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SWI/SNF chromatin remodeling complex BAF subunit (SMARCB1)-deficient lung carcinoma is rare and may be suitable for immunotherapy. This article presents the case of an elderly male who presented with "a persistent dry cough for 3 months without obvious inducement", and was initially diagnosed with non-small cell lung carcinoma following bronchoscopic biopsy. Immunohistochemical staining for SMARCB1 (-), SMARCA4 (+), PD-L1(22C3) (tumor proportion score is 60%), Ki-67 (+) with a positive index of 60%. The final diagnosis is SMARCB1-deficient lung carcinoma. He was subsequently treated once with a combination of chemotherapy and immunotherapy, and has been followed up for 34 months. Currently, the patient continues to survive with tumor.

ATP-dependent chromatin remodeling complexes play crucial roles in various biological processes including enhancing local DNA accessibility, regulating gene transcription, and facilitating DNA replication and repair. Based on their functional structural domains, these complexes may be categorized into four families, including SWI/SNF, ISWI, CHD and INO80. Such families are vital factors for regulating gene expression and play pivotal roles in developmental processes. Variants of genes encoding the components of such complexes have been closely associated with human developmental disorders and neurodevelopmental genetic syndromes. This review has summarized the classification, fundamental functions and underlying mechanism of the ATP-dependent chromatin remodeling complexes, as well as common neurological genetic disorders due to variants of genes encoding the subunits of such complexes.

Structural maintenance of chromosomes (SMC), including cohesin, condensin and the SMC5/6 complex, are protein complexes which maintain the higher structure and dynamic stability of chromatin. Such circular complexes, with similar structures, play pivotal roles in chromatid cohesion, chromosomal condensation, DNA replication and repair, as well as gene transcription. Despite extensive research on the functions of the SMCs, our understanding of the SMC5/6 complex has remained limited compared with the other two complexes. This article has reviewed the architecture and crucial physiological roles of the SMCs, and explored the associated phenotypes resulting from mutations of the SMC components such as Cornelia de Lange syndrome (CdLS) and microcephaly, with an aim to provide insights into their functions in eukaryotic cells and implications for human diseases.

Twelve DEK-NUP214 fusion gene-positive patients with acute myeloid leukemia and on allo-HSCT treatment at the Hematology Hospital of the Chinese Academy of Medical Sciences from November 2016 to August 2022 were included in the study, and their clinical data were retrospectively analyzed. The patients comprised five men and seven women with a median age of 34 (16-52) years. At the time of diagnosis, all the patients were positive for the DEK-NUP214 fusion gene. Chromosome karyotyping analysis showed t (6;9) (p23;q34) translocation in 10 patients (two patients did not undergo chromosome karyotyping analysis), FLT3-ITD mutation was detected in 11 patients, and high expression of WT1 was observed in 11 patients. Nine patients had their primary disease in the first complete remission state before transplantation, one patient had no disease remission, and two patients were in a recurrent state. All patients received myeloablative pretreatment, five patients received sibling allogeneic hematopoietic stem cell transplantation, and seven patients received haploid hematopoietic stem cell transplantation. The median number of mononuclear cells in the transplant was 10.87 (7.09-17.89) ×10(8)/kg, and the number of CD34(+) cells was 3.29 (2.53-6.10) ×10(6)/kg. All patients achieved blood reconstruction, with a median time of 14 (10-20) days for neutrophil implantation and 15 (9-27) days for platelet implantation. The 1 year transplant-related mortality rate after transplantation was 21.2%. The cumulative recurrence rates 1 and 3 years after transplantation were 25.0% and 50.0%, respectively. The leukemia free survival rates were (65.6±14.0) % and (65.6±14.0) %, respectively. The overall survival rates were (72.2±13.8) % and (72.2±13.8) %, respectively.

Objective: To investigate the clinicopathological, immunohistochemical and molecular genetic characteristics of gastric carcinoma with NTRK-rearrangement/amplification. Methods: The clinicopathological data of gastric carcinoma cases with NTRK-rearrangement/amplification diagnosed from January 2011 to September 2020 at the Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School, China, were collected. The clinicopathological, immunophenotypic and molecular pathological features were analyzed. The relevant literature was reviewed. Results: There were 4 cases of gastric carcinoma with NTRK-rearrangement/amplification. All 4 patients were male, aged 57-67 years (average, 63 years). Tumor sizes ranged from 3.5 to 5.2 cm (average, 4.8 cm). All tumors were in the antrum. All 4 patients underwent radical gastrectomy and were followed up after the surgery. Morphologically, all tumors showed histological features with enteroblastic-differentiated gastric carcinoma. Tumor cells showed predominantly tubular/papillary architecture, with conspicuous vesicular nuclei and pale staining or transparent cytoplasm. Immunohistochemistry showed pan-TRK expression in all cases, with various degrees of positivity in the cytoplasm. All cases were subject to NTRK1/2/3 detection using fluorescence in situ hybridization. There were NTRK translocations in 2 cases and NTRK amplifications in 2 cases. These cases were further verified by RNAseq next generation sequencing which confirmed that NTRK1 gene translocation (TPM3-NTRK1) and NTRK2 gene translocation (NTRK2-SMCHD1) occurred in two cases, respectively. Conclusions: NTRK mutation occurs less frequently in gastric cancer. In this study, the cases mainly occur in the antrum. The morphology has the characteristics of enteroblastic differentiation. The tumors have unique histological, immunophenotypic and molecular characteristics, which require much attention from pathologists to effectively guide clinicians to choose the best treatment.

