中国实验血液学杂志最新文献

{"title":"[Analysis of genetic diagnosis results of 1 501 suspected Cases of thalassemia patients from 2020 to 2022].","authors":"Xue-Li Yang, Zhen-Yu Liu, Jun-Ning Zhang, Guang-Yu Wang, Ji-Ming Li, Chun-Hong Li, Xian-Liang Hou","doi":"10.19746/j.cnki.issn.1009-2137.2024.06.032","DOIUrl":"https://doi.org/10.19746/j.cnki.issn.1009-2137.2024.06.032","url":null,"abstract":"<p><strong>Objective: </strong>To explore the genotypes and frequency distribution of thalassemia in Lingui District, Guilin City, and provide reference for the prevention and control of thalassemia in this area.</p><p><strong>Methods: </strong>The results of genetic testing for thalassemia in 1 501 suspected cases at the Second Affiliated Hospital of Guilin Medical University were analyzed retrospectively. The deletional mutations of α-thalassemia were detected by gap-PCR, the non-deletional mutations of α-thalassemia and β-thalassemia mutations were detected by PCR-reverse dot blot (PCR-RDB).</p><p><strong>Results: </strong>In 1 501 samples, a total of 678 cases of thalassemia carriers were detected, with a detection rate of 45.17%. Among them, 379 cases were α-thalassemia (including deletional α-thalassemia and non-deletional α-thalassemia), with a detection rate of 25.25%, the most common genotype was -- ^<i>SEA</i>/αα (227 cases, 15.12%), followed by -α^3.7/αα (53 cases, 3.53%). 270 cases of β-thalassemia were detected, with a detction rate of 17.99%, and β^{<i>CD41-42</i>} /β^<i>N</i> (144 cases, 9.59%) was the main genotypes, followed by β^<i>CD17</i>/β^<i>N</i> (66 cases, 4.40%) . In addition, there were 29 cases of αβ compound thalassemia, accounting for 1.93%, and the most common genotype was --^<i>SEA</i>/αα complex β^{<i>CD41-42</i>} /β^<i>N</i> (5 cases, 0.33%).</p><p><strong>Conclusion: </strong>Lingui District in Guilin City is a high-incidence area of thalassemia, and the genotypes of carriers are complex and diverse, with genetic heterogeneity. The results of this study provide a scientific basis for genetic counseling and prenatal diagnosis in this area.</p>","PeriodicalId":35777,"journal":{"name":"中国实验血液学杂志","volume":"32 6","pages":"1848-1851"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915095","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Analysis of Gene Mutation and Clinical Characteristics Related to Myelodysplastic Syndrome].","authors":"Yu-Feng Wang, Yan-Li Yang, Ying-Hua Geng","doi":"10.19746/j.cnki.issn.1009-2137.2024.06.025","DOIUrl":"https://doi.org/10.19746/j.cnki.issn.1009-2137.2024.06.025","url":null,"abstract":"<p><strong>Objective: </strong>To explore the characteristics of gene mutation in patients with myelodysplastic syndrome (MDS) and its correlation with clinical features.</p><p><strong>Methods: </strong>From January 2017 to December 2021, 172 patients with MDS in The First Affiliated Hospital of Bengbu Medical University were analyzed retrospectively. Fourteen high frequency genes related to MDS were detected, and the relationship between gene mutation and clinical characteristics of patients as well as revised International Prognostic Scoring System (IPSS-R) was analyzed. The impact of gene mutations on prognosis was explored.