Background:The growth of the B-cell lymphoma subtype,Hodgkin lymphoma(HL),is associated with increased autophagy.A mycobacterial antigen,Ag85,has been reported to inhibit cell autophagy under a variety of conditions.W...Background:The growth of the B-cell lymphoma subtype,Hodgkin lymphoma(HL),is associated with increased autophagy.A mycobacterial antigen,Ag85,has been reported to inhibit cell autophagy under a variety of conditions.Whether Ag85 could inhibit autophagy in HL is unknown.Methods:Lymph node samples from patients with HL and healthy controls were collected to assess proliferation and autophagy.The human HL cell line,L-428,was cultured and subjected to Ag85B treatment.Autophagy in L-428 cells was evaluated through western blotting analysis,immunohistochemistry,and transmission electron microscopy.Apoptosis in these cells was measured using flow cytometry and western blotting.The associated signaling pathways were also analyzed utilizing western blotting.The in vivo impact of Ag85B was studied using BALB/c Nude mice xenografted with L-428 cells.Results:We observed increased proliferation and autophagy in primary lymphoma tissues of patients.Administration of Ag85B inhibited the proliferation and autophagy of HL cell lines.Moreover,Ag85B promoted apoptotic pathway activation in vitro,which might be associated with mitochondrial dysfunction.Mechanistically,Ag85B inhibits autophagy by activating the phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B/mechanistic target of rapamycin kinase(PI3K/AKT/mTOR)and mitogen-activated protein kinase(MAPK)pathways.Ag85B also inhibited lymphoma growth in mice xenografted with HL cell lines,but no potential toxicity was observed.Conclusion:Altogether,these results suggest that Ag85B inhibits HL growth via autophagy regulation.Current treatments for HL are associated with adverse events;therefore,Ag85B-mediated autophagy inhibition might be a promising strategy in to treat HL.展开更多
OBJECTIVE： To explore the effect of kidney-rein- forcing, blood-activating and stasis-removing recipes on adhesion molecule expression of bone mar- row mesenchymal stem cells （MSCs） from patients with chronic aplas...OBJECTIVE： To explore the effect of kidney-rein- forcing, blood-activating and stasis-removing recipes on adhesion molecule expression of bone mar- row mesenchymal stem cells （MSCs） from patients with chronic aplastic anemia （CAA）. METHODS： We used three Traditional Chinese Medicine recipes, namely a kidney-reinforcing recipe （KRR）, blood-activating and stasis-removing recipe （BASRR）, and kidney-reinforcing, blood-activating and stasis-removing recipe （KRBASFIR）, and a nor- mal saline control to prepare herbal medicine se- rum in Sprague Dawley rats. Thirty CAA patients were enrolled in the experimental group, including 17 kidney-Yang deficient patients and 13 kidney-Yin deficient patients. Ten healthy individuals were included in the control group. MSCs were isolated from bone marrow samples, and the cell density was observed to measure their proliferation ability by microscopy on days 2, 7, and 14 after isolation. In addition, the expression of adhesion molecules of bone marrow MSCs （CD106, CD49d, CD31 and CD44） were detected by flow cytometry after 48 h of treatment with the four different herbal medi- cine serums. RESULTS： The proliferation of MSCs from kid- ney-Yang deficient and kidney-Yin deficient pa- tients was weaker than that of MSCs from the con- trol group. The expression of all adhesion mole- cules of bone marrow MSCs from CAA patients was obviously lower than that in the control group （P〈 0.01）. The expression of CD49d and CD31 in MSCs from patients with a kidney-Yin deficiency was low- er than in those with a kidney-yang deficiency （P〈 0.05 and P〈O.01, respectively）. For kidney-Yang defi- cient patients, CD31 expression in the KRBASRR group was significantly higher than that in the BASRR group （P〈O.01）, while CD44 in the KRBASRR group was significantly higher than that in both KRR and BASRR groups （P〈O.01）. For kidney-Yin defi- cient patients, CD106 and CD49d expression in the KRBASRR group was obviously higher than that in the KRR group （P〈0.05）, while CD31 and CD44 ex- pression in the KRBASRR group was significantly higher than that in both KRR and BASRR groups （P〈 0.05 and P〈0.01, respectively）. CONCLUSION： The bone marrow microenviron- ment in CAA patients is abnormal. The effect of KRBASRR may be better than that of KRR and BASRR for kidney-Yang deficient and kidney-Yin de- ficient patients by improving the expression levels of MSC adhesion molecules.展开更多
