Mycoplasma pneumoniae infection and macrolide resistance of children hospitalized in Ningbo Area， Zhejiang Province in 2019‒2021 / 上海预防医学

ObjectiveTo investigate Mycoplasma pneumoniae （MP） infection and macrolide resistance of hospitalized children in Ningbo Area in 2019‒2021. MethodsA total of 6 782 respiratory throat swab specimens were collected from hospitalized pediatric patients with pneumoniae， admitted in Ningbo Women and Children's Hospital from January， 2019 to December 2021. MP and its mutations in 23S rRNA were detected by real-time polymerase chain reaction. ResultsAmong 6 782 respiratory throat swab specimens from 2019‒2021， 1 290 cases （19.02%） were MP positive， and the positive rate decreased year by year （P<0.05）. The positive rate in 2019 was 28.12%， higher than that in 2020 （7.16%） and 2021 （5.16%） （all P<0.017）. The mutation of 23S rRNA occurred in 947 cases， with a mutation rate of 73.41%. The mutation rate in 2020 was 84.04%， higher than that in 2019 （73.01%） and 2021 （66.23%）. The differences of positive rate and mutation rate in different seasons were significant （P<0.05） （all P<0.008）. The positive rate was the highest in summer （25.00%）， and the mutation rate was the highest in winter （78.89%）. The positive rate of female children was 20.52%， higher than that of male children （17.82%） （P<0.05）， and the mutation rates of female and male children were 74.93% and 71.77% （P>0.05）， respectively. The difference of positive rate among the different age was significant （P<0.05）. The positive rates in the 5‒ and 8‒ years groups were 27.24% and 26.38%， higher than those in the 1‒ and 2‒ years groups， respectively. The difference of mutation rate among the four groups in age was not significant （P>0.05）. ConclusionThe infection rate of MP in children decreases in Ningbo Area from 2019 to 2021. MP infection may be related with gender， seasonal distribution， age， and the resistance rate of MP to macrolide is high.

Selection and Validation of Appropriate Reference Genes of Cinnamomum cassia and Cinnamomum cassia var. macrophyllum / 中国实验方剂学杂志

Objective:To screen out stable internal reference genes suitable for real-time quantitative polymerase chain reaction（Real-time PCR） analysis of different parts of<italic> Cinnamomum cassia</italic> and <italic>C. cassia</italic> var. <italic>macrophyllum</italic>，in order to provide stable internal reference genes for gene expression analysis of three different parts of and <italic>C. cassia</italic> var.<italic> macrophyllum</italic> branches and leaves. Method:With 6 different tissues and organs， such as bark，branches and leaves of two plants of <italic>C. cassia</italic> and <italic>C. cassia</italic> var. <italic>macrophyllum</italic> as experimental materials，Real-time PCR technology was used to detect the five internal reference genes， namely glyceraldehyde-3-phosphate dehydrogenase（GAPDH），actin，ubiquitin-ligase enzymes（UBE），histone and tubin（TUB）. The analysis of the expression of the data. Furthermore， three commonly used internal reference gene analysis software，namely geNorm，NormFinder and BestKeeper，was used to analyze and evaluate the stability of the candidate internal reference gene. Result:The internal five reference genes were expressed in the bark，branches and leaves of the two plants，but with differences in stability. Comprehensive analysis showed that the expression stability of candidate internal reference genes was in the order of GAPDH>actin>UBE>histone>TUB. The internal reference genes of the two plants were analyzed separately，and the optimal internal reference gene was still GAPDH，indicating that GAPDH was the most suitable internal reference gene. TUB and histone ranked low in the three software，and should be eliminated in the screening of reference genes. They were not suitable for gene expression analysis of <italic>C. cassia </italic>and <italic>C. cassia</italic> var. <italic>macrophyllum</italic>. Conclusion:The most suitable internal reference gene for different parts of cinnamon，branches，and leaves of <italic>C. cassia</italic> and <italic>C. cassia</italic> var. <italic>macrophyllum</italic> was GAPDH. In this study，a screening system for internal reference genes of Real-time PCR of <italic>C. cassia</italic> and <italic>C. cassia</italic> var. <italic>macrophyllum</italic> was established to provide theoretical basis for studying functional regulation and expression of genes during the accumulation of effective components in different parts.

