1.Injury in myocardial cells induced by citreoviridin.
Mi-feng LIU ; Xin JIANG ; Hong-ju YAO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2006;24(3):177-178
Animals
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Apoptosis
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drug effects
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Aurovertins
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toxicity
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Cells, Cultured
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DNA Damage
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drug effects
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Female
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Male
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Myocytes, Cardiac
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drug effects
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Rats
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Rats, Wistar
2.Primary culture of rat cerebellar granule neurons in vitro
Hong-ju, YAO ; Ling-wang, ZHOU ; Jun-rui, PEI ; Xiao-na, LIU ; Jing, WANG
Chinese Journal of Endemiology 2013;(1):38-41
Objective To establish a stable primary culture of rat cerebellar granule neurons in vitro for further study the toxic effects of chronic arsenic exposure on cerebellar cells.Methods Cerebellar cortices were taken from brain of Wistar rat 5-7 day old after born under stereoscopic microscope.Single cell suspension was acquired after digestion and washing with trypsin (0.25%) and DNase Ⅰ solution,respectively.Granule cells were purified from other cells by differential velocity adherence method for two times.Rat cerebellar granule neurons were seeded in culture plate pre-coated with poly-L-lysine.Neurons growth,development and synaptic connections were observed daily.The neurons were identified by neuron specific enolase (NSE) immunofluorescence technique.Results The neurons were affixed to the culture plate in 24 hours,in reticular arrangement observed under contrast microscope.Granule cells gradually turned round from oval and outlines became clearer in 2-3 days.In 4-6 days,there were a wide range of synaptic connections among the neurons and a mature nerve cell network formed.A large quantity of cerebellar granule neurons was seen by NSE identification.Few bigger cells such as purkinjes cells and glial cell outlines were also seen in the same visual field.Conclusions This is a successful primary culture method for acquirement of rat cerebellar granule neurons.The method can provide experimental basis for future studies the toxic effects of chronic arsenic exposure on cerebellar cells.
3.Analysis of impact factors for post-thaw embryo survival rate and clinical pregnancy rate of frozen-thawed embryo transfer program
Ning, YAO ; Ju-fen, ZHENG ; Zu-qiong, XIANG ; Lei-wen, ZHAO ; Xiao-ming, ZHAO ; Yun, SUN ; Yan, HONG ; Pei, CHEN
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(6):729-732
Objective To explore the impact factors for post-thaw embryo survival rate and clinical pregnancy rate in frozen-thawed embryo transfer program. Methods The clinical data of 573 cycles of frozen-thawed embryo transfers were retrospectively analysed. Groups were divided according to the pre-freeze embryo quality, pre-freeze embryonic developmental stage, frozen-thawed embryo quality and cryopreservation technique, respectively, and post-thaw embryo survival rates and/or clinical pregnancy rates were compared among groups. Results The clinical pregnancy rate of high quality pre-freeze embryo was significantly higher than that of low quality pre-freeze embryo (31.8% vs 20.0%) (P< 0.05). There was no significant difference in the post-thaw survival rates and clinical pregnancy rates between embryos frozen at day 2 of ferrtilization and those frozen at day 3 of ferrtilization(79. 1% vs 82.9% and 25.5% vs 31.2%, respectively) (P>0.05). The clinical pregnancy rates of the transfer cycles only with fully intact embryos and with mixed embryos were significantly higher than that only with partially damaged embryos(36.7% vs 24.1% and 29.2% vs 24.1%, respectively)(P<0.05). The post-thaw survival rate and post-thaw high-quality embryo rate were significantly higher in those processed with modified cryopreservation technique than in those processed with original cryopreservation technique (82.0% vs 66.3% and 50.0% vs 27.5%, respectively)(P<0.05). Conclusion Pre-freeze embryo quality, post-thaw embryo survival rate and post-thaw embryo quality have a positive correlation to subsequent clinical pregnancy rate. Favorable cryopreservation technique may ensure the success of post-thaw embryo recovery and transfer.
