks(DSB)has not been determined.In addition,no previous study has addressed if DNA breaks preferentially reside in specific genome regions such as those forming the toroid linker regions,or are rather spread throughout the regions linked to protamines.The main aim of the present work,therefore,was to elucidate the type and localization of the DNA damage generated by cryopreservation and to evaluate its impact on artificial insemination outcomes in cattle.Results The incidence of SSB and DSB was evaluated in 12 ejaculates before and after cryopreservation with the Comet assay,and the localization of the DNA breaks was assessed using pulsed-field gel electrophoresis(PFGE).Before cryopreservation,the incidence of SSB was 10.99%±4.62%and involved 20.56%±3.04%of sperm cells,whereas these figures significantly(P<0.0001)increased up to 34.11%±3.48%and 53.36%±11.00%in frozen-thawed sperm.In contrast,no significant differences in the incidence of DSB were observed(P>0.990)before and after cryopreservation(before:incidence of 13.91%±1.75%of sperm DNA affecting 56.04%±12.49%of sperm cells;after:incidence of 13.55%±1.55%of sperm DNA involving 53.36%±11.00%of sperm cells).Moreover,PFGE revealed that the percentage of sperm DNA fragments whose length was shorter than a toroid(<31.5 kb)was greater(P<0.0001)after(27.00%±4.26%)than before freeze-thawing(15.57%±4.53%).These differences indicated that the DNA breaks induced by cryopreservation affect the regions condensed in protamines,which are structured in toroids.On the other hand,in vivo fertility rates were associated to the incidence of SSB and DSB in frozen-thawed sperm(P=0.032 and P=0.005),but not with the size of the DNA fragments resulting from these breaks(P>0.05).Conclusion Cryopreservation of bovine sperm generates single-strand DNA breaks,which are mainly located in protamine-condensed toroidal regions.The incidence of DNA breaks in cryopreserved sperm has an impact on cat-tle fertility,regardless of the size of generated fragments.
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: Cystinosis is a multisystemic autosomal recessive deficiency of the lysosomal membrane transporter protein (cystinosin) caused by mutations in CTNS gene. Objective: This study summarizes the Portuguese experience in the diagnosis and management of patients with this rare disease over the past few years and reports recurrent mutations in the CTNS gene. Methods: Unrelated patients from different pediatric and adult hospitals all over Portugal with non-nephrotic proteinuria, hypercalciuria, hypokalemia impaired proximal reabsorption of amino acids, glycosuria and hypophosphatemia, suggestive of a Fanconi syndrome and ocular problems, were studied. Intra-leukocyte cystine levels were determined and molecular analysis was performed, to determine the presence or absence of the 57-kb deletion in CTNS, followed by direct sequencing of the coding exons of CTNS. Results: From 1998 to 2017, twenty-one cystinotic patients were biochemically diagnosed. From the remaining seventeen (four deceased), eleven were studied for CTNS gene. Five out of eleven patients were homozygous for the 57-kb deletion (10/22;45.5%), and other five were compound heterozygous for this variant (15/22;68.2%). The other mutations found were p.Q128X (c.721 C>T;2/22), p.S139F (c.755 C>T;4/22) and c.18-21delGACT (p.T7FfsX7;1/22). All of these seventeen cystinotic patients are in treatment. Approximately 84% are adults, 16% are young children, and 54.5% are kidney transplant recipient. Conclusions: The authors would like to emphasize the importance of first screening for the 57-kb deletion since it is very common in our population. This genetic study is the first in our country and it could be the basis for future genetic counseling in Portuguese population.
基金This study was supported by Hebei Natural Science Foundation(H2022206019)Science and Technology(S and T)Program of Hebei(21377721D)+1 种基金Hebei Province Medical Technology Tracking Project(GZ2021028)Medical Science Research Project of Hebei Province(20170084,and 20211494).
文摘The sperm DNA fragmentation index(DFI)is a metric used to assess DNA fragmentation within sperm.During in vitro fertilizationembryo transfer(IVF-ET),high sperm DFI can lead to a low fertilization rate,poor embryo development,early miscarriage,etc.A kinase anchoring protein(AKAP)is a scaffold protein that can bind protein kinase A(PKA)to subcellular sites of specific substrates and protects the biophosphorylation reaction.Sperm protein antigen 17(SPA17)can also bind to AKAP.This study intends to explore the reason for the decreased fertilization rate observed in high sperm DFI(H-DFI)patients during IVF-ET.In addition,the study investigates the expression of AKAP,protein kinase A regulatory subunit(PKARIl),and SPA17 between H-DFI and low sperm DFI(L-DFI)patients.SPA17 at the transcriptional level is abnormal,the translational level increases in H-DFI patients,and the expression of AKAP4/PKARIl protein decreases.H,O,has been used to simulate oxidative stress damage to spermatozoa during the formation of sperm DFI.It indicates that H,O,increases the expression of sperm SPA17 protein and suppresses AKAP4/PKARIl protein expression.These processes inhibit sperm capacitation and reduce acrosomal reactions.Embryo culture data and IVF outcomes have been documented.The H-DFI group has a lower fertilization rate.Therefore,the results indicate that the possible causes for the decreased fertilization rate in the H-DFI patients have included loss of sperm AKAP4/PKARIl proteins,blocked sperm capacitation,and reduced occurrence of acrosome reaction.