Copy number variants (CNVs) are pervasive in the human genome and are responsible for many Mendelian diseases and genomic disorders. The detection of CNVs is an essential element of a complete mutation screening strat...Copy number variants (CNVs) are pervasive in the human genome and are responsible for many Mendelian diseases and genomic disorders. The detection of CNVs is an essential element of a complete mutation screening strategy. Many techniques have been developed for gene dosage testing. Multiplex ligation-dependent probe amplification (MLPA) is a robust, easy and flexible technique that can detect both deletions and duplications for more than 40 loci in one assay. It has been widely used in research and diagnostic laboratories. We routinely develop our own MLPA assays for quick validation of array comparative genomic hybridization (CGH) findings. Here we discuss the general principles and critical aspects of MLPA assay development and validation using all synthetic MLPA probes. We believe that MLPA will play important roles in the rapid detection of genomic disorders associated with genomic imbalances, the confirmation of pathogenic mutations involving exonic deletions/duplications, CNV genotyping and population frequency analysis of CNVs.展开更多
Mitochondrial diseases are caused by variants in both mitochondrial and nuclear genomes.A nuclear gene HPDL(4-hydroxyphenylpyruvate dioxygenase-like),which encodes an intermembrane mitochondrial protein,has been recen...Mitochondrial diseases are caused by variants in both mitochondrial and nuclear genomes.A nuclear gene HPDL(4-hydroxyphenylpyruvate dioxygenase-like),which encodes an intermembrane mitochondrial protein,has been recently implicated in causing a neurodegenerative disease characterized by pediatric-onset spastic movement phenotypes.Here,we report six Chinese patients with bi-allelic HPDL pathogenic variants from four unrelated families showing neuropathic symptoms of variable severity,including developmental delay/intellectual disability,spasm,and hypertonia.Seven different pathogenic variants are identified,of which five are novel.Both fibroblasts and immortalized lymphocytes derived from patients show impaired mitochondrial respiratory function,which is also observed in HPDL-knockdown(KD)He La cells.In these He La cells,overexpression of a wild-type HPDL gene can rescue the respiratory phenotype of oxygen consumption rate.In addition,a decreased activity of the oxidative phosphorylation(OXPHOS)complex II is observed in patient-derived lymphocytes and HPDL-KD He La cells,further supporting an essential role of HPDL in the mitochondrial respiratory chain.Collectively,our data expand the clinical and mutational spectra of this mitochondrial neuropathy and further delineate the possible disease mechanism involving the impairment of the OXPHOS complex II activity due to the bi-allelic inactivations of HPDL.展开更多
Human genetic variants have long been known to play an important role in both Mendelian disorders and common diseases. Notably, pathogenic variants are not limited to single-nucleotide variants. It has become apparent...Human genetic variants have long been known to play an important role in both Mendelian disorders and common diseases. Notably, pathogenic variants are not limited to single-nucleotide variants. It has become apparent that human diseases can also be caused by copy number variations （CNVs）, especially patient- specific novel CNVs （lafrate et al., 2004; Sebat et al., 2004; Redon et al., 2006; LuDski, 2007; Zhan~ et al.. 2009： Wu et al.. 2015）.展开更多
Short stature is clinically defined as a standing height less than two standard deviations below the mean height at the same age,ethnicity,and sex.As a typical complex symp-tom,height has a high heritability of 80%,wh...Short stature is clinically defined as a standing height less than two standard deviations below the mean height at the same age,ethnicity,and sex.As a typical complex symp-tom,height has a high heritability of 80%,which is affected by multiple genes and geneegene interactions.A genome-wide association study(GWAS)revealed that 23.3%of the heritability of short stature could be explained by 697 in-dependent variants.1 Whole exome sequencing of a large sample size demonstrated that 83 rare and low-frequency variants could explain 1.7%heritability of height.展开更多
