1.Construction of pGL3-Basic-SREBP-1c-promoter reporter gene vector and detection of its function
Xiaojun LIU ; Xingxing KONG ; Rui WANG ; Di SHAO ; Aijun QIAO ; Yongsheng CHANG ; Fude FANG
Basic & Clinical Medicine 2006;0(05):-
Objective To construct human SREBP-1c-promoter reporter gene vector and to detect its function.Methods Human blood genome DNA was extracted and pGL3-Basic-SREBP-1c-promoter reporter gene vector was constructed.Furthermore,the function of SREBP-1c-promoter was confirmed by dual-luciferase reporter assay.ResultspGL3-Basic-SREBP-1c-promoter reporter gene vector was successfully constructed and the promoter activity was obviously repressed by co-transfection FoxO1.Overexpression FoxO1 inhibited the SREBP-1c protein expression.Conclusion FoxO1 repressed the SREBP-1c protein expression through inhibition the SREBP-1c transcription.
2.Effects of nonylphenol and cadmium on sperm acrosome reaction in vitro in mice.
Zhu-Xiong SHAO ; Hong-Tao JIANG ; Fang LIANG ; Bao-Chang ZHU
National Journal of Andrology 2011;17(4):318-321
OBJECTIVETo study the effects of nonylphenol and cadmium on acrosome reaction in vitro in mouse spermatozoa.
METHODSSperm were collected from the vas deferens of mice, capacitated in vitro and stimulated with A23187 at 30 micromol/L to induce acrosome reaction. Then the sperm suspension was treated with nonylphenol at 10, 20, 30, 60 and 100 micromol/L or cadmium at 500, 2500 and 5 000 micromol/L, and the control group treated with the carrier solvent. Acrosome reaction of the sperm was analyzed by FITC-PSA staining.
RESULTSCompared with the control group, nonylphenol significantly inhibited acrosome reaction at the concentration of > 60 micromol/L (P < 0.01), but not at < 30 micromol/L (P > 0.05), and the sperm survival rate was reduced with increased concentration of nonylphenol. However, cadmium exhibited no significant influence on either acrosome reaction (P > 0.05) or sperm survival rate at 500 - 5 000 micromol/L.
CONCLUSIONNonylphenol and cadmium affect the spermatogenesis of mice in different ways; the former directly inhibits sperm acrosome reaction, while the latter has no direct effect on it.
Acrosome ; drug effects ; Acrosome Reaction ; drug effects ; Animals ; Cadmium ; pharmacology ; In Vitro Techniques ; Male ; Mice ; Mice, Inbred C57BL ; Phenols ; pharmacology ; Spermatozoa ; drug effects
3.Identification of deletion/duplication mutations in DMD gene by multiple ligation probe amplification and denaturing high-performance liquid chromatography.
Ben-Chang SHEN ; Cheng ZHANG ; Xiao-Fang SUN ; Shao-Ying LI
Acta Academiae Medicinae Sinicae 2007;29(1):83-86
OBJECTIVETo compare the effectiveness of using multiple ligation probe amplification (MLPA) and denaturing high-performance liquid chromatography (DHPLC) in screening the exon deletions and duplications of the DMD gene.
METHODSMLPA technique was applied to detect exon deletions and duplications previously confirmed by denaturing high-performance liquid chromatography (DHPLC).
RESULTSFrom October 2004 to October 2005, 22 unrelated DMD probands and their possible female relatives with clinical diagnosis with dystrophinopathy at our hospital entered this study. Both DHPLC and MPLA detected DMD gene depletion in 11 probands and DMD duplications in 3 probands. MLPA detected deletions and duplications in 2 probands, which were not detected by DHPLC. MLPA also successfully identified the carriage status of the potential female carriers of the probands.
CONCLUSIONCompared with DHPLC and traditional PCR techniques, MLPA is a superior tool to analyze the deletions and duplications in affected males as well as in the identification of the carriage status of potential females carriers.
