AIM: To detect the pathogenetic mutations responsible for nonsyndromic autosomal recessive retinitis pigmentosa(RP) in 2 nonconsanguineous Chinese families. METHODS: The clinical data, including detailed medical histo...AIM: To detect the pathogenetic mutations responsible for nonsyndromic autosomal recessive retinitis pigmentosa(RP) in 2 nonconsanguineous Chinese families. METHODS: The clinical data, including detailed medical history, best corrected visual acuity(BCVA), slit-lamp biomicroscope examination, fundus photography, optical coherence tomography, static perimetry, and full field electroretinogram, were collected from the members of 2 nonconsanguineous Chinese families preliminarily diagnosed with RP. Genomic DNA was extracted from the probands and other available family members;wholeexome sequencing was conducted with the DNA samples provided by the probands, and all mutations detected by whole-exome sequencing were verified using Sanger sequencing in the probands and the other available family members. The verified novel mutations were further sequenced in 192 ethnicity matched healthy controls.RESULTS: The patients from the 2 families exhibited the typical symptoms of RP, including night blindness and progressive constriction of the visual field, and the fundus examinations showed attenuated retinal arterioles, peripheral bone spicule pigment deposits, and waxy optic discs. Whole-exome sequencing revealed a novel nonsense mutation in FAM161 A(c.943 A>T, p.Lys315*) and compound heterozygous mutations in RP1 L1(c.56 C>A, p.Pro19 His;c.5470 C>T, p.Gln1824*). The nonsense c.5470 C>T, p.Gln1824* mutation was novel. All mutations were verified by Sanger sequencing. The mutation p.Lys315* in FAM161A co-segregated with the phenotype, and all the nonsense mutations were absent from the ethnicity matched healthy controls and all available databases.CONCLUSION: We identify 2 novel mutations in genes responsible for autosomal recessive RP, and the mutation in FAM161A is reported for the first time in a Chinese population. Our result not only enriches the knowledge of the mutation frequency and spectrum in the genes responsible for nonsyndromic RP but also provides a new target for future gene therapy.展开更多
Background：Dysferlinopathy is caused by mutations in the dysferlin （DYSF） gene.Here,we described the genetic features of a large cohort of Chinese patients with this disease.Methods：Eighty-nine index patients were...Background：Dysferlinopathy is caused by mutations in the dysferlin （DYSF） gene.Here,we described the genetic features of a large cohort of Chinese patients with this disease.Methods：Eighty-nine index patients were included in the study.DYSF gene analysis was performed by Sanger sequencing in 41 patients and targeted next generation sequencing （NGS） in 48 patients.Multiplex ligation-dependent probe amplification （MLPA） was performed to detect exon duplication/deletion in patients with only one pathogenic mutation.Results：Among the 89 index patients,79 patients were demonstrated to carry two disease-causing （73 cases） or possibly disease-causing mutations （6 cases）,including 26 patients with homozygous mutations.We identified 105 different mutations,including 59 novel ones.Notably,in 13 patients in whom only one pathogenic mutation was initially found by Sanger sequencing or NGS,3 were further identified to carry exon deletions by MLPA.The mutations identified in this study appeared to cluster in the N-terminal region.Mutation types included missense mutations （30.06%）,nonsense mutations （1 7.18%）,frameshift mutations （30.67%）,in-frame deletions （2.45%）,intronic mutations （17.79%）,and exonic rearrangement （1.84%）.No genotype-phenotype correlation was identified.Conclusions：DYSF mutations in Chinese patients clustered in the N-terminal region of the gene.Exonic rearrangements were found in 23% of patients with only one pathogenic mutation identified by Sanger sequencing or NGS.The novel mutations found in this study greatly expanded the mutational spectrum of dysferlinopathy.展开更多