Integrative molecular characterization of Chinese prostate cancer specimens / 亚洲男科学杂志(英文版)

Prostate cancer (PCa) exhibits epidemiological and molecular heterogeneity. Despite extensive studies of its phenotypic and genetic properties in Western populations, its molecular basis is not clear in Chinese patients. To determine critical molecular characteristics and explore correlations between genomic markers and clinical parameters in Chinese populations, we applied an integrative genetic/transcriptomic assay that combines targeted next-generation sequencing and quantitative real-time PCR (qRT-PCR) on samples from 46 Chinese patients with PCa. Lysine (K)-specific methyltransferase 2D (KMT2D), zinc finger homeobox 3 (ZFHX3), A-kinase anchoring protein 9 (AKAP9), and GLI family zinc finger 1 (GLI1) were frequently mutated in our cohort. Moreover, a clinicopathological analysis showed that RB transcriptional corepressor 1 (RB1) deletion was common in patients with a high risk of disease progression. Remarkably, four genomic events, MYC proto-oncogene (MYC) amplification, RB1 deletion, APC regulator of WNT signaling pathway (APC) mutation or deletion, and cyclin-dependent kinase 12 (CDK12) mutation, were correlated with poor disease-free survival. In addition, a close link between KMT2D expression and the androgen receptor (AR) signaling pathway was observed both in our cohort and in The Cancer Genome Atlas Prostate Adenocarcinoma (TCGA-PRAD) data. In summary, our results demonstrate the feasibility and benefits of integrative molecular characterization of PCa samples in disease pathology research and personalized medicine.

Applicable Value of AMSS-PCR in Lung Cancer Gene Mutation Detection / 中国肺癌杂志

BACKGROUND@#The detection of driver oncogenes of lung cancer is of great importance. There are various gene detection techniques nowadays which are different from each other. We carried out this study to investigate the specificity and sensitivity of assay panels based on an Amplification Refractory Mutation System-polymerase chain reaction (ARMS-PCR) technique of Amplification Mutation Specific System (AMSS) in detection of lung cancer gene mutation. To estimate the applicable value of assay panels in clinical settings.@*METHODS@#We collected cancer tissue specimens or fluid specimens from 309 patients. Mutation results were presented for those samples previously detected by ARMS-PCR. In comparison, we carried out AMSS-PCR using (epidermal growth factor receptor, EGFR) assay panel and Six-Alliance assay panel as well as Sanger sequencing. Software SPSS 22.0 (SPSS IBM) was used for statistical analysis.@*RESULTS@#The rates of consistency between the results by assay panels and Sanger sequencing or ARMS-PCR were 97.41% and 97.73%, respectively. Besides, EGFR assay panel had higher consistency rates with other detection methods than Six-Alliance assay panel. As for consistency test, the Kappa values of assay panels with Sanger sequencing, assay panels with ARMS-PCR, and ARMS-PCR with Sanger sequencing were 0.946, 0.953, and 0.913, respectively. The receiver operating characteristic curve (ROC) area under curve (AUC) of assay panels was 0.976 referring to Sanger sequencing, and 0.975 as ARMS-PCR was referred to.@*CONCLUSIONS@#AMSS-PCR can make an optimal cancer gene mutation detection method for clinical settings.

Study of Bacteria Associated with Dental Caries Using a 3 Tone Disclosing Agent / 대한소아치과학회지

The aim of this study was to determine the efficacy of a 3 tone plaque disclosing gel in assessing the risk of caries related to the population of Streptococcus mutans, Streptococcus sobrinus, and Lactobacillus spp. quantified using a quantitative real-time polymerase chain reaction (qRT-PCR).15 healthy children of ages 9 – 12 years were randomly examined. The 3 tone plaque disclosing gel was applied on teeth surfaces, which changed the color to pink or red, blue or purple and light blue. Plaque was divided into 3 groups based on staining. Genomic DNA from each sample was subjected to a qRT-PCR assay for quantitative detection of target bacteria. The Kruskal-Wallis test was conducted for correlation between the color of plaque and the number of bacterial species.The levels of S. mutans, S. sobrinus, and Lactobacillus spp. were significantly different in the plaque samples of the 3 groups (p < 0.05). The proportion of S. sobrinus to S. mutans showed correlation to the color of plaque.The different color-dyed plaque was related to the number of acidogenic bacteria. The 3 tone plaque disclosing gel could be used as one of the indicators to assess the clinical risk of caries associated with the population of S. mutans, S. sobrinus, and Lactobacillus spp.