</p><p><strong>Results: </strong>Among 172 patients, there were 101 males and 71 females, with a median age of 67 (15-89) years old, and gene mutations were detected in 88 cases (51.2%). The genes with mutation incidence >5% were arranged in descending order as follows: <i>TET2 (16.9%), RUNX1 (12.8%), ASXL1 (12.2%), CEBPA (8.1%), TP53 (7.0%)</i> and <i>DNMT3A (6.4%)</i>. According to biological functional classification, the genes with the highest mutation frequency were epigenetic regulatory genes (36.6%). The proportion of primitive bone marrow cells in mutation group was higher than that in non-mutation group (<i>P</i> < 0.001). The incidence of gene mutation varied in different MDS subtypes, and the difference was statistically significant (<i>P</i> < 0.05). The mutation incidence in IPSS-R higher risk group (IPSS-R score >3.5) was 65.7%, which was significantly higher than 30.0% in IPSS-R lower risk group (IPSS-R score ≤3.5) (<i>P</i> < 0.05), and there was a statistically significant difference in the incidence of <i>TP53</i> gene mutation between the two groups (<i>P</i> < 0.05). Multivariate Cox survival analysis showed that <i>TP53, NPM1</i> and <i>TET2</i> gene mutation were independent risk factors affecting prognosis.</p><p><strong>Conclusion: </strong>MDS patients are prone to gene mutation, and the increasing number of mutations and the presence of <i>TP53, NPM1</i> and <i>TET2</i> gene mutation may be factors affecting the prognosis.</p>","PeriodicalId":35777,"journal":{"name":"中国实验血液学杂志","volume":"32 6","pages":"1798-1806"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915160","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Application Analysis of Screening for Thalassemia in the Population of Childbearing Age in Quanzhou].","authors":"Mei-Zhen Yan, Xiao-Long Liu, Yuan-Bai Wang, Yu-Ying Jiang, Jian-Long Zhuang, Geng Wang, Qian-Mei Zhuang","doi":"10.19746/j.cnki.issn.1009-2137.2024.06.031","DOIUrl":"https://doi.org/10.19746/j.cnki.issn.1009-2137.2024.06.031","url":null,"abstract":"<p><strong>Objective: </strong>To analyze the application value of MCV, MCH and HbA₂ in screening for thalassemia in the population of childbearing age in Quanzhou area, and to determine the optimal screening cut-off value of relevant indicators in this area.</p><p><strong>Methods: </strong>2 725 couples of childbearing age were included in the study and underwent routine blood test, capillary hemoglobin electrophoresis, and α and β thalassemia gene test. Statistical methods were used to analyze the distribution of thalassemia genotypes, and compare the performance of MCV, MCH, and HbA₂ in screening various types of thalassemia. According to the ROC curve, the best cut-off values of MCV, MCH and HbA₂ in screening for thalassemia in this area were determined.</p><p><strong>Results: </strong>In this study, a total of 1 801 thalassemia carriers were detected, including 1 341 cases of α-thalassemia, 420 cases of β-thalassemia, and 40 cases of αβ compound thalassemia. The most common genotypes of α-thalassemia and β-thalassemia were -- ^<i>SEA</i>/αα and β^<i>654</i> /β^<i>N</i> , respectively. ROC curves were drawn to evaluate the performance of MCV, MCH and HbA₂ in screening for α-thalassemia, mild β-thalassemia, αβ compound thalassemia, silent α-thalassemia, mild α-thalassemia, and intermediate α-thalassemia. The maximum areas under the curves (AUC) were 0.747, 0.865, 0.724, 0.486, 0.812, 0.841; 0.747, 0.846, 0.703, 0.479, 0.796, 0.903; 0.613, 0.980, 0.909, 0.465, 0.674, 0.996, respectively; and the best cut-off values corresponding to the three screening indicators were 76.15fl, 71.95fl, 77.35fl, 86.15fl, 75.41fl, 61.15fl; 24.35pg, 21.51pg, 25.45pg, 28.65pg, 24.01pg, 20.51pg; 2.45%, 3.05%, 3.55%, 3.25%, 2.45%, 1.65%, respectively.</p><p><strong>Conclusion: </strong>The levels of MCV, MCH and HbA₂ are correlated with the phenotype of thalassemia, and the detection of these indicators is of great significance for the prevention and control of thalassaemia.