Objective: To investigate the clinical features, treatment regime, and outcome of pediatric acute myeloid leukemia (AML) with DEK-NUP214 fusion gene. Methods: The clinical data, genetic and molecular results, treatment process and survival status of 7 cases of DEK-NUP214 fusion gene positive AML children admitted to the Pediatric Blood Diseases Center of Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences from May 2015 to February 2022 were analyzed retrospectively. Results: DEK-NUP214 fusion gene positive AML accounted for 1.02% (7/683) of pediatric AML diagnosed in the same period, with 4 males and 3 females. The age of disease onset was 8.2 (7.5, 9.5) years. The blast percentage in bone marrow was 0.275 (0.225, 0.480), and 6 cases were M5 by FAB classification. Pathological hematopoiesis was observed in all cases except for one whose bone marrow morphology was unknown. Three cases carried FLT3-ITD mutations, 4 cases carried NRAS mutations, and 2 cases carried KRAS mutations. After diagnosis, 4 cases received IAE induction regimen (idarubicin, cytarabine and etoposide), 1 case received MAE induction regimen (mitoxantrone, cytarabine and etoposide), 1 case received DAH induction regimen (daunorubicin, cytarabine and homoharringtonine) and 1 case received DAE induction regimen (daunorubicin, cytarabine and etoposide). Complete remission was achieved in 3 cases after one course of induction. Four cases who did not achieved complete remission received CAG (aclarubicin, cytarabine and granulocyte colony-stimulating factor), IAH (idarubicin, cytarabine and homoharringtonine), CAG combined with cladribine, and HAG (homoharringtonine, cytarabine and granulocyte colony-stimulating factor) combined with cladribine reinduction therapy, respectively, all 4 cases reached complete remission. Six patients received hematopoietic stem cell transplantation (HSCT) after 1-2 sessions of intensive consolidation treatment, except that one case was lost to follow-up after complete remission. The time from diagnosis to HSCT was 143 (121, 174) days. Before HSCT, one case was positive for flow cytometry minimal residual disease and 3 cases were positive for DEK-NUP214 fusion gene. Three cases accepted haploid donors, 2 cases accepted unrelated cord blood donors, and 1 case accepted matched sibling donor. The follow-up time was 20.4 (12.9, 53.1) months, the overall survival and event free survival rates were all 100%. Conclusions: Pediatric AML with DEK-NUP214 fusion gene is a unique and rare subtype, often diagnosed in relatively older children. The disease is characterized with a low blast percentage in bone marrow, significant pathological hematopoiesis and a high mutation rate in FLT3-ITD and RAS genes. Low remission rate by chemotherapy only and very high recurrence rate indicate its high malignancy and poor prognosis. Early HSCT after the first complete remission can improve its prognosis.

Objective: To explore the clinical and genetic features of a child with autosomal dominant mental retardation type 40 (MRD40) due to variant of the CHAMP1 gene., Methods: Clinical characteristics of the child were analyzed. Genetic testing was carried out by low-depth high-throughput and whole genome copy number variant sequencing (CNV-seq) and whole exome sequencing (WES). A literature review was also carried out for the clinical phenotype and genetic characteristics of patients with MRD40 due to CHAMP1 gene variants., Results: The child, a 11-month-old girl, has presented with intellectual and motor developmental delay. CNV-seq revealed no definite pathogenic variants. WES has detected the presence of a heterozygous c.1908C>G (p.Y636*) variant in the CHAMP1 gene, which was carried by neither parent and predicted to be pathogenic. Literature review has identified 33 additional children from 12 previous reports. All children had presented with developmental delay and mental retardation, and most had dystonia (94.1%), delayed speech and/or walking (85.2%, 82.4%) and ocular abnormalities (79.4%). In total 26 variants of the CHAMP1 gene were detected, with all nonsense variants being of loss-of-function type, located in exon 3, and de novo in origin., Conclusion: The heterozygous c.1908C>G (p.Y636*) variant of the CHAMP1 gene probably underlay the WRD40 in this child. Genetic testing should be considered for children featuring global developmental delay, mental retardation, hypertonia and facial dysmorphism.

Objective: To investigate the dynamic change and clinical impact of DEK-NUP214 fusion gene in patients with acute myeloid leukemia (AML) receiving allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods: Real-time quantitative polymerase chain reaction (RQ-PCR) and multicolor flow cytometry (FCM) were used to detect DEK-NUP214 gene expression and leukemia-associated immunophenotype (LAIP) in 15 newly diagnosed patients with positive DEK-NUP214 and receiving allo-HSCT from September 2012 to September 2017 at Peking University People's Hospital. The clinical outcome was analyzed using Kaplan-Meier survival curves. The impact of DEK-NUP214 expression was analyzed by log-rank test. Results: The subjects were followed-up with a median period of 657 (62-2 212) days. The median DEK-NUP214 expression level at diagnosis was 488% (274%-1 692%). Thirteen patients achieved complete remission before allo-HSCT. Thirteen patients had a residual DEK-NUP214 expression of 0.38% (0.029%-738.9%) before allo-HSCT. After allo-HSCT, DEK-NUP214 expression in 9/13 patients remained positive, which dropped by around 500 folds (5.7-5 663.0 folds) within a month post-transplant. Five patients died and 2 patients relapsed. The 3-year cumulative incidence of relapse in patients with positive DEK-NUP214 before transplant was 17.5%±11.3% and the 3-year overall survival was 60.5%±13.8%. After allo-HSCT, DEK-NUP214-negative patients had a better outcome. Conclusion: Quantitative monitor of DEK-NUP214 fusion gene could be a sensitive indicator of MRD status after allo-HSCT.