In the last two decades, great efforts have been made in the treatment of metastatic colorectal cancer(m CRC) due to the approval of new target agents for cytotoxic drugs. Unfortunately, a large percentage of patients...In the last two decades, great efforts have been made in the treatment of metastatic colorectal cancer(m CRC) due to the approval of new target agents for cytotoxic drugs. Unfortunately, a large percentage of patients present with metastasis at the time of diagnosis or relapse after a few months. The complex molecular heterogeneity of this disease is not completely understood; to date, there is a lack of predictive biomarkers that can be used to select subsets of patients who may respond to target drugs. Only the RAS-mutation status is used to predict resistance to anti-epidermal growth factor receptor agents in patients with m CRC. In this review, we describe approved targeted therapies for the management of metastatic m CRC and discuss new candidate targets on the horizon.展开更多
AIM: To discover the molecular pathogenic basis of the blepharophimosis, ptosis, and epicanthus inversus syndrome(BPES), and to predict the clinical subtype according to in vitro experiments, which is significant to t...AIM: To discover the molecular pathogenic basis of the blepharophimosis, ptosis, and epicanthus inversus syndrome(BPES), and to predict the clinical subtype according to in vitro experiments, which is significant to the prognosis.METHODS: A 3-year-old sporadic female patient with typical clinical manifestations of BPES was enrolled. The coding region of forkhead box L2(FOXL2) gene was sequenced, and the functional assays were performed in vitro by Western blotting, subcellular localization experiment, luciferase reporter assay, and quantitative realtime polymerase chain reaction.RESULTS: A novel FOXL2 point pathogenic variant(c.274G>T) was detected, resulting in a truncated protein(p.E92*). Functional studies demonstrated that the FOXL2 pathogenic variant induced the subcellular mislocalization and the abnormal transcriptional activity on promoters of the steroidogenic acute regulatory protein(StAR or STARD1) gene and the odd-skipped related 2 transcription factor(OSR2) gene.CONCLUSION: A novel pathogenic variant is identified to expand the spectrum of the known FOXL2 mutations. The in vitro experiments provide reference data and more insights to the molecular pathogenesis of BPES. The predicted high risk of ovarian insufficiency makes it significant for the patient enrolled to have further follow-up and therapy concerning female endocrinology.展开更多
目的研究黏多糖贮积症ⅣA型（mucopolysaccharidosis type ⅣA，MPSⅣA）患者发病的分子遗传学机制，揭示其基因型与表现型的相互关系，为产前基因诊断等创造必要的前提条件。方法采用尿糖胺聚糖（glycosaminoglycans，GAGs）定性检测...目的研究黏多糖贮积症ⅣA型（mucopolysaccharidosis type ⅣA，MPSⅣA）患者发病的分子遗传学机制，揭示其基因型与表现型的相互关系，为产前基因诊断等创造必要的前提条件。方法采用尿糖胺聚糖（glycosaminoglycans，GAGs）定性检测法对疑似MPSIVA型的先证者进行初诊，然后采用PCR及扩增产物直接测序法对先证者及其家庭成员进行突变检测。在检出GALNS基因c．1567T〉G新突变后，先后建立XspI酶切鉴定法和扩增阻碍突变系统（amplification refractory mutation system，ARMS）快速特异鉴定法，对随机采集的110名正常对照与先证者及其家庭成员的GALNS基因第14外显子进行序列分析，同时采用生物信息学方法对蛋白质二级、三级结构进行预测，以及直接测定患儿GALNS酶活性的方法，对该新突变进行致病性鉴定。结果先证者尿检呈弱阳性GAGs（±），其GALNS基因第14外显子内存在杂合的c．1567T〉G终止密码突变，第4外显子存在杂合的c．374C〉T错义突变，为两种突变的复合杂合子。其妹突变类型与先证者完全相同，其母仅在第14外显子存在杂合的终止密码突变，为该病的携带者，其父仅在第4外显子存在杂合的错义突变，也为杂合子；第14外显子的PCR产物经XspI酶切后，正常对照组切出28bp、120bp、399bp3条带，而患者和携带者的母亲均切出28bp、120bp、148bp和399bp4条带；用ARMS特异引物扩增后，正常对照组均扩增阴性，而患者及携带者均扩增阳性；蛋白质二级、三级结构预测结果显示：c ．1567T〉G变异导致终止密码（TAG）突变为谷氨酸（GAG），使多肽链延长了92个氨基酸残基，导致蛋白质二、三级结构发生明显改变，而正常对照无此变化。酶活性测定的结果显示：患者的GALNS酶活性仅为8．3nmol／17h／mgpr，明显低于正常值（正常参考值为41．9～92．1nmol／17h，／mgpr）。结论c．1567T�展开更多
Listed examples of virus transmission epidemics that can be strongly transmitted through the air<span "=""> </span><span style="font-family:Verdana;">caused by sunspot change...Listed examples of virus transmission epidemics that can be strongly transmitted through the air<span "=""> </span><span style="font-family:Verdana;">caused by sunspot change cycle</span><span style="font-family:Verdana;">, </span><span style="font-family:Verdana;">analyzed the mechanism that promotes the generation of new viruses. From the schematic diagram of the changes in the combined force of the hydrodynamic effect of the sun sweeping the earth and the sweeping force, </span><span style="font-family:Verdana;">we </span><span style="font-family:Verdana;">obtain the </span><span style="font-family:Verdana;">places that are prone to light vortices are 30 degrees north latitude and 30 degrees</span><span "=""> </span><span style="font-family:Verdana;">south latitude on the east coast of the mainland creatively</span><span style="font-family:Verdana;">. </span><span style="font-family:Verdana;">The curved continental lines are perfect, the range of the light vortex generated is more obviously, and the effect is stronger. And the curved continental lines are perfect, the range of the light vortex generated is more obviously, and the effect is stronger. It is inferred that the light vortex produces