</p>","PeriodicalId":35777,"journal":{"name":"中国实验血液学杂志","volume":"32 6","pages":"1841-1847"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915682","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[The Role of Matrix Metalloproteinase-13 in Multiple Myeloma].","authors":"Hai-Ying Jia, Shu-Li Guo, Liang Yu, Guo-Hong Huang, Chang-Min Wang","doi":"10.19746/j.cnki.issn.1009-2137.2024.06.022","DOIUrl":"https://doi.org/10.19746/j.cnki.issn.1009-2137.2024.06.022","url":null,"abstract":"<p><strong>Objective: </strong>To investigate the role of matrix metalloproteinase-13 (<i>MMP-13</i> ) in the development, diagnosis and prognosis of multiple myeloma (MM).</p><p><strong>Methods: </strong>Blood samples of 57 MM patients and 45 normal controls were collected, and real-time PCR was performed to detect the <i>MMP-13</i> mRNA expression level in the study subjects, and the difference of <i>MMP-13</i> mRNA level between MM patients and normal controls was compared. The correlations of <i>MMP-13</i> with MM bone disease and its severity, ISS stage, DS stage, and treatment efficacy were analyzed.</p><p><strong>Results: </strong>The <i>MMP-13</i> mRNA in patients with MM was significantly higher than that in normal controls (<i>P</i> < 0.05). The <i>MMP-13</i> mRNA in MM patients with bone disease was significantly higher than that in patients without bone disease, and the more severe the bone disease, the more obvious the increase in <i>MMP-13</i> mRNA (<i>P</i> < 0.05). The <i>MMP-13</i> gene expression level was also significantly different between ISS and DS stage Ⅰand stage Ⅲ patients (<i>P</i> < 0.05), and the <i>MMP-13</i> mRNA level was significantly decreased after treatment (<i>P</i> < 0.05).</p><p><strong>Conclusion: </strong>The <i>MMP-13</i> mRNA expression level is related to the occurrence and development of MM, and it has certain guiding significance in disease diagnosis and prognosis evaluation.</p>","PeriodicalId":35777,"journal":{"name":"中国实验血液学杂志","volume":"32 6","pages":"1776-1780"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915750","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[The Value of IgG Anti-A/Anti-B Antibody Titers after Absorption of IgG Anti-AB Antibodies in Predicting ABO Fetal Neonatal Hemolytic Disease].","authors":"Chen Cheng, Yi Zhang, Yi-Jing Chen, Qun Luo, Hai-Long Zhuo","doi":"10.19746/j.cnki.issn.1009-2137.2024.06.041","DOIUrl":"https://doi.org/10.19746/j.cnki.issn.1009-2137.2024.06.041","url":null,"abstract":"<p><strong>Objective: </strong>To analyze the diagnostic value of IgG anti-A/anti-B antibody titer in the serum of type O pregnant women after absorption of IgG anti-AB antibody for ABO hemolytic disease of fetus and newborn (ABO-HDFN).</p><p><strong>Methods: </strong>From February 2020 to September 2020, 235 samples of neonatal hemolytic disease whose mother's blood type O from Beijing Blood Center were selected. The titer of IgG anti-A/anti-B antibody in mother's serum before and after absorption of IgG anti -AB antibody was detected by microcolumn gel card, and the incidence of ABO-HDFN was statistically analyzed. The titer level of IgG anti-A/ant-B antibody and the incidence of ABO-HDFN were compared before and after the absorption of IgG anti-AB antibody, and the diagnostic efficacy of the titer level of IgG anti -A/anti-B antibody in the serum of type O pregnant women after the absorption of IgG anti-A and B antibodies on the incidence of ABO-HDFN was analyzed using the receiver's work characteristic (ROC) curve.