2.CpG Island Methylation Regulates BNIP3 Gene Expression in Gastric Cancer Cells
Wei SHEN ; Kun LIU ; Lu SUI ; Dan ZOU ; Jinyao HU
Journal of China Medical University 2015;(3):221-225
Objective To detect the methylation status of the promoter of BNIP3 gene in gastric cancer cell lines MKN1,and to explore the mecha?nism of DNA methylation regulating the expression of BNIP3 in gastric cancer cells. Methods The methylation status of BNIP3 promoter was de?tected by bisulfate sequencing PCR. Reverse transcription PCR was used to evaluate BNIP3 mRNA expression. MKN1 cells were treated with 5?Aza?2′?deoxycytidine(5?Aza?CdR),and after the treatment,the methylation status and BNIP3 mRNA expression were observed. Chromatin immuno?precipitation(ChIP)was used to determine the combination of BNIP3 with DNA methyltransferase 1(DNMT1). Results The promoter DNA of BNIP3 in MKN1 cells was in state of hypermethylation. Compared to the control group,methylation status and mRNA expression of BNIP3 in the drug treatment group(the 5?Aza?CdR concentration was 10μmol/L)were reversed,which showed statistical differences(P<0.05). 5?Aza?CdR inhibited the combination of BNIP3 with DNMT1. Conclusion CpG island methylation regulates BNIP3 gene expression in MKN1 cells. DNA methylation is related with the binding between the promoter of BNIP3 and DNMT1.
3.Exposure level of total N-nitroso compounds in residents of high-and low-risk areas for esophageal cancer in southern.
Kun LIN ; Wenying SHEN ; Yongning WU ; Shixin LU
Chinese Journal of Preventive Medicine 2002;36(6):386-389
OBJECTIVETo assess the exposure level of total N-nitroso compounds (TNOCs) in the residents of high- and low-risk areas for esophageal cancer in southern China.
METHODSSamples of duplicate plate diets and 12 hr overnight urine were collected from 120 male adults in each of the 2 areas, a high-risk area (Nanao county) and a low-risk area (Lufeng county) for esophageal cancer. The 240 male healthy subjects (35 - 64 years old) were selected by a 3-stage random cluster sampling procedure. Levels of TNOC, N-nitrosamino acids (NAAs) and volatile N-nitroso compounds (VNOC) in the samples were measured by Thermo Energy Analyzer.
RESULTSThe detectable rate (95%) of diet TNOC, daily dietary TNOC intake (4.25 +/- 0.84) micromol/day, 12-hr urinary TNOC excretion levels (1.76 +/- 0.23 ng/12 h) and daily dietary intake of VNOC (266 +/- 31.2 microg/day) in the high-risk area were all significantly higher than those of the low-risk area. Oesophageal cancer mortality rates were positively and significantly associated with daily dietary TNOC intake and 12-hr urinary TNOC excretion. Urinary NAAs excretion levels were not different in the two areas.
CONCLUSIONThe results suggest that TNOCs may be implicated in the etiology of esophageal cancer in southern China.
Adult ; China ; Esophageal Neoplasms ; etiology ; mortality ; Humans ; Male ; Middle Aged ; Nitroso Compounds ; administration & dosage ; adverse effects ; urine
4.Diagnosis and Treatment of 19 Children with Pulmonary Sequestration
gen, LU ; kun-ling, SHEN ; ying-hui, HU ; jin-jin, ZENG
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To summarize the clinical characteristics,diagnosis and treatment of pulmonary sequestration(PS) in children.Methods The clinical data of 19 children with PS confirmed by operation and(or) imaging examination from Mar.2003 to May.2008 were analyzed retrospectively.Thirteen cases were male,and 6 cases were female.One case was newborn infant.Seventeen cases received operation.Intralober type had a pulmonary lobectomy and extralobar type had a sequestrectomy.Chest X-ray and CT scan examinations were performed on the patients before operation.Results Seventeen cases got complete cure by operation,and the mean age at operation was 5.2 years.Chest enhanced CT indicated abnormal feeding arteries.Lobectomy was performed in 13 cases of intralobar PS,4 cases of extralobar PS were resected in the separated lung tissue,and all patients had unilateral lesions.No late deaths occurred in this group except 1 case who was complicated with malformation,and the postoperative follow-up showed an excellent recovery.Conclusions The main diagnostic methods of PS are CT and angiography.The diagnosis of PS can be confirmed when systemic feeding arteries are indicated on enhanced CT scans.Surgical resection is the main choice of treatment in all cases of PS in order to prevent recurrent infection and hemoptysis.The excellent results can be obtained by surgery.
