Ming Guan

Publications of Ming Guan

• Correlation of DLC1 gene methylation with oncogenic PIK3CA mutations in extramammary Paget's disease.

  Authors: Zhihua Kang, Feng Xu, Qiao-An Zhang, Jinran Lin, Zhiyuan Wu, Xinju Zhang, Yan Luo, Jinhua Xu, Ming Guan

  Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc. 04/2012;

  Extramammary Paget's disease is a rare cutaneous malignant neoplasm. The genetic and epigenetic mechanisms underlying its pathology remain unknown. In this study, we investigated the expressionExtramammary Paget's disease is a rare cutaneous malignant neoplasm. The genetic and epigenetic mechanisms underlying its pathology remain unknown. In this study, we investigated the expression levels, and mutation and methylation status of a common tumor suppressor gene, deleted in liver cancer 1 (DLC1), and an oncogene, PIK3CA, in tumor (n=132) and normal tissues (n=20) from unrelated patients. The presence of epigenetic and genetic lesions was then correlated to the patient pathology data to determine the potential role of these genes in extramammary Paget's disease etiology and progression. The DLC1 gene was found to be downregulated in 43 (33%) tumors, as compared with immunohistochemistry results from normal tissues. Methylation-sensitive, high-resolution melting analysis indicated that the DLC1 promoter was hypermethylated in 51 (39%) extramammary Paget's disease tumors. This hypermethylation was associated with significantly decreased DLC1 levels (P=0.011), and had a strong positive correlation with advanced age (P=0.002). PIK3CA mutations were detected by direct sequencing in 32 (24%) tumors, the majority of which were invasive. Furthermore, PIK3CA mutations significantly correlated with DLC1 hypermethylation. Thus, aberrant DLC1 methylation and PIK3CA mutations may have important roles in extramammary Paget's disease pathogenesis, and may represent potential molecular targets for therapy.Modern Pathology advance online publication, 20 April 2012; doi:10.1038/modpathol.2012.65.

• Coexistence of hepatitis B surface antigen and anti-HBs in Chinese chronic hepatitis B virus patients relating to genotype C and mutations in the S and P gene reverse transcriptase region.

  Authors: Weiwei Liu, Tingting Hu, Xinyu Wang, Yuming Chen, Minying Huang, Chao Yuan, Ming Guan

  Archives of virology. 01/2012;

  We aimed to determine the prevalence of the coexistence of HBsAg and anti-HBs and to analyze the clinical and virological features of infection, including amino acid (aa) patterns of the S gene andWe aimed to determine the prevalence of the coexistence of HBsAg and anti-HBs and to analyze the clinical and virological features of infection, including amino acid (aa) patterns of the S gene and reverse transcriptase (RT) region in Chinese chronic hepatitis B (CHB) patients. Fifty-four (2.90%) CHB patients who were positive for both HBsAg and anti-HBs were tested, and sequences were obtained from 52 of them as well as 48 patients from a control group. S gene and RT region sequences were amplified and sequenced using in-house protocols. There was no significant difference between patients with and without anti-HBs with regard to age, gender, alanine aminotransferase level, and the proportion positive for HBeAg and HBcAb. The occurrence of genotype C (P = 0.001) and anti-HBeAb positivity (P = 0.027) was significantly higher in HBsAg+/anti-HBs+ individuals. In the S gene, the number of mutated residues in the HBsAg+/anti-HBs+ group was markedly higher than in control patients (1.88 versus 1.02 substitutions per 100 amino acids, P = 0.022). The amino acid exchange occurred mostly within the N-terminal region (2.15 versus 0.87 substitutions per 100 amino acids, P = 0.023) and the "a" determinant (3.61 versus 1.56 substitutions per 100 amino acids, P = 0.049) in the two groups. In the RT region, the mean number of substitution per 100 aa showed a tendency to be significantly higher in HBsAg+/anti-HBs+ patients than in controls (2.34 versus 1.46, P = 0.040). This study showed a prevalence of coexistence of anti-HBs in HBsAg-positive patients and an increased frequency of genotype C and aa variability within both HBsAg and RT involving functionally important regions of those proteins.

• Copy number variations of interleukin-17F, interleukin-21, and interleukin-22 are associated with systemic lupus erythematosus.

  Authors: Bo Yu, Ming Guan, Yinghui Peng, Yong Shao, Chao Zhang, Xupeng Yue, Jie Zhang, Hong Yang, Hejian Zou, Wencai Ye, Jun Wan, Wei Zhang

  Arthritis and rheumatism. 11/2011; 63(11):3487-92.

