The laying quail is a worldwide breed which exhibits high economic value. In our current study, the vas- oactive intestinal peptide receptor-1 （VIPR-1） was selected as the candidate gene for identifying traits of eg...The laying quail is a worldwide breed which exhibits high economic value. In our current study, the vas- oactive intestinal peptide receptor-1 （VIPR-1） was selected as the candidate gene for identifying traits of egg produc- tion. A single nucleotide polymorphism （SNP） detection was performed in 443 individual quails, including 196 quails from the H line, 202 quails from the L line, and 45 wild quails. The SNPs were genotyped using a polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）. Two mutations （G373T, A313G） were detected in all the tested quail populations. The associated analysis showed that the SNP genotypes of the VIPR-1 gene were sig- nificantly linked with the egg weight of G373T and A313G in 398 quails. The quails with the genotype GG always exhibited the largest egg weight for the two mutations in the H and L lines. Linkage disequilibrium （LD） analysis in- dicated that G373T and A313G loci showed the weakest LD. Seven main diplotypes from the four main reconstructed haplotypes were observed, indicating a significant association of diplotypes with egg weight. Quails with the hlh2 （GGGT） diplotype always exhibited the smallest egg weight and largest egg number at 20 weeks of age. The overall results suggest that the alterations in quails may be linked with potential major loci or genes affecting reproductive traits.展开更多
Improved yield potential is the goal of barley domestication and cultivation.During this process,two-and six-rowed barley types emerged and have been utilised in breeding and production.The six-rowed type could produc...Improved yield potential is the goal of barley domestication and cultivation.During this process,two-and six-rowed barley types emerged and have been utilised in breeding and production.The six-rowed type could produce three times as many grains as its ancestral two-rowed forms,thus dominating barley cultivation for thousands of years.The deficiens form of the two-rowed type,characterised by extremely suppressed lateral spikelets,has gained dominance over the past few decades in barley-growing regions worldwide.We hypothesised that the absence of lateral spikelets in deficiens barley affects spike architecture and spike-related traits,contributing to its superior yield potential of deficiens barley cultivation.Currently,a deficiens barley variety,RGT Planet,is the most popular barley variety in the world.In this study,we used two F_(2) populations derived from crossing RGT Planet with two canonical two-rowed barley and identified the functional allele Vrs1.t1 associated with deficiens morphology.We observed that the Vrs1.t1 allele may contribute to high yield potential by optimising spike architecture through increased spikelet length,grain number,and grain size.Phylogenetic analysis suggests that the deficiens mutation was likely present from the early stages of barley cultivation in the Fertile Crescent and spread to Ethiopia and beyond with agricultural expansion.We conclude that the ancient deficiens allele Vrs1.t1 has been a critical driver for the recent success of modern barley improvement by optimising spike architecture.展开更多