Chromatography, High Pressure Liquid ; Female ; Gene Deletion ; Gene Duplication ; Genetic Predisposition to Disease ; Humans ; Male ; Muscular Dystrophy, Duchenne ; genetics ; Mutation ; Nucleic Acid Amplification Techniques ; methods
4.CT features of colloid carcinomas of the pancreas.
Fang-yuan REN ; Cheng-wei SHAO ; Chang-jing ZUO ; Jian-ping LU
Chinese Medical Journal 2010;123(10):1329-1332
BACKGROUNDColloid carcinomas of the pancreas have better prognosis than ordinary ductal adenocarcinoma, and preoperative distinction of colloid carcinoma from other pancreatic tumors is valuable for patient therapeutic planning and prognosis assessment. However, data about CT features of colloid carcinoma are very limited. This study aimed to investigate the CT features of this tumor.
METHODSInstitutional review board approval was obtained for this study. Seven patients with pathologically proven colloid carcinoma of the pancreas were included. Unenhanced and dynamic enhanced CT was performed in all the patients. CT features were analyzed retrospectively and correlations with pathological findings were evaluated.
RESULTSMean age of the patients was 59.8 years (41 - 76 years). Five tumors were located in the pancreatic head, and the other two in body and tail respectively. The maximum mean diameter of the tumors on axial scanning was 3.9 cm (3.0 - 6.7 cm). Tumors were round (n = 5) and lobular (n = 2). Tumors appeared slight hyp-attenuation on unenhanced CT, and peripheral and internal meshlike progressive delayed enhancement with great percent of cystic areas on enhanced CT. Calcification and gas in the tumor was seen in one patient whose duodenum was invaded by the tumor.
CONCLUSIONSColloid carcinomas of the pancreas appear as round or labular masses with great percent of cystic areas and slight hyp-attenuation on unenhanced CT and peripheral and internal meshlike progressive delayed enhancement on enhanced CT.
Adenocarcinoma, Mucinous ; diagnostic imaging ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Neoplasms ; diagnostic imaging ; Tomography, X-Ray Computed ; methods
5.Serological survey on viral hepatitis B in the population of Shanxi province.
Ru-fang ZHAI ; Ming GUANG ; Shao-ying CHANG ; Jian-hui AN ; Tai-sheng LI ; Fu-min ZHAO ; Hai-jiao WANG ; Hong LI ; Xiao-fang WANG
Chinese Journal of Epidemiology 2010;31(4):479-480
Adolescent
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Adult
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Child
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Child, Preschool
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China
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epidemiology
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Hepatitis B
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epidemiology
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Humans
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Infant
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Middle Aged
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Seroepidemiologic Studies
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Young Adult
6.Current situation researching of methylation in tumor.
Di SHAO ; An-fang CUI ; Liu-luan ZHU ; Ai-jun QIAO ; Xing-xing KONG ; Xiao-jun LIU ; Yong-sheng CHANG ; Fu-de FANG
Acta Academiae Medicinae Sinicae 2009;31(6):786-790
The disorders of DNA and histone methylation have a close relationship with the development and progression of tumors. Epigenetic regulation is critical in maintaining the stability and integrity of the expression profiles of different cell types by modifying DNA methylation and histone methylation. However, the abnormal changes of methylation often result in the development and progression of tumors. This review summarized the theory of tumor genomic and histone methylation, detection methods of methylation and their applications, and the clinical application of methylation as biological markers and drug targets.
DNA Methylation
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Histones
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metabolism
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Humans
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Methylation
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Neoplasms
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genetics
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metabolism
7.Research advances in Sirt1 gene.
Ai-jun QIAO ; Jin ZHIAO ; Xiao-jun LIU ; Di SHAO ; Liu-luan ZHU ; An-fang CUI ; Xing-xing KONG ; Fu-de FANG ; Yong-sheng CHANG
Acta Academiae Medicinae Sinicae 2009;31(6):782-785
As the most homologic homologue of silent information regulator 2 of yeast, Sirt1 gene is extensively expressed in mature tissues, and is rich in early embryo and reproductive cells. It is involved in the regulation of gene transcription, energy metabolism and cell aging. It promotes fat mobilization in adipocytes and glucose production in liver and regulates insulin secretion in islet beta cell. Furthermore, Sirt1 gene is an essential endogenous apoptosis inhibitor. In future, it may be used as new drug targets or applied in other disease management modalities.
