1.Regulation of histone acetylation and apoptosis by trichostatin in HL-60 cells.
Xingang, LI ; Weikai, CHEN ; Junxia, GU ; Guohui, CUI ; Yan, CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(6):572-4
In order to examine the strong anticancer action and low toxicity of Trichostatin A (TSA), the effect of TSA was examined on the growth inhibition, acetylation of histone H3 and apoptosis in HL-60 cells by employing MTT, immunocytochemical techniques, and Annexin-V-FITC/ PI assay. Our results showed that TSA could inhibit proliferation of HL- 60 cells in a time- and dose-dependent manner, and the IC50 at the 36th h was 100 ng/ml. The apoptosis-inducing effect of TSA on HL-60 cells was also time- and dose-dependent. But it didn't demonstrate apparent apoptosis induction in NPBMNCs within specific dose and time range. Both of the acetylation of histone H3 in HL-60 cells and NPBMNCs increased significantly (P<0.05) after treated with 100 ng/ml TSA for 4 h. However, there was no significant differences between the two groups (P>0.05). It is concluded that TSA can inhibit growth and induce apoptosis of HL-60 cells in a time- and dose-dependent manner, and is able to selectively induce apoptosis in HL-60 cells but does not respond in NPBMNCs under the same conditions. The difference of TSA between HL-60 cells and NPBMNCs can't be explained by the regulation of histone acetylation.
Acetylation
;
Antineoplastic Agents/pharmacology
;
Apoptosis/*drug effects
;
HL-60 Cells
;
Histone Deacetylases/antagonists & inhibitors
;
Histone Deacetylases/*chemistry
;
Hydroxamic Acids/*pharmacology
2.The management of emergency materials
Shufang LI ; Yuesheng CHEN ; Xiuxia ZHOU ; Weikai CHEN ; Minxia LI
Chinese Journal of Practical Nursing 2006;0(12):-
Objective To make out the effective method of emergency materials management. Methods According to the native regular emergency diseases and the orders of superior section make out a general method for emergency materials management. Results This general management method can effective solve the problems which have existed in the aspect of emergency materials management. Conclusion By using general management method could advance the quality of first aid.
3.A genetic and clinical study in a family with familial hypercholesterolemia
Hong LAI ; Jinbo FENG ; Tong WANG ; Kehua ZHOU ; Weikai HOU ; Li CHEN
Chinese Journal of Internal Medicine 2011;50(2):120-123
Objective To investigate the low density lipoprotein receptor (LDLR)gene and apolipoprotein (Apo) B gene mutation in a Chinese family with familial hypercholesterolemia(FH) and give the kindrids clinical check-ups. Methods After physical examination, the kindreds underwent ECG and ultrasound checks. Blood samples were tested for lipid profiles. The promoter and all eighteen exons of LDLR gene were investigated by using PCR and agarose gel electrophoresis in combination with DNA sequence analysis. The results were compared with the normal sequences in GenBank and FH database ( www. ucl. ac. uk/fh ) to find mutations. In addition, the apolipoprotein B100 gene for known mutations (R3500Q,R3531C,R3501W and R3480W)that cause familial defective ApoB100 (FDB)was also tested using the same method. Results A novel homozygous G > A mutation at the 1581 bp of exon 10 was detected in the proband and his siblings. It caused a substitution of amimo acid Glu to Gly at codon 496. A novel heterozygous G >A mutation at the 1581 bp of exon 10 was detected in his parents. No mutations of R3500Q,R3531C,R3501W and R3480W of ApoB100 were observed. ECGs were normal. Atherosclerosis were found in all family members by ultrasound checks. Conclusions The homozygous G > A mutation at the 1581 bp of exon 10 was first determined in our country. The change of amino acid Glu to Gly is responsible for FH of the family. The type of the gene mutation was not found in the FH database( www. ucl.ac. uk/ih). It's a new type of LDLR mutation.
4.Construction of Sox9 gene eukaryotic expression vector and its inductive effects on directed differentiation of bone marrow stromal cells into precartilaginous stem cells in rats.
Weihua, HU ; Fengjing, GUO ; Feng, LI ; Hui, HUANG ; Weikai, ZHANG ; Anmin, CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(3):291-5
Sox9 gene was cloned from immortalized precartilaginous stem cells and its eukaryotic expression vector constructed in order to explore the possibility of bone marrow-derived stromal cells differentiation into precartilaginous stem cells induced by Sox9. A full-length fragment of Sox9 was obtained by RT-PCR, inserted into pGEM-T Easy clone vector, and ligated with pEGFP-IRES2 expression vector by double digestion after sequencing. The compound plasmid was transfected into born marrow-derived stromal cells by Lipofectamine 2000, and the transfection efficacy and the expression of Sox9 and FGFR-3 were observed. Flow cytometry was used to identify the cell phenotype, and MTT was employed to assay proliferative viability of cells. Sequencing, restrictive endonuclease identification and RT-PCR confirmed that the expansion of Sox9 and construction of Sox9 expression vector were successful. After transfection of the recombinant vector into bone marrow-derived stromal cells, the expression of Sox9 and FGFR-3 was detected, and proliferative viability was not different from that of precartilaginous stem cells. It was concluded that Sox9 gene eukaryotic expression vector was successfully constructed, and the transfected bone marrow-derived stromal cells differentiated into the precartilaginous stem cells.
