Deficiency of IRG1/ itaconate aggravates endotoxemia-induced acute lung injury by inhibiting autophagy in mice.

Itaconate, produced by aconitate decarboxylase 1 (ACOD1), which is encoded by immune-responsive gene 1 (Irg1), is one of the metabolites derived from the tricarboxylic acid cycle. It has been reported that exogenous itaconate plays an anti-inflammatory role in the progression of multiple diseases and pathological processes, including activated macrophage, ischemia-reperfusion injury, and acute lung injury. However, the role and specific mechanism of endogenous itaconate in endotoxemia-induced acute lung injury (ALI) remain unclear. The animal model of ALI in wild-type and Irg1-/- mice was constructed by LPS intraperitoneal injection. Ultrahigh-performance liquid chromatography-tandem mass spectroscopy (UPLC-MS/MS) analysis was performed to measure the quantity of endogenous itaconate. The protective effect of itaconate was investigated by the behavioral assessment and the levels of inflammatory cytokines. Acute lung injury was assessed by hematoxylin and eosin staining, total protein in BALF, and Evans blue leakage. Western blotting was used to detect the IRG1 expression and autophagic protein in the lung. We demonstrated that IRG1 was highly expressed in ALI and that endogenous itaconate was produced simultaneously and was 100 times higher. Using Irg1-/- mice, we found that endogenous itaconate was likely to exert an anti-inflammatory effect by activating NRF2 and promoting autophagy. Furthermore, autophagy was restrained by LPS but enhanced by 4-octyl itaconate (4-OI) pretreatment. Our study illustrated that a deficiency of IRG1/Itaconate aggravates ALI and that the IRG1/itaconate pathway protects against ALI. The protective mechanisms could be related to the facilitation of autophagy. Such findings may provide a theoretical foundation for the treatment of endotoxemia-induced ALI.

[Inhibitory effect of polyphyllin â on the proliferation of prostate cancer PC3 cells via ERK1/2/P65/DNMT1 and its molecular mechanism].

OBJECTIVE: To explore the inhibitory effect of polyphyllin Ⅰ (PPⅠ) on the proliferation of castration-resistant prostate cancer PC3 cells and its molecular mechanism. METHODS: We cultured human prostate cancer PC3 cells in vitro and treated them with PPⅠ at the concentrations of 0 (blank group), 0.4, 0.8, 1.2, 1.6, 2.0, and 2.4 µmol/L for 24, 48, and 72 hours, respectively. Then we detected the proliferation of the cells by MTT assay, measured their apoptosis by flow cytometry, and determined the expressions of p-ERK1/2, ERK1/2, NF-κB/p65 and DNMT1 proteins as well as the level of NF-κB/p65 in the cells additionally treated with the ERK1/2 inhibitor SP600125 by Western blot. RESULTS: Compared with the blank control group, the PPⅠ-treated PC3 cells showed a concentration- and time-dependent reduction of the survival rate (1.00 ± 0.00 vs 0.85 ± 0.05, P < 0.01) at 0.4 µmol/L after 48 hours of intervention, concentration-dependent early apoptosis at 0.8 µmol/L (4.83 ± 0.95 vs 13.83 ± 2.97, P < 0.01), time-dependent increase of the expressions of p-ERK1/2 (1.00 ± 0.00 vs 1.73 ± 0.17, P < 0.01) and ERK1/2 (1.00 ± 0.00 vs 1.36 ± 0.12, P < 0.01) at 2 hours, and concentration-dependent decrease of the expressions of NF-κB/p65 and DNMT1 at 1.2 µmol/L (1.00 ± 0.00 vs 0.78 ± 0.10 and 0.63 ± 0.06, P < 0.01) and 1.6 µmol/L (1.00 ± 0.00 vs 0.67 ± 0.11 and 0.52 ± 0.09, P<0.01). Inhibition of ERK1/2 phosphorylation with PD98059 markedly reversed PPⅠ-induced decrease of the NF-κB/p65 expression as compared with that in the PPⅠ group (0.86 ± 0.18 vs 0.43 ± 0.09, P < 0.05). CONCLUSIONS: PPⅠ induces the early apoptosis and suppresses the proliferation of PC3 cells, probably by activating the ERK1/2 pathway and inhibiting the expressions of the NF-κB/p65 and DNMT1 proteins.

Curcumin reduces lung inflammation via Wnt/ß-catenin signaling in mouse model of asthma.