  Systemic lupus erythematosus (SLE) represents the classic prototype of systemic autoimmune disease. The identification of the Th17 cell subset has provided new understanding regarding the underlyingSystemic lupus erythematosus (SLE) represents the classic prototype of systemic autoimmune disease. The identification of the Th17 cell subset has provided new understanding regarding the underlying mechanisms of autoimmunity. Copy number variations (CNVS) have been discovered to have phenotypic consequences and are associated with various types of diseases. We undertook this study to explore a possible association between CNVS of Th17 cell-related genes and the risk of SLE. We extracted genomic DNA and RNA from 938 SLE patients and 1,017 healthy controls. We examined CNVS of Th17 cell-related genes, including retinoic acid receptor-related orphan nuclear receptor γt, STAT-3, interleukin-6 (IL-6), transforming growth factor β, tumor necrosis factor α, IL-17A, IL-17F, IL-21, IL-22, IL-23A, CCL20, and CCR6, and levels of messenger RNA (mRNA) for IL-17F, IL-21, and IL-22. Genotype and allele frequencies for copy number amplifications of IL-17F, IL-21, and IL-22 were found to be significantly higher in SLE patients than in healthy controls. CNVS of IL-17F, IL-21, and IL-22 had no synergistic contribution to SLE. The mRNA expression of IL-17F, IL-21, and IL-22 in the samples with >2 copies of DNA was significantly higher than that in those with 2 copies of DNA. Our findings indicate that CNVS of IL-17F, IL-21, and IL-22 are associated with the risk of SLE.

• Single-nucleotide polymorphisms of MAMDC1 are associated with rash and photosensitivity, but not disease risk, of systemic lupus erythematosus in Chinese mainland population.

  Authors: Qi Wu, Bo Yu, Yuewen Chen, Yong Shao, Jie Zhang, Qili Zhong, Xi Peng, Hong Yang, Xiaoping Hu, Bancheng Chen, Ming Guan, Jun Wan, Wei Zhang

  Clinical rheumatology. 06/2011; 30(10):1373-8.

  Systemic lupus erythematosus (SLE) is a complex immune disease affected by both genetic dispositions and environmental factors. Recently, the polymorphisms in MAMDC1 gene have been reported toSystemic lupus erythematosus (SLE) is a complex immune disease affected by both genetic dispositions and environmental factors. Recently, the polymorphisms in MAMDC1 gene have been reported to associate with disease risk of SLE in European population. However, whether this association is replicated in Chinese population is unknown yet. A total of 491 SLE patients and 533 controls were recruited. Unlabeled probe-based high-resolution melting analysis (HRMA) was used in genotyping. HRMA with unlabeled probe successfully distinguished all genotypes. SNP rs961616 was associated with rash [P = 0.015, odds ratio (OR) = 0.73, 95% confidence interval (CI) = 0.57-0.94] and photosensitivity (P = 0.001, OR = 0.63, 95%CI = 0.48-0.84), but not the disease risk (P = 0.133, OR = 0.88, 95%CI = 0.74-1.04), of SLE in Chinese population. Polymorphisms of rs961616 in MAMDC1 gene were associated with rash and photosensitivity, but not disease risk, of systemic lupus erythematosus in Chinese population.

• The association between single-nucleotide polymorphisms of NCF2 and systemic lupus erythematosus in Chinese mainland population.

  Authors: Bo Yu, Yuewen Chen, Qi Wu, Ping Li, Yong Shao, Jie Zhang, Qili Zhong, Xi Peng, Hong Yang, Xiaoping Hu, Bancheng Chen, Ming Guan, Jun Wan, Wei Zhang

  Clinical rheumatology. 04/2011; 30(4):521-7.

  Systemic lupus erythematosus (SLE) is a complex immune disease. The genetic variation in the NCF2 gene was found to associate with SLE in US and European populations. However, the association ofSystemic lupus erythematosus (SLE) is a complex immune disease. The genetic variation in the NCF2 gene was found to associate with SLE in US and European populations. However, the association of rs10911363 with SLE was not extensively studied in Chinese mainland population. A total of 488 SLE patients and 380 controls were recruited. Unlabeled probe-based high-resolution melting analysis (HRMA) was used in genotyping. HRMA with unlabeled probe successfully distinguished all genotypes. Neither genotype nor allele frequencies of single-nucleotide polymorphism (SNP) rs10911363 showed statistically significant differences between SLE patients and controls. The association of SNP rs10911363 with the diagnostic criteria of SLE was also examined. Minor allele (G) of rs10911363 was found to significantly associate with the incidence of arthritis (p = 0.024, odds ratio (OR) = 1.35, and 95% confidence interval (CI) = 1.04-1.75) and increased abnormalities of antinuclear antibody (p = 0.002, OR = 1.51, and 95%CI = 1.17-1.95) and anti-DNA (p = 0.013, OR = 1.40, and 95%CI = 1.07-1.82). Polymorphisms of rs13277113 in NCF2 gene were associated with arthritis and autoantibody production, but not disease risk, of SLE in Chinese population.

