1.Proportion of Coptidis rhizoma and Evodiae fructus in the compound preparation: its effect in inducing apoptosis of SGC-7901 cells.
Qiu-xian PENG ; Da-jian YANG ; Jue SHI ; Hong-bing CAI ; Zhi-xian MO
Journal of Southern Medical University 2011;31(2):226-229
OBJECTIVETo investigate the effect of water extracts of Coptidis Rhizoma and Evodiae Fructus (CREF) on the proliferation and apoptosis of human gastric carcinoma cells (SGC-7901) and determine the optimal proportion of Coptidis rhizoma to Evodiae fructus.
METHODSThe growth inhibition of SGC-7901 cells treated with the water extracts of CREF of varying proportions was tested with MTT assay. The cell apoptotic rate and mitochondrial membrane potential were analyzed with flow cytometry.
RESULTSThe water extract of CREF with Coptidis Rhizoma: Evodiae Fructus proportions at 1:6, 2:5, 3:4, 4:3, 5:2, and 6:1 all significantly inhibited the growth of SGC-7901 cells after a 24-h or 48-h treatment (P<0.05). The growth inhibition and cell death ratio both exhibited a dose-dependent pattern of Coptidis Rhizoma. Flow cytometry analysis showed that, after treatment of the cells with CREF at the proportions of 1:6, 2:5, 3:4, 4:3, 5:2, and 6:1, the apoptotic rate were (8.50 ∓ 1.59)%, (9.90 ∓ 1.01)%, (17.15∓1.68)%, (21.55 ∓ 1.97)%, (34.10 ∓ 1.06)% and (34.40 ∓ 1.02)%, respectively, all significantly higher than that in the control group [(1.69 ∓ 1.91)%, P<0.05]. JC-1 Kit staining showed that mitochondrial membrane potential of SGC-7901 cells was decreased and the ratio of green to red fluorescence increased significantly after incubation with CREF.
CONCLUSIONCREF can inhibit the growth and induce apoptosis of SGC-7901 cells, and the strongest effect is achieved at the optimal proportion of Coptidis Rhizoma and Evodiae Fructus at 6:1.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Chemistry, Pharmaceutical ; Drug Compounding ; Drugs, Chinese Herbal ; pharmacology ; Evodia ; chemistry ; Humans ; Stomach Neoplasms ; pathology
2.Lymphadenectomy adjacent to inferior mesenteric artery root during radical operation and prognosis in rectal cancer.
Peng DENG ; Dong-Qiu DAI ; Jun-Qing CHEN ; Hui-Mian XU ; Shu-Bao WANG ; Ji-Xian SHAN
Chinese Journal of Gastrointestinal Surgery 2008;11(3):241-245
OBJECTIVETo investigate the effect of lymphadenectomy adjacent to inferior mesenteric artery root on the prognosis of rectal cancer.
METHODSClinicopathological data of 260 cases with rectal cancer undergone radical operation were analyzed retrospectively. The patients were divided into two groups. Group D(2): the lymph nodes adjacent to mesenteric artery root were not excised (n=188). Group D(3): the lymph nodes adjacent to mesenteric artery root were excised (n=72). Prognosis of two groups was compared during the follow-up period.
RESULTSIn group D(2), the 1-, 3-, 5-year total survival rates (TS) were 97.3%, 87.2% and 77.1%, and tumor-free survival rates (TFS) were 93.1%, 83.0% and 76.8% respectively. In group D(3 ), the 1-, 3-, 5-year total survival rates (TS) were 94.4%, 79.2% and 73.6%, and tumor-free survival rates (TFS) were 86.1%, 76.4% and 71.0% respectively. The differences of TS and TFS between two groups were not significant according to Kaplan-Meier analysis (P>0.05). Multivariate analysis revealed that the excision of lymph nodes adjacent to mesenteric artery root was not statistically correlated with the recurrence, metastasis and survival time after radical operation of rectal cancer.
CONCLUSIONExcision of lymph nodes adjacent to inferior mesenteric artery root has no significant impact on prognosis and it is unnecessary in the radical operation of rectal cancer.
Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lymph Node Excision ; methods ; mortality ; Lymph Nodes ; surgery ; Lymphatic Metastasis ; Male ; Mesenteric Artery, Inferior ; surgery ; Middle Aged ; Prognosis ; Rectal Neoplasms ; mortality ; pathology ; surgery ; Survival Rate ; Treatment Outcome
3.Inhibitory effect of short hairpin RNA targeting survivin gene on human ectopic endometrial cells.
Dong-xian PENG ; Yuan-li HE ; Li-wen QIU
Journal of Southern Medical University 2010;30(4):859-862
OBJECTIVETo construct a lentiviral expression vector for short hairpin RNA (shRNA) of human survivin gene, and assess its gene silencing effect in human ectopic endometrial cells.
METHODSHuman survivin gene shRNA sequence was designed using a software available on-line. The synthesized shRNA sequence was cloned into the pGCL-GFP vector to construct LV-survivin shRNA, which was confirmed by PCR and DNA sequence analysis. The packaging 293T cells were cotransfected with LV-survivin shRNA, pHelper 1.0 and pHelper 2.0, and the titer of the lentivirus was determined. The recombinant lentivirus was injected into human ectopic endometrial cells and the survivin mRNA expression was examined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) in comparison with that in the non-transfected and blank vector-transfected human ectopic endometrial cells.
RESULTSPCR analysis and DNA sequencing confirmed correct insertion of the shRNA sequence into the lentiviral vector. The titer of virus after packaging was 8x10(8) U/ml. Survivin mRNA expression in human ectopic endometrial cells transfected by LV-survivin shRNA was significantly inhibited compared with those in the non-transfected and empty vector transfected human ectopic endometrial cells (P<0.01), and no significant difference was found between the latter two groups.
CONCLUSIONThe lentiviral shRNA vector of survivin gene constructed can effectively inhibit the expression of survivin gene in human ectopic endometrial cells in vitro. This vector provides a tool for investigating the role of survivin gene in the occurrence and progression of endometriosis and for searching new therapeutic targets.
Cells, Cultured ; Endometriosis ; genetics ; pathology ; Female ; Gene Targeting ; Genetic Vectors ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; biosynthesis ; genetics ; Lentivirus ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection
4.Inhibitory effect of interleukin-6 on NMDA-stimulated neuronal firing activity and possible mechanism involved in the effect.
Xian-Feng ZHAN ; Bing LI ; Bei WU ; Yu-Ping PENG ; Yi-Hua QIU
Chinese Journal of Applied Physiology 2010;26(3):365-369
OBJECTIVETo study the effect and the possible mechanism of IL-6 on NMDA-excited neuronal discharges of rats in vitro.
METHODSThe cerebellar slices were prepared and spontaneous discharges of single cerebellar interposed nuclear (IN) neurons were recorded by extracellular recordings. The cerebellar slices were perfused with artificial cerebral spinal fluid (ACSF) containing N-methyl-D-aspartate (NMDA), IL-6, JAK inhibitor AG490. The changes in firing activities of the neurons treated with the drugs were recorded. The levels of phosphorylation at serine 897 site of NMDA receptor subunit 1 (NR1) in the neurons treated with various drugs mentioned above were detected by Western blot.
RESULTSThe discharge rates of the neurons that were treated with IL-6 together with NMDA were significantly lower than those of the neurons treated with NMDA alone. AG490 partially blocked the inhibitory effect of IL-6 on the NMDA-stimulated neuronal firing activity. The treatment of the neurons with IL6 and NMDA led to a concentration-dependent suppression of the phospho-NR1 expression relative to those neurons treated with NMDA alone. AG490 blocked the effect of the IL-6-induced depression of phospho-NR1 expression.
CONCLUSIONIL-6 inhibits NMDA-stimulated neuronal firing activity, and simultaneously down-regulates the phosphorylation of NR1 at serine 897 site.