the special amplified energy so</span><span style="font-family:Verdana;"> that can</span><span style="font-family:Verdana;"> make the virus mutate to produce a new highly infectious novel coronavirus. The earliest known place and time of the novel coronavirus origin are consistent with the reasoning of the new theory. Because the radius and frequency of the light vortex are different, the resulting virus strains are also different. Moreover, the fatality rate in the light vortex area is much higher than that in the non-light vortex area, indicating that the virus</span><span style="font-family:Verdana;">’</span><span style="font-family:Verdana;">s toxicity and lethality are higher in the light vortex area, so it can explain why Russia, India, and countries in the African equatorial region mortality are much lower than展开更多
基金Supported by the National Natural Science Foundation of China(No.81360154)
文摘AIM: To detect the pathogenetic mutations responsible for nonsyndromic autosomal recessive retinitis pigmentosa(RP) in 2 nonconsanguineous Chinese families. METHODS: The clinical data, including detailed medical history, best corrected visual acuity(BCVA), slit-lamp biomicroscope examination, fundus photography, optical coherence tomography, static perimetry, and full field electroretinogram, were collected from the members of 2 nonconsanguineous Chinese families preliminarily diagnosed with RP. Genomic DNA was extracted from the probands and other available family members;wholeexome sequencing was conducted with the DNA samples provided by the probands, and all mutations detected by whole-exome sequencing were verified using Sanger sequencing in the probands and the other available family members. The verified novel mutations were further sequenced in 192 ethnicity matched healthy controls.RESULTS: The patients from the 2 families exhibited the typical symptoms of RP, including night blindness and progressive constriction of the visual field, and the fundus examinations showed attenuated retinal arterioles, peripheral bone spicule pigment deposits, and waxy optic discs. Whole-exome sequencing revealed a novel nonsense mutation in FAM161 A(c.943 A>T, p.Lys315*) and compound heterozygous mutations in RP1 L1(c.56 C>A, p.Pro19 His;c.5470 C>T, p.Gln1824*). The nonsense c.5470 C>T, p.Gln1824* mutation was novel. All mutations were verified by Sanger sequencing. The mutation p.Lys315* in FAM161A co-segregated with the phenotype, and all the nonsense mutations were absent from the ethnicity matched healthy controls and all available databases.CONCLUSION: We identify 2 novel mutations in genes responsible for autosomal recessive RP, and the mutation in FAM161A is reported for the first time in a Chinese population. Our result not only enriches the knowledge of the mutation frequency and spectrum in the genes responsible for nonsyndromic RP but also provides a new target for future gene therapy.
文摘Background：Dysferlinopathy is caused by mutations in the dysferlin （DYSF） gene.Here,we described the genetic features of a large cohort of Chinese patients with this disease.Methods：Eighty-nine index patients were included in the study.DYSF gene analysis was performed by Sanger sequencing in 41 patients and targeted next generation sequencing （NGS） in 48 patients.Multiplex ligation-dependent probe amplification （MLPA） was performed to detect exon duplication/deletion in patients with only one pathogenic mutation.Results：Among the 89 index patients,79 patients were demonstrated to carry two disease-causing （73 cases） or possibly disease-causing mutations （6 cases）,including 26 patients with homozygous mutations.We identified 105 different mutations,including 59 novel ones.Notably,in 13 patients in whom only one pathogenic mutation was initially found by Sanger sequencing or NGS,3 were further identified to carry exon deletions by MLPA.The mutations identified in this study appeared to cluster in the N-terminal region.Mutation types included missense mutations （30.06%）,nonsense mutations （1 7.18%）,frameshift mutations （30.67%）,in-frame deletions （2.45%）,intronic mutations （17.79%）,and exonic rearrangement （1.84%）.No genotype-phenotype correlation was identified.Conclusions：DYSF mutations in Chinese patients clustered in the N-terminal region of the gene.Exonic rearrangements were found in 23% of patients with only one pathogenic mutation identified by Sanger sequencing or NGS.The novel mutations found in this study greatly expanded the mutational spectrum of dysferlinopathy.