• High-resolution melting facilitates mutation screening of rpsL gene associated with streptomycin resistance in Mycobacterium tuberculosis.

  Authors: Feifei Wang, Hongbo Shen, Ming Guan, Ying Wang, Yun Feng, Xinhua Weng, Honghai Wang, Wenhong Zhang

  Microbiological research. 02/2011; 166(2):121-8.

  Drug resistance remains a serious threat to tuberculosis control worldwide. As one of the important first-line antitubercular agents, resistance to streptomycin (SM) and its derivatives has increasedDrug resistance remains a serious threat to tuberculosis control worldwide. As one of the important first-line antitubercular agents, resistance to streptomycin (SM) and its derivatives has increased in recent years and has become one of the characteristics of extensively drug-resistant tuberculosis (XDR-TB). A novel rapid assay to screen for rpsL gene mutations associated with SM resistance in Mycobacterium tuberculosis (M. tuberculosis), was developed using high-resolution melting (HRM) analysis. The HRM results of 134 SM-resistant clinical isolates and 20 SM-susceptible clinical isolates of M. tuberculosis for rpsL gene mutations were perfectly matched with those of DNA sequencing. SM resistance was highly associated with rpsL mutations in M. tuberculosis. HRM technique represented an inexpensive, highly sensitive and high-throughput method to facilitate the screening of large numbers of clinical samples for epidemiological studies of drug-resistance of M. tuberculosis, especially in developing countries.

• Loss of DMBT1 expression in human prostate cancer and its correlation with clinical progressive features.

  Authors: Junhua Du, Ming Guan, Jie Fan, Haowen Jiang

  Urology. 02/2011; 77(2):509.e9-13.

  To investigate alterations of DMBT1 in prostate cancer and determine the correlation of its alterations to the clinicopathologic features of prostate cancer. DMBT1 has been proposed as a candidateTo investigate alterations of DMBT1 in prostate cancer and determine the correlation of its alterations to the clinicopathologic features of prostate cancer. DMBT1 has been proposed as a candidate tumor suppressor gene for epithelial cancer. The alterations of DMBT1 expression after treatment with DNA methyltransferase inhibitor in 2 prostate cancer cell lines (LNCaP and PC-3) were analyzed by genome microarray and real-time polymerase chain reaction (PCR). A total of 36 prostate cancer tissues and 16 benign prostatic tissues were evaluated with reverse transcription-PCR, Western blot, and immunohistochemistry for DMBT1 expression. Treatment with 5-aza-2'-deoxycytidine reactivated expression of DMBT1 in PC3 cells, but not in LNCaP cells. Downregulation or loss of DMBT1 mRNA and protein expression was observed in prostate cancer, but not in benign tissues. Immunostaining analysis showed DMBT1 protein was absent in 14 cancer samples with Gleason score of 8-10 and weakly stained in 16 cancer samples with Gleason score of 4-7, compared with strong immunostaining in all 15 benign prostatic tissues. Loss of DMBT1 expression was correlated with local invasion (P = .048) and bone metastasis (P = .039) but was not correlated with patient age, prostate-specific antigen level, or tumor grade at diagnosis. Our study provides evidence that loss of DMBT1 expression is associated with prostate cancer, suggesting that DMBT1 may function as a tumor suppressor gene in prostate carcinogenesis.

• Double-stranded RNA induces melanocyte death via activation of Toll-like receptor 3.

  Authors: Ning Yu, Shujie Zhang, Tao Sun, Kefei Kang, Ming Guan, Leihong Xiang

  Experimental dermatology. 02/2011; 20(2):134-9.