目的探讨星状神经节阻滞(SGB)通过长链非编码RNA(lncRNA)牛磺酸上调基因1(TUG1)-NOD样受体热蛋白结构域相关蛋白3(NLRP3)轴在体外脑缺血再灌注模型中对炎症反应和自噬溶酶体形成的调节作用。方法培养大鼠海马神经元细胞系H19-7,并将细...目的探讨星状神经节阻滞(SGB)通过长链非编码RNA(lncRNA)牛磺酸上调基因1(TUG1)-NOD样受体热蛋白结构域相关蛋白3(NLRP3)轴在体外脑缺血再灌注模型中对炎症反应和自噬溶酶体形成的调节作用。方法培养大鼠海马神经元细胞系H19-7,并将细胞分为8组:(i)正常对照组:正常培养的神经元细胞;(ii)氧-糖剥夺/复氧(OGD/R)组:采用氧-糖剥夺/复氧法模拟脑缺血再灌注损伤;(iii)OGD/R+SGB组:OGD/R联合麻醉药0.5%布比卡因用于体外模拟SGB;(iv)OGD/R+SGB+TUG1过表达阴性对照组:OGD/R联合布比卡因并联合TUG1过表达阴性对照质粒转染细胞;(v)OGD/R+SGB+TUG1过表达组:OGD/R联合布比卡因并联合TUG1过表达质粒转染细胞;(vi)OGD/R+SGB+TUG1过表达+MCC950组:OGD/R联合布比卡因、TUG1过表达质粒转染及NLRP3抑制剂MCC950处理细胞;(vii)OGD/R+TUG1过表达组:OGD/R联合TUG1过表达质粒转染细胞;(viii)OGD/R+MCC950组:OGD/R联合NLRP3抑制剂MCC950处理细胞。进一步通过实时定量PCR(Quantitative Real Time PCR,qRTPCR)实验检测细胞中lncRNATUG1的表达;利用Western blot法检测细胞中NLRP3、微管相关蛋白1轻链3(LC3)-I、LC3-II、自噬相关基因5(Atg5)、苄氯素1(beclin1)、自噬接头蛋白(p62)、溶酶体相关膜蛋白1(LAMP1)的表达水平;利用透射电镜(TEM)检测自噬溶酶体的数量;并用酶联免疫吸附测定(ELISA)法检测细胞培养上清中白细胞介素(IL)-1β、IL-6、IL-18、肿瘤坏死因子(TNF)-α的含量。结果与正常对照组相比,OGD/R组lncRNA TUG1、NLRP3、Atg5、beclin1、p62、LAMP1的表达水平以及LC3-II/I比值均显著上调(^(均)P<0.05),自噬溶酶体数量增加(P<0.05),IL-1β、IL-6、IL-18、TNF-α的含量显著升高(^(均)P<0.05)。与OGD/R组相比,OGD/R+SGB组的上述指标均显著下调(P<0.05)。与OGD/R+SGB+TUG1过表达阴性对照组相比,OGD/R+SGB+TUG1过表达组的lncRNA TUG1、NLRP3、Atg5、beclin1、p62、LAMP1的表达水平以及LC3-II/I比值均显著上调(^(均)P<0.05),自噬溶酶体数量增加(P<0.05),IL-1β、IL-6、IL-18、TNF-α的含量显著升高(^(均)P<0.05),然而,加入NLRP3的抑制剂MCC950后,除lncRNA TUG1外其余指标均显著下调(^(均)P<0.05)。另外,与OGD/R组比,OGD/R+TUG1过表达组的上述指标进一步上调(^(均)P<0.05)。与OGD/R组比,OGD/R+MCC950组则抑制了除lncRNA TUG1外的其余指标(^(均)P<0.05)。结论星状神经节阻滞通过调节lncRNA TUG1-NLRP3轴有效减轻体外脑缺血再灌注损伤引起的炎症反应和自噬溶酶体形成,提示其可能作为治疗缺血性脑损伤的潜在策略。展开更多
Polyembryony has posed a significant impediment to the advancement of citrus hybrid breeding.FhRWP is widely regarded as a pivotal factor governing asexual reproduction in citrus,and prior research has demonstrated th...Polyembryony has posed a significant impediment to the advancement of citrus hybrid breeding.FhRWP is widely regarded as a pivotal factor governing asexual reproduction in citrus,and prior research has demonstrated that FhARID1,acting as an upstream regulator,modulates FhRWP expression.In this study,we performed a genome-wide characterization of the ARID-HMG-related genes using the short juvenile minicitrus Fortunella hindsii.A total of 20 ARID-HMG-related genes were identified.Protein interaction network and enrichment analysis suggested that ARID-HMG-related proteins might might be involved in chromatin remodeling complexes.Knockout of FhARID1 in F.hindsii did not induce the conversion from polyembryony to monoembryony.However,fharid1 plants in T1 generation exhibited abnormal proliferation at axillary buds,which is similar to phenotype of fhrwp plants.Expression analysis of fharid1 ovary tissues revealed the downregulation of FhRWP.The results indicated that FhARID1,as an upstream regulator of FhRWP,has an effect on the development of citrus axillary buds.Expression analysis of overexpressed leaves of FhARID1 lines showed that no significant up-regulation of FhRWP,indicating that FhARID1 is not the sole upstream regulatory factor of FhRWP.Only FhARID2 showed a correlation in expression with FhARID1 among the ARID-related genes,further supporting the notion that this gene may be involved in complex formation rather than acting alone.Yeast two-hybrid and MS/MS spectra further indicated that FhARID1 function requires casein kinase II-mediated post-transcriptional phosphorylation.This study elucidated the function of FhARID1 in citrus apomixis and axillary bud development,providing a fundamental basis for understanding the role of ARID-HMG-related genes.展开更多