4.Overexpression of TLR2,TLR4 and MyD88 is associated with inflammation in C3H mice during Chlamydia respiratory infection
Gao-Ju PANG ; Li-Da SUN ; Nan YAO ; Xiao-Yu ZHA ; Ju-You LIANG ; Lu TAN ; Hong ZHANG ; Sai QIAO ; Hong BAI
Chinese Journal of Immunology 2018;34(5):737-740
Objective:To investigate the mechanism of excessive inflammation in the lung of C3H/HeN(C3H) mice following Chlamydia muridarum(Cm) airway infection.Methods:Chlamydial pneumonitis was induced in C3H and C57BL/6(C57) mice by intranasal inoculation with 1×103IFU (inclusion forming unites) of Cm strains.The expression of TLR2,TLR4 and MyD88 mRNA in the lung at different time point post-infection was measured by RT-PCR.Results:Cm infection induced Toll-like receptors expression in two strains of mice.The expression of TLR2 and TLR4 mRNA,especially TLR2 mRNA(P<0.001 or P<0.05),were significantly higher in highly susceptible C3H mice on day 7 and day 14 d post-infection compared with C57 mice.Further studies showed that the expression of MyD88 mRNA was also significantly higher in C3H mice on day 7 post-infection,and maintained high expression untill the day 14.Conclusion:Cm lung infection induced high level of TLR2,TLR4 and MyD88 mRNA expression in C3H mice,which may associate with excessive inflammation in C3H mice.
5.Value of in vivo monitoring of abdominal aortic atherosclerosis by high field magnetic resonance imaging in apoE-/- mice fed a high fat diet or infused with angiotensin II.
Rui ZHAO ; Yu-yu YAO ; Gang DENG ; Sheng-hong JU ; Zhong-juan WANG ; Song WEN ; Jun CHEN ; Hui JIN
Chinese Journal of Cardiology 2010;38(9):823-828
OBJECTIVEto explore the value of in vivo dynamic monitoring of abdominal aortic atherosclerosis (AS) by high field magnetic resonance (MR) imaging (MRI) in apoE-/- mice fed a high fat diet or infused with angiotensin.
METHODShigh fat diet or angiotensin II infusion was applied to apoE-/- mice for establishment of abdominal aortic atherosclerosis model. Abdominal aorta MRI was performed at 3 time points (baseline, 3 and 6 months) in 13 high fat diet fed apoE-/- mice aged 10-12 months and 3 wild-type control mice; 10 apoE-/- mice aged 6 months were infused with angiotensin II (1000 or 500 ng × kg(-1)× min(-1), n = 5 each) or saline for 14 d through Osmotic minipump. The abdominal aortic artery MRI was performed at baseline and 14 d after infusion. Black blood sequences of FLASH T1 weighted images and Proton density weighted-T2 weighted dual echo images were obtained. At each observation time post MRI, mice (n = 3, 5 and 5 for high fat diet group and n = 5 and 5 for angiotensin II infusion group) were sacrificed for pathological examination of the abdominal artery.
RESULTS(1) the abdominal aorta atherosclerosis was identified in both high fat diet and angiotensin II treated apoE-/- mice but in WT controls. Lesion progression was documented in high fat diet fed apoE-/- mice characterized by significantly increased vessel wall (a marker of atherosclerotic burden, F = 29.94, P < 0.05) and gradually increased plaque signal in PDW and T2W images. Results derived from MRI corresponded histopathology findings in high fat diet fed apoE-/- mice (correlative coefficient = 0.84, 0.95, 0.90, P < 0.05, respectively). Both MRI and histology showed increased lipid composition and decreased fibrotic composition in these mice. (2) The vessel wall area increased significantly [(1.21 ± 0.21) mm(2) vs. (2.65 ± 0.48) mm(2), P < 0.05] and the abdominal aortic dissection aneurysms was identified in apoE-/- mice infused with high angiotensin II. The vessel wall area also increased [(0.85 ± 0.11) mm(2) vs. (1.01 ± 0.17) mm(2), P < 0.05] in low angiotensin II infused apoE-/- mice and the coefficient between MR and histopathology is 0.934.
CONCLUSIONabdominal aortic unstable plaque model could be established by both high fat diet and angiotensin II infusion in apoE mice, angiotensin II infusion can transiently accelerate the progression of AS and can induce abdominal aortic dissection. Serial MR black blood sequences could demonstrate the development and progression of atherosclerosis in mouse abdominal aorta with excellent agreement to histopathology finding in terms of atherosclerotic burden and plaque composition. Thus, MRI appears to be a useful tool for in vivo AS plaque dynamic monitoring in mice.
Angiotensin II ; administration & dosage ; Animals ; Aorta, Abdominal ; Apolipoproteins E ; Arteriosclerosis ; Diet ; Dietary Fats ; administration & dosage ; Disease Models, Animal ; Magnetic Resonance Imaging ; methods ; Male ; Mice ; Mice, Knockout
6.Study on pharmacokinetics of emodin in Rhizoma Polygontum Cuspidatum and its compound.