OBJECTIVES: Asthma is a chronic inflammatory, heterogeneous airway disease affecting millions of people around the world. Curcumin has been found to have anti-inflammatory and antifibrosis effects. Researchers reported that curcumin regulated Wnt/ß-catenin signaling in lots of cells. However, whether curcumin regulates the levels of Wnt/ß-Catenin signaling in lung tissues and DCs (dendritic cells) remains unclear. In this study, we assessed the effects of curcumin on DCs and asthma. METHODS: C57BL/6 mice immunized with OVA (ovalbumin) were challenged thrice with an aerosol of OVA every second day for 8 days. Dexamethasone or curcumin was administered intraperitoneally to OVA-immunized C57BL/6 mice on day 24 once a day for 9 days. Mice were analyzed for effects of curcumin on asthma, inflammatory cell infiltration and cytokine levels in lung tissue. DCs were isolated from mouse bone morrow. The surface markers CD40, CD86 and CD11c of DCs was detected by FACS (fluorescence activated cell sorting) and the function of DCs was detected by mixed lymphocyte reaction. The expression of GSK-3ß and ß-catenin was detected by Western Blot. RESULTS: Results showed that OVA increased the number of inflammatory factors in BALF (bronchoalveolar lavage fluid), elevated lung inflammation scores in mice. Curcumin dose-dependently reversed the alterations induced by OVA in the asthmatic mice. Curcumin activated Wnt/ß-catenin signaling pathway in DCs and asthmatic mouse lungs. CONCLUSIONS: Curcumin could influence the morphology and function of DCs, ease asthma symptom and inflammatory reaction through the activation of Wnt/ß-catenin signaling. These results provide new evidence new evidence for application of curcumin on asthma.

[Clinical significance of serum carbohydrate antigen 125 in acute exacerbation of chronic obstructive pulmonary disease].

OBJECTIVE: To study the serum level of carbohydrate antigen 125 (CA125) in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and its relation with pulmonary hypertension. METHODS: Forty-six patients with AECOPD complicated by pulmonary hypertension, 46 with AECOPD and 38 healthy control subjects were examined for their clinical data, pulmonary function, echocardiographic findings, and serum levels of lung tumor markers and brain natriuretic peptide (BNP). RESULTS: Compared with the healthy control group, COPD patients with or without pulmonary hypertension showed significantly decreased pulmonary function (P<0.05), especially in those with AECOPD and concurrent pulmonary hypertension (P<0.05). Serum CA125 level was obviously higher in AECOPD group than in the healthy control group, and further increased in AECOPD patients with pulmonary hypertension (P<0.05). The levels of lung tumor markers (CEA, NSE, CYFRA and PROGRP) were similar among the 3 groups (P>0.05). The serum level of BNP in patients with AECOPD and concurrent pulmonary hypertension was significantly higher than that in patients with AECOPD (P<0.05). Pearson linear correlation analysis showed that serum CA125 was positively correlated with pulmonary artery systolic pressure and BNP in AECOPD patients with pulmonary hypertension (P<0.01). CONCLUSION: Serum CA125 may serve as a serological index to identify AECOPD patients with pulmonary hypertension.

[Kidney-tonifying Chinese medicine for male infertility: A systematic review of randomized controlled trials].

OBJECTIVE: To systematically evaluate the efficacy and safety of kidney-tonifying traditional Chinese medicine in the treatment of male infertility. METHODS: Based on the principles and methods of Cochrane systematic reviews, we searched CNKI, VIP, and Wanfang databases from inception to December 2012 for randomized controlled clinical trials addressing the treatment of male infertility with kidney-tonifying traditional Chinese medicine. According to the inclusion and exclusion criteria and retrieval strategies, we extracted the data, evaluated the quality of the included literature, and conducted meta-analysis using the RevMan 5. 2 software. RESULTS: Twenty trials involving 2,272 patients were included, and the sample size of each study was from 60 to 270 cases. All the studies were graded as of poor quality, with Jadad scores of no more than 3 points. The results of meta-analysis showed that the total effectiveness rate of traditional Chinese medicine versus Western medicine on male infertility was RR = 1.71, 95% CI 1.19-2.47, and that of Chinese-Western combined therapy versus Western medicine was RR = 1.15, 95% CI 1.01-1.30. Both traditional Chinese medicine and Chinese-Western combined therapy showed a significantly better total effectiveness than Western medicine alone in improving the pregnancy rate without serious adverse reactions. CONCLUSION: Due to the poor methodological quality and high heterogeneity of the included studies, the evidence for the efficacy and safety of kidney-tonifying traditional Chinese drugs in the treatment of male infertility is of but limited value, and further validation is needed by more high-quality studies.