  As cutaneous pigment-producing cells, melanocytes can become targets of primary and secondary immune response as can be seen in diseases like vitiligo and Vogt-Koyanagi-Harada (VKH) syndrome. ViralAs cutaneous pigment-producing cells, melanocytes can become targets of primary and secondary immune response as can be seen in diseases like vitiligo and Vogt-Koyanagi-Harada (VKH) syndrome. Viral infections have previously been implicated as a possible precipitating factor in the destruction of melanocytes in these disorders. During viral replication, double-stranded RNA (dsRNA) is produced as an intermediate metabolite, which induces antiviral and inflammatory responses through Toll-like receptor 3 (TLR3) in cells of innate immune system. The functional responses of melanocytes to dsRNA, however, remain unclear. Herein, we demonstrated that human melanocytes expressed TLR3 at a constitutive and inducible level. Stimulation with poly(I:C), a synthetic dsRNA analogue, triggered apoptosis of melanocytes. The apoptosis-inducing effect was shown by RNA interference to be largely dependent on TLR3, but occurred independently of NF-κB activation since treatment with specific NF-κB inhibitor Bay 11-7082 failed to prevent the process. In contrast, IFN-β neutralizing Ab blocked the apoptosis-inducing effect of dsRNA, indicating the involvement of IFN-β autocrine signalling. Furthermore, studies on the intracellular signal transduction pathways revealed that dsRNA induces the activation of p38, ERK1/2 and JNK1/2 in melanocytes. Using specific inhibitors, we demonstrated that activation of p38 and ERK1/2 controlled both IFN-β secretion and IFN-β mediated cell death. Taken together, these data suggest that viral dsRNA stimulates TLR3 in human melanocytes and triggers the cellular apoptosis through autocrine of IFN-β.

• Expression of pigment epithelium-derived factor and tumor necrosis factor-α is correlated in bladder tumor and is related to tumor angiogenesis.

  Authors: Chen-Chen Feng, Pao-Hsun Wang, Qiang Ding, Ming Guan, Yuan-Fang Zhang, Hao-Wen Jiang, Hui Wen, Zhong Wu

  Urologic oncology. 02/2011;

  OBJECTIVE: Angiogenesis is a pivotal process on which solid tumor growth is substantially dependent. Pigment epithelium-derived factor (PEDF) is the most potent natural anti-angiogenic factor, whichOBJECTIVE: Angiogenesis is a pivotal process on which solid tumor growth is substantially dependent. Pigment epithelium-derived factor (PEDF) is the most potent natural anti-angiogenic factor, which has seldom been studied in bladder tumor, and whose functioning pathway remains unclear. We have thus investigated PEDF expression in relation to tumor necrosis factor-α (TNF-α) and microvessel density (MVD) with immunohistochemistry. METHODS: Antibodies of PEDF and TNF-α were examined by Western blotting before immunohistochemistry. Sixty-four urothelial tumor sections and 23 normal controls were stained and expression of PEDF, TNF-α, and MVD were studied. RESULTS: Decreased PEDF expression and increased TNF-α expression was noticed in tumorous tissue compared with healthy urothelium. Lower PEDF expression was related to higher tumor grade but stage. Increased TNF-α expression was noticed in recurrent, larger tumors as well as in tumors with progression in grade and stage. Expression of PEDF and TNF-α was correlated in bladder tumor. PEDF or TNF-α was correlated with MVD negatively or positively, respectively, in cancerous tissue and tumorous grouping without correlation in papillary urothelial neoplasm of low malignant potential. CONCLUSION: Expressional change of PEDF and TNF-α is in relation to angiogenesis of bladder tumor, especially in bladder cancer development.

• Quantitative assessment of AKAP12 promoter methylation in human prostate cancer using methylation-sensitive high-resolution melting: correlation with Gleason score.

  Authors: Weiwei Liu, Jian Gong, Jinghui Hu, Tingting Hu, Yaofei Sun, Junhua Du, Chuanyu Sun, Ming Guan, Haowen Jiang, Yuan Lu

  Urology. 02/2011; 77(4):1006.e1-7.

  To quantitatively investigate the A kinase anchoring protein 12 (AKAP12) gene promoter methylation and its association with clinicopathologic variables in human prostate cancer (PCa). The AKAP12 geneTo quantitatively investigate the A kinase anchoring protein 12 (AKAP12) gene promoter methylation and its association with clinicopathologic variables in human prostate cancer (PCa). The AKAP12 gene has shown reduced expression and marked hypermethylation in a variety of cancers. The percentage levels of DNA methylation were measured in 78 PCa, 22 benign prostatic hyperplasia, and 22 normal adjacent tissue samples using an AKAP12 methylation-sensitive high-resolution melting assay. AKAP12 gene expression was also examined in 4 human prostate carcinoma cell lines, PC-3, DU145, LNCaP, and 22RV1, using quantitative reverse transcriptase-polymerase chain reaction and methylation-sensitive high-resolution melting analysis and after DNA methyltransferase inhibition with 5-aza-2'-deoxycytidine. Methylation (>1%) of the AKAP12 promoter region was present in 47 (60.2%) of the 78 PCa, 5 (22.7%) of the 22 benign prostatic hyperplasia, and 2 (9.1%) of the 22 adjacent normal tissue samples. AKAP12 methylation was significantly greater in the PCa than in the benign prostatic hyperplasia or adjacent tissue samples (P < .01). AKAP12 methylation was significantly greater in the PCa samples with higher Gleason scores (P = .03); however, no correlation was found with age, pT category, or serum prostate-specific antigen level. Reverse transcriptase-polymerase chain reaction demonstrated that PC-3 and DU-145 cells expressed AKAP12 RNA and LNCaP and 22RV1 did not. The AKAP12 locus was methylated in the LNCaP and 22RV1 cells. Treatment of LNCaP cells with 5-aza-2'-deoxycytidine markedly decreased the methylation levels and increased the expression of AKAP12. The results of the present study have demonstrated that AKAP12 promoter methylation is a frequent event in human PCa. AKAP12 methylation represents a potential molecular biomarker for predicting the malignancy of PCa.