BACKGROUND Wolfram syndrome is a rare autosomal recessive genetic disorder characterized by early-onset diabetes and progressive neurodegeneration,most notably sensorineural hearing loss and optic atrophy.Because its ...BACKGROUND Wolfram syndrome is a rare autosomal recessive genetic disorder characterized by early-onset diabetes and progressive neurodegeneration,most notably sensorineural hearing loss and optic atrophy.Because its initial manifestations are usually similar to those of type 1 diabetes,the diagnosis may be delayed until other manifestations appear.Pathogenic variations of the WFS1 gene can disrupt endoplasmic reticulum function and cellular homeostasis,but the complete mutation spectrum of WFS1 has not been fully determined.Early identification of monogenic diabetes caused by Wolfram syndrome is of vital importance,as it enables the provision of targeted multidisciplinary care and genetic counseling for affected families.CASE SUMMARY A 2-year-old Han Chinese girl was admitted with a 1-month history of polydipsia,polyuria,and polyphagia,and was diagnosed with diabetic ketoacidosis and impaired insulin secretion.Sensorineural hearing loss was also detected.Wholeexome sequencing identified a previously unreported heterozygous mutation,WFS1 c.986T>C(p.Phe329Ser),in the patient and her father,confirming the diagnosis of Wolfram syndrome.Bioinformatic analysis supported the likely pathogenicity of this mutation.In silico pathogenicity predictors(REVEL,SIFT,Poly-Phen-2,MutationTaster,and GERP+)supported a deleterious effect on wolframin structure and function.The patient was initially treated with intravenous insulin and fluid resuscitation,then transitioned to a basal–bolus insulin regimen.Glycemic control was subsequently maintained with continuous subcutaneous insulin infusion and intermittent subcutaneous injections.At the 1-and 6-month follow-ups,blood glucose remained well controlled(hemoglobin A1c:5.89%and 6.58%,respectively),with no evidence of organ dysfunction or further complications.CONCLUSION This case identifies WFS1 c.986T>C(p.Phe329Ser)as a novel pathogenic variant causing Wolfram syndrome.It highlights the importance of early genetic testing in pediatric patients with atypical diabetes presentations to enable timely diagnosis,individualized therapy,and comprehensive family support.展开更多
本研究通过同源比对和结构域分析预测了水稻中38个PR1基因的理化性质、细胞定位和启动子结构特征,然后根据已有基因表达信息分析了PR1基因在生物胁迫中的响应表达情况,最后分析了这38个基因在水稻感染白叶枯病菌后的表达水平。结果发现3...本研究通过同源比对和结构域分析预测了水稻中38个PR1基因的理化性质、细胞定位和启动子结构特征,然后根据已有基因表达信息分析了PR1基因在生物胁迫中的响应表达情况,最后分析了这38个基因在水稻感染白叶枯病菌后的表达水平。结果发现38个PR蛋白中有35个蛋白的长度约为160 aa,有26个为酸性蛋白,32个为亲水性蛋白,27个可分泌到胞外。启动子顺式作用元件预测发现PR1基因启动子普遍含有MYB和茉莉酸响应结合元件。有20个PR1基因定位于7号染色体上,且多数PR1基因在染色体上呈现串联重复样排列。有37个PR基因含有CAP结构域。白叶枯病菌侵染水稻后,前3 d PR1-13未有表达,但3 d后表达显著提高;PR1-33在接种后持续性高表达;PR1-2、PR1-3、PR1-8、PR1-32、PR1-34、PR1-35、PR1-37的表达受病原菌抑制。以上结果暗示PR1基因在染色体上的串联排列可能有助于水稻快速响应病原菌的侵染,但PR1基因响应生物胁迫的表达与染色体定位无关,可能主要受启动子的顺式作用元件调控。本研究为分析PR1在抗病中的作用及机制提供了理论依据。展开更多
Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-asso...Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa.展开更多