An Improved LC-MS/MS Method for Simultaneous Determination of the Eleven Bioactive Constituents for Quality Control of Radix Angelicae Pubescentis and Its Related Preparations.

An improved LC-MS/MS method was developed for simultaneous determination of eleven bioactive constituents of Radix Angelicae Pubescentis and its related preparations. It was the first report on the quantification of bioactive constituents in different preparations of Radix Angelicae Pubescentis by LC-MS/MS analytical method. These samples were separated with an Agilent Zorbax Extend reversed-phase C18 column (1.8 µm, 4.6 × 100 mm) by linear gradient elution using aqueous ammonium acetate and acetonitrile as mobile phase. The flow rate was 0.3 mL min(-1). The eleven bioactive constituents showed good regression (R > 0.990) within test ranges and the recoveries were in the range of 87.1-110%. The limit of detections and quantifications for most of the major constituents were less than 0.5 and 1.0 ng mL(-1), respectively. All results indicated that the developed method could be readily utilized as a suitable quality control method for Radix Angelicae Pubescentis and related preparations.

Simultaneous determination of scopoletin, psoralen, bergapten, xanthotoxin, columbianetin acetate, imperatorin, osthole and isoimperatorin in rat plasma by LC-MS/MS for pharmacokinetic studies following oral administration of Radix Angelicae Pubescentis extract.

A rapid and sensitive bioassay based on liquid chromatography tandem mass spectrometry (LC-MS/MS) has been developed and validated for the simultaneous determination of eight coumarins in rat plasma. The liquid-liquid extraction method with ethyl acetate was used to prepare the plasma samples after addition of warfarin as an internal standard (IS). Chromatographic separation was performed on an Eclipse plus C18 column (100mm×4.6mm, 1.8µm) using gradient elution when 1mM ammonium acetate aqueous solution - acetonitrile was used as the mobile phase. The lower limit of quantitation (LLOQ) of each coumarin was lower than 2.16ngmL(-1). Intra-day and inter-day precisions were less than 15%. The accuracies were in the range of 88.9-117%. The mean recoveries of coumarins and IS were higher than 84%. The method was successfully applied to a pharmacokinetic study of eight coumarins in rats after oral administration of radix angelicae pubescentis.

The experimental study of Astragalus membranaceus on meridian tropsim: the distribution study of astragaloside IV in rat tissues.

According to Traditional Chinese Medicine (TCM) theories, TCM with different meridian tropism have different therapeutic effects. In view of the meridian tropism of Astragalus membranaceus (Huangqi), astragaloside IV, one of the effective phytochemicals of Huangqi, was appointed and observed its distribution in rat tissues following a single intravenous (i.v.) dose. A simple and accurate LC-ESI-MS/MS method was developed and validated for astragaloside IV quantification in heart, liver, spleen, lung and kidney using warfarin as an internal standard (IS). Chromatographic separation was performed on a Eclipse plus C18 (4.6mm×100mm, 1.8µm) when the flow rate was set at 0.300mLmin(-1) and ammonium acetate aqueous solution - acetonitrile was used as mobile phase. The intra- and inter-day precisions of the quality control samples were within 15% and accuracies were within 90.0-110%. The recoveries were more than 90.0% at high, medium and low concentrations, respectively. This method was successfully applied for distribution of astragaloside IV after intravenous (i.v.) dose of 4mgkg(-1) astragaloside IV in rats. Astragaloside IV concentration was highest in liver and kidney and remained much higher than that in other tissues over the experiment course. Lung, heart and spleen were also detected to contain astragaloside IV. The results clearly demonstrated that astragaloside IV was one of the material bases of the meridian tropism of Huangqi.

Simultaneous determination of catechin, epicatechin and epicatechin gallate in rat plasma by LC-ESI-MS/MS for pharmacokinetic studies after oral administration of Cynomorium songaricum extract.