Animals ; Cerebellum ; drug effects ; metabolism ; In Vitro Techniques ; Interleukin-6 ; pharmacology ; N-Methylaspartate ; pharmacology ; Nerve Growth Factors ; metabolism ; Neurons ; drug effects ; metabolism ; physiology ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism
5.Clinical value of genome-wide chromosome microarray technique in diagnosis of fetal cerebral ventriculomegaly.
Yi-Xian PENG ; Yu-Wen QIU ; Qing-Xian CHANG ; Yan-Hong YU ; Mei ZHONG ; Kun-Rui LI
Journal of Southern Medical University 2018;38(3):353-357
OBJECTIVETo investigate the clinical value of gnome-wide chromosome microarray (CMA) technique in genetic etiological diagnosis of fetal cerebral ventriculomegaly.
METHODSA retrospective analysis was conducted in 109 women with singleton pregnancy, who were admitted in Nanfang Hospital with the diagnosis of cerebral ventriculomegaly in the fetuses by ultrasound between January, 2014 and December, 2016. Routine karyotype analysis and chromosome microarray analysis were performed to identify the chromosomal abnormalities in the fetuses.
RESULTSKaryotype analysis detected chromosomal abnormalities at a rate of 12.84% in these fetuses, significantly lower than the rate of 26.60% with CMA technique (P=0.004); the combined detection rate of the two techniques was 28.44%. In 17 cases, karyotype analysis yielded normal results while CMA microarray showed abnormalities with an extra abnormal detection rate of 15.60%. Among the 17 fetuses with chromosomal abnormalities, 6 had micro-deletion, 9 had micro-duplication, 1 had both micro-deletion and micro-duplication, and 1 had heterozygous loss of single parent diploid.
CONCLUSIONCMA technique can be used to detect abnormal chromosomal copy numbers in fetuses with cerebral ventriculomegaly to increase the detection rate of chromosomal abnormalities and facilitate prenatal consultation and prognostic evaluation.
6.Screen of inflammatory genes regulated by heat shock factor 1 and corroboration with SOCS3 gene.
Feng-xiu YU ; Hua-li ZHANG ; Guang-wen CHEN ; Qiu-peng WANG ; Yong-zhong SHI ; Ying LIU ; Qiu-juan LIANG ; Xian-zhong XIAO
Journal of Central South University(Medical Sciences) 2006;31(2):167-173
OBJECTIVE:
To screen the inflammatory mediators genes regulated by HSF1, and explore the mechanism of downstream genes regulated by HSF1.
METHODS:
HSF-/- and HSF1+/+ mice were injected with 15 mg/kg LPS intraperitoneally (ip), respectively, and were treated as previous after HSR. The total RNA of lung tissues were extracted and filtrated by SuperArray gene Microarry. The promoter of candidate genes were analyzed by transcription element search software to search for heat shock element (HSE). Select the suppressor of cytokine signaling 3 (SOCS3) with HSE. Macrophage cells were stimulated with 400 ng/mL LPS, and were treated as previous after HSR, then the total RNA was extracted respectively. RT-PCR and northern blot assay were performed to detect the expression levels of SOCS3 mRNA.
RESULTS:
Fifteen genes were repressed by HSF1, including 9 genes with complete HSE. Eleven genes were accelerated by HSF1 possibly, including 8 genes with complete HSE. The promoter of SOCS3 gene contained one complete HSE. LPS stimulation obviously increased the levels of SOCS3 mRNA in macrophages of RAW264.7 mice, which was inhibited by HSR and over-expression of HSF1.
CONCLUSION
HSR or HSF1 inhibits LPS induced expression of SOCS3 mRNA; HSF1 might inhibit LPS-induced expression of SOCS3 mRNA by binding to HSE in the promoter of SOCS3 gene.