Objective To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress.Methods Twenty male SD rats were ra...Objective To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress.Methods Twenty male SD rats were randomized equally into control group and heat stress group.After exposure to 32℃for 2 weeks in the latter group,the rats were examined for histopathological changes and Bmal1 expression in the thoracic aorta using HE staining and immunohistochemistry.In the cell experiments,cultured rat thoracic aortic endothelial cells(RTAECs)were incubated at 40℃for 12 h with or without prior transfection with a Bmal1-specific small interfering RNA(si-Bmal1)or a negative sequence.In both rat thoracic aorta and RTAECs,the expressions of Bmal1,the cell cycle proteins CDK1,CDK4,CDK6,and cyclin B1,and apoptosis-related proteins Bax and Bcl-2 were detected using Western blotting.TUNEL staining was used to detect cell apoptosis in rat thoracic aorta,and the changes in cell cycle distribution and apoptosis in RTAECs were analyzed with flow cytometry.Results Compared with the control rats,the rats exposed to heat stress showed significantly increased blood pressures and lowered heart rate with elastic fiber disruption and increased expressions of Bmal1,cyclin B1 and CDK1 in the thoracic aorta(P<0.05).In cultured RTAECs,heat stress caused significant increase of Bmal1,cyclin B1 and CDK1 protein expression levels,which were obviously lowered in cells with prior si-Bmal1 transfection.Bmal1 knockdown also inhibited heat stress-induced increase of apoptosis in RTAECs as evidenced by decreased expression of Bax and increased expression of Bcl-2.Conclusion Heat stress upregulates Bmal1 expression and causes alterations in expressions of cyclins to trigger apoptosis of rat thoracic aorta endothelial cells,which can be partly alleviated by suppressing Bmal1 expression.展开更多
基金supported by the Open Project of Hubei Key Laboratory of Animal Embryo and Molecular Breeding(No.2015ZD146),China
文摘The laying quail is a worldwide breed which exhibits high economic value. In our current study, the vas- oactive intestinal peptide receptor-1 （VIPR-1） was selected as the candidate gene for identifying traits of egg produc- tion. A single nucleotide polymorphism （SNP） detection was performed in 443 individual quails, including 196 quails from the H line, 202 quails from the L line, and 45 wild quails. The SNPs were genotyped using a polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）. Two mutations （G373T, A313G） were detected in all the tested quail populations. The associated analysis showed that the SNP genotypes of the VIPR-1 gene were sig- nificantly linked with the egg weight of G373T and A313G in 398 quails. The quails with the genotype GG always exhibited the largest egg weight for the two mutations in the H and L lines. Linkage disequilibrium （LD） analysis in- dicated that G373T and A313G loci showed the weakest LD. Seven main diplotypes from the four main reconstructed haplotypes were observed, indicating a significant association of diplotypes with egg weight. Quails with the hlh2 （GGGT） diplotype always exhibited the smallest egg weight and largest egg number at 20 weeks of age. The overall results suggest that the alterations in quails may be linked with potential major loci or genes affecting reproductive traits.