</p><p><strong>Results: </strong>Of the 235 neonatal hyperbilirubinemia samples with maternal blood type O, 127 were blood type A, 38 of which were diagnosed as ABO-HDFN; 108 were blood type B, of which 31 were diagnosed as ABO-HDFN. Before and after absorption of IgG anti-AB antibody, there was a significant difference in the titer of IgG anti-A/anti-B antibody (<i>P</i> < 0.05). Among the 69 confirmed cases, the incidence of ABO-HDFN increased with the increase of IgG anti-A/anti-B antibody with or without the IgG anti-AB antibody, but the anti-A/anti-B antibody titer ≥1∶512 before the absorption of IgG anti-AB antibody, while the anti-A/anti-B antibody titer decreased significantly, decreasing by three titers, all≤1∶512. The ROC curve shows that the titers of IgG anti-A/anti-B antibodies before and after absorption of IgG anti-AB antibodies can be used as the efficacy indicators for the diagnosis of ABO-HDFN. However, there was a significant difference in the potency of IgG anti-A/anti-B antibody titer for the diagnosis of ABO-HDFN before and after the absorption of IgG anti-AB antibody (<i>P</i> < 0.05). The AUC values were greater than before absorption, indicating that the IgG anti-A/anti-B antibody after the absorption of IgG anti-AB antibody was better than before absorption (<i>P</i> < 0.05).</p><p><strong>Conclusion: </strong>The higher the titer of IgG anti-A/anti-B antibody measured after absorbing IgG anti-AB antibody, the higher the incidence of ABO-HDFN. In addition, the efficacy of IgG anti-A/anti-B antibody titer to diagnose ABO-HDFN after absorption of IgG anti-AB antibody is higher than that before absorption.</p>","PeriodicalId":35777,"journal":{"name":"中国实验血液学杂志","volume":"32 6","pages":"1903-1908"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915754","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Molecular Mechanism of Protein C Deficiency Caused by Mutations of <i>PROC</i> Gene N355S, G392E, T314A].","authors":"Tian-Yi Li, Miao Jiang, Lu-Lu Huang, Jing-Jing Han, Zhen-Ni Ma, Xia Bai, Li-Jun Xia","doi":"10.19746/j.cnki.issn.1009-2137.2024.06.030","DOIUrl":"https://doi.org/10.19746/j.cnki.issn.1009-2137.2024.06.030","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Objective: &lt;/strong&gt;To study the molecular mechanism of functional defect of protein C (PC) caused by point mutations of human protein C gene (&lt;i&gt;PROC&lt;/i&gt; ) N355S , G392E and T314A.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;The wild-type and mutant plasmids (PC&lt;sub&gt;WT&lt;/sub&gt;, PC&lt;sub&gt;N355S&lt;/sub&gt;, PC&lt;sub&gt;G392E&lt;/sub&gt;, PC&lt;sub&gt;T314A&lt;/sub&gt;) of &lt;i&gt;PROC&lt;/i&gt; gene were constructed and transiently transfected into HEK293 cells. The expression of mutant proteins in vitro were tested. The mRNA level changes of wild-type and mutant PC after 24 h of transfection were detected by real-time PCR. Western blot and ELISA were used to detect the changes of intracellular and extracellular protein levels of wild-type and mutant PC. The supernatant of cells transfected for 24-48 h was concentrated by ultrafiltration. The protein in the concentrated solution was quantified, and PC activation and enzyme kinetics tests were performed. Clustal Omega multiple sequence alignment was used to analyze the conservation of amino acid mutation sites. The effect of mutation on PC protein structure was analyzed by PyMOL software.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;The relative expression abundances of &lt;i&gt;PROC&lt;/i&gt; mRNA in PC&lt;sub&gt;N355S&lt;/sub&gt;, PC&lt;sub&gt;G392E&lt;/sub&gt; and PC&lt;sub&gt;T314A&lt;/sub&gt; groups were 1.14±0.46, 0.96±0.08 and 1.08±0.17, respectively, and there were no significant differences compared with 1.02±0.24 in PC&lt;sub&gt;WT&lt;/sub&gt; group (&lt;i&gt;P&lt;/i&gt; &gt;0.05). Western blot analysis of the lysates of transfected cells showed that the content of PC&lt;sub&gt;T314A&lt;/sub&gt; recombinant protein slightly decreased and the band became relatively