5.A study on the application of CT perfusion and CT angiography with 64-slice spiral CT in the evaluation of internal carotid artery and middle cerebral artery stenosis and occlusion
Yan GAO ; Kun-Cheng LI ; Xiang-Ying DU ; Jie LU ; Jia-Bin LIU ; Yan-Hui YANG ; Yun SHEN ;
Chinese Journal of Radiology 1999;0(10):-
0.05),the MTT values were(5.2?1.1)and(10.9?2.6)s, respectively(t=7.24,P
6.Surgical management of early breast cancer.
Gen-hong DI ; Jiong WU ; Ke-da YU ; Jin-song LU ; Kun-wei SHEN ; Zhen-zhou SHEN ; Zhi-min SHAO
Chinese Journal of Oncology 2007;29(1):62-65
OBJECTIVETo evaluate the available surgical treatment modalities so as to explore the optimal strategy of managing early breast cancer.
METHODSThe clinical data of 2173 consecutive early-stage breast cancer patients treated by surgery treatments were retrospectively reviewed in order to clarify the indications and contraindications of different modalities. Therapeutic outcome of different surgical treatment modes were compared in terms of recurrence-free survival ( RFS) , disease-free survival ( DFS) , overall survival (OS). The cosmetic results of breast conservation and reconstruction were also evaluated .
RESULTSThe median age of these patients was 51 years ranging from 18 to 91. Of 2173 patients, 547 had stage 0- I lesions and 1626 stage II , and 1155 (53. 2% ) premenopausal. The proportion of patients who received radical surgery, breast conservation and reconstruction after mastectomy was 83. 6% (1817/2173), 10. 5% (229/2173) and 2. 5% (55/2173) , respectively. Younger and premenopausal patients prefer conservative and reconstructive surgeries, which are reasonable for stage 0-I and non-invasive breast cancer patients. Conservative surgery was not suitable for Paget's disease of breast (P = 0. 004) , mastectomy followed by reconstruction in this type of cancer was up to 38. 5%. The recurrence and metastasis rate of conservation or mastectomy were similar with a comparable 3-year RFS of 97. 4% and 95. 4% , respectively; there were also no significant differences in RFS(P =0. 2435) , DFS( P =0. 1395) and OS(P =0. 9406) after having been followed for 3 to 64 months. Similarly, immediate reconstruction did not show any negative effects with only 1 recurrence and 1 metastasis. Aesthetic outcomes were assessed as excellent or good in 90. 0% of breast conservation surgery, and the acceptability of reconstruction was 94. 5%.