Carrier screening had been demonstrated as a powerful practice in preventing selected severe genetic disorders. This practice is expanding its scope and impact in the era of next-generation sequencing. Empirical and t...Carrier screening had been demonstrated as a powerful practice in preventing selected severe genetic disorders. This practice is expanding its scope and impact in the era of next-generation sequencing. Empirical and theoretical data support the utility of expanded carrier screening. The authors propose a comprehensive carrier screening program as a main component of the first-tier measure in preventing severe genetic disorders and birth defects in China. We discussed the key principles and important aspects to ensure the success of such a program. The authors believe this program will play a pivotal role in our endeavor for a healthier nation.展开更多
The traditional approach of clinical genetic practice,known as the phenotype-first approach,usually starts with a thorough review of electronic medical records of the patient and a complete evaluation of patient's...The traditional approach of clinical genetic practice,known as the phenotype-first approach,usually starts with a thorough review of electronic medical records of the patient and a complete evaluation of patient's clinical presentation,as well as a survey of family history.Often more than one experienced clinical geneticists will order a variety of additional laboratory or imaging tests to obtain relevant information in order to reach a reasonable clinical diagnosis.Finally,molecular testing often helps to confirm the clinical diagnosis.This practice requires the availability of electronic medical records,well trained clinical geneticists who are able to perform relevant clinical evaluation,order appropriate tests and understand the molecular reports.These critical components of clinical genetics are mostly lacking in China today,1 as a consequence,most of patients with genetic condition do not receive a proper evaluation by clinical geneticists,only a small percent of patients received a clinical diagnosis and the majority remains undiagnosed for life.展开更多
基金supported by the Natural Science Foundation of Xinjiang Uygur Autonomous Region of China(grant number 2022D01C739)the National Natural Science Foundation of China(grant numbers 82160031,82071276)the Key Research and Development Program of Xinjiang Uyghur Autonomous Region of China(grant number 2024B03038-1).
文摘Background:The growth of the B-cell lymphoma subtype,Hodgkin lymphoma(HL),is associated with increased autophagy.A mycobacterial antigen,Ag85,has been reported to inhibit cell autophagy under a variety of conditions.Whether Ag85 could inhibit autophagy in HL is unknown.Methods:Lymph node samples from patients with HL and healthy controls were collected to assess proliferation and autophagy.The human HL cell line,L-428,was cultured and subjected to Ag85B treatment.Autophagy in L-428 cells was evaluated through western blotting analysis,immunohistochemistry,and transmission electron microscopy.Apoptosis in these cells was measured using flow cytometry and western blotting.The associated signaling pathways were also analyzed utilizing western blotting.The in vivo impact of Ag85B was studied using BALB/c Nude mice xenografted with L-428 cells.Results:We observed increased proliferation and autophagy in primary lymphoma tissues of patients.Administration of Ag85B inhibited the proliferation and autophagy of HL cell lines.Moreover,Ag85B promoted apoptotic pathway activation in vitro,which might be associated with mitochondrial dysfunction.Mechanistically,Ag85B inhibits autophagy by activating the phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B/mechanistic target of rapamycin kinase(PI3K/AKT/mTOR)and mitogen-activated protein kinase(MAPK)pathways.Ag85B also inhibited lymphoma growth in mice xenografted with HL cell lines,but no potential toxicity was observed.Conclusion:Altogether,these results suggest that Ag85B inhibits HL growth via autophagy regulation.Current treatments for HL are associated with adverse events;therefore,Ag85B-mediated autophagy inhibition might be a promising strategy in to treat HL.