Animals
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Humans
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Sirtuin 1
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genetics
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metabolism
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physiology
9.Impact of imipenem treatment on colonic mycobiota in rats with double-hit sepsis.
Jun GUAN ; Shao-Ze LIU ; Zhao-Fen LIN ; Wen-Fang LI ; Xue-Feng LIU ; De-Chang CHEN
Chinese Medical Journal 2013;126(10):1850-1854
BACKGROUNDBroad-spectrum antibiotic administration promotes intestinal colonization of exogenous fungal pathogens in healthy animals and has been recognized as one of the risk factors of invasive fungal infection in clinical settings. It is unclear whether broad-spectrum antibiotic treatment would change the intestinal mycobiota without exogenous fungal challenge in the context of sepsis.
METHODSWe established a rat model of double-hit sepsis using burn injury and endotoxin challenge. Rats with burn injury or double-hit sepsis received imipenem treatment for 3 days or 9 days, and their colon contents were sampled for selective fungal culture and isolation counts.
RESULTSImipenem treatment promoted the overgrowth of the commensal fungus Geotrichum capitatum in rats with burn injury. Imipenem treatment also promoted colon colonization by exogenous fungi in rats with burn injury and double-hit sepsis, including Trichosporon cutaneum, Candida albicans, Candida krusei, and Candida glabrata. A longer duration of imipenem treatment had a stronger impact on colon colonization by exogenous fungi.
CONCLUSIONImipenem treatment facilitates the overgrowth of commensal fungi and colonization by exogenous, potentially pathogenic fungi in the colons of rats with burn injury or double-hit sepsis.
Animals ; Anti-Bacterial Agents ; therapeutic use ; Burns ; complications ; microbiology ; Candida ; pathogenicity ; Colon ; microbiology ; Imipenem ; therapeutic use ; Male ; Rats ; Rats, Sprague-Dawley ; Sepsis ; drug therapy ; microbiology ; Trichosporon ; pathogenicity
10.Identification of disease-causing point mutations in DMD patients' dystrophin gene without large deletions/duplications.
Ben-chang SHEN ; Cheng ZHANG ; Song-lin CHEN ; Xiao-fang SUN ; Shao-ying LI ; Xiao-li YAO ; Shu-hui WANG ; Xi-lin LU
Chinese Journal of Medical Genetics 2006;23(4):392-396
OBJECTIVETo detect the disease-causing point mutations in the dystrophin gene of Duchenne muscular dystrophy (DMD) patients.
METHODSThe approach of denaturing high performance liquid chromatography (DHPLC) coupling with sequencing was used to screen the point mutations of 79 exons and the untranslated regions of dystrophin gene without large deletions/duplications, which was in 6 unrelated DMD probands from 6 DMD families.
RESULTSFive disease-causing mutations, 697-698insGT, C616T, G1255T, C4279T, and C2302T, were ides created the new stop codons in downstream sites of mutations, respectively. In addition to the disease-causing point mutations, a point mutation T5586+61A in intron 39 was also found at patient 3, and a missense mutation A694T in exon 8 was detected at patient 5. Four point mutations, C2168+13T, 5740-13dupG, G5234A and C5280T, were also detected at patient 6 whose causative point mutation was unavailable. Seven point mutations have not been reported previously. Bi-directional PCR amplification of specific alleles (Bi-PASA) method was established to distinguish the haplotypes of heterozygote or homozygote in a single PCR reaction.
CONCLUSIONVia automated DHPLC screening or detecting the subexonic mutations in dystrophin gene is feasible to clinical laboratories, and also is a superior method in terms of sensitivity and efficiency.
Base Sequence ; Chromatography, High Pressure Liquid ; DNA Mutational Analysis ; Dystrophin ; genetics ; Gene Duplication ; Humans ; Male ; Muscular Dystrophy, Duchenne ; genetics ; Point Mutation ; Polymerase Chain Reaction ; Sequence Deletion