Animals
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DNA-Binding Proteins
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genetics
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pharmacology
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Endotoxemia
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chemically induced
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genetics
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Heat Shock Transcription Factors
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Heat-Shock Proteins
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genetics
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Inflammation
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genetics
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Lipopolysaccharides
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Macrophages
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metabolism
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Mice
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Mice, Inbred BALB C
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Mice, Knockout
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Mutation
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RNA, Messenger
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biosynthesis
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genetics
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Signal Transduction
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Suppressor of Cytokine Signaling 3 Protein
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Suppressor of Cytokine Signaling Proteins
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biosynthesis
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genetics
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Transcription Factors
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genetics
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pharmacology
7.The effect of cell killing and apoptosis by human herpes simplex virus- thymidine kinase/ganciclovir system combined with allitride in BIU87 cells.
Shao-peng QIU ; Xiao-peng MAO ; Kai-yuan CAO ; Xian-jing CHEN ; Guang-qing YUAN ; Lin XU ; Xiao-rong HUANG
Chinese Journal of Surgery 2005;43(6):382-386
OBJECTIVETo study the killing effect of human herpes simplex virus-thymidine kinase/ganciclovir (HSV-TK/GCV) system combined with allitride and the possible apoptosis mechanism in BIU87 cells.
METHODSThe cytotoxicity after combination were estimated by theamine blue tetrazolium bromide (MTT). The morphological changes were observed with inverted microscope and in-situ cell apoptosis detection kit. Changes of apoptosis rate and cell cycle were assessed by flow cytometry. B-cell lymphoma-2 (bcl-2), bax, caspase-3 (cysteine aspartate specific proteinase) mRNA changes were detected by reverse transcriptase polymerase chain reaction, and caspase-3 activity was estimated with colorimetry.
RESULTSFor combination group, the cell killing rate was raised to 72.50% to compare with 35.00% of GCV and 37.00% of allitride separately and there was a synergistic effect between these two drugs. The cell apoptosis was induced in all three groups and for the combination group the time of S-phase and G(2)-phase arrest were earlier than other two groups. Both drugs could inhibit the expression of bcl-2 and promote the expression and activity of caspase-3.
CONCLUSIONSThe combination of HSV-TK/GCV system with allitride can inhibit the proliferation of BIU87 cells congenerously through apoptosis, which may be correlated with S- and G(2)-phase arrest, down-regulation of bcl-2 and increased caspase-3 expression and its activity.
Apoptosis ; drug effects ; Drug Synergism ; Ganciclovir ; pharmacology ; Genetic Therapy ; Herpesvirus 1, Human ; enzymology ; genetics ; Humans ; In Vitro Techniques ; Sulfinic Acids ; pharmacology ; Thymidine Kinase ; genetics ; Transfection ; Urinary Bladder Neoplasms ; pathology ; therapy
8.Association between gene mutation of cytochrome P450 1A1 in exon 7 A4889G locus and susceptibility to endometriosis.
Dong-xian PENG ; Yuan-li HE ; Li-wen QIU ; Fang YANG ; Jing-ming LIN
Chinese Journal of Medical Genetics 2003;20(4):284-286
OBJECTIVETo assess the possible association between gene mutation of cytochrome P450 1A1(CYP1A1) in exon 7 A4889G locus and the susceptibility to endometriosis (EM).
METHODSAllele specific-polymerase chain reaction method was used to analyze gene mutation in exon 7 A4889G locus of CYP1A1 in 76 patients with endometriosis and 80 healthy controls.
RESULTSThe frequency of allele G on A4889G locus of CYP1A1 gene showed a significant difference between the study cohort and the control group (Chi2=7.498, P<0.01), with an odds ratio of 1.957. Statistically significant difference in the frequencies of genotypes AA, AG and GG was observed between the two groups (Chi2=6.915, P<0.05). Individuals with homozygotes for G allele were at higher risk of suffering from EM when compared against those with homozygotes for A allele, the odds ratio being 3.437 (Chi2=5.430, P<0.05).
CONCLUSIONThe above results suggest that gene mutation of CYP1A1 in exon 7 A4889G locus might be a genetic susceptible factor of endometriosis. The mutation allele of CYP1A1 gene appears to increase the risk of endometriosis.
Alleles ; Cytochrome P-450 CYP1A1 ; genetics ; Endometriosis ; genetics ; Exons ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Mutation ; Polymerase Chain Reaction ; Pregnancy
9.Estradiol regulates miR-135b and mismatch repair gene expressions via estrogen receptor-beta in colorectal cells.