• Expression of pigment epithelium-derived factor in bladder tumour is correlated with interleukin-8 yet not with interleukin-1α.

  Authors: Chenchen Feng, Ming Guan, Qiang Ding, Yuanfang Zhang, Haowen Jiang, Hui Wen, Paohsun Wang, Zhong Wu

  Journal of Huazhong University of Science and Technology. Medical sciences = Hua zhong ke ji da xue xue bao. Yi xue Ying De wen ban = Huazhong keji daxue xuebao. Yixue Yingdewen ban. 02/2011; 31(1):21-5.

  Pigment epithelium-derived factor (PEDF) is an antiangiogenic factor which is effective in tumour inhibition in a variety of tumours and has not yet been studied in bladder tumour before. In thisPigment epithelium-derived factor (PEDF) is an antiangiogenic factor which is effective in tumour inhibition in a variety of tumours and has not yet been studied in bladder tumour before. In this study the expression of PEDF, interleukin-1α (IL-1α) and -8 (IL-8) in bladder tumours was investigated. Immunohistochemistry was performed on 64 bladder tumour and 23 normal uroepithelium samples. Expression change of the factors was compared with clinicopathological parameters. Correlations between PEDF, IL-1α and IL-8 were analyzed. None of the factors was in relation to gender, tumour occurrence, and size or onset pattern. PEDF (P=0.014) and IL-1α (P=0.049) expression was down-regulated with grade progression. PEDF expression was lower in normal uroepithelium than in papillary urothelial neoplasm of low malignant potential (PUNLMP) (P=0.000) and carcinoma (P=0.009) whilst IL-1α (P=0.000 and P=0.000 respectively) and IL-8 (P=0.000 and P=0.023 respectively) expression was higher in the same grouping. PEDF expression had a negative correlation with IL-8 in PUNLMP (P=0.049, r=-0.578) as well as in tumour grouping (P=0.033, r=-0.276). Deranged expressional change of PEDF, IL-1α and IL-8 could be in relation to loss of differentiation from normal uroepithelium to papillary lesion and eventually to carcinoma.

• Copy number variations of EphA3 are associated with multiple types of hematologic malignancies.

  Authors: Ming Guan, Li Liu, Xiaosu Zhao, Qi Wu, Bo Yu, Yong Shao, Hong Yang, Xinrong Fu, Jun Wan, Wei Zhang

  Clinical lymphoma, myeloma & leukemia. 02/2011; 11(1):50-3.

  EphA3 is a component of the Eph receptor family, the largest subgroup of the receptor tyrosine kinase (RTK) family. A recent array-based study implicated the presence of copy-number variations (CNVs)EphA3 is a component of the Eph receptor family, the largest subgroup of the receptor tyrosine kinase (RTK) family. A recent array-based study implicated the presence of copy-number variations (CNVs) of EphA3 in the genomes of acute myelogenous leukemia. CNVs are present in the general population at varying degrees, and have been found to associate with various types of diseases including hematologic malignancies. However, most of the current studies focused on the genome-wide screening of CNVs, and the functional impact of such regions needs to be extensively investigated in large number of clinical samples. In our study, we collected 617 bone marrow samples from multiple types of hematologic malignancies as well as healthy controls. DNA copy numbers and mRNA levels of EphA3 in these samples were examined. We found significant association between the CNVs of EphA3 and these hematologic malignancies including acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), and myelodysplastic syndrome (MDS). We also observed a positive correlation between the relative mRNA level and gene dosage of EphA3. The CNVs of EphA3 were associated with multiple types of hematologic malignancies including ALL, AML, CLL, CML, MM, and MDS.

• Re-expression of AKAP12 inhibits progression and metastasis potential of colorectal carcinoma in vivo and in vitro.