Characterization of the fecal microbiota differs between age groups in Koreans

BACKGROUND/AIMS: Tens of trillions of microorganisms constitute the gut microbiota of the human body. The microbiota plays a critical role in maintaining host immunity and metabolism. Analyses of the gut microbial composition in Korea are limited to a few studies consisting of small sample sizes. To investigate the gut microbial community in a large sample of healthy Koreans, we analyzed the 16S ribosomal RNA of 4 representative bacterial genera Lactobacillus, Bifidobacterium, Bacteroides, and Clostridium. METHODS: A total of 378 DNA samples extracted from 164 infants and 214 adults were analyzed using quantitative real-time polymerase chain reaction. RESULTS: Analysis of 16S ribosomal RNA of 4 representative bacterial genera Lactobacillus, Bifidobacterium, Bacteroides, and Clostridium showed that the gut microbiota in infants had higher relative abundances of Bifidobacterium and Lactobacillus than that in adults, which was dominated by Bacteroides and Clostridium. CONCLUSIONS: To the best of our knowledge, this was the first study evaluating the distinct characteristics of the microbial community of Korean infants and adults. The differences between the 2 populations suggest that external factors such as age, diet, and the environment are important contributing factors to the change in gut microbial composition during development.

Application of polyhydroxy acrylic acid and Van-clear in the detection of EGFRgene mutations in non small-cell lung cancer by qRT-PCR method / 西安交通大学学报(医学版)

Objective To compare two different methods to detect the differences of gene mutation rate, sensitivity, specificity and coincidence rate of epidermal growth factor receptor (EGFR) in non-small-cell lung cancer (NSCLC) so as to assess the clinical value of qRT-PCR method and its environmental-friendly technologyplatforms.One uses environmental fixative poly hydroxyl acrylic acid and green transparent liquid dewaxing Van-clear alone or in combination to replace the traditional fixative 4％ (volume fraction) neutral buffered formalin and the traditional transparent dewaxing liquid xylene in application of quantitative real-time polymerase chain reaction (qRT-PCR).The other uses traditional reagents in direct sequencing.Methods We selected 91 cases of primary NSCLC specimens resected between May 2013 and March 2016 in Zhongshan Bo`ai Hospital and Zhongshan Hospital of Traditional Chinese Medicine.Five samples were taken from the same tumor lesion.We used a random number table to randomly divide these samples into Groups A, B , C, D, and E.Group A received direct sequencing method in detection of EGFR gene mutations.Besides, during the experiment, 4％ neutral buffered formalin was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group B received qRT-PCR method to detect EGFR gene mutations.Meanwhile, during the experiment, 4％ neutral buffered formalin was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group C received qRT-PCR method in detection of EGFR gene mutations.At the same time, during the experiment, polyhydroxy acrylic acid was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group D received qRT-PCR method to detect EGFR gene mutations.In the meantime, 4％ neutral buffered formalin was used for fixing, Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.Group E received qRT-PCR method in detection of EGFR gene mutations.In addition, during the experiment, polyhydroxy acrylic acid was used for fixing, and Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.In addition, during the experiment, polyhydroxy acrylic acid was used for fixing, and Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.The mutations of Exons 18, 19, 20, and 21 in EGFR genes were respectively determined in the five groups of NSCLC.Results ① Groups B, C, D, E and A did not significantly differ in the percentage of people with mutations or target site mutation rates of EGFR genes in NSCLC (P> 0.05).② The detection results of EGFR target site mutation in Groups B, C, D, E and A had good sensitivity, strong specificity, and high compliance rate.Conclusion The green transparent liquid dewaxing Van-clear alone or in combination to replace the traditional fixative 4％ neutral buffered formalin and the traditional transparent dewaxing liquid xylene in the application of qRT-PCR so as to detect EGFR gene mutations in NSCLC has good consistent results compared with the method that uses traditional reagents in direct sequencing.It has the significance and value in clinical application.