Objective: To construct RNA interference (RNAi) lentiviral expression vector of DEK gene, and to explore its effect on the biological behavior of liver cancer cells. Methods: Double-stranded oligo DNAs were annealed and synthesized according to the interference sequence of DEK gene by RNAi technology. Small interfering RNA expression vector pLKO.1 was cloned after enzymatic digestion. The recombinant lentiviral pLKO.1-sh hDEK was constructed, and then the virus supernatant was collected, packed and infected by 293T cells. Real-time reverse transcription PCR (RT-PCR) and Western blot were used to detect DEK expression in human liver cancer cells Bel-7402, Hu-7, SmMC-7721 and HepG2, and DEK knockdown efficiency in each group of lentivirus-infected cells. Cell proliferation ability, cloning ability, apoptosis and migration ability were detected by cell counting kit-8 (CCK8), flow cytometry and scratch test, respectively. The t-test was used to compare the mean between the two groups, and one-way ANOVA was used to compare the multiple groups. Results: Enzymatic digestion and DNA sequencing results confirmed that the recombinant lentiviral vectors pLKO.1-sh hDEK1 and pLKO.1-sh hDEK3 were successfully constructed. RT-PCR and Western blot results showed that the expression of DEK in human liver cancer cells BEL-7402 and Huh7 cells was higher, and pLKO.1-sh hDEK3 was more effective in inhibiting the DEK gene expression ( P < 0.05). Therefore, pLKO.1-sh hDEK3 was selected to infect BEL-7402 and Huh7 cells for subsequent functional experiments. CCK8 cell proliferation test result showed that the cell proliferation ability of BEL-7402 and Huh7 cells infected with recombinant lentivirus was weakened when compared with blank control and negative control group ( P < 0.05). Apoptosis results showed that the apoptosis rate of knockdown group was higher than that of blank and the negative control group ( P < 0.05). Cell scratch test result showed that the wound healing rate of knockdown group was lower than that of blank control and negative control group ( P < 0.05), and the difference was statistically significant; however, there was no statistically significant difference between blank control and negative control group. Conclusion: Targeting DEK expression in silent liver cancer cells can inhibit the cell proliferation, migration ability, and induce apoptosis, which lays the foundation for further study of the role of DEK gene in liver cancer.

A boy, aged 6 months, had the manifestations of intellectual and motor developmental delay, head instability, general weakness, unawareness of grasping objects by hands, and unusual facies (slightly wide eye distance, epicanthus, esotropia, mouth-opening appearance, short philtrum, and low-set ears). Gene detection results showed a de novo heterozygous frameshift mutation of the CHAMP1 gene at the chromosomal location of chr13:115089847, and nuclear acid was changed to c.530delCinsTTT, resulting in a change in amino acid to p.S177Ffs*2. Therefore, the boy was diagnosed with autosomal dominant intellectual disability-40 caused by the mutation in the CHAMP1 gene. This case report suggests that for children with unexplained intellectual disability, especially those with generalized hypotonia and severe language disorder, the possibility of CHAMP1 gene mutation should be considered, and genetic testing should be performed as early as possible.

Objective: To construct a eukaryotic expression vector of bromodomain-containing protein 7 (BRD7) with deletion of bromodomain (BRD7△brd) using the homologous recombination and reverse PCR amplification techniques., Methods: The linear DNA fragments of bromodomain-deleted mutation of BRD7 (pIRES2-EGFP- 3Flag/BRD7△brd) were amplified by one pair of reverse PCR primers using high-fidelity enzyme, and then these fragments were transformed into E.coli to obtain the eukaryotic expression vector expressing BRD7△brd protein based on homologous recombination and plasmid cyclization., Results: Bromodomain-deleted clones were identified by digestion with restrictive enzymes, and then the sequence and protein expression were further confirmed by sequencing and Western blot assays. The results suggest that pIRES2-EGFP-3Flag/BRD7△brd was successfully constructed., Conclusion: We establish a simple and quick method to construct plasmids with pIRES2-EGFP- 3Flag/BRD7△brd using reverse PCR amplification and homologous recombination techniques. We also found that the concentration of template in PCR reaction system is one of the critical factors that affect the rate of homologous recombination. Of all, this improved technique could be widely used in the construction of gene mutations.