基金Funding for this research was provided by the Australia Grain Research and Development Corporation(9176507)the Western Crop Genetics Alliance.Jingye Cheng thanks The University of Tasmania,Australia for the scholarship(495802)。
文摘Improved yield potential is the goal of barley domestication and cultivation.During this process,two-and six-rowed barley types emerged and have been utilised in breeding and production.The six-rowed type could produce three times as many grains as its ancestral two-rowed forms,thus dominating barley cultivation for thousands of years.The deficiens form of the two-rowed type,characterised by extremely suppressed lateral spikelets,has gained dominance over the past few decades in barley-growing regions worldwide.We hypothesised that the absence of lateral spikelets in deficiens barley affects spike architecture and spike-related traits,contributing to its superior yield potential of deficiens barley cultivation.Currently,a deficiens barley variety,RGT Planet,is the most popular barley variety in the world.In this study,we used two F_(2) populations derived from crossing RGT Planet with two canonical two-rowed barley and identified the functional allele Vrs1.t1 associated with deficiens morphology.We observed that the Vrs1.t1 allele may contribute to high yield potential by optimising spike architecture through increased spikelet length,grain number,and grain size.Phylogenetic analysis suggests that the deficiens mutation was likely present from the early stages of barley cultivation in the Fertile Crescent and spread to Ethiopia and beyond with agricultural expansion.We conclude that the ancient deficiens allele Vrs1.t1 has been a critical driver for the recent success of modern barley improvement by optimising spike architecture.
文摘目的探讨星状神经节阻滞(SGB)通过长链非编码RNA(lncRNA)牛磺酸上调基因1(TUG1)-NOD样受体热蛋白结构域相关蛋白3(NLRP3)轴在体外脑缺血再灌注模型中对炎症反应和自噬溶酶体形成的调节作用。方法培养大鼠海马神经元细胞系H19-7,并将细胞分为8组:(i)正常对照组:正常培养的神经元细胞;(ii)氧-糖剥夺/复氧(OGD/R)组:采用氧-糖剥夺/复氧法模拟脑缺血再灌注损伤;(iii)OGD/R+SGB组:OGD/R联合麻醉药0.5%布比卡因用于体外模拟SGB;(iv)OGD/R+SGB+TUG1过表达阴性对照组:OGD/R联合布比卡因并联合TUG1过表达阴性对照质粒转染细胞;(v)OGD/R+SGB+TUG1过表达组:OGD/R联合布比卡因并联合TUG1过表达质粒转染细胞;(vi)OGD/R+SGB+TUG1过表达+MCC950组:OGD/R联合布比卡因、TUG1过表达质粒转染及NLRP3抑制剂MCC950处理细胞;(vii)OGD/R+TUG1过表达组:OGD/R联合TUG1过表达质粒转染细胞;(viii)OGD/R+MCC950组:OGD/R联合NLRP3抑制剂MCC950处理细胞。进一步通过实时定量PCR(Quantitative Real Time PCR,qRTPCR)实验检测细胞中lncRNATUG1的表达;利用Western blot法检测细胞中NLRP3、微管相关蛋白1轻链3(LC3)-I、LC3-II、自噬相关基因5(Atg5)、苄氯素1(beclin1)、自噬接头蛋白(p62)、溶酶体相关膜蛋白1(LAMP1)的表达水平;利用透射电镜(TEM)检测自噬溶酶体的数量;并用酶联免疫吸附测定(ELISA)法检测细胞培养上清中白细胞介素(IL)-1β、IL-6、IL-18、肿瘤坏死因子(TNF)-α的含量。结果与正常对照组相比,OGD/R组lncRNA TUG1、NLRP3、Atg5、beclin1、p62、LAMP1的表达水平以及LC3-II/I比值均显著上调(^(均)P<0.05),自噬溶酶体数量增加(P<0.05),IL-1β、IL-6、IL-18、TNF-α的含量显著升高(^(均)P<0.05)。与OGD/R组相比,OGD/R+SGB组的上述指标均显著下调(P<0.05)。与OGD/R+SGB+TUG1过表达阴性对照组相比,OGD/R+SGB+TUG1过表达组的lncRNA TUG1、NLRP3、Atg5、beclin1、p62、LAMP1的表达水平以及LC3-II/I比值均显著上调(^(均)P<0.05),自噬溶酶体数量增加(P<0.05),IL-1β、IL-6、IL-18、TNF-α的含量显著升高(^(均)P<0.05),然而,加入NLRP3的抑制剂MCC950后,除lncRNA TUG1外其余指标均显著下调(^(均)P<0.05)。另外,与OGD/R组比,OGD/R+TUG1过表达组的上述指标进一步上调(^(均)P<0.05)。与OGD/R组比,OGD/R+MCC950组则抑制了除lncRNA TUG1外的其余指标(^(均)P<0.05)。结论星状神经节阻滞通过调节lncRNA TUG1-NLRP3轴有效减轻体外脑缺血再灌注损伤引起的炎症反应和自噬溶酶体形成,提示其可能作为治疗缺血性脑损伤的潜在策略。
基金funded by the National Key Research and Development Program of China(Grant No.2022YFF1003100)Modern Citrus Industry Technology System of China(Grant No.CARS-26).