CONCLUSIONBreast conserving surgery not only has comparable survival as mastectomy, but also has better cosmetic outcomes. Immediate breast reconstruction can be a suitable option without compromising survival. It is very important in the management for early breast cancer by selecting the most suitable surgery mode for every individual patient not only to cure her disease but also to satisfy the patient psychologically. Conservation should be preferred prior to reconstruction whenever possible.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; pathology ; surgery ; Carcinoma, Ductal, Breast ; pathology ; surgery ; Carcinoma, Intraductal, Noninfiltrating ; pathology ; surgery ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Mastectomy ; methods ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Paget's Disease, Mammary ; pathology ; surgery ; Reconstructive Surgical Procedures ; Retrospective Studies
7.Proteasomal inhibitor induces PINK1 aggresome formation and aggregating features
Yu-Hu ZHANG ; Bei-Sha TANG ; Lu WEN ; Bo XU ; Jian-Guang TANG ; Ji-Feng GUO ; Kun XIA ; Lu SHEN ; Hong JIANG ;
Chinese Journal of Neurology 2000;0(05):-
Objective To study the PINK1 aggresome formation and it's features in response to proteasomal inhibition.Methods Full-length PINK1 cDNA were amplified by polymerase chain reaction (PCR)from fetus brain cDNA library and subcloned into the EcoR I and BamH I sites of the vector pEGFP- N1.The integrity of the constructs was confirmed by sequencing.COS-7 cells were transiently transfected with PINK1-pEGFP-N1 using Lipofectamine 2000.Cells were treated by MG-132 in order to test the effect of proteasome inhibition on aggregation formation.The protein level of wild-type PINK1 with or without MG-132 treatment was confirmed by Western blot analysis.The formation of PINK1 aggregates was tested by fluorescence and the presence of ubiquitin,and ?-synuclein in PINK1 aggregates was examined by immunofluorescence and confocal microscopy.Results The expression level of PINK1 was significant increased into the form of aggregate in cells treated with MG-132;immunostaining for endogenous ubiquitin and ?-synuclein revealed a co-localization of both proteins in PINK1-positive aggregates.Conclusions In the presence of MG-132,overexpressed PINK1 forms into aggregates,whose components are ubiquitin and ?-synuclein.
8.Intake of volatile N-nitrosamines and their ability to exogenously synthesize in the diet of inhabitants from high-risk area of esophageal cancer in southern China.
Kun LIN ; Zhong-Ying SHEN ; Shih-Hsin LU ; Yong-Ning WU
Biomedical and Environmental Sciences 2002;15(4):277-282
OBJECTIVENan'ao County in Guandong Province is a high-risk area of esophageal cancer in Southern China. Of the suspected etiological factors in the environment, N-nitrosamines and their precursors have received the greatest attention.
METHODSSixty samples of the diet ingested by the inhabitants were collected and detected for volatile N-nitrosamines and their precursors. Five N-nitrosamines detected by Gas Chromatography-Thermal Energy Analyzer were N-nitrosodimethylamine, N-nitrosodiethylamine, N-nitrosopyrrolidine, N-nitrosopiperidine and N-nitrosomethyl-benzylamine.
RESULTSThe average content of 5 volatile N-nitrosamines in the diet was 312.0 micrograms/kg (median). The daily intake of the nitrosamines was 286.5 micrograms/head/day. Only the ability to exogenously synthesize N-nitrosopiperidine was powerful among 5 volatile N-nitrosamines. By a computerized stepwise regression analysis and curve fitting, we studied the correlation among the nitrosamines, the precursors and the major food items in the samples.
CONCLUSIONIt demonstrated that a relatively high content of volatile N-nitrosamines was present in the diet collected in the area.
Adult ; China ; epidemiology ; Diet ; Esophageal Neoplasms ; epidemiology ; etiology ; Female ; Humans ; Male ; Middle Aged ; Nitrosamines ; administration & dosage ; adverse effects ; Risk Factors ; Volatilization
9.Frequency of spinocerebellar ataxia types 1, 2, 3, 6, 7, 8, 10, 12, 17 and dentatorubral-pallidoluysian atrophy in Chinese Han population