To explore the association between single nucleotide polymorphisms (SNPs) in the promoter region of the inner centromere protein (INCENP) gene and bovine semen quality, the haplotypes in 250 Chinese Holstein bulls were detected using PCR-RFLP method in this study. Two SNPs (g.-556 G>T, rs 136823901 and g.-692 C>T, rs 211010999) and three haplotypes (CG, TT, TG) were identified in the promoter region of INCENP. The genotype frequency and allele frequency of these two SNPs as well as the correlation between different SNP haplotype combinations and bovine semen quality were then analyzed. Our results showed that fresh sperm motility of the GT genotype was significantly higher than that of the GG genotype (P<0.05) at the SNP site g.-556 G>T, while fresh and frozen-thawed sperm motilities of the haplotype combinations H1H1(CCGG), H1H3(CTGT), H2H3(TTGT) and H3H3(TTTT) were significantly higher than that of H1H2 (P<0.05). To further study the possible mechanisms by which g.-556 G>T and g.-692 C>T affect semen quality, three haplotype plasmids were respectively transfected into MLTC-1 cells. The TG haplotype demonstrated the highest luciferase activity, suggesting that g.-556 G>T and g.-692 C>T are functional mutations which could regulate INCENP gene expression by affecting promoter activity and thus affect semen quality.

Objective: To study the immunophenotype and molecular genetics of epithelioid sarcoma (ES), INI1 expression and its role in differential diagnosis., Methods: Twenty cases of ES were retrieved from the archival files and selected for immunohistochemical study, DNA sequencing and fluorescence in-situ hybridization. The clinical and pathologic features were also reviewed., Results: The age of patients ranged from 16 to 75 years (mean = 40.2 years). The median age of patients in classic ES and proximal-type was 37.9 years and 42.0 years, respectively. The male-to-female ratio was 1.2: 1.0. Classic ES mostly occurred in the extremities while proximal-type ES often affected the perineum and external genitalia and trunk. Histologically, granuloma-like structures, consisting of aggregates of epithelioid and spindly tumor cells with central necrosis, were observed in classic ES. The epithelioid tumor cells contained abundant eosinophilic cytoplasm, merged with spindly cells at the periphery and admixed with collagen fibers. In proximal-type ES, the tumor cells showed prominent epithelioid and/or rhabdoid features, had marked cytologic atypia and grew in multinodular or diffuse patterns. In 2 cases of proximal-type ES studied, the "rhabdoid" tumor cells demonstrated a diffuse sheet-like growth pattern, mimicking malignant rhabdoid tumor. Immunohistochemical study showed that vimentin was positive in all cases. Pan-cytokeratin, CK8, CK7, epithelial membrane antigen and CD34 were expressed in 16, 15, 1, 18 and 13 cases, respectively. The staining for S-100 protein was focal and weak in 5 cases. None of the cases studied expressed CD31 and HMB45. Loss of INI1 was demonstrated in 10 of the 13 classic ES cases and 5 of the 7 proximal-type ES cases. The mutation of INI1 gene was detected in 1 of the 6 cases. Deletion of INI1 gene including heterozygous deletion, homozygous deletion and haploid was observed in 8 of the 11 cases., Conclusions: Owing to the histologic heterogeneity, pitfalls in diagnosis of ES sometimes are encountered. INI1 is lost in most cases of ES. Immunohistochemical study, including staining for INI1, provides useful clues in pathologic diagnosis. Instead of INI1 mutation, inactivation of INI1 gene related deletion is not uncommon.

Objective: To investigate the changes in the gene expression profiles in a multiple myeloma cell line after knocking-down the expression of SNF5, a core component of the SWI/SNF chromatin remodeling complex., Methods: The total RNA were extracted from tetracycline-inducible SNF5 knockdown (KD) cell line derived from KM3 cells for gene expression profiling by affymetrix microarray and bioinformatic analysis., Results: Knockdown of SNF5 inhibited KM3 cell proliferation. A total of 545 genes were found to be differentially expressed in the cells with SNF5 knockdown, among which 214 were up-regulated and 331 were down-regulated., Conclusions: SNF5 is essential for the growth of multiple myeloma cells and can regulate the expression of the genes associated with cell growth and apoptosis.

BEX2 (Brain expressed X-linked protein 2), a 13 kDa protein, is highly expressed in brain and testis. It is reported that the protein expression of BEX2 dramatically alters during the embryo development, but little is known about its function. By means of yeast two-hybrid screening, we isolated that INI1/hSNF5 was a binding partner for BEX2, a key component of SWI/SNF chromosome remolding complex. GST Pull-down experiment interaction is physical and specific. Further analysis using truncated mutations demonstrated that the two partner for BEX2, a key component of SWI/SNF chromosome remolding complex. GST Pull-down experiment confirmed that BEX2 can interact with INI1/hSNF5 directly and specifically. Truncated mutations analysis further demonstrated that the two conserved reverse repeats sequences within INI1/hSNF5 were necessary for the interaction. Sub-cellular localization showed that both BEX2 and INI1/hSNF5 mainly localized in cell nucleus, which indicated that the interaction may be involved in the regulation of gene expression. Our experiments also showed that co-overexpressing of the two proteins affected cell cycle by increasing the cells in S phase, indicating that BEX2 could regulate cell cycle by interacting with INI1/hSNF5.