lighter. The ELISA results of the concentrated cell culture supernatants showed that the PC:Ag levels of PC&lt;sub&gt;N355S&lt;/sub&gt; and PC&lt;sub&gt;G392E&lt;/sub&gt; were 98.8%±2.4% and 101.4%±3.1%, respectively, with no significant differences compared with PC&lt;sub&gt;WT&lt;/sub&gt;, while PC&lt;sub&gt;T314A&lt;/sub&gt; decreased compared with PC&lt;sub&gt;WT&lt;/sub&gt; (PC:Ag: 88.6%±3.2%) (&lt;i&gt;P&lt;/i&gt; &lt; 0.05). The results of enzyme kinetics test showed that APC&lt;sub&gt;N355S&lt;/sub&gt; (K&lt;sub&gt;m&lt;/sub&gt;=338.3±43.2, V&lt;sub&gt;max&lt;/sub&gt;=2.015±0.12), APC&lt;sub&gt;G392E&lt;/sub&gt; (K&lt;sub&gt;m&lt;/sub&gt;=292.2±28.4, V&lt;sub&gt;max&lt;/sub&gt;=1.893±0.07) and APC&lt;sub&gt;T314A&lt;/sub&gt; (K&lt;sub&gt;m&lt;/sub&gt;=299.5±24.6, V&lt;sub&gt;max&lt;/sub&gt;=1.775±0.06) showed an increase in K&lt;sub&gt;m&lt;/sub&gt; and a decrease in V&lt;sub&gt;max&lt;/sub&gt; compared with APC&lt;sub&gt;WT&lt;/sub&gt; (K&lt;sub&gt;m&lt;/sub&gt;=238.2±4.58, V&lt;sub&gt;max&lt;/sub&gt;=3.205±0.06). Multiple sequence alignment suggested that the three mutations be highly conservative in different species. The structural model suggested that the amino acid substitutions of N355S, G392E and T314A mutations collide with the surrounding amino acid groups, causing distortion of the surrounding structure, which may have adverse effects on the folding and biological function of PC.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusion: &lt;/strong&gt;The N355S, G392E and T314A mutations in the &lt;i&gt;PROC&lt;/i&gt; gene cause functional defects in PC by weakening the binding between PC and substrate. These three mutations have caused se","PeriodicalId":35777,"journal":{"name":"中国实验血液学杂志","volume":"32 6","pages":"1834-1840"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915808","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Correlation of <i>BRAF V600E</i> Mutation with Clinical Features and Prognosis of Langerhans Cell Histiocytosis in Cildren].","authors":"Xi Li, Li Xiao, Ming-Zhu Luo, Xiao-Ying Lei, Hai-Yan Liu, Xin-Yuan Yao, Yu-Xia Guo, Ying Dou, Jie Yu","doi":"10.19746/j.cnki.issn.1009-2137.2024.06.043","DOIUrl":"https://doi.org/10.19746/j.cnki.issn.1009-2137.2024.06.043","url":null,"abstract":"<p><strong>Objective: </strong>To explore the gene mutations of Langerhans cell histiocytosis in children, and to analyze the correlation of <i>BRAF V600E</i> mutation with clinical features and prognosis of LCH, so as to provide reference for clinical diagnosis and treatment.</p><p><strong>Methods: </strong>Fluorescence PCR was used to detect gene mutations in paraffin-embedded tissue samples from 78 children with LCH, and the correlation of <i>BRAF V600E</i> mutation with clinical characteristics and prognosis of LCH in children was analyzed.</p><p><strong>Results: </strong>Among the 78 children, 41 cases (52.6 %) had <i>BRAF V600E</i> mutation, 8 cases (10.3 %) had <i>MAP2K1</i> mutation, 1 case (1.3 %) had <i>BRAF Exon 12</i> mutation, 1 case (1.3 %) had <i>ARAF</i> mutation, and 1 case (1.3%) had <i>PIK3CA</i> mutation. <i>BRAF V600E</i> mutation was not significantly correlated with sex, age, multisystem involvement, risk-organ involvement, CNS-risk lesions, and early treatment response in children with LCH (<i>P</i> >0.05), and it was also not significantly correlated with the recurrence and event-free survival (EFS) of children with LCH (<i>P</i> >0.05).</p><p><strong>Conclusion: </strong>LCH is an inflammatory myeloid tumor. <i>BRAF V600E</i> mutation is not correlated with clinical features, early treatment response, recurrence and prognosis of LCH.