Establishment of a real-time PCR method to identify Ekpoma virus gene in blood sample of a returnee from Angola / 中华实验和临床病毒学杂志

Objective@#To establish quantitative real-time PCR (qPCR) method based on Taqman probe for detecting Ekpoma virus (EKV).@*Methods@#According to the conserved region of gene in EKV genome from GenBank, primers and probe for qPCR were designed. Validity and sensitivity were evaluated in this study. Both whole blood and serum of a returnee from Angola were tested by the established EKV-1 and EKV-2 qPCR method .@*Results@#Sensitivity of EKV-1 and EKV-2 qPCR method was respectively 41 copies/μl and 70 copies/μl. Coefficient of variance (CV) was respectively 1.27%, 0.20%, 0.82%; 2.12%, 1.74%, and 1.40%. EKV-2 gene was detected in both whole blood and serum of a returnee from Angola.@*Conclusions@#The first EKV-2 gene was confirmed in both whole blood and serum of a returnee from Angola by real-time RT-PCR..

Detection of minimal residual disease in children with acute lymphoblastic leukemia and its application / 中华实用儿科临床杂志

Childhood acute lymphoblastic leukemia (ALL) is a kind of abnormal proliferation of malignant tumor diseases originated in lymphocytes,which is the most common malignant tumor in children.Although stratified treatment has significantly improved the efficacy of ALL in children,but 15％-20％ of the patients still have ultimately relapsed due to the minimal residual disease(MRD).MRD refers to the leukemia patients after induction chemotherapy complete remission (or bone marrow transplant),in vivo residual morphology which could not be detected in trace amounts of leukemia cells.The detection methods of MRD mainly include flow cytometry,quantitative real-time polymerase chain reaction and immune repertoire sequencing.MRD level detection is of great important to judge the prognosis,and to guide the risk grouping and individual treatment,and so on.Now,the progress in the clinical application of MRD in children with ALL in recent years was reviewed.

Clinical significance of differential Drosha expression between endometrial cancer tissue and other tissues / 天津医药

Objective To detect the different expressions of Drosha between endometrial cancer (EC) tissue and other tissues and to explore correlation between Drosha mRNA transcription level and protein expression level with clinicopatho?logical characteristics of EC. Methods The mRNA transcription and protein expression levels of Drosha were examinaned by q-PCR and Western blot respectively in normal endometrial tissues (25 cases), atypical hyperplasia of endometrial tis?sues (20 cases) and endometrial cancer tissues (40 cases). Correlations between Drosha mRNA transcription and protein ex?pression with clinicopathological characteristics of EC were analyzed. Results The levels of Drosha mRNA and protein lev?els in EC were obviously lower than those in endometrial atypical hyperplasia and normal endometrium (P0.05). The protein expression levels of Drosha were consistent with transcription of mRNA transcription levels. Drosha mRNA expression does not differ significantly with differentiation, histological type, myometrial invasion, lymphatic metasta?sis and FIGO stages of EC (P>0.05). Conclusion The expression levels of Drosha in EC tissues were down-regulated, therefore the reduction of Drosha may contributed to tumorigenesis of EC.

Detecting EB virus to determine curative effect in extranodal natu-ral killer/T-cell lymphoma / 中国肿瘤临床

Objective:To investigate the clinical significance of detecting Epstein Barr virus (EBV) infection in evaluating recent curative and long-term effects in patients with extranodal natural killer (NK)/T-cell lymphoma. Methods:The EBV-DNA copies in the plasma of 109 patients, who were pathologically and immunohistochemically diagnosed with extranodal natural killer/T-cell lymphoma in the First Affiliated Hospital of Zhengzhou University between January 2011 and April 2014, were monitored via quantitative re-al-time polymerase chain reaction. Subsequently, the difference in recent curative and long-term effects between EBV positive and EBV negative patients was compared. Results:Among the 109 patients with extranodal NK/T-cell lymphoma, 34 (64.2%) cases of EBV posi-tive patients were at the advanced stage (Ⅲ~Ⅳ stages), and 22 (39.3%) cases of EBV negative patients were at the terminal stage (Ⅲ~Ⅳstages). EBV positive patients who accompanied by B symptoms were 33 (62.3%) , and there were 21 (37.5%) cases with B symptoms in EBV negative patients, the differences between stages and B symptoms were statistically significant. The attained objec-tive response rate of the EBV-DNA negative patients (34, 60.7%) was significantly higher than that of the EBV-DNA positive patients (22, 41.5%) (P<0.05). Similarly, the 2-year progression-free survival (PFS) rate of EBV negative patients was better than that of EBV positive patients (P<0.05). Conclusion:Detecting EBV in plasma has clinical significance in evaluating the recent curative effect and the 2-year PFS rate in patients diagnosed with extranodal NK/T-cell lymphoma.