Objective: To explore the role of centromere protein H (CENP-H) in the proliferation of human gastric cancer cells., Methods: RT-PCR and Western blot analysis were employed to examine the mRNA and protein expressions of CENP-H in 7 human gastric cancer cell lines and immortalized human gastric epithelial cells (GES-1). The cells were infected with the retrovirus vectors pMSCV-CENP-H or CENP-H-RNAi to establish stable cell lines with high CENP-H expression or CENP-H expression interference. MTT assay and colony formation assay were used to examine the changes in the cell proliferation after the infection., Results: CENP-H was over-expressed in gastric cancer cell lines AGS, BGC823, SGC-7901, MKN45, HGC27, MGC-803 and MKN28 at both mRNA and protein levels. The established AGS/CENP-H cell line with increased CENP-H expression showed enhanced proliferative activity, while the cell line MGC-803/CENP-H-RNAi with CENP-H expression interference showed an obviously lowered proliferation ability., Conclusion: CENP-H promotes the proliferation of human gastric cancer cells, suggesting its important role in the occurrence and development of gastric cancer.

Objective: To localize and define the region of nucleus export signal (NES) on BRD7, and determine the role of this region in nucleus export of the external protein., Methods: Based on an in vitro expressed model of green fluorescence protein (GFP), we performed DNA walking analysis to set BRD7 into several sections according to the structural characteristics of BRD7, investigated the effect of different sections of BRD7 on nucleus export of GFP, defined the region of nucleus export signal sequence of BRD7, and further ascertained the content of amino acids in BRD7 and potential localization of BRD7 NES by bioinformatics., Results: B7C1 fragments ranged from aa219 to aa450 in BRD7 were found to target the external protein GFP into the cytoplasm detected by GFP direct fluorescence, which could be inhibited by NES inhibitor Leptomycin B (LMB). This region was rich in hydrophobic amino acid residues but no typical NES with characteristics of leucine-rich sequence by bioinformatics., Conclusion: The region from aa219 to aa450 is primarily defined as an atypical NES in BRD7.

Objective: To determine the correlation between the expression of CARMA1 mRNA and MUM1 protein, as well as its effects on clinicopathological features and prognosis of diffuse large B cell lymphoma (DLBCL)., Methods: The immunophenotype (CD20, CD79a, CD10, MUM1, Bcl6) and proliferation index of DLBCL cells were examined by immunohistochemistry (IHC). CARMA1 mRNA was detected by RT-PCR., Results: CARMA1 mRNA was detected in 76 of 89 (85.40%) cases with DLBCL. The level of CARMA1 mRNA was higher in MUM1-postive group than in MUM1-negative group. No correlation was found in the expression intensity between the two molecules (P = 0.084). Ki67 positive rate was higher in MUM1(+) cases than in MUM1(-) ones (P = 0.030). There was no difference between MUM1(+) and MUM1(-) cases in sex, median age, staging, primary site and other clinicopathological features. In 58 CARMA1 mRNA positive cases, low expression cases showed more in earlier stage and more males. No difference in survival status was identified between cases with and without MUM1 expression, over- and low-expression of CARMA1 mRNA, as well as over- and low-expression of CARMA1 mRNA among 58 cases with MUM1 expression., Conclusion: The expression of CARMA1 mRNA is likely associated with the expression of MUM1 and shows male predominance in DLBCL. The expression of CARMA1 may be involved with pathogenesis and progression of ABC-like DLBCL. The two molecules correlated somewhat with some clinicopathological features, but not with survival of DLBCL.

This study was purposed to investigate the relationship between brd7 gene and differentiation of leukemia cells and the role of brd7 gene in differentiation of leukemia cells. The HL-60 and K562 cell lines were induced by all-trans retinoic acid (ATRA) for 7 days, then the cell morphologic change was observed under inverted microscope with Wright-Giema staining, the expression level of CD11b was detected by flow cytometry for evaluating cell differentiation level, the expression changes of BRD7 protein before inducing differentiation and in process of cell differentiation were determined by Western blot. The results showed that ATRA could inhibit the proliferation and induce differentiation of HL-60 cells, but no differentiation in K562 cells was induced by ATRA. The level of CD11b expression in HL-60 cells was up-regulated gradually during ATRA-induced cell differentiation. The expression of BRD7 protein increased markedly along with differentiation of HL-60 cells towards granulocytes. However, BRD7 protein did not significantly alter in K562 cells in which inducing differentiation was not found. It is concluded that brd7 gene expression enhances as the HL-60 cells differentiate, underlying which the mechanism remains to clarify.

In Drosophila melanogasler,the wings apart-like (wapl) gene encodes a protein that regulates heterochromatin structure. Human wapl gene is human homolognue of wapl,they have same function. HWAPL is a cohesion-binding protein that affect chromosomal segregation. It has a character of oncoproteins and linked to cervical carcinogenesis and tumor progression.