  Authors: Weiwei Liu, Ming Guan, Tingting Hu, Xiaoye Gu, Yuan Lu

  PloS one. 01/2011; 6(8):e24015.

  AKAP12/Gravin (A kinase anchor protein 12) is one of the A-kinase scaffold proteins and a potential tumor suppressor gene in human primary cancers. Our recent study demonstrated the highly recurrentAKAP12/Gravin (A kinase anchor protein 12) is one of the A-kinase scaffold proteins and a potential tumor suppressor gene in human primary cancers. Our recent study demonstrated the highly recurrent loss of AKAP12 in colorectal cancer and AKAP12 reexpression inhibited proliferation and anchorage-independent growth in colorectal cancer cells, implicating AKAP12 in colorectal cancer pathogenesis. To evaluate the effect of this gene on the progression and metastasis of colorectal cancer, we examined the impact of overexpressing AKAP12 in the AKAP12-negative human colorectal cancer cell line LoVo, the single clone (LoVo-AKAP12) compared to mock-transfected cells (LoVo-CON). pCMV6-AKAP12-mediated AKAP12 re-expression induced apoptosis (3% to 12.7%, p<0.01), migration (89.6±7.5 cells to 31.0±4.1 cells, p<0.01) and invasion (82.7±5.2 cells to 24.7±3.3 cells, p<0.01) of LoVo cells in vitro compared to control cells. Nude mice injected with LoVo-AKAP12 cells had both significantly reduced tumor volume (p<0.01) and increased apoptosis compared to mice given AKAP12-CON. The quantitative human-specific Alu PCR analysis showed overexpression of AKAP12 suppressed the number of intravasated cells in vivo (p<0.01). These results demonstrate that AKAP12 may play an important role in tumor growth suppression and the survival of human colorectal cancer.

• The association between the copy-number variations of ZMAT4 and hematological malignancy.

  Authors: Jun Wan, Yan Gao, Xiaosu Zhao, Qi Wu, Xinrong Fu, Yong Shao, Hong Yang, Ming Guan, Bo Yu, Wei Zhang

  Hematology (Amsterdam, Netherlands). 01/2011; 16(1):20-3.

  Copy-number variations (CNVs) have been found in association with various types of diseases, including hematological malignancies. A recent array-based study implicated the presence of CNVs of ZMAT4Copy-number variations (CNVs) have been found in association with various types of diseases, including hematological malignancies. A recent array-based study implicated the presence of CNVs of ZMAT4 in the genome of acute myelogenous leukemia. In our study, we collected 617 bone marrow samples from multitypes of hematological malignancies as well as healthy controls. We found significant association between the CNVs of ZMAT4 and these hematological malignancies, including acute lymphoblastic leukemia, acute myelogenous leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, multiple myeloma, and myelodysplastic syndrome. We also examined the expression of ZMAT4 mRNA in the samples with 1 or 2 copies of DNA, and observed a weak yet positive correlation between the relative expression level and gene dosage. In conclusion, the CNVs of ZMAT4 have the potential to serve as a diagnostic indicator, alone or in combination with other markers, for hematological malignancies.

• Copy number variations of Interleukin-12B and T-bet are associated with systemic lupus erythematosus.

  Authors: Bo Yu, Yong Shao, Xupeng Yue, Jie Zhang, Ming Guan, Jun Wan, Wei Zhang

  Rheumatology (Oxford, England). 01/2011; 50(7):1201-5.

  Th1 cells have been implicated as the causal agents in the pathogenesis of autoimmunity. SLE represents the classical prototype of systemic autoimmune disease. Copy number variations (CNVs) have beenTh1 cells have been implicated as the causal agents in the pathogenesis of autoimmunity. SLE represents the classical prototype of systemic autoimmune disease. Copy number variations (CNVs) have been discovered to have phenotypic consequences and associate with various types of diseases. The current study aims to explore a possible association between CNVs of Th1 cell-related genes and the risk of SLE. Genomic DNA and RNA from 532 SLE patients and 576 healthy controls were extracted. CNVs of Th1 cell-related genes (T-bet, Stat4, IL-12A, IL-12B, IFN-γ, IP-10 and CXCR3) as well as Th2 and Treg cell-related genes (c-Mef, GATA3, Foxp3, IL-6 and TGF-β) were examined, and mRNA levels of IL-12B and T-bet were examined. Frequencies of IL-12B and T-bet CNVs in SLE patients were significantly higher than those in healthy controls. CNVs of IL-12B and T-bet had no synergistic contribution to SLE. The mRNA levels of IL-12B and T-bet in the samples with more than two copies of DNA were significantly higher than those with two copies of DNA. CNVs of IL-12B and T-bet are associated with the risk of SLE.