Shu-Kun YAO ; Ye JIANG ; Xiao-Hua HAO ; Hong-Ju LIU ; Shao-Hao JANG ; Wei-Na LIU
China Journal of Chinese Materia Medica 2005;30(6):463-465
OBJECTIVETo study the difference in the pharmacokinetics of emodin in Zhiganning capsules and Rhizoma Polygontum Cuspidatum by nonaqueous RP-HPLC.
METHODThe rats were orally administered with the extraction of Rhizoma Polygontum Cuspidatum and Zhiganning capsules. After hydrolysis and extraction, the content of emodin in the plasma is determined by Nonaqueous RP-HPLC.
RESULTThe concentration-time profiles of emodin fit two-compartment model. The pharmacokinetics parameters including, t1/2alpha, AUC(0-infinity), CL(s) and C(max) of emodin in the group of Rhizoma Polygontum Cuspidatum were significantly different from these in the group of its compounds.
CONCLUSIONThere is a significant difference in pharmacokinetics of emodin between zhiganning capsules and the extraction of Rhizoma Polygontum Cuspidatum.
Animals ; Area Under Curve ; Capsules ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Emodin ; isolation & purification ; pharmacokinetics ; Female ; Male ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Rats ; Rats, Sprague-Dawley ; Rhizome ; chemistry
7.Detection of genetic variations of regulator of G-protein signaling 2 in hypertensives by sequencing.
Zhang JU-HONG ; Li NAN-FANG ; Yan ZHI-TAO ; Yao XIAO-GUANG ; Wang HONG-MEI ; Zhang DE-LIAN ; Wang XIN-LING
Acta Academiae Medicinae Sinicae 2011;33(2):200-204
OBJECTIVETo investigate the new genetic variations of regulator of G-protein signalling 2 (RGS2) gene in Kazakh hypertensives.
METHODSTotally 94 Kazakh patients with essential hypertension were enrolled and genomic DNA was extracted from their peripheral blood leukocytes. All the exon regions and their flanking sequences of RGS2 were directly sequenced.
RESULTSWe identified 13 variants including 5 common- single nucleotide polymorphisms with a minor allele frequency over 5%single nucleotide polymorphisms and 8 novel variations in 94 Kazakh hypertensives. Among these variations, 2 were in the introns and 7 in the promoter region. One subject had a G-to-C substitution at nucleotide 54 in exon 1, which lead to an amino acid substitution from K-to-N at position 18; another individual had an A-to-G substitution at nucleotide 2422 in exon 5, resulting in an amino acid from Y-to-C at position 178. Among eight common single nucleotide polymorphisms, -638A>G, -395G>C, 1891-1892TC I/D, and 2971G>C,and -43A>T and 2297A>G were in tight linkage disequilibrium with an r-square of more than 0.8, respectively.
CONCLUSIONSThe variants and their frequencies in RGS2 gene in Kazakh hypertensives may have ethnic differences when compared with other populations. The frequencies of the mutations are low in this population, and whether they influence blood pressure regulation requires further functional experiments.
Adult ; China ; Female ; Genetic Variation ; Humans ; Hypertension ; genetics ; Linkage Disequilibrium ; Male ; Middle Aged ; Minority Groups ; RGS Proteins ; genetics
8.A common variation within the STEAP4 gene exons is associated with obesity in Uygur general population.
Yan-ying GUO ; Nan-fang LI ; Ling ZHOU ; Xiao-guang YAO ; Hong-mei WANG ; Ju-hong ZHANG ; Wen-li LUO
Chinese Medical Journal 2011;124(14):2096-2100
BACKGROUNDCoordinated regulation of nutrient and inflammatory responses by six transmembrane epithelial antigen of prostate 4 (STEAP4) was essential for metabolic homeostasis. STEAP4 expression in human white adipose tissue was associated with obesity. This study aimed to evaluate association between STEAP4 genetic polymorphisms and obesity in Uygur Chinese general population.
METHODSThe functional regions of STEAP4 gene were sequenced in 96 Uygur with obesity (body mass index (BMI) > 30 kg/m²). Representative variations were selected according to the function and linkage disequilibrium and genotyped in 1507 obesity (BMI ≥ 25 kg/m²) and 825 non-obesity control (BMI < 25 kg/m²), all of whom were selected from epidemiology study of obesity-related diseases during January to February 2007 among Uygur population in Hetian area of Xinjiang Uygur Autonomous Region.