A rapid and valid method was developed for simultaneous determination catechin, epicatechin and epicatechin gallate in rat plasmas using scopoletin (103 ng mL(-1)) as an internal standard (IS). The separation was performed on Eclipse plus C18 column (100 mm × 4.6 mm, 1.8 µm) at a flow rate of 0.3 mL min(-1), and acetonitrile-0.1% formic acid was used as mobile phase. The recoveries of three analytes and IS were more than 78.9%. The lower limits of quantitation (LLOQ) in rat plasma were 2.14, 2.38 and 2.08 ng mL(-1) respectively for catechin, epicatechin and epicatechin gallate. Intra-day and inter-day precisions were within 12%. The accuracies were more than 85%. After single oral administration of 15.25 g kg(-1) Cynomorium songaricum extract, C(max) of catechin, epicatechin and epicatechin gallate in rat plasma were respectively 86.69±38.65, 32.57±15.00 and 36.93±12.62 ng mL(-1) while T(max) values were respectively 0.15±0.09, 0.20±0.10 and 0.20±0.13 h. The results demonstrated that the present LC-MS/MS method was sensitive enough for pharmacokinetic study of catichins following oral administration of C. songaricum extract.

Differential expression of Epstein-Barr virus-encoded RNA and several tumor-related genes in various types of nasopharyngeal epithelial lesions and nasopharyngeal carcinoma using tissue microarray analysis.

Studies have revealed that Epstein-Barr virus (EBV) infection, genetic aberration, and environmental factors are of importance in the development of nasopharyngeal carcinoma (NPC), although the definite mechanism remains to be fully elucidated. The aim of our study is to investigate using tissue microarray analysis whether differential expression of EBV-encoded small RNA-1 (EBER-1) and several tumor-related genes were associated with NPC carcinogenesis. Immunohistochemistry and in situ hybridization were performed on tissue microarrays containing 148 NPCs and 164 noncancerous nasopharyngeal epithelia (NPE) with different morphologic features. We found that overexpressions of EBER-1 hybridization signals, p53, p21ras, and bcl-2 proteins and loss expressions of p16 and p27 proteins were significantly increased in NPC tissues compared with normal NPE and hyperplastic NPE (P

The transcriptional regulation role of BRD7 by binding to acetylated histone through bromodomain.

Studies showed that the bromodomain binds to acetyl-lysines on histone tails, which is involved in deciphering the histone codes. BRD7, a novel bromodomain gene, is the first described bromodomain gene involved in nasopharyngeal carcinoma (NPC). Previous studies showed that ectopic expression of BRD7 inhibited cell growth and cell cycle progression from G1 to S phase in HNE1 cells (a NPC cell line) by transcriptionally regulating some cell cycle related genes including E2F3 gene. In the present study, we revealed the co-localization between acetylated H3 and BRD7 and found that the bromodomain of BRD7 is required for this co-localization. More importantly, wild-type BRD7 interacted with H3 peptide acetylated at Lys14, while the bromodomain deleted mutant lost this ability. We also found that the mutant BRD7 failed to regulate E2F3 promoter activity and inhibit cell cycle progression. These results indicated that the transcriptional regulation role of BRD7 was achieved by binding to acetylated histone H3 and that the bromodomain was essential for this role. In addition, no obvious changes were observed in the acetylated level of histone H3 after transfection with BRD7, indicating that chromatin remodeling, not chromatin modification, is the major mechanism of BRD7 mediated gene transcription. Taken together, the present work shed light on the fact that a novel bromodomain gene, BRD7, is of importance in transcriptional regulation and cellular events including cell cycle.

Profiling of differentially expressed genes in LRRC4 overexpressed glioblastoma cells by cDNA array.

Our previous study has shown that LRRC4 is a novel member of the leucine-rich repeat (LRR) superfamily and has the potential to suppress brain tumor growth. In order to further analyze the functions of LRRC4 on the maintenance of normal function and suppression of tumorigenesis in the central nervous system, we investigated alterations in gene expression related to neurobiology by the Atlas array in two inducible dual-stable LRRC4-overexpressing cell lines. Seventeen of 588 genes spotted on the Atlas membrane showed altered expression levels in LRRC4 transfected U251MG Tet-on cells, which are involved in cell proliferation and cell cycle progression, tumor invasion and metastasis, and neurotransmitter synthesis and release. In addition, cell invasion assay results showed that LRRC4 can inhibit the U251MG cell migration. These studies represent the first cDNA array analysis of the effects of LRRC4 on the involvement of different neurobiological genes in U251MG glioblastoma cells and provide new insights into the function of LRRC4 in glioma.