Junling WANG ; Qian XU ; Lifang LEI ; Lu SHEN ; Hong JIANG ; Xiaohui LI ; Yafang ZHOU ; Jiping YI ; Jie ZHOU ; Xinxiang YAN ; Qian PAN ; Kun XIA ; Beisha TANG
Chinese Journal of Neurology 2009;42(10):672-675
Objective To assess the frequency of different subtype of spinocerebellar ataxias (SCAs) in Chinese Han population. Methods The nueleotide repeat mutations of SCA1, SCA2, SCA3/ MJD, SCA6, SCAT, SCA8, SCA10, SCA12, SCA17 and dentatorubral-pallidoluysian atrophy (DRPLA) were detected by the polymerase chain reaction (PCR), denaturing polyacrylamide gel electrophoresis (PAGE), Southern blot, recombinant DNA technology by T-vector cloning and direct sequencing technique in a cohort of 559 Mainland Chinese patients affected by spinocerebellar ataxia, including 363 families with autosomal dominant SCA (AD-SCA) and 196 sporadic cases. Results Among the 363 AD-SCA families, 15 families (4. 13%) were positive for SCA1, 26 (7. 16%) for SCA2, 187 (51.52%) for SCA3/MJD, 6 (1.65%) for SCA6, 7 (1.93%) for SCA7, 1 (0. 28%) for SCA12 and 1 (0. 28%) positive for SCA17; 120(33. 06%) were negative for all the tested SCAs. There were 2 (1.02%) SCAI, 3 (1.53%) SCA2, 15 (7. 65%) SCA3/MJD, 3 (1.53%) SCA6 and 173 (88.27%) not identified in the 196 sporadic SCA patients. None of the SCA8, SCA10 and DRPLA mutation was found. Conclusions SCA3/MJD is a substantially common subtype of AD-SCAs and sporadic SCA in Chinese Han patients with SCAs, subsequently followed by SCA2, SCA1, SCAT and SCA6; SCA12 and SCA17 are uncommon subtypes, while SCA8, SCA10, and DRPLA are rare, if not absent. SCA17 subtype was initially identified in mailand China. Some other genes might be causative in those unidentified AD-SCA pedigrees, and other etiological factors besides genetic cause might contribute for those sporadic cases.
10.Spinocerebellar ataxias in mainland China: an updated genetic analysis among a large cohort of familial and sporadic cases
Junling WANG ; Lu SHEN ; Lifang LEI ; Qian XU ; Jie ZHOU ; Yutao LIU ; Wenjuan GUAN ; Qian PAN ; Kun XIA ; Beisha TANG ; Hong Junling WANG
Journal of Central South University(Medical Sciences) 2011;36(6):482-489
Objective To undertake an updated genetic spectrum analysis in patients with hereditary spinocerebellar ataxia (SCA) in mainland China. Methods SCA 1, 2, 3, 6, 7, 8, 10, 12, 17 and dentatorubral-pallidoluysian atrophy (DRPLA) nucleotide repeat mutations were detected in 430 families with autosomal dominant SCA (ADCA) and 237 patients with sporadic ataxias by PCR and DNA sequencing. Subsequently, point and Indel (Insertion/deletion) mutation analyses of SCA5, SCA11, SCA13, SCA14, SCA15/16/29, SCA27, SCA31 and SCA35 were detected in 91 families with ADCA and 196 patients with sporadic ataxias excluded from SCA1, 2, 3, 6, 7, 8, 10, 12, 17 and DRPLA genotypes via PCR and Denaturing High Performance Liquid Chromatography (PCR-DHPLC), Multiplex ligation-dependent probe amplification and DNA direct sequencing analysis. Results Among the 430 ADCA families, there were 25 SCA1 (5.81%), 27 SCA2 (6.28%), 267 SCA3/MJD (62.09%), 8 SCA6 (1.86%), 8 SCA7 (1.86%), 1 SCA12 (0.23%), 1 SCA17 (0.23%) and 2 SCA35 (0.47%), and the remaining 91 families (21.16%) were genetically unidentified. Among the 237 sporadic SCA patients, there were 6 SCA1 (2.53%), 9 SCA2 (3.80%), 23 SCA3/MJD (9.70%) and 3 SCA6 (1.27%), and the remaining 196 (82.7%) were genetically unidentified. No pathogenic point mutation causing SCA5, SCA11, SCA13, SCA14, SCA27 or SCA31 subtypes was found. Conclusion SCA3/MJD is substantially the most common subtype in patients with ADCA and sporadic forms in mainland China, followed by SCA2, SCA1, SCA6 and SCA7. While SCA12, SCA17 and SCA35 are seldom found, SCA5, SCA8, SCA10, SCA11, SCA13, SCA27, SCA31 and DRPLA are very rare. The high proportion of genetically unidentified cases further verify that SCAs are of highly genetic heterogeneity, suggesting that other disease-causing genes might be involved in the negative ADCA pedigrees, and other etiological factors may involve in those sporadic cases other than genetics.