RESULTSFourteen novel and 6 known single nucleotide polymorphism (SNPs), including 2 nonsynonymous SNPs (nsSNPs), in the STEAP4 gene were identified. Of the 3 representative SNPs, the nsSNP rs1981529 (Gly75Asp, 224A/G) was significantly associated with obesity phenotype (additive P/Pc = 0.001/0.006, dominant P/Pc = 0.003/0.018, odds ratio (OR) and 95% confidence interval (CI) adjusted for age, gender and drinking 0.755 (0.641 - 0.890) and 0.750 (0.621 - 0.907), respectively). By the multiple linear regression analysis, the quantitative phenotypes of BMI (P/Pc = 0.002/0.004) and waist circumference (P/Pc = 0.004/0.008) were found to be significantly associated with the genotypes of rs1981529 (Gly75Asp, 224A/G) in Uygur general population, and effect size (beta value) of one allele G of rs1981529 (Gly75Asp, 224A/G) was - 0.553 kg/m² for BMI and - 1.311 cm for waist circumference after controlling age, gender and drinking factors.
CONCLUSIONSThe present study shows an association of the common variation rs1981529 (Gly75Asp, 224A/G) in the STEAP4 gene with obesity in Uygur general population. Further studies should replicate the results using larger populations.
Adult ; Asian Continental Ancestry Group ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Linkage Disequilibrium ; genetics ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Obesity ; genetics ; Oxidoreductases ; genetics ; Polymorphism, Single Nucleotide ; genetics ; Waist Circumference ; genetics
9.Therapy-related chronic myelomonocytic leukemia secondary to acute promyelocytic leukemia in remission for 15 years: one case report.
Yun Ju MA ; Wen hong SHEN ; Xiao Wen TANG ; Hai Ping DAI ; Hong Jie SHEN ; Ting Ting TAO ; Dan Dan LIU ; Li YAO ; Xia Ming ZHU ; De Pei WU
Chinese Journal of Hematology 2018;39(8):628-628
10.Expression of recombination-activating genes and T cell receptor gene recombination in the human T cell leukemia cell line.
Hong-yun ZOU ; Li MA ; Min-jie MENG ; Xin-sheng YAO ; Ying LIN ; Zhen-qiang WU ; Xiao-wei HE ; Ju-fang WANG ; Xiao-ning WANG
Chinese Medical Journal 2007;120(5):410-415
BACKGROUNDRecent studies have suggested that mature T cells can change their specificity through reexpression of recombination-activating genes (RAG) and RAG-mediated V(D)J recombination. This process is named receptor revision and has been observed in mature peripheral T cells from transgenic mice and human donors. However, whether thebreceptor revision in mature T cells is a random or orientated process remains poorly understood. Here we used the Jurkathuman T cell line, which represents a mature stage of T cell development, as a model to investigate the regulation of Tcell receptor (TCR) gene recombination.
METHODSTCR Dbeta-Jbeta signal joint T cell receptor excision DNA circles (sjTRECs) were determined by nested and seminested PCR. Double-strand DNA breaks at recombination signal sequences (RSSs) in the TCRVbeta chain locus were detected by ligation-mediated-PCR. Further analysis of the complementarity-determining region 3 (CDR3) size of the TCRVbeta chain was examined by the TCR GeneScan technique.
RESULTSRAG1, RAG2, and three crucial components of the nonhomologous DNA end-joining (NHEJ) pathway were readily detected in Jurkat. Characteristics of junctional diversity of Dbeta2-Jbeta2 signal joints and ds RSS breaks associated with the Dbeta2 5' and Dbeta 2 3' sites were detected in DNA from Jurkat cells. CDR3 size and the gene sequences of the TCRVbeta chain did not change during cell proliferation.
CONCLUSIONSRAG1 and RAG2 and ongoing TCR gene recombination are coexpressed in Jurkat cells, but the ongoing recombination process may not play a role in modification of the TCR repertoire.However, the results suggest that Jurkat could be used as a model for studying the regulation of RAGs and V(D)J recombination and as a "special" model of the coexistence of TCR gene rearrangements and "negative" receptor revision.
Antigens, Nuclear ; genetics ; Base Sequence ; Complementarity Determining Regions ; DNA Breaks ; DNA-Binding Proteins ; genetics ; Genes, RAG-1 ; Genes, T-Cell Receptor ; Humans ; Jurkat Cells ; Ku Autoantigen ; Leukemia, T-Cell ; genetics ; Molecular Sequence Data ; Nuclear Proteins ; genetics ; Recombination, Genetic