• [Influence of MDR1 gene C3435T on peripheral white blood cell counts in workers exposed to benzene].

  Authors: Jian-shu Huang, Xin-jü Zhang, Xiao Xu, Ming Guan, Yuan-ling Zhou, Ling Lü, He-jian Zou

  Zhonghua lao dong wei sheng zhi ye bing za zhi = Zhonghua laodong weisheng zhiyebing zazhi = Chinese journal of industrial hygiene and occupational diseases. 01/2011; 29(1):20-3.

  To explore the effects of MDR1 C3435T on the peripheral white blood cell counts in workers exposed to benzene. One hundred and twenty-one benzene-exposed workers and 110 healthy controls withoutTo explore the effects of MDR1 C3435T on the peripheral white blood cell counts in workers exposed to benzene. One hundred and twenty-one benzene-exposed workers and 110 healthy controls without benzene exposure were enrolled in this study. White blood cell counts influenced by the polymorphism of MDR1 gene were analyzed. The frequency of MDR1 3435 C/C, C/T, T/T in healthy controls was 37.27%, 46.36%, 16.37%, respectively, and it was 38.84%, 41.33%, 19.83% in the benzene-exposed workers, respectively. The frequency of the MDR1 gene was also not significantly different between benzene exposed workers and controls. Subjects exposed to benzene with MDR1 3435 mutation genotype (T/T) had the significantly lower WBC [(5.46 ± 1.51) × 10(9)/L] than those carrying wild type (C/C) and heterozygous (C/T), whose WBC were (6.08 ± 1.28) × 10(9)/L (P = 0.044). P-glycoprotein encoded by MDR1 gene may be implicated into the hematotoxicity of benzene. Subjects carrying MDR1 3435 T/T genotype may have a higher risk of benzene poisoning.

• Development and inter-laboratory validation of unlabeled probe melting curve analysis for detection of JAK2 V617F mutation in polycythemia vera.

  Authors: Zhiyuan Wu, Hong Yuan, Xinju Zhang, Weiwei Liu, Jinhua Xu, Wei Zhang, Ming Guan

  PloS one. 01/2011; 6(10):e26534.

  JAK2 V617F, a somatic point mutation that leads to constitutive JAK2 phosphorylation and kinase activation, has been incorporated into the WHO classification and diagnostic criteria of myeloidJAK2 V617F, a somatic point mutation that leads to constitutive JAK2 phosphorylation and kinase activation, has been incorporated into the WHO classification and diagnostic criteria of myeloid neoplasms. Although various approaches such as restriction fragment length polymorphism, amplification refractory mutation system and real-time PCR have been developed for its detection, a generic rapid closed-tube method, which can be utilized on routine genetic testing instruments with stability and cost-efficiency, has not been described. Asymmetric PCR for detection of JAK2 V617F with a 3'-blocked unlabeled probe, saturate dye and subsequent melting curve analysis was performed on a Rotor-Gene® Q real-time cycler to establish the methodology. We compared this method to the existing amplification refractory mutation systems and direct sequencing. Hereafter, the broad applicability of this unlabeled probe melting method was also validated on three diverse real-time systems (Roche LightCycler® 480, Applied Biosystems ABI® 7500 and Eppendorf Mastercycler® ep realplex) in two different laboratories. The unlabeled probe melting analysis could genotype JAK2 V617F mutation explicitly with a 3% mutation load detecting sensitivity. At level of 5% mutation load, the intra- and inter-assay CVs of probe-DNA heteroduplex (mutation/wild type) covered 3.14%/3.55% and 1.72%/1.29% respectively. The method could equally discriminate mutant from wild type samples on the other three real-time instruments. With a high detecting sensitivity, unlabeled probe melting curve analysis is more applicable to disclose JAK2 V617F mutation than conventional methodologies. Verified with the favorable inter- and intra-assay reproducibility, unlabeled probe melting analysis provided a generic mutation detecting alternative for real-time instruments.

• [High-resolution melting: a analysis for genotyping of MDR1 C3435T in benzene-exposed workers].

  Authors: Jian-shu Huang, Xin-ju Zhang, Xiao Xu, Ming Guan, Yuan-ling Zhou, Ling Lü, He-jian Zou

  Zhonghua lao dong wei sheng zhi ye bing za zhi = Zhonghua laodong weisheng zhiyebing zazhi = Chinese journal of industrial hygiene and occupational diseases. 01/2011; 29(1):70-2.