文摘BACKGROUND Cholangiocarcinoma(CCA)is a highly malignant cancer,characterized by frequent mucin overexpression.MUC1 has been identified as a critical oncogene in the progression of CCA.However,the comprehensive understanding of how the mucin family influences CCA progression and prognosis is still incomplete.AIM To investigate the functions of mucins on the progression of CCA and to establish a risk evaluation formula for stratifying CCA patients.METHODS Single-cell RNA sequencing data from 14 CCA samples were employed for elucidating the roles of mucins,complemented by bioinformatic analyses.Subse-quent validations were conducted through spatial transcriptomics and immuno-histochemistry.The construction of a risk evaluation model utilized the least absolute shrinkage and selection operator regression algorithm,which was further confirmed by independent cohorts and diverse data types.RESULTS CCA tumor cells with elevated levels of MUC1 and MUC4 showed activated nucleotide metabolic pathways and increased invasiveness.MUC5AC-high cells were found to promote CCA progression through WNT signaling.MUC5B-high cells exhibited robust cellular oxidation activities,leading to resistance against antitumoral treatments.MUC13-high cells were observed to secret chemokines,recruiting and transforming macrophages into the M2-polarized state,thereby suppressing antitumor immunity.MUC16-high cells were found to promote tumor progression through interleukin-1/nuclear factor kappa-light-chain-enhancer of activated B cells signaling upon interaction with neutrophils.Utilizing the expression levels of these mucins,a risk factor evaluation formula for CCA was developed and validated across multiple cohorts.CCA samples with higher risk factors exhibited stronger metastatic potential,chemotherapy resistance,and poorer prognosis.CONCLUSION Our study elucidates the functional mechanisms through which mucins contribute to CCA development,and provides tools for risk stratification in CCA.
文摘Egyptian species of the leafhopper genus Exitianus Ball,1929,E.capicola(St?l,1855),E.nanus(Distant,1908),and E.pondus Ross,1968 are reviewed.Illustrations,morphological descriptions,and a key for their identification are provided.In this study,we used molecular techniques to confirm morphological identification and detect phylogeny among the three Exitianus species.The partial nucleotide sequences of the amplified products obtained were determined by Macrogen Korea.The nucleotide sequences of 28S rDNA and COX genes of the EGY-ARC-9,EGY-ARC-4,and EGY-ARC-5 were determined by Macrogen Korea,blasted into BLAST at the National Center for Biotechnology Information website(NCBI)and compared with those deposited in the GenBank DNA database.The results represented the homology percentage between the partial sequences of the 28S rRNA and COX genes from each species and related species obtained from GenBank DNA database.The morphological identifications of the three Exitianus species were confirmed by molecular characterization and sequencing of 28S rDNA and COX genes and identified as E.capicola,E.nanus,and E.pondus.Also,their sequences of 28srDNA and COX genes were deposited in GenBank with an accession number(LC670610,LC670607,and OQ196105)for 28srDNA gene and(LC775357,LC775358,and LC775359)for the COX gene.
基金Supported by Higher Education Commission of Pakistan(NRPU#2835)Pakistan Science Foundation Project No.Biotech 101,funded to Professor Dr.Ali Muhammad Waryah.