The diagnosis value of combined detection of serum mir-125b and alpha-fetoprotein for primary hepatocellular carcinoma / 中国肿瘤临床

Objective:To investigate the possibility of miR-125b in serum as a novel tumor marker for primary hepatocellular car-cinoma (HCC) and the diagnosis value of combined detection of miR-125b and alpha-fetoprotein (AFP) for HCC. Methods:We detected serum miR-125b expression of 65 cases of HCC patients and 30 cases of healthy controls by real-time quantitative PCR. Moreover, we analyzed the significance of miR-125b and explored the relationship between miR-125b and clinical pathological factors. Results:The level of miRNA-125b was down regulated in serum of HCC patients compared with healthy controls which showed significant differences (P0.05). The expression level of miRNA-125b correlated the difference with liver Cirrhosis, tumor size and tumor node metastasis (TNM) stages, which were considered significant differences (P<0.05). The receiver operating characteristic (ROC) curve analysis of serum miR-125b for the diagnosis of HCC yielded AUC of 0.917(95%CI:0.851~0.960, P<0.001)with 85.9%sensitivity and 93.5%specificity. The ROC curve analysis of combined miR-125b and AFP for HCC detection yielded AUC of 0.951(95%CI:0.894~0.982, P<0.001)with 92.9%sensitivity and 93.5%specificity. The ROC curve analysis of serum miR-125b as biomarkers for the group (AFP<20μg/L) of HCC yielded AUC of 0.889(95%CI:0.776~0.957, P<0.001)with 84.0%sensitivity and 87.1%specificity. Conclusion:Serum miRNA-125b combined with AFP has considerable clinical value for the early diagnosis of primary HCC.

Cloning and expression of Schistosoma japonicum VCP gene and its mRNA expression levels in different stages / 中国血吸虫病防治杂志

Objective To prokaryotically express the valosin-containing protein(VCP)of Schistosoma japonicum,and ana-lyze its VCP mRNA expressions in the cercaria,schistosomulum,adult worm(female and male worms)and egg. Methods RNA of S. japonicum eggs were extracted,and reversely transcribed into cDNA. The VCP gene of S. japonicum was amplified by using polymerase chain reaction(PCR),and subcloned into the prokaryotically expressed vector pET15b. The recombined plasmid was transformed into BL21 cells,and the expression of the target gene was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG). The recombinant protein was yielded through the purification of inclusion body,and identified by using sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The RNA(s)of cercaria,schistosomulum,female adult worm,male adult worm,and egg of S. japonicum were extracted,digested with DNase,purified,and reversely transcribed into cDNA. The mRNA expressions of the VCP gene in various developmental stages of S. japonicum were determined by using fluorescence-based quantitative real-time PCR. Results The VCP gene of S. japonicum was yielded by PCR amplification,and the recombinant pro-tein was obtained through recombinant plasmid expression and purification of inclusion body. The highest VCP mRNA expression in S. japonicum cercaria was detected by the fluorescence-based quantitative real-time PCR,while low expressions were found in the schistosomulum,egg,female and male adult worms. Conclusion The recombinant protein encoded by the VCP gene of S. ja-ponicum is successfully obtained,and the VCP mRNA expression is determined in various developmental stages of S. japonicum.

Expression of SOCS1 and SOCS3 genes in human graft-versus-host disease after allogeneic hematopoietic stem cell transplantation