Objective: To evaluate the expression level of BRD7 gene in bone marrow mononuclear cells (BMNCs) in patients with acute leukemia (AL) and to analyze BRD7 single nucleotide polymorphism(SNP)., Methods: RT-PCR was used to detect BRD7 expression in patients with AL and normal bone marrow subjects. Single-strand conformation polymorphism and DNA sequence analysis were also used to identify BRD7 mutation or SNP to investigate the relation between BRD7 and AL., Results: BRD7 mRNA in BMNCs from 52 patients with AL and 30 control subjects was expressed. The mRNA relative expression of BRD7 in patients with AL was higher than that of the control group (P=0.001). Three SNPs (C657A,C495T and A737G) in BRD7 gene coding region (447 approximately 844 bp) were found, and A737G was coupled with C495T . The allele frequencies of SNP C657A were not significantly different between AL and the control group. The genotype and the allele frequencies of the 2 coupled SNPs were significantly different (P<0.01). But there was no significant discrepancy among the mRNA expression levels of AA, AG, and GG genotypes in the leukemia group (P>0.05)., Conclusion: Expression of BRD7 gene is up-regulated in AL cells. The 2 coupled SNPs (C495T and A737G ) in BRD7 gene coding region (447 approximately 844 bp) are correlated with AL, indicating that SNPs may be one of the genetic susceptibility factors of AL.

There is obvious allele disequilibrium in nasopharyngeal carcinoma at chromosome 3p, 9p, 6q, 11q, 13q and 14q. Nasopharyngeal carcinoma (NPC) susceptibility/suppressor gene candidates were obtained by molecular biology methods,such as cDNA representational difference ana-lysis. The functional research of NPC susceptibility/ suppressor gene candidates indicated: (1) The increased expression of Cx contributed to obstacles of gap junctional intercellular communication (GJIC), and resulted an aberration of GJIC; (2) BRD7, a transcript factor, was associated with cell cycle regulation; (3) NAG7,an estrogen receptor repressor, inhibited the invasive potential of human NPC cells by regulating ERalpha expression and the H-ras/p-c-Raf and JNK/AP-1/MMP1 signaling pathways; (4) NGX6, a metastasis-associated protein, can negative-regulate EGF/Ras/MAPK signaling transduction pathway, and interact with ezrin protein to inhibit invasion and metastasis of NPC cells; (5) SPLUNC1, a secreted protein, can inhibit the bacterium clone formation, and is an innate immune molecule. These data will lay an important foundation for the NPC mechanism.

Objective: To study the influence of siRNA inhibition of CENP-A expression on the biological behavior of HepG2 cells., Methods: Three pairs of 21 bp reverse repeated motifs of CENP-A target sequence with 9 spacer were synthesized and inserted into vector pSilencer 2.1-U6 neo to generate siRNA eukaryotic expression plasmids. After stable transfection into HepG2 cells, cell growth, apoptosis, cell cycles and plate clone forming efficiency were investigated. Expressions of CENP-A mRNA was monitored by the reverse transcriptase polymerase chain reaction (RT-PCR). The protein expression of CENP-A, bcl-2, Bax, p53, p21waf1 and mdm2 were detected by Western-blotting., Results: Two eukaryotic expression plasmids with significant siRNA specific inhibition to the CENP-A gene were created. Compared with control cells, HepG2 cells transfected with the constructs showed G1 phase delay (P < 0.01) and cell number decrease in the S phase (P < 0.001), along with an increased apoptotic rate (P = 0.003), significant increase of Bax expression and decreased bcl-2 expression (P< or =0.001). The protein expressions of p21waf1 was higher and mdm2 was lower than those of the control groups. However, the wild type p53 protein expression was not effected by CENP-A siRNA., Conclusions: An altered expression of CENP-A may be related to the proliferation of hepatocellular carcinoma through cell cycle regulation involving an altered bcl-2/Bax expression, that may be p53 independent.

Objective: To study the expression of centromere protein A (CENP-A) and its significance in hepatocellular carcinoma (HCC) and adjacent non-neoplastic liver tissue., Methods: The expression levels of CENP-A mRNA in 20 samples of HCC and adjacent non-neoplastic liver tissue were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and real-time quantitative polymerase chain reaction (qRT-PCR). Immunohistochemical study for CENP-A and p53 proteins was also performed on tissue microarrays containing 80 samples of HCC and adjacent liver tissue., Results: The expression level of CENP-A mRNA in HCC (0.64 +/- 0.18) was higher than that in adjacent non-neoplastic liver tissue (0.09 +/- 0.09) (t = 12.78, P < 0.01). Of the 80 samples of HCC, 57 cases (71.25%) and 60 cases (75%) expressed CENP-A and p53 proteins respectively. The positivity rates of CENP-A and p53 proteins in non-neoplastic liver tissue were 43.75% (35/80) and 16.25% (13/80) respectively. There was a statistically significant difference in CENP-A and p53 protein expression between HCC and non-neoplastic liver tissue (P < 0.01). The coincident rate between CENP-A and p53 expression was 88.75% (71/80). Expression of CENP-A protein showed a positive correlation with that of p53 protein (r = 0.57, P < 0.01)., Conclusion: The over-expression of CENP-A occurs at transcriptional level and may be related to malignant proliferation of HCC via possible interaction with p53 gene.