Study of a novel brain relatively specific gene LRRC4 involved in glioma tumorigenesis suppression using the Tet-on system.

LRRC4 is a novel relatively specific gene, which displays significant down-regulation in primary brain tumor biopsies and has the potential to suppress brain tumor growth. In this study, we investigated the growth inhibitory effect of LRRC4 on tumorigencity in vivo and on cell proliferation in vitro by a tetracycline-inducible expression system. Results showed that LRRC4 significantly reduced the growth and malignant grade of xenografts arising from glioblastoma U251MG cells. Cell proliferation was markedly inhibited after U251MG Tet-on-LRRC4 cell induction with doxycycline. Flow cytometry and Western blot analysis demonstrated that LRRC4 mediated a delay of the cell cycle in late G1, possibly through up-regulating the expressions of p21Waf1/cip1 and p27Kip1 and down-regulating the expressions of cyclin-dependent kinase 2, retinoblastoma protein and epidermal growth factor receptors. Together, these findings provide clues to the function of LRRC4 as a negative regulator of cell growth and underscore a link between the above-mentioned cyclins, cyclin-associated molecules and tumorigenicity.

[Preliminary biomarker related to nasopharyngeal carcinoma filtered from the whole genome expression profiling involved in microdissection nasopharyngeal tissues].

OBJECTIVE: To filter biomarkers of nasopharyngeal carcinoma (NPC) by constructing the homogenesis tissue gene expression profiling with the whole human genome GeneChip. METHODS: The epithelium cells of the homogenesis NPC and the pure nasopharyngeal normal tissues microdissected from nasopharyngeal biopsy which was preserved in the RNAlater were used to isolate RNA and then to harvest the aRNA through in vitro transcription, and aRNA prober was labled to hybridize to HG-U133. plus 2.0, so the expression profiling of each homogenesis tissue could be constructed. RESULTS: Some candidate biomarker genes related to the tumorigenesis of NPC had been filtered by comparing the expression profiling of NPC samples with the expression profiling of normal nasopharyngeal epithelia samples. Any genes regarding the metastasis of NPC might have been selected by comparing the expression profiling of no-metastasis samples with those of the metastasis samples. CONCLUSION: Using the whole genome GeneChip to construct the expression profiling for the microdissected homogenesis tissue is effective to filter the candidate biomarker genes.

A susceptibility locus at chromosome 3p21 linked to familial nasopharyngeal carcinoma.

Nasopharyngeal carcinoma (NPC) poses one of the serious health problems in southern Chinese, with an incidence rate ranging from 15 to 50/100,000. Chromosome translocation t(1;3) and frequent loss of heterogeneity on short arms of chromosome 3 and 9 have been reported to be associated with NPC, and a genome-wide scan identified an NPC susceptibility locus on chromosome 4p15.1-q12 recently. In our study, we collected samples from 18 families at high risk of NPC from the Hunan province in southern China, genotyped with a panel of polymorphic markers on short arms of chromosomes 3, 9, and 4p15.1-q12. A locus on 3p21 was identified to link to NPC with a maximum logarithm of odds for linkage score of 4.18. Fine mapping located the locus to a 13.6-cM region on 3p21.31-21.2, where a tumor suppressor gene cluster resided. Our findings identified a novel locus for NPC and provided a map location for susceptibility genes candidates. In contrast to a recent study, no significant evidence for NPC linkage to chromosomes 4 and 9 was observed.

[Studies on the relationship between D6S1581, a high frequency allele imbalance locus, and genetic susceptibility to nasopharyngeal carcinoma].

OBJECTIVE: To investigate the relationship of nasopharyngeal carcinoma (NPC) with the high frequency allele imbalance locus D6S1581, and the NPC associated gene FBXO30 which is located near D6S1581. METHODS: Genescan was used to genotype D6S1581 of 12 NPC pedigrees, 85 sporadic NPC patients and 181 normal volunteers. Then parametric/nonparametric linkage analysis and association analysis were performed. RESULTS: D6S1581 was linked with NPC, a Lod score of 2.611436 (P=0.00245) was obtained, and a significant difference in allele frequency was observed between familial NPC and control (P<0.005). CONCLUSION: These results suggest that D6S1581 is highly associated with NPC, and there may be one or more NPC associated genes near D6S1581, including FBXO30.

Effects of NPC-associated gene NAG7 on cell cycle and apoptosis in nasopharyngeal carcinoma cells.