文摘AIM:To find out the association of secreted protein acidic and rich in cysteine(SPARC)-related modular calcium binding 2(SMOC2)gene variants rs2255680 and rs13208776 with genotypic and phenotypic characteristics in both familial and non-familial primary open angle glaucoma(POAG)patients.METHODS:A total of 212 POAG patients,comprising 124 familial and 88 non-familial,were enrolled.For genotyping the SMOC2 variant rs2255680,amplification refractory mutation system(ARMS)-polymerase chain reaction(PCR)method and PCR-restriction fragment length polymorphism(PCR-RFLP)were utilized for analyzing rs13208776 variant.RESULTS:The mean age of familial POAG patients was 50.92±9.12y,with 78 males and 46 females.The mean age of non-familial POAG patients was 53.14±13.44y,with 52 males and 36 females.The SMOC2 gene variant rs13208776 showed the significant association with POAG between familial and non-familial groups.The homozygous G/G variant was frequent among non-familial(60.2%)whereas the heterozygous G/A variant was more frequent in familial POAG patients(46%).There were significant differences in G/A variant between familial and non-familial glaucoma patients,and the risk was decreased to 0.53-fold in non-familial glaucoma patients[odds ratio(OR):0.53;95%confidence interval(CI):0.29-0.94;P=0.033]in codominant model.The risk was further reduced to 0.49-fold(95%CI:0.28-0.86;P=0.012)in dominant model for non-familial patients.No significant association of SMOC2 gene variant rs2255680 between familial and non-familial glaucoma patients was found in our population.The haplotype analysis showed the decreased risk for TA[OR:0.48(95%CI:0.29-0.79);P=0.004]and an increased risk for TG[OR=2.28(95%CI:1.22-4.25);P=0.01]haplotypes.CONCLUSION:Current findings show significant association of SMOC2 gene variant rs13208776 with POAG between familial and non-familial Pakistani patients.
文摘Although climate changes are predicted to be an increasingly dominant threat to plant biodiversity, the degradation of ecosystems witnessed to date has been largely driven by factors such as human-induced habitat loss and fragmentation, overexploitation, pollution and the introduction of invasive species. Given the evidence that climate changes and anthropogenic pressures have greatly increased the extinction of natural populations of species, we can recognize that human-induced land use and climate changes are perhaps the greatest threats to terrestrial biodiversity. In this context, effective prioritization of conservation efforts is critical for the sustainability of biodiversity, as current environmental changes are likely to continue in the future. Countries with limited financial resources for conservation projects may be at greater risk from habitat loss, direct harvesting and invasive species, and may also lead to unsustainable exploitation of resources, further accelerating species loss through direct harvesting and causing rapid loss of biodiversity. In this context, the protection of biodiversity is an important issue that concerns the entire world population. Causes such as anthropogenic pressures, great fires, introduction of new species from different regions, invasion of cultivars and dominant species cause a dramatic impact on plant biodiversity as well as an increase in the number of threatened species. Plant biodiversity constitutes the natural source of products used in the food and pharmaceutical industries and also provides basic different raw materials. On the other hand, plant biodiversity is important in the development of species and more productive species that are more resistant to biological and environmental stresses, and in providing new genetic information for feeding programs. Advances in plant biotechnology, particularly in vitro cultures and molecular biology, have been a powerful tool in the control and conservation of plant biodiversity. Today, biotechnological methods include the most suitable methods for the pathogen-free short-, medium- and long-term preservation of ornamental plants, medicinal and aromatic plants and woody species that are in danger of extinction. In vitro conservation strategies are especially important in the protection of plant species that are vegetatively propagated and have seeds that are intolerant to desiccation. In addition, in vitro techniques provide a reliable platform for the international exchange of plant material, enable the creation of large collections using minimal space, enable the acquisition of valuable materials for wild species recovery, and facilitate molecular research and ecological studies.
基金supported by Princess Nourah bint Abdulrahman University Researchers Supporting Project number(PNURSP2024R318)Princess Nourah bint Abdulrahman University,Riyadh,Saudi Arabia.The authors extend their appreciation to the Deanship of Research and Graduate Studies at King Khalid University for funding this work through Large Research Project under grant number RGP2/342/45supported by the Deanship of Scientific Research,Vice Presidency for Graduate Studies and Scientific Research,King Faisal University,Saudi Arabia(KFU241870).