  Using high resolution melting (HRM) to analysis MDR1 C3435T in people exposed to benzene. Restriction fragment length polymorphism (RFLP) was utilized to detect the polymorphism of MDR1 3435 in 121Using high resolution melting (HRM) to analysis MDR1 C3435T in people exposed to benzene. Restriction fragment length polymorphism (RFLP) was utilized to detect the polymorphism of MDR1 3435 in 121 benzene-exposed workers, and the results were compared with the HRM in 10% samples and were confirmed with direct sequencing for six people in them. By direct sequencing, consistent results of benzene-exposed workers with RFLP or HRM were got. The new high resolution melting curve analysis is more efficient, more convenient, and cheaper than RFLP. High-resolution melting analysis provides a valid approach to efficiently detect DNA genetic diagnosis, which is suitable for detect susceptible genes in occupational surveillance.

• Examination of copy number variations of CHST9 in multiple types of hematologic malignancies.

  Authors: Xiaosu Zhao, Qi Wu, Xinrong Fu, Bo Yu, Yong Shao, Hong Yang, Ming Guan, Xiaojun Huang, Wei Zhang, Jun Wan

  Cancer genetics and cytogenetics. 12/2010; 203(2):176-9.

  Carbohydrate N-acetylgalactosamine 4-0 sulfotransferase 9 (CHST9) belongs to the N-acetylgalactosamine 4-sulfotransferase (GalNAc4ST) family. A recent array-based study implicated the presence ofCarbohydrate N-acetylgalactosamine 4-0 sulfotransferase 9 (CHST9) belongs to the N-acetylgalactosamine 4-sulfotransferase (GalNAc4ST) family. A recent array-based study implicated the presence of copy-number variations (CNV) of the region encompassing CHST9 in the genomes of acute myelogenous leukemia. Most of the current studies, however, focused on the genome-wide screening of CNV, and the functional impact of such regions needs to be extensively investigated in large amounts of clinical samples. In our study, we collected 617 bone marrow samples from multi-types of hematologic malignancies, as well as healthy controls, and detected the CNV of CHST9 by real-time polymerase chain reaction (PCR). We found significant association between the CNV of CHST9 and these hematologic malignancies including acute lymphoblastic leukemia, acute myelogenous leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, multiple myeloma, and myelodysplastic syndrome. We also examined CHST9 mRNA expression in the samples with one or two copies of DNA, and observed a weak yet positive correlation between the relative expression level and gene dosage. In general, the CNV of CHST9 have been shown to associate with hematologic malignancies. The functional consequences of CNV, however, need to be investigated extensively in the future.

• Identification of BANK1 polymorphisms by unlabelled probe high resolution melting: association with systemic lupus erythematosus susceptibility and autoantibody production in Han Chinese.

  Authors: Ming Guan, Bo Yu, Jun Wan, Xinju Zhang, Zhiyuan Wu, Qili Zhong, Wei Zhang, Hejian Zou

  Rheumatology (Oxford, England). 11/2010; 50(3):473-80.

  The three functional SNPs of BANK1 (rs10516487, rs17266594 and rs3733197) have been shown to be associated with SLE in Caucasian populations. The aim of this study was to investigate whether theThe three functional SNPs of BANK1 (rs10516487, rs17266594 and rs3733197) have been shown to be associated with SLE in Caucasian populations. The aim of this study was to investigate whether the association of BANK1 polymorphisms with SLE could be replicated in a Chinese population and whether the autoantibody production is relevant to BANK1 polymorphisms. Genotyping of three variants in BANK1 was carried out by unlabelled probe high resolution melting (HRM) assay in 264 SLE cases and 268 controls in a Chinese Han population living in Shanghai region. The genotype frequencies of the detected polymorphisms were analysed in relation to the production of autoantibodies (ANA, anti-dsDNA, anti-RNP, anti-SSA, anti-SSB and anti-Smith) in SLE patients. Samples with the target genotypes were accurately detected and easily distinguishable by unlabelled probe HRM assay. The frequencies of the rs10516487 C allele and the rs17266594 T allele were significantly increased compared with the controls (C allele: 88.6 vs 83.2%, P = 0.011; T allele: 88.3 vs 83.2%, P = 0.019). However, the frequencies of the rs3733197 G allele were not associated with SLE (G allele: 79.9 vs 79.1%, P = 0.741). The rs10516487 and rs17266594 polymorphisms were significantly associated with high-titre ANA (≥1 : 320) and production of anti-SSA antibodies in SLE patients compared with the control subjects. Genotyping using unlabelled probes is a rapid, accurate and cost-effective closed-tube method. This study implies that rs10516487 and rs17266594 polymorphisms might contribute to individual susceptibility to SLE and influence the ANA/SSA autoantibody response in SLE patients in Chinese population.