</p>","PeriodicalId":35777,"journal":{"name":"中国实验血液学杂志","volume":"32 6","pages":"1917-1922"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Generation and Evaluation of Human Umbilical Cord Derived Mesenchymal Stem Cells with Antioxidant Capacity].","authors":"Xiao-Yu Zhang, Pei-Lin Li, Jie Tang, Zhi-Ling Li, Rui-Cong Hao, Xiao-Tong Li, Wen-Jing Zhang, Shi-Rong Zhao, Li Ding, Wen-Qing Wu, Heng Zhu","doi":"10.19746/j.cnki.issn.1009-2137.2024.06.039","DOIUrl":"https://doi.org/10.19746/j.cnki.issn.1009-2137.2024.06.039","url":null,"abstract":"<p><strong>Objective: </strong>To prepare mesenchymal stem cells with antioxidant capacity (AO-MSC) from human umbilical cords and evaluate its cell biological properties.</p><p><strong>Methods: </strong>In control group, mesenchymal stem cells (MSC) were isolated by digesting human umbilical cord Wharton's Jelly tissues with 0.2% collagenase II, and the released cells were collected and cultured in an animal serum-free culture medium. In AO-MSC group, incompletely collagenase II-digested tissue debris were allowed to adhere to flusk flat bottoms and the AO-MSC was harvested by adherent culture. The conventional digestion and culture method was used as control. MSC colony forming ability was evaluated by fibroblast colony forming assay (CFU-F). MSC proliferative capacity was evaluated by CCK-8 assay. The MSC surface markers were detected by using flow cytometry and immunofluorescence staining. The adipogenic and osteogenic capacity of MSC was evaluated by multi-differentiation <i>in vitro</i>, and the mRNA expression of genes that control adipogenic and osteogenic differentiation was detected by real-time fluorescence quantitative PCR (RT-qPCR); Moreover, the mRNA expression of antioxidant substances such as <i>SOD-1, GSH, GAT</i>, and <i>NQO1</i> in MSC was also evaluated by RT-qPCR.</p><p><strong>Results: </strong>The AO-MSC isolated by this strategy reached a confluence of 80%-90% at around 18 days and grew in a swirling pattern. Flow cytometry and immunofluorescence staining assays showed that CD73, CD29, CD105, CD90 were highly expressed and CD31, CD45, HLA-DR were scarcely expressed in AO-MSC. AO-MSC exhibited stronger self-renewal and differentiation ability compared to MSC. However, the <i>in vitro</i> adipogenic-osteogenic capacity of MSC in the control group was stronger than that of AO-MSC. RT-qPCR assay showed that AO-MSC expressed higher mRNA levels of antioxidant substances compared to MSC.</p><p><strong>Conclusion: </strong>Human AO-MSC is successfully prepared from human umbilical cord without animal serum.</p>","PeriodicalId":35777,"journal":{"name":"中国实验血液学杂志","volume":"32 6","pages":"1888-1895"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915867","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[<i>Ku80</i> Inhibition Affects the Chemotherapeutic Sensitivity of T-Acute Lymphoblastic Leukemia Cell Line Jurkat].","authors":"Zhuo-Yi Fan, Ai-Bin Liang","doi":"10.19746/j.cnki.issn.1009-2137.2024.06.009","DOIUrl":"https://doi.org/10.19746/j.cnki.issn.1009-2137.2024.06.009","url":null,"abstract":"<p><strong>Objective: </strong>To investigate the influence of <i>Ku80</i> inhibition on the chemotherapeutic sensitivity of the T-acute lymphoblastic leukemia(T-ALL) cell line Jurkat, and to explore the potential mechanism.</p><p><strong>Methods: </strong>The transcription and expression level of <i>Ku80</i> in 6 hematological malignant cell lines were detected by RT-qPCR and Western blot, respectively. The expression of Ku80 in Jurkat cells was detected by Western blot after transfection with the recombinant sh<i>Ku80</i> lentiviral vector. The proliferation capacity of Jurkat cells was explored by CCK-8 after <i>Ku80</i> inhibition. The colony formation ability, apoptosis, and γH2AX(a protein marker of DNA damage) expression in Jurkat cells were investigated after <i>Ku80</i> silencing and co-treated with etoposide(VP16) for 4 hours through soft agar assay, flow cytometry and Western blot, respectively.