文摘In the last two decades, great efforts have been made in the treatment of metastatic colorectal cancer(m CRC) due to the approval of new target agents for cytotoxic drugs. Unfortunately, a large percentage of patients present with metastasis at the time of diagnosis or relapse after a few months. The complex molecular heterogeneity of this disease is not completely understood; to date, there is a lack of predictive biomarkers that can be used to select subsets of patients who may respond to target drugs. Only the RAS-mutation status is used to predict resistance to anti-epidermal growth factor receptor agents in patients with m CRC. In this review, we describe approved targeted therapies for the management of metastatic m CRC and discuss new candidate targets on the horizon.
基金Supported by Funds of Plastic Surgery Hospital of the Chinese Academy of Medical Sciences,Peking Union Medical College (No.YS202010)。
文摘AIM: To discover the molecular pathogenic basis of the blepharophimosis, ptosis, and epicanthus inversus syndrome(BPES), and to predict the clinical subtype according to in vitro experiments, which is significant to the prognosis.METHODS: A 3-year-old sporadic female patient with typical clinical manifestations of BPES was enrolled. The coding region of forkhead box L2(FOXL2) gene was sequenced, and the functional assays were performed in vitro by Western blotting, subcellular localization experiment, luciferase reporter assay, and quantitative realtime polymerase chain reaction.RESULTS: A novel FOXL2 point pathogenic variant(c.274G>T) was detected, resulting in a truncated protein(p.E92*). Functional studies demonstrated that the FOXL2 pathogenic variant induced the subcellular mislocalization and the abnormal transcriptional activity on promoters of the steroidogenic acute regulatory protein(StAR or STARD1) gene and the odd-skipped related 2 transcription factor(OSR2) gene.CONCLUSION: A novel pathogenic variant is identified to expand the spectrum of the known FOXL2 mutations. The in vitro experiments provide reference data and more insights to the molecular pathogenesis of BPES. The predicted high risk of ovarian insufficiency makes it significant for the patient enrolled to have further follow-up and therapy concerning female endocrinology.
文摘Listed examples of virus transmission epidemics that can be strongly transmitted through the air<span "=""> </span><span style="font-family:Verdana;">caused by sunspot change cycle</span><span style="font-family:Verdana;">, </span><span style="font-family:Verdana;">analyzed the mechanism that promotes the generation of new viruses. From the schematic diagram of the changes in the combined force of the hydrodynamic effect of the sun sweeping the earth and the sweeping force, </span><span style="font-family:Verdana;">we </span><span style="font-family:Verdana;">obtain the </span><span style="font-family:Verdana;">places that are prone to light vortices are 30 degrees north latitude and 30 degrees</span><span "=""> </span><span style="font-family:Verdana;">south latitude on the east coast of the mainland creatively</span><span style="font-family:Verdana;">. </span><span style="font-family:Verdana;">The curved continental lines are perfect, the range of the light vortex generated is more obviously, and the effect is stronger. And the curved continental lines are perfect, the range of the light vortex generated is more obviously, and the effect is stronger. It is inferred that the light vortex produces the special amplified energy so</span><span style="font-family:Verdana;"> that can</span><span style="font-family:Verdana;"> make the virus mutate to produce a new highly infectious novel coronavirus. The earliest known place and time of the novel coronavirus origin are consistent with the reasoning of the new theory. Because the radius and frequency of the light vortex are different, the resulting virus strains are also different. Moreover, the fatality rate in the light vortex area is much higher than that in the non-light vortex area, indicating that the virus</span><span style="font-family:Verdana;">’</span><span style="font-family:Verdana;">s toxicity and lethality are higher in the light vortex area, so it can explain why Russia, India, and countries in the African equatorial region mortality are much lower than