文摘Polyembryony has posed a significant impediment to the advancement of citrus hybrid breeding.FhRWP is widely regarded as a pivotal factor governing asexual reproduction in citrus,and prior research has demonstrated that FhARID1,acting as an upstream regulator,modulates FhRWP expression.In this study,we performed a genome-wide characterization of the ARID-HMG-related genes using the short juvenile minicitrus Fortunella hindsii.A total of 20 ARID-HMG-related genes were identified.Protein interaction network and enrichment analysis suggested that ARID-HMG-related proteins might might be involved in chromatin remodeling complexes.Knockout of FhARID1 in F.hindsii did not induce the conversion from polyembryony to monoembryony.However,fharid1 plants in T1 generation exhibited abnormal proliferation at axillary buds,which is similar to phenotype of fhrwp plants.Expression analysis of fharid1 ovary tissues revealed the downregulation of FhRWP.The results indicated that FhARID1,as an upstream regulator of FhRWP,has an effect on the development of citrus axillary buds.Expression analysis of overexpressed leaves of FhARID1 lines showed that no significant up-regulation of FhRWP,indicating that FhARID1 is not the sole upstream regulatory factor of FhRWP.Only FhARID2 showed a correlation in expression with FhARID1 among the ARID-related genes,further supporting the notion that this gene may be involved in complex formation rather than acting alone.Yeast two-hybrid and MS/MS spectra further indicated that FhARID1 function requires casein kinase II-mediated post-transcriptional phosphorylation.This study elucidated the function of FhARID1 in citrus apomixis and axillary bud development,providing a fundamental basis for understanding the role of ARID-HMG-related genes.
基金Supported by Beijing Holistic Integrative Medicine Association Clinical Research Funding Program,No.ZHKY-2024-2209.
文摘BACKGROUND Wolfram syndrome is a rare autosomal recessive genetic disorder characterized by early-onset diabetes and progressive neurodegeneration,most notably sensorineural hearing loss and optic atrophy.Because its initial manifestations are usually similar to those of type 1 diabetes,the diagnosis may be delayed until other manifestations appear.Pathogenic variations of the WFS1 gene can disrupt endoplasmic reticulum function and cellular homeostasis,but the complete mutation spectrum of WFS1 has not been fully determined.Early identification of monogenic diabetes caused by Wolfram syndrome is of vital importance,as it enables the provision of targeted multidisciplinary care and genetic counseling for affected families.CASE SUMMARY A 2-year-old Han Chinese girl was admitted with a 1-month history of polydipsia,polyuria,and polyphagia,and was diagnosed with diabetic ketoacidosis and impaired insulin secretion.Sensorineural hearing loss was also detected.Wholeexome sequencing identified a previously unreported heterozygous mutation,WFS1 c.986T>C(p.Phe329Ser),in the patient and her father,confirming the diagnosis of Wolfram syndrome.Bioinformatic analysis supported the likely pathogenicity of this mutation.In silico pathogenicity predictors(REVEL,SIFT,Poly-Phen-2,MutationTaster,and GERP+)supported a deleterious effect on wolframin structure and function.The patient was initially treated with intravenous insulin and fluid resuscitation,then transitioned to a basal–bolus insulin regimen.Glycemic control was subsequently maintained with continuous subcutaneous insulin infusion and intermittent subcutaneous injections.At the 1-and 6-month follow-ups,blood glucose remained well controlled(hemoglobin A1c:5.89%and 6.58%,respectively),with no evidence of organ dysfunction or further complications.CONCLUSION This case identifies WFS1 c.986T>C(p.Phe329Ser)as a novel pathogenic variant causing Wolfram syndrome.It highlights the importance of early genetic testing in pediatric patients with atypical diabetes presentations to enable timely diagnosis,individualized therapy,and comprehensive family support.