基金Supported by 2011 Zhejiang province key science and technology innovation team(No.2011R09042-02)Special Item of Important Disease of Zhejiang Province TCM Sci-Tech Innovation Platform(No.2009ZDJB01,2009ZDJB01-08)
文摘OBJECTIVE： To explore the effect of kidney-rein- forcing, blood-activating and stasis-removing recipes on adhesion molecule expression of bone mar- row mesenchymal stem cells （MSCs） from patients with chronic aplastic anemia （CAA）. METHODS： We used three Traditional Chinese Medicine recipes, namely a kidney-reinforcing recipe （KRR）, blood-activating and stasis-removing recipe （BASRR）, and kidney-reinforcing, blood-activating and stasis-removing recipe （KRBASFIR）, and a nor- mal saline control to prepare herbal medicine se- rum in Sprague Dawley rats. Thirty CAA patients were enrolled in the experimental group, including 17 kidney-Yang deficient patients and 13 kidney-Yin deficient patients. Ten healthy individuals were included in the control group. MSCs were isolated from bone marrow samples, and the cell density was observed to measure their proliferation ability by microscopy on days 2, 7, and 14 after isolation. In addition, the expression of adhesion molecules of bone marrow MSCs （CD106, CD49d, CD31 and CD44） were detected by flow cytometry after 48 h of treatment with the four different herbal medi- cine serums. RESULTS： The proliferation of MSCs from kid- ney-Yang deficient and kidney-Yin deficient pa- tients was weaker than that of MSCs from the con- trol group. The expression of all adhesion mole- cules of bone marrow MSCs from CAA patients was obviously lower than that in the control group （P〈 0.01）. The expression of CD49d and CD31 in MSCs from patients with a kidney-Yin deficiency was low- er than in those with a kidney-yang deficiency （P〈 0.05 and P〈O.01, respectively）. For kidney-Yang defi- cient patients, CD31 expression in the KRBASRR group was significantly higher than that in the BASRR group （P〈O.01）, while CD44 in the KRBASRR group was significantly higher than that in both KRR and BASRR groups （P〈O.01）. For kidney-Yin defi- cient patients, CD106 and CD49d expression in the KRBASRR group was obviously higher than that in the KRR group （P〈0.05）, while CD31 and CD44 ex- pression in the KRBASRR group was significantly higher than that in both KRR and BASRR groups （P〈 0.05 and P〈0.01, respectively）. CONCLUSION： The bone marrow microenviron- ment in CAA patients is abnormal. The effect of KRBASRR may be better than that of KRR and BASRR for kidney-Yang deficient and kidney-Yin de- ficient patients by improving the expression levels of MSC adhesion molecules.
文摘Copy number variants (CNVs) are pervasive in the human genome and are responsible for many Mendelian diseases and genomic disorders. The detection of CNVs is an essential element of a complete mutation screening strategy. Many techniques have been developed for gene dosage testing. Multiplex ligation-dependent probe amplification (MLPA) is a robust, easy and flexible technique that can detect both deletions and duplications for more than 40 loci in one assay. It has been widely used in research and diagnostic laboratories. We routinely develop our own MLPA assays for quick validation of array comparative genomic hybridization (CGH) findings. Here we discuss the general principles and critical aspects of MLPA assay development and validation using all synthetic MLPA probes. We believe that MLPA will play important roles in the rapid detection of genomic disorders associated with genomic imbalances, the confirmation of pathogenic mutations involving exonic deletions/duplications, CNV genotyping and population frequency analysis of CNVs.