BACKGROUND & OBJECTIVE: NPC-associated gene NAG7 was a novel candidate tumor suppressor gene associated with nasopharyngeal carcinoma cloned in our laboratory. This study was designed to investigate the potential effect of NAG7 on the cell cycle and apoptosis of nasopharyngeal carcinoma cell line HNE1 and its molecular mechanism. METHODS: NAG7 gene was introduced into HNE1 cells using lipofectin transfection technique. The expression level of NAG7 gene was analyzed by Northern blot. Cell cycle, cyclins, and cell apoptosis were detected by flow cytometry, and the expressions of cyclin D1 and cyclin E were detected by Western blot. RESULTS: NAG7 gene was re-expressed in NAG7 transfected HNE1 cells. Compared with HNE1 cells and vector transfected HNE1 cells, NAG7 transfected HNE1 cells arrested in G0/G1 phase increased (P < 0.05) and cells in S phase decreased (P < 0.05), the apoptosis cells increased (P < 0.05), and the levels of cyclins of A, B1, D1, and E decreased. Furthermore, the expression of cyclin D1 and E decreased in NAG7 transfected HNE1 cells. CONCLUSION: NAG7 gene re-expression could inhibit overproliferation of NPC cell by delaying the progression of G1 into S in cell cycle and inducing cell apoptosis.

[Expression of connexin43 and connexin45 in nasopharyngeal carcinoma].

BACKGROUND AND OBJECTIVE: The gap junction plays an important role in the exchange of nutrients, ions, and regulatory molecules between cells. It will result in an uncontrolled cell proliferation if it is abnormal. It was reported that some cancer tissues and cancer cells had abnormal gap junction and restoration of the gap junction function could revert the cancer cells to a normal phenotype. Therefore, to investigate the expression of connexin(cx) in human nasopharyngeal carcinoma, may provide a new way for clinical diagnosis and the pathogenesis of nasopharyngeal carcinoma. METHOD: The expressions of Cx43 and Cx45 in the biopsies of nasopharyngeal carcinoma(NPC) and chronic nasopharyngitis tissues were determined by using the immunohistochemical staining. RESULTS: 1. Cx43 and Cx45 expressed differentially in NPC and chronic nasopharyngitis tissues(P < 0.01). In NPC, expression of Cx43 and Cx45 were 44.8% and 46.6%, respectively, while in columnar cells of chronic nasopharyngitis tissues, their expressions were 86.5% and 100% respectively, which were higher than those in NPC, 2. The percentage in squamous epithelial cells of chronic nasopharyngitis tissues was 29.7%, which was lower than that in NPC, (56.9%). 3. The expressions of Cx43 and Cx45 in paracancer columnar epithelial cells were lower than that in paracancer squamous epithelial cells (P < 0.001), but higher than that in NPC cells (P < 0.01). CONCLUSIONS: The abnormal expression of Cx43 and Cx45 in nasopharynx tissues may be associated with cancerization and squamatization of human nasopharynx tissue.

Analysis of bromodomain of BRD-7 gene and its prokaryotic expression.

BACKGROUND & OBJECTIVE: BRD-7 is a novel gene (AF: 152604), containing a bromodomain, was cloned in our lab. Previous studies showed that BRD-7 plays an obviously suppressive role on NPC cell growth. In order to clarify the function mechanism of this gene, we investigate an important motif of BRD-7, the bromodomain. METHODS: The bromodomain of BRD-7 was analyzed by homology-based amino sequence and secondary structure analysis. In addition, we constructed a prokaryotic expression vector of bromodomain. Western blot analysis was used to confirm the expression of the bromodomain protein in Escherichia coli. RESULTS: Homology-based sequence analysis revealed that the bromodomain of BRD-7 possibly contains four alpha helices (Z, A, B and C), and a hydrophobic pocket which is an important structure to recognize acetylated histone peptide. This bromodomain encoding a 12.8 kD protein, was introduced into Escherichia coli using the pGEX-4T-2 expression vector. After isopropyl beta-D-thiogalactopyranoside(IPTG) induction, a new anticipated protein of 38.8 kD appeared on SDS-PAGE and the result was confirmed by Western blot analysis. CONCLUSION: BRD-7 of bromodomain protein is similar to three proteins containing known structural bromodomain motif by bio-informatics analysis, suggesting the bromodomain of BRD-7 should belong to co-activator subgroup and may have similar function that can selectively interact with acetylated histone peptide.