Base Sequence
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Bone Marrow Cells/*cytology
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Cartilage/*cytology
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Cell Differentiation/genetics
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Cells, Cultured
;
Cloning, Molecular
;
Genetic Vectors/genetics
;
Molecular Sequence Data
;
Receptor, Fibroblast Growth Factor, Type 3/metabolism
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Recombinant Proteins/biosynthesis
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Recombinant Proteins/genetics
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SOX9 Transcription Factor/biosynthesis
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SOX9 Transcription Factor/*genetics
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Stem Cells/*cytology
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Stromal Cells/*cytology
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Transfection
5.Effect of trichostatin A on histone acetylation level and apoptosis in HL-60 cells.
Wei-Kai CHEN ; Yan CHEN ; Jun-Xia GU ; Guo-Hui CUI
Journal of Experimental Hematology 2004;12(3):324-328
In order to explore the underlying mechanism of high effects and low toxicity of trichostatin A (TSA), the effect of TSA on growth inhibition, histone acetylation level and apoptosis in HL-60 cells and normal human peripheral blood mononuclear cells (NPBMNC) were examined using MTT method, immunocytochemistry technology, and Annexin-V-FITC/PI double staining flow cytometry. The results showed that TSA inhibited growth of HL-60 in time- and dose-dependent manners, and the IC(50) of 36 hours was 100 ng/ml. The apoptosis induction effect of TSA in HL-60 cells was also time- and dose-dependent. Besides, the dose of TSA showing significant apoptotic cytotoxicity in HL-60 cells did not demonstrate apparent apoptosis induction in NPBMNC within definite dose and time range. The histone acetylation level in HL-60 cells and NPBMNC both showed remarkable increase (P < 0.05) after incubated with 100 ng/ml TSA for 4 hours without statistical difference between them is detected (P > 0.05). It is concluded that TSA shows effects of definite and significant growth inhibition and apoptosis induction on HL-60 cells in time- and dose-dependent manners. TSA is able to selectively induce apoptosis in HL-60 cells with low toxicity in NPBMNC at the same time. The mechanism of this selectivity can not be ascribed to the differential regulation of histone acetylation level between HL-60 cells and NPBMNC.
Acetylation
;
Apoptosis
;
drug effects
;
Cell Division
;
drug effects
;
DNA-Binding Proteins
;
HL-60 Cells
;
drug effects
;
Histone Deacetylases
;
physiology
;
Histones
;
metabolism
;
Humans
;
Hydroxamic Acids
;
pharmacology
;
RNA, Messenger
;
analysis
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Telomerase
;
genetics
6.Regulation of histone acetylation and apoptosis by trichostatin in HL-60 cells.
Xingang LI ; Weikai CHEN ; Junxia GU ; Guohui CUI ; Yan CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(6):572-574
In order to examine the strong anticancer action and low toxicity of Trichostatin A (TSA), the effect of TSA was examined on the growth inhibition, acetylation of histone H3 and apoptosis in HL-60 cells by employing MTT, immunocytochemical techniques, and Annexin-V-FITC/ PI assay. Our results showed that TSA could inhibit proliferation of HL- 60 cells in a time- and dose-dependent manner, and the IC50 at the 36th h was 100 ng/ml. The apoptosis-inducing effect of TSA on HL-60 cells was also time- and dose-dependent. But it didn't demonstrate apparent apoptosis induction in NPBMNCs within specific dose and time range. Both of the acetylation of histone H3 in HL-60 cells and NPBMNCs increased significantly (P<0.05) after treated with 100 ng/ml TSA for 4 h. However, there was no significant differences between the two groups (P>0.05). It is concluded that TSA can inhibit growth and induce apoptosis of HL-60 cells in a time- and dose-dependent manner, and is able to selectively induce apoptosis in HL-60 cells but does not respond in NPBMNCs under the same conditions. The difference of TSA between HL-60 cells and NPBMNCs can't be explained by the regulation of histone acetylation.
Acetylation
;
Antineoplastic Agents
;
pharmacology
;
Apoptosis
;
drug effects
;
HL-60 Cells
;
Histone Deacetylase Inhibitors
;
Histone Deacetylases
;
chemistry
;
Humans
;
Hydroxamic Acids
;
pharmacology
7.Research Progress of Genes Related to Treatment and Prognosis of Alpha-fetoprotein Producing Gastric Cancer
Shunli LU ; Jianping YU ; Weikai CHEN ; Andong LI ; Qingyuan HE ; Chao CHEN ; Xiaopeng HAN
Cancer Research on Prevention and Treatment 2022;49(7):738-744
Alpha-fetoprotein producing gastric cancer (AFPGC) is a special type of gastric cancer. AFPGC is considered to be the most highly invasive tumor with a high degree of malignancy and prone to metastasis. As a consequence, it usually causes unsatisfied treatment effect and the prognosis is poor. At present, treatment methods and monitoring indicators have limited effect on AFPGC. VEGF, HER2, AFP, GPC3 and SALL4 are cogently associated with tumor genesis and development. If we can reasonably guide the treatment and prognosis of AFPGC patients, it will greatly improve the situation of patients and improve the survival of patients. This article reviews the research progress of the genes related to the treatment and prognosis of AFPGC.