Objective: Rett syndrome (RTT) is a neurodevelopmental disorder which causes severe mental retardation. This study aimed at elucidating clinical features of 66 Chinese RTT cases diagnosed by The Department of Pediatric Neurology, Peking University First Hospital since 1987, and at analysis of the MeCP2 genotype / phenotype correlation., Methods: Sixty-six RTT cases were followed up every one to two years to get the information of their clinical manifestations and the response to the L-carnitine treatment which was administered to the patients at a dose of 80-100 mg/(kg d). MeCP2 mutation analysis by PCR and sequencing were performed on 39 cases., Results: In this cohort of cases, the onset of the disease occurred between 3 and 38 months of age, 89% of the cases lost their purposeful hand use at 7 months to six years of age, all the cases had stereotype hand movement which presented at 1 to 5 years of age, 85% of the cases lost language ability at 11 months to eight years of age, 21% of the cases lost the ability of walking at ages of 2 years and 9 months to 15 years. The symptoms/signs such as small head circumference, seizures, breathing irregularities, teeth grinding, scoliosis/ kyphosis were presented in many of the cases. The clinical manifestations were improved in 6 cases after L-carnitine treatment. MeCP2 gene mutation was found in 64% of the cases. Two cases with non-sense mutation C502t (amino acid change R168X) died, two cases with missense mutation C397T (amino acid change R133C) and one case with missense mutation A398T (amino acid change R133H) preserved several words., Conclusion: Deceleration of the head growth, loss of acquired purposeful hand use, stereotype hand movement and language deterioration were the main characteristics of RTT. L-carnitine could improve the clinical manifestation of some cases. There are some correlations between MeCP2 genotype and phenotype.

At present, a point that cell biologists and medicine scientists focus their close attention on is the mechanisms of cell proliferation and carceration. Breast cancer, one of the frequently occurring cancers, is often been studied intensively. Centromere constitutive molecules, related to various regulatory factors, play an important role in cell proliferation check point regulation. Cell cycle engine molecules, oncogenes, anti-oncogenes and other molecules conform a cell proliferation network. The basic courses of all tumors are associated to this network. However, there are still many problems to be resolved in the analyses of cancer related genes which cause tumors and tumor gene markers. In the current study, using Northern blot, 31 samples of breast cancer tissues and their normal (not cancerous) tissues a little far away from them in the same individuals showed that, in the majority of the tests (87.1%), the mRNA of centromere protein CenpB over expressed in breast cancer tissues, and moreover, tissue in situ hybridization also revealed that all of the CenpB-over-expressed cancer tissues, having identified with Northern blot, over expressed CenpB mRNA. Analyzing the same samples by means of Western blot, the result was highly consistent to the studies in the RNA level. A conclusion was drawn that the over expression of CenpB gene probably relates to malignant cell proliferation in breast gland. It has been testified by researchers that a few of CenpB homogenous proteins are co-operative, the loss of their genes resulting in chromosomes' separating abnormally and cell growth's slowing down. Having transfected HeLa (Tet-Off) cells with anti-sense Cenp in a previous experiment, we ever got a result that cellular duplicating time was prolonged for another 32.8 h, and together with the inhibition of centromere assembly, the mitotic index dropped sharply. In another research, we drew a conclusion that CenpG may be related to cancer, and its differential expressing probably relates to malignant cell proliferation. Combined with these researches, the results obtained from the current study are beneficial to further recognition of the mechanism of cancer.

BRD7 was isolated through cDNA representational difference analysis (RDA) (GenBank accession No. AF152604). Previous studies showed that BRD7 gene was down-regulated in nasopharyngeal carcinoma (NPC) cells and tissues, and three cSNPs (coding-region single nucleotide polymorphisms) were found on BRD7. In addition, six BRD7-interacting proteins were identified by yeast two-hybrid system. To study the function of this gene on carcinogenesis in NPC, BRD7 gene was transfected into HNE1 cells low-expressed BRD7 by using liposomes and a stable cell line over-expressing BRD7 was established. After two-dimensional gel electrophoresis(2-DE), twenty differentially expressed proteins were identified by MALDI-TOF-MS including argininosuccinate lyase, metalloproteinase inhibitor-2 precursor, proteaseome activator28 beta subunit, thyroid transcriptional factor-1, cyclinH (MO15-associated protein), and so on. These differentially expressed proteins are related to cell cycling, transcription regulation, signaling pathway etc. Therefore, BRD7 may exert its functions by mediating differential expression of these proteins.

Objective: To explore the correlation between expression of surviving gene in acute leukemic cells and its clinical effects., Methods: By using semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) technique, surviving gene expression in 50 previously untreated acute leukemia (AL) patients was analysed. The apoptosis of primary leukemia cells cultured in vitro was assayed with terminal deoxyribonucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL)., Results: Surviving gene expression levels in cells of AL patients at diagnosis were significantly higher than that in normal bone marrow mononuclear cells (MNCs) (82.0% vs 33.3%, P < 0.05). The expression level was higher in ALL cells than in ANLL cells (89.5% vs 75.0%). Among 22 cases of ANLL, bone marrow remission (BMR) rate was higher in surviving gene negative expression cells from patients accepted a course of chemotherapy than in positive expression cells (83.3% vs 25.0%, P = 0.023). Among 13 ANLL patients received a course of HA regimen chemotherapy, the BMR was higher in patients surviving mRNA negative expression cells than in positive cells (100.0% vs 27.3%). Patients with surviving/beta-actin ratio>0.6 attained lower BMR., Conclusion: Higher expression level of surviving mRNA in AL cells may be one of the reasons that leukemic cells are insensitive to chemotherapy.