文摘Developing high-yield maize hybrids is critical for sustaining maize production,especially in the face of rapid climate changes and the growing global population.Exploring the genetic diversity and combining ability in parental inbreds is needed for developing such high-yielding hybrids.Consequently,this study aimed at evaluating parental genetic diversity employing simple sequence repeats(SSR)markers,estimating effects of general(GCA)and specific(SCA)combining abilities for grain yield and yield contributing characters,identifying high yielding hybrids,and evaluating the association of SCA effects and performance of hybrids with genetic distance.Half-diallel mating scheme was utilized to develop 21 F_(1) hybrids from seven diverse maize inbred lines.The F_(1) hybrids along with check hybrid(SC-10),were investigated in a field trial over two growing seasons under arid conditions.The assessed F_(1) hybrids displayed significant genetic variations across all recorded traits.The inbreds P_(1) and P_(3) were detected as effective combiners to develop early maturing hybrids.Additionally,P_(3) and P_(4) were recognized as better combiners for improving grain yield and yield attributed characters.The hybrids P_(1)×P_(5) and P_(4)×P_(7) displayed significant SCA effects coupled with favorable agronomic performance.These hybrids are recommended for further evaluation and release as variety for arid environments to increase total maize production and contribute to food security.The alleles per locus differed between 2 and 5,with average of 3.5 alleles/locus.The polymorphic information content(PIC)altered between 0.21 to 0.74,with a mean of 0.56.Unweighted neighbor-joining tree grouped the inbred lines into three clusters,providing a valuable tool to decrease the crosses needed to be assessed in the trial field.Parental genetic distance varied from 0.63 to 0.90,averaging 0.79.The relationship between genetic diversity assessed through SSR markers and SCA effects was insignificant for all considered traits.Otherwise,SCA demonstrated a significant correlation with hybrid performance,suggesting that SCA serves as a reliable predictor for hybrid performance.The assessed maize inbred lines and developed hybrids revealed substantial genetic variability,offering valuable resources for enhancing maize productivity under arid conditions.The identified promising inbred lines(P_(1),P_(3),and P_(4))might be regarded as effective combiners for developing early-maturing genotypes and excellent combiners for enhancing yield attributes.Notably,the developed hybrids P_(1)×P_(5) and P_(4)×P_(7) possessed significant SCA alongside superior yield traits.SCA demonstrated a significant correlation with hybrid performance,suggesting its potential as a reliable predictor for the performance of developed hybrids.
基金supported by the National Natural Science Foundation of China(81790643,82121003,and 82271105)the Sichuan Science and Technology Program(2019YFS0368,2021YFS0404,2021YFS0369,2021YFS0388,2021ZYD0082,2021YFS0033,and 2022NSFSC1352)+2 种基金the CAMS Innovation Fund for Medical Sciences(2019-12M-5-032)Sichuan Provincial People's Hospital(2021QN13)2021 open project of Sichuan Provincial Key Laboratory for Human Disease Genetics Research(2021kflx008).
文摘Dear Editor,Genome-wide association studies(GWASs)have reported a genetic association between certain populations and the severity of COVID-19.In severe patients,changes in the locus 3p21.31(including SLC6A20,LZTFL1,CCR9,FYCO1,CXCR6,and XCR1)are associated with COVID-19 hospitalization(Ellinghaus et al.,2020).Moreover,variations in 9p34.2 region(including genes related to the ABO blood grouping)can affect both susceptibility and severity of COVID-19;in particular,individuals with blood group A may have an increased risk of COVID-19 infection,while those with blood group O may have a protective effect(Ellinghaus et al.,2020;Shelton et al.,2021).
基金Supported by Ratchadaphiseksomphot Fund,Graduate Affairs,Faculty of Medicine,Chulalongkorn University,No.GA66/020Ratchadaphiseksomphot Fund,Chulalongkorn University,No.RCU_H_64_007_30.
文摘BACKGROUND Thus far,genetic analysis of patients clinically diagnosed with glycogen storage diseases(GSDs)in Thailand has not been reported.AIM To evaluate the clinical and biochemical profiles,molecular analysis and long-term outcomes of Thai children diagnosed with hepatic GSD.METHODS Children aged<18 years diagnosed with hepatic GSD and followed up at King Chulalongkorn Memorial Hospital were recruited.Whole-exome sequencing(WES)was performed to identify the causative gene variants.Medical records were assessed.RESULTS All eight children with histopathologically confirmed diagnosis were classified by WES into subtypes Ia(n=1),III(n=3),VI(n=3),and IX(n=1).A total number of 10 variants were identified including G6PC(n=1),AGL(n=4),PYGL(n=5),and PHKA2(n=1).AGL had two novel variants.The clinical manifestations were hepatomegaly(n=8),doll-like facies(n=3),wasting(n=2),and stunting(n=5).All patients showed hypoglycemia,transaminitis,and dyslipidemia.The mainstay of treatment was cornstarch supplementation and high-protein and low-lactosefructose diet.After a median follow-up time of 9.59 years,height turned to normal for age in 3/5 patients and none had malnutrition.Liver enzymes,blood sugar,and lipid profiles improved in all.CONCLUSION Hepatomegaly,transaminitis,and hypoglycemia are the hallmarks of GSD confirmed by liver histopathology.Molecular analysis can confirm the diagnosis or classify the subtype that might benefit from personalized treatment,prognosis,and long-term care.