基金funded by the Precision Medical Research of National Key Research and Development Program(2018YFC1002200,2019YFC1005100 to Y.Yu,2018YFC1002400 to Y.Sun,and 2018YFC1002501 to Y.Shen)National Natural Science Foundation of China(81873633 and 82071276 to Y.Shen,81830071 to J.Lyu,81873724 to Y.Sun,and 82070914 and 81873671 to Y.Yu)+7 种基金Shanghai Shen Kang Hospital Development Center(SHDC12017109 to Y.Yu)the Shanghai Science and Technology Commission(19140904500 to Y.Yu)Jiaotong University Cross Biomedical Engineering(YG2017MS72 to Y.Yu)the“Eastern Scholar”Fundthe“Guangxi Bagui Scholar”fund(to Y.Shen)the Major Research Plan of the Provincial Science and Technology Foundation of Guangxi(AB16380214 to Y.Shen)Foundation of Shanghai Municipal Health Commission(shslczdzk05702,to Y.Yu and Y.Sun)Municipal Education Commission-Gaofeng Clinical Medicine Grant Support(20191908,to Y.Yu)。
文摘Mitochondrial diseases are caused by variants in both mitochondrial and nuclear genomes.A nuclear gene HPDL(4-hydroxyphenylpyruvate dioxygenase-like),which encodes an intermembrane mitochondrial protein,has been recently implicated in causing a neurodegenerative disease characterized by pediatric-onset spastic movement phenotypes.Here,we report six Chinese patients with bi-allelic HPDL pathogenic variants from four unrelated families showing neuropathic symptoms of variable severity,including developmental delay/intellectual disability,spasm,and hypertonia.Seven different pathogenic variants are identified,of which five are novel.Both fibroblasts and immortalized lymphocytes derived from patients show impaired mitochondrial respiratory function,which is also observed in HPDL-knockdown(KD)He La cells.In these He La cells,overexpression of a wild-type HPDL gene can rescue the respiratory phenotype of oxygen consumption rate.In addition,a decreased activity of the oxidative phosphorylation(OXPHOS)complex II is observed in patient-derived lymphocytes and HPDL-KD He La cells,further supporting an essential role of HPDL in the mitochondrial respiratory chain.Collectively,our data expand the clinical and mutational spectra of this mitochondrial neuropathy and further delineate the possible disease mechanism involving the impairment of the OXPHOS complex II activity due to the bi-allelic inactivations of HPDL.
基金supported by the National Basic Research Program of China(No.2012CB944600)the National Key Research and Development Program of China(No.2016YFC0905100)+1 种基金the National Natural Science Foundation of China(Nos.31521003,31625015,31571297,31601046,31525014 and 91331204)the Science and Technology Commission of Shanghai Municipality(No.16YF1413900)
文摘Human genetic variants have long been known to play an important role in both Mendelian disorders and common diseases. Notably, pathogenic variants are not limited to single-nucleotide variants. It has become apparent that human diseases can also be caused by copy number variations （CNVs）, especially patient- specific novel CNVs （lafrate et al., 2004; Sebat et al., 2004; Redon et al., 2006; LuDski, 2007; Zhan~ et al.. 2009： Wu et al.. 2015）.
基金supported by the National Key R&D Program of China(No.2021YFC2701100 to H.W.)the National Natural Science Foundation of China(No.81930036,8215008 to H.W.)the Commission for Science and Technology of Shanghai Municipality,China(No.20JC1418500 to H.W.).
文摘Short stature is clinically defined as a standing height less than two standard deviations below the mean height at the same age,ethnicity,and sex.As a typical complex symp-tom,height has a high heritability of 80%,which is affected by multiple genes and geneegene interactions.A genome-wide association study(GWAS)revealed that 23.3%of the heritability of short stature could be explained by 697 in-dependent variants.1 Whole exome sequencing of a large sample size demonstrated that 83 rare and low-frequency variants could explain 1.7%heritability of height.
文摘Carrier screening had been demonstrated as a powerful practice in preventing selected severe genetic disorders. This practice is expanding its scope and impact in the era of next-generation sequencing. Empirical and theoretical data support the utility of expanded carrier screening. The authors propose a comprehensive carrier screening program as a main component of the first-tier measure in preventing severe genetic disorders and birth defects in China. We discussed the key principles and important aspects to ensure the success of such a program. The authors believe this program will play a pivotal role in our endeavor for a healthier nation.
文摘The traditional approach of clinical genetic practice,known as the phenotype-first approach,usually starts with a thorough review of electronic medical records of the patient and a complete evaluation of patient's clinical presentation,as well as a survey of family history.Often more than one experienced clinical geneticists will order a variety of additional laboratory or imaging tests to obtain relevant information in order to reach a reasonable clinical diagnosis.Finally,molecular testing often helps to confirm the clinical diagnosis.This practice requires the availability of electronic medical records,well trained clinical geneticists who are able to perform relevant clinical evaluation,order appropriate tests and understand the molecular reports.These critical components of clinical genetics are mostly lacking in China today,1 as a consequence,most of patients with genetic condition do not receive a proper evaluation by clinical geneticists,only a small percent of patients received a clinical diagnosis and the majority remains undiagnosed for life.