BACKGROUND: Suppressor of cytokine signaling genes (SOCS) are regarded as pivotal negative feedback regulators of cytokine signals, including the interferon-gamma (IFN-gamma), granulocyte-colony stimulating factor, and interleukin families, released by T cells. A detailed understanding of the involvement of SOCS genes in graft-versus-host disease (GVHD) is critical to effectively manage GVHD, yet their expression patterns among recipients remain largely unexplored. METHODS: Expression levels of SOCS1 and SOCS3 were determined by real-time quantitative reverse transcription PCR (qRT-PCR) in patients with acute GVHD (aGVHD) and chronic GVHD (cGVHD), in a severity-dependent manner, after allogeneic hematopoietic stem cell transplantation (HSCT). A total of 71 recipients with AML (N=40), ALL (N=12), myelodysplastic syndromes (MDS; N=10), chronic myelogenous leukemia (CML; N=2), severe aplastic anemia (SAA; N=5), or others (N=2), who received allogeneic HSCT from human leukocyte antigen-identical siblings or unrelated donors between 2009 and 2011, were included in the present study. RESULTS: Overall, the expression levels of SOCS1 decreased in recipients with grade II to IV aGVHD and cGVHD when compared to normal donors and non-GVHD recipients. Interestingly, the expressions of SOCS1 decreased significantly more in cGVHD than in aGVHD recipients (P=0.0091). In contrast, SOCS3 expressions were similarly reduced in all the recipients. CONCLUSION: This is the first study to show that SOCS1 and SOCS3 are differentially expressed in recipients following allogeneic HSCT, suggesting a prognostic correlation between SOCS genes and the development of GVHD. This result provides a new platform to study GVHD immunobiology and potential diagnostic and therapeutic targets for GVHD.

Expression of SOCS1 and SOCS3 genes in human graft-versus-host disease after allogeneic hematopoietic stem cell transplantation

BACKGROUND: Suppressor of cytokine signaling genes (SOCS) are regarded as pivotal negative feedback regulators of cytokine signals, including the interferon-gamma (IFN-gamma), granulocyte-colony stimulating factor, and interleukin families, released by T cells. A detailed understanding of the involvement of SOCS genes in graft-versus-host disease (GVHD) is critical to effectively manage GVHD, yet their expression patterns among recipients remain largely unexplored. METHODS: Expression levels of SOCS1 and SOCS3 were determined by real-time quantitative reverse transcription PCR (qRT-PCR) in patients with acute GVHD (aGVHD) and chronic GVHD (cGVHD), in a severity-dependent manner, after allogeneic hematopoietic stem cell transplantation (HSCT). A total of 71 recipients with AML (N=40), ALL (N=12), myelodysplastic syndromes (MDS; N=10), chronic myelogenous leukemia (CML; N=2), severe aplastic anemia (SAA; N=5), or others (N=2), who received allogeneic HSCT from human leukocyte antigen-identical siblings or unrelated donors between 2009 and 2011, were included in the present study. RESULTS: Overall, the expression levels of SOCS1 decreased in recipients with grade II to IV aGVHD and cGVHD when compared to normal donors and non-GVHD recipients. Interestingly, the expressions of SOCS1 decreased significantly more in cGVHD than in aGVHD recipients (P=0.0091). In contrast, SOCS3 expressions were similarly reduced in all the recipients. CONCLUSION: This is the first study to show that SOCS1 and SOCS3 are differentially expressed in recipients following allogeneic HSCT, suggesting a prognostic correlation between SOCS genes and the development of GVHD. This result provides a new platform to study GVHD immunobiology and potential diagnostic and therapeutic targets for GVHD.

Optimized Internal Control and Gene Expression Analysis in Epstein-Barr Virus-Transformed Lymphoblastoid Cell Lines

The Epstein-Barr virus-transformed lymphoblastoid cell line (LCL) is one of the major genomic resources for human genetics and immunological studies. Use of LCLs is currently extended to pharmacogenetic studies to investigate variations in human gene expression as well as drug responses between individuals. We evaluated four common internal controls for gene expression analysis of selected hematopoietic transcriptional regulatory genes between B cells and LCLs. In this study, the expression pattern analyses showed that TBP (TATA box-binding protein) is a suitable internal control for normalization, whereas GAPDH (glyceraldehyde-3-phosphate dehydrogenase) is not a good internal control for gene expression analyses of hematopoiesis-related genes between B cells and LCLs at different subculture passages. Using the TBP normalizer, we found significant gene expression changes in selected hematopoietic transcriptional regulatory genes (downregulation of RUNX1 , RUNX3 , CBFB , TLE1, and NOTCH2; upregulation of MSC and PLAGL2) between B cells and LCLs at different passage numbers. These results suggest that these hematopoietic transcriptional regulatory genes are potential cellular targets of EBV infection, contributing to EBV-mediated B-cell transformation and LCL immortalization.