基金Acknowledgment This work was supported by a grant from the National Nature Science Foundation of China （No. 39970374）. The authors wish to thank Professor Yi-Pong Hu, Second Military Medical University of China, for his kindness in providing us the recombinant plasmid （pBR322-HBV）. We wish to thank Mr. Michael Talion of Shantou University Medical College, English Language Training Section for his assistance in proofreading this manuscript. We gratefully acknowledge the support of the leaders of Shantou University Medical College.
文摘Aim： To detect the expression of hepatitis B virus （HBV） genes （HB S and C genes） in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fertilization （IVF） technique. Methods： Human sperm-mediated HBV genes were delivered into zona-free hamster oocytes by the IVF method. Polymerase chain reaction （PCR） was used to detect HB S and pre-Core/Core （pre-C/C） coding genes both in one- and two-cell embryos. Reverse transcription-PCR （RT-PCR） analysis was used to study the expression of the two genes. Fluorescence in situ hybridization （FISH） analysis using the full-length HBV DNA as the hybridization probe was performed to confirm the integration of viral DNA in the host embryonic genome. Results： Both HB S and pre-C/C coding genes are present and transcribed in one- and two-cell embryos originated from hamster ova IVF with human spermatozoa carrying HBV DNA sequences. Conclusion： Sperm-mediated HBV genes are able to replicate and express themselves in early embryonic cells. These results provide direct evidence that HBV DNA could transmit vertically to the next generation via the male germ line.
基金supported by the National Research Foundation, China (No. 20921140094)the Chinese Academy of Sciences, Knowledge innovation Project(No. KSCX2-YW-G-054, KZCX2-YW-JC407)CASTWAS Postdoctoral Fellowships 2009
文摘The combination of PCR amplification of 16S rRNA genes with denaturing gradient gel electrophoresis （DGGE） analysis was used to reveal the compositions and dynamics of bacterial communities in a sewage treatment plant with two systems, i.e., an anoxic- anaerobic-aerobic system （inverted A2O） and an anaerobic-anoxic-aerobic one （conventional A2O） over a period from February to July 2009, during which both systems experienced serious sludge bulking problems. The DGGE patterns showed that there were many common bands in both systems, suggesting the high similarity of bacterial communities of the two systems. Meanwhile, the moving window correlation analysis showed that the two systems experienced different microbial community structure changes during the period, which might be related with the different situations of the occurrence and disappearance of sludge bulking, as being reflected by sludge volume index （SVI） values. Major bands of DGGE patterns of sludge samples were further sequenced. Phylogenetic affiliation indicated that the majority of the sequences obtained were affiliated with Actinobacteria, Firmicutes, Bacteroidetes/Chlorobi group and α- and β-Proteobacteria. Two sequences showed high similarities to typical filamentous bacteria Microthrix parvicella and Nostocoida limicola I, indicating that these bacterial species have been involved in the sludge bulking problems.
基金Supported by A grant from Stem Cell Organization:www.stem cell.ir
文摘AIM:To improve hepatic differentiation of human mesenchymal stem cell(MSC)using insulin growth factor 1(IGF-Ⅰ),which has important role in liver development,hepatocyte differentiation and function.METHODS:Bone marrow of healthy donors was aspirated from the iliac crest.The adherent cells expanded rapidly and were maintained with periodic passages until a relatively homogeneous population was established.The identification of these cells was carried out by immunophenotype analysis and differentiation potential into osteocytes and adipocytes.To effectively induce hepatic differentiation,we designed a protocol based on a combination of IGF-Ⅰ and liver specificfactors(hepatocyte growth factor,oncostatin M and dexamethasone).Morphological features,hepatic functions and cytological staining were assessed to evaluate transdifferentiation of human marrow-derived MSCs.RESULTS:Flow cytometric analysis and the differentiation potential into osteoblasts and adipocytes showed that more than 90% of human MSCs which were isolated and expanded were positive by specif ic markers and functional tests.Morphological assessment and evaluation of glycogen storage,albumin and α-feto protein expression,as well as albumin and urea secretion revealed a statistically signif icant difference between the experimental groups and control.CONCLUSION:In vitro differentiated MSCs using IGF-Ⅰwere able to display advanced liver metabolic functions,supporting the possibility of developing them as potential alternatives to primary hepatocytes.
文摘We have yet to develop a fundamental understanding of the molecular complexities of human spermatozoa.This encompasses the unique packaging and structure of the sperm genome along with their paternally derived RNAs in preparation for their delivery to the egg.The diversity of these transcripts is vast,including several anti-sense mol- ecules resembling known regulatory micro-RNAs.The field is still grasping with its delivery to the oocyte at fertiliza- tion and possible significance.It remains tempting to analogize them to maternally-derived transcripts active in early embryo patterning.Irrespective of their role in the embryo,their use as a means to assess male factor infertility is promising.