Yu Qi HE ; Jian Qiu SHENG ; Xian Long LING ; Lei FU ; Peng JIN ; Lawrence YEN ; Jianyu RAO
Experimental & Molecular Medicine 2012;44(12):723-732
Estrogen has anti-colorectal cancer effects which are thought to be mediated by mismatch repair gene (MMR) activity. Estrogen receptor (ER) expression is associated with microRNA (miRNA) expression in ER-positive tumors. However, studies of direct link between estrogen (especially estradiol E2), miRNA expression, and MMR in colorectal cancer (CRC) have not been done. In this study, we first evaluated the effects of estradiol (E2) and its antagonist ICI182,780 on the expression of miRNAs (miR-31, miR-155 and miR-135b) using COLO205, SW480 and MCF-7 cell lines, followed by examining the association of tissue miRNA expression and serum E2 levels using samples collected from 18 colorectal cancer patients. E2 inhibited the expressions of miRNAs in COLO205 cells, which could be reversed by E2 antagonist ICI 182.780. The expression of miR-135b was inversely correlated with serum E2 level and ER-beta mRNA expression in CRC patients' cancer tissues. There were significant correlations between serum E2 level and expression of ER-beta, miR-135b, and MMR in colon cancer tissue. This study suggests that the effects of estrogen on MMR function may be related to regulating miRNA expression via ER-beta, which may be the basis for the anti-cancer effect in colorectal cells.
Adaptor Proteins, Signal Transducing/genetics/metabolism
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Adult
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Aged
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Cell Line, Tumor
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Colorectal Neoplasms/*genetics/metabolism
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DNA Mismatch Repair/*genetics
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Estradiol/analogs & derivatives/blood/*pharmacology
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Estrogen Antagonists/pharmacology
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Estrogen Receptor beta/genetics/*metabolism
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Female
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*Gene Expression Regulation, Neoplastic
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Humans
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Male
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MicroRNAs/genetics/*metabolism
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Middle Aged
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MutS Homolog 2 Protein/genetics/metabolism
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Nuclear Proteins/genetics/metabolism
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RNA, Messenger/biosynthesis
10.Overexpression of CYP46A1 has anti-Alzheimer's disease like effects
Min ZHAO ; Yan-Ying KONG ; Hua-Cheng YAN ; Le-Bin LIU ; Jian-Xin SU ; Zhi-Jian ZHOU ; De-Xian YU ; Qiu-Ju PENG ; Li XIE
Medical Journal of Chinese People's Liberation Army 2018;43(4):271-277
Objective To investigate the effect of CYP46A1 on the pathogenesis of Alzheimer's disease.Methods Recombinant lentiviral vectors which including anthropogenic CYP46A1 were injected into bilateral hippocampus of 3-monthold male 5XFAD transgenic mice,while empty vectors were injected into the corresponding position of the control group.After two months,the ability of learning and memory were tested by Morris water maze and T maze experiments,and amyloid plaque and inflammatory infiltration in the brain were detected by immunohistochemical staining and ELISA respectively.Results Compared with the control group,CYP46A1 virus injection significantly increased the CYP46A1 mRNA and protein expression in hippocampus.In addition,CYP46A1 overexpression significantly decreased the latency to find the platform in Morris water maze test and increased the correct rate to choose in T maze test.Aβ immunohistochemical staining and plaques area statistics demonstrated that the amyloid plaque area of hippocampus in CYP46A1 overexpression mice was significantly reduced,and there was a significantly decrease of hippocampal astrocytes expression by means of GFAP staining.Furthermore,hippocampal CYP46A1 overexpression significantly decreased the expression level of Aβ40,Aβ42,IL-1β and TNF-α,while compare with the control group.Conclusion CYP46A1 overexpression in hippocampus can promote the cognitive impairment,as well as ameliorate the brain inflammatory infiltration in 5XFAD transgenic mice,suggesting that CYP46A1 has anti-Alzheimer's disease like effects.