</p><p><strong>Results: </strong>The mRNA level and protein expression of Ku80 were both highest in Jurkat among 6 hematological malignant cell lines. <i>Ku80</i> expression was successfully down regulated in Jurkat cells after relative plasmid transfected. The proliferative ability of cells was significantly decreased after <i>Ku80</i> inhibition(<i>P</i> < 0.05). The colony formation capacity of Jurkat cells was obviously reduced and the cells apoptosis and γH2AX expression were increased after <i>Ku80</i> inhibition, with or without VP16 incubation.</p><p><strong>Conclusion: </strong>Targeted silencing of <i>Ku80</i> could enhance the sensitivity of VP16 in Jurkat cells, which might be associated with the elevated level of DNA damage accumulation.</p>","PeriodicalId":35777,"journal":{"name":"中国实验血液学杂志","volume":"32 6","pages":"1689-1695"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915034","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[The Impact of <i>SRSF1</i> -Mediated Alternative Splicing of <i>RBBP6</i> on the Proliferation of Multiple Myeloma Cells].","authors":"Wei-Min Zhang, Sha Song, Wen-Zhuo Zhuang, Bing-Zong Li","doi":"10.19746/j.cnki.issn.1009-2137.2024.06.016","DOIUrl":"https://doi.org/10.19746/j.cnki.issn.1009-2137.2024.06.016","url":null,"abstract":"<p><strong>Objective: </strong>To investigate the effect of different isoforms of <i>RBBP6</i> on the proliferation of multiple myeloma (MM) cells after alternative splicing mediated by splicing factor <i>SRSF1</i> .</p><p><strong>Methods: </strong>RT-PCR was used to detect the expression levels of <i>RBBP6</i> mRNA splicing isoforms regulated by <i>SRSF1</i> . The GEO database was used to analyze the changes of <i>RBBP6</i> isoform 1 in the progression of plasma cell disease, and survival analysis was used to evaluate the value of this gene in the prognosis of MM patients. <i>in vitro</i> loss-of-function and gain-of-function experiments were conducted by transfecting control siRNA, <i>RBBP6</i> isoform 1 siRNA, empty vector (EV), OE-<i>RBBP6</i> isoform 3 into MM.1S cells, then the expression levels of <i>RBBP6</i> isoform 1 and <i>RBBP6</i> isoform 3 were detected by real-time PCR and Western blot. CCK-8 assay was performed to detect the cell proliferation ability.</p><p><strong>Results: </strong>Knockdown of <i>SRSF1</i> increased the expression of <i>RBBP6</i> isoform 3 and decreased the expression of <i>RBBP6</i> isoform 1. <i>RBBP6</i> isoform 1 was closely related to the progression of plasma cell disease, and the high expression of <i>RBBP6</i> isoform 1 was associated with poor prognosis in patients with MM. Downregulation of <i>RBBP6</i> isoform 1 expression and overexpression of <i>RBBP6</i> isoform 3 both reduced the proliferation ability of MM cells. And after downregulating the expression of <i>RBBP6</i> isoform 1, the level of p53 protein in MM cells was significantly increased (<i>P</i> < 0.05).</p><p><strong>Conclusion: </strong>In MM, splicing factor <i>SRSF1</i> can cause alternative splicing abnormalities in <i>RBBP6</i> . The <i>RBBP6</i> isoform 1 promotes MM cell proliferation, while the <i>RBBP6</i> isoform 3 inhibits MM cell proliferation, and the mechanism may be related to regulating the p53 pathway.</p>","PeriodicalId":35777,"journal":{"name":"中国实验血液学杂志","volume":"32 6","pages":"1738-1743"},"PeriodicalIF":0.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}