8.Effects of Yangyin Huoxue Prescription on the imbalance of Treg/Th17 and atherosclerosis in ApoE-/- mice induced by high-fat diet
Liyu ZHOU ; Yuanjing MA ; Yuting SUN ; Weikai CHEN ; Runze QIU ; Dongping YUAN ; Jun LONG
Journal of China Pharmaceutical University 2020;51(1):60-67
Atherosclerosis(AS), characterized with the accumulation of lipids on the vessel wall, is an immune-related inflammatory disease which promotes the progression of cardiovascular diseases(CVD). The imbalance of Treg/Th17 accelerates the progression of AS. Yangyin Huoxue Prescription(YHF)is an efficient traditional Chinese medicine used in the treatment of AS, but the effects of YHF on the balance of immunity have still not been clarified. This project was designed to investigate the effects of YHF on the imbalance of Treg/Th17 and AS in ApoE-/- mice induced by high-fat diet(HFD). ApoE-/- mice were given HFD to induce AS and administered low-dose YHF(18 g/kg)or high-dose YHF(36 g/kg)for 20 weeks. Atherosclerotic plaque area was analyzed by oil red O staining. Serum lipids were measured by biochemical kits. Treg or Th17 cells in peripheral blood were detected by flow cytometry. mRNA and protein expression of Foxp3 and RORγt of aortas were determined by qRT-PCR, Western blot and immunohistochemistry. Splenic CD4+T cells of mice were isolated and activated by anti-CD3/CD28, and then treated with lipopolysaccharide(LPS)and YHF. The expression of mRNA and protein of Foxp3 and RORγt were detected by qRT-PCR and immunofluorescence. It was found that YHF reduced the plaque area, decreased lipid level and increased the ratio of Treg cells in peripheral blood. Moreover, YHF increased mRNA or protein expression of Foxp3 in aortas in vivo or CD4+T cells in vitro while decreasing mRNA or protein expression of RORγt. These results suggested that YHF can regulate the imbalance of Treg/Th17 in ApoE-/- mice induced by HFD, and reduce the inflammatory stimulation of LPS on CD4+T cells, thereby improving AS.
9.Analysis of epidemic characteristics of respiratory tract pathogens among children before and after COVID-19 epidemic in Lanzhou
Bin YAN ; Xilong CHEN ; Luwei YAN ; Bingying ZHOU ; Weikai WANG
International Journal of Pediatrics 2022;49(11):773-776
Objective:To investigate the distribution and epidemic characteristics of respiratory pathogens in children before and after COVID-19 epidemic in Lanzhou.Methods:Two hundred and eighty-six children hospitalized with acute upper respiratory tract infection in Central Hospital of Gansu Province and Gansu Maternal and Child Health Hospital from October to November of 2020 and October to November of 2021 were selected respectively as the research objects, and a retrospective analysis was made.IgM antibodies of nine pathogens, including influenza virus A(IVA), influenza virus B(IVB), parainfluenza virus(PIV), adenovirus(ADV), mycoplasma pneumoniae(MP), chlamydia pneumoniae(CP), respiratory syncytial virus(RSV), echovirus(ECHO)and coxsackie virus B(CVB), were detected, and the basic information and epidemic characteristics were statistically analyzed.Results:The total positive rates of IgM antibodies of nine pathogens before and after the epidemic in COVID-19 were 31.8%(91/286)and 5.9%(17/286)respectively, after the epidemic, the detection rates dropped significantly, and there was significant difference among them( χ2= 62.505, P<0.05); After the epidemic, the detection rates of ADV, MP and CVB were all lower than those before the epidemic, and there were significant differences among these groups( χ2= 39.281, 12.167, 10.155, all P<0.05). The positive detection rates in the age group of 1 month ~1 year, ~3 years, ~6 years and>6 years before the outbreak were 37.4%(37/99), 38.3%(36/94), 16.7%(12/72)and 28.6%(6/21)respectively, and there were significant differences among these groups( χ2=34.055, P<0.05); Among them, the detection rates of MP in the age group 1 month ~1 year, ~3 years, ~6 years and>6 years were 16.2%(6/37), 25.0%(9/36), 16.7%(2/12)and 100%(6/6)respectively, and there were significant differences among these groups( χ2=10.289, P<0.05); CVB was not detected in>6 years group, the positive detection rates of CVB were 16