Objective: To investigate the role and molecular mechanism of surviving, an anti-apoptosis gene, in cell growth and transformation., Methods: Coding sequence of surviving was amplified from Daudi cell mRNA by RT-PCR and then cloned into prokaryotic and eukaryotic vectors. The vectors with surviving were transfected into BL21 cells of Escherichia coli and human embryonic kidney 293 cells. The cells were cultured. Protein was extracted from the cells and examined by gel electrophoresis. Suspension of 293 cells was cultured and the number of cells was counted every other day, thus a growth curve was drawn. Another suspension of 293 cells was cultured in soft agar to observe the number of colonies. Cells transfected with plasmids void of surviving were used as controls., Results: The anti-apoptosis gene surviving was well expressed in BL21 cells and 293 cells. The growth curve showed that the proliferation rate of 293 cells was slightly faster than that of control cells, however, without significant difference. Soft agar assay showed that the colonies formed by surviving-transfected 293 cells were of greater size and with greater number. Western blotting showed overexpression of cyclin D1 and c-myc, two important cancer proteins, in cells transfected with surviving., Conclusion: The anti-apoptosis gene surviving promotes cell transformation. These effects may depend on the functions of cyclin D1 and c-myc.

Objective: To study the expression of a novel inhibitor gene of apoptosis, survivin, in cervical carcinoma and its relationship with the expression of bcl-2, p53 genes., Methods: Using streptavidin-biotin peroxidase (SP) method, we examined the expression of survivin, bcl-2 and p53 genes in 10 normal cervical tissues and 59 cervical carcinoma tissues., Results: Survivin was expressed in 41 of 59 (69.5%) cases of cervical carcinomas. In contrast, normal cervical epithelium did not express survivin. Overexpression of survivin was related to the tumor grade and clinical stage. Survivin positive cases were strongly associated with bcl-2 expression (36/45, 80.0% versus 5 of 14 cases, P < 0.005) and p53 expression (33/43, 76.7% versus 8 of 16 cases, P < 0.05)., Conclusions: Apoptosis inhibition by survivin, alone or in cooperation with bcl-2 and p53 may participate in the onset and progression of cervical carcinoma. Survivin could be a new diagnostic/therapeutic target in cervical cancer.

Objective: To study the expression of surviving and its relationship with expression of P53, c-myc, k-ras in non-small-cell lung cancer (NSCLC)., Methods: Expression of the surviving mRNA was evaluated by reverse transcriptase polymerase chain reaction (RT-PCR) in 76 NSCLC tumor samples, 20 benign phymatoid lesion and 21 adjacent normal lung tissue samples. Immunohistochemical assay was to detect the expression of P53, c-myc, k-ras proteins., Results: Expression of surviving gene was detected in a significantly greater proportion of NSCLC (61%) than phymatoid lesion (30%) and adjacent normal lung tissue (19%) (P < 0.001). There was no relationship between surviving gene expression and histologic subtype, differentiation, TNM stages, or lymph node metastases. The expression of surviving gene correlated with P53 or c-myc expression, but not k-ras expression., Conclusion: (1) The up-regulation expression of surviving gene in NSCLC suggested that surviving may play a role in the pathway of carcinogenesis and surviving may be identified as a potential therapeutic target in NSCLC. (2) surviving, de-activation of antioncogene P53 and up-regulation of oncogene c-myc might play synergetic roles in the process of carcinogenesis of NSCLC. But surviving and k-ras may be independently involved in the pathogenesis of lung cancer.

The centromere/kinetochore is a specialized structure at the primary constriction of mammalian chromosomes. It participates in and is necessary for the mitotic chromosome movement. Centromere Protein B(CENP-B) is a highly conserved protein located at the centromere/kinetochore region. In this article, we explore the relationship between CENP-B expression and cell proliferation. HeLa cells were synchronized at different phases of the cell cycle and the synchronized cells were examined by flow cytometry and 3H-TdR labelling. ACA immunostaining showed the discrete single spots in nuclei of the cells at G1 and S phase, and spots in pairs mostly in those at G2 phase. Dot blot and Northern blot indicated that CENP-B gene was expressed at all phases of the cell cycle, but the expression level very much different with the highest at G2 phase and the lowest at S phase. Interestingly relatively high expression of CENP-B gene was also found in M phase, showing the continuity of the CENP-B gene expression during the cell cycle. This continuity implies a possibility that the assembly of the new centromere/kinetochore can not occur until centromere proteins reach a critical concentration when cells enter S phase and G 2 phase. Also, the continuous expression of centromere proteins may be necessary for the centromere/kinetochore function. In addition, the relationship between CENP-B gene expression and the nuclear skeleton was investigated. The nuclear skeleton-associated DNA extracted after Bam HI digestion were hybridized with 32P-labelled cDNA of CENP-B gene by means of Southern blot technique. The results that there stronger positive hybridization reaction in G 2 phase than in S phase indicated that CENP-B gene was more closely associated with the nuclear skeleton in G 2 phase cells than in S phase cells, which was in consistence with the level of the CENP-B gene expression at G 2 and S phase cells.