文摘本研究通过同源比对和结构域分析预测了水稻中38个PR1基因的理化性质、细胞定位和启动子结构特征,然后根据已有基因表达信息分析了PR1基因在生物胁迫中的响应表达情况,最后分析了这38个基因在水稻感染白叶枯病菌后的表达水平。结果发现38个PR蛋白中有35个蛋白的长度约为160 aa,有26个为酸性蛋白,32个为亲水性蛋白,27个可分泌到胞外。启动子顺式作用元件预测发现PR1基因启动子普遍含有MYB和茉莉酸响应结合元件。有20个PR1基因定位于7号染色体上,且多数PR1基因在染色体上呈现串联重复样排列。有37个PR基因含有CAP结构域。白叶枯病菌侵染水稻后,前3 d PR1-13未有表达,但3 d后表达显著提高;PR1-33在接种后持续性高表达;PR1-2、PR1-3、PR1-8、PR1-32、PR1-34、PR1-35、PR1-37的表达受病原菌抑制。以上结果暗示PR1基因在染色体上的串联排列可能有助于水稻快速响应病原菌的侵染,但PR1基因响应生物胁迫的表达与染色体定位无关,可能主要受启动子的顺式作用元件调控。本研究为分析PR1在抗病中的作用及机制提供了理论依据。
基金supported by the National Natural Science Foundation of China,Nos.82071008(to BL)and 82004001(to XJ)Medical Science and Technology Program of Health Commission of Henan Province,No.LHGJ20210072(to RQ)Science and Technology Department of Henan Province,No.212102310307(to XJ)。
文摘Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa.
文摘Objective To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress.Methods Twenty male SD rats were randomized equally into control group and heat stress group.After exposure to 32℃for 2 weeks in the latter group,the rats were examined for histopathological changes and Bmal1 expression in the thoracic aorta using HE staining and immunohistochemistry.In the cell experiments,cultured rat thoracic aortic endothelial cells(RTAECs)were incubated at 40℃for 12 h with or without prior transfection with a Bmal1-specific small interfering RNA(si-Bmal1)or a negative sequence.In both rat thoracic aorta and RTAECs,the expressions of Bmal1,the cell cycle proteins CDK1,CDK4,CDK6,and cyclin B1,and apoptosis-related proteins Bax and Bcl-2 were detected using Western blotting.TUNEL staining was used to detect cell apoptosis in rat thoracic aorta,and the changes in cell cycle distribution and apoptosis in RTAECs were analyzed with flow cytometry.Results Compared with the control rats,the rats exposed to heat stress showed significantly increased blood pressures and lowered heart rate with elastic fiber disruption and increased expressions of Bmal1,cyclin B1 and CDK1 in the thoracic aorta(P<0.05).In cultured RTAECs,heat stress caused significant increase of Bmal1,cyclin B1 and CDK1 protein expression levels,which were obviously lowered in cells with prior si-Bmal1 transfection.Bmal1 knockdown also inhibited heat stress-induced increase of apoptosis in RTAECs as evidenced by decreased expression of Bax and increased expression of Bcl-2.Conclusion Heat stress upregulates Bmal1 expression and causes alterations in expressions of cyclins to trigger apoptosis of rat thoracic aorta endothelial cells,which can be partly alleviated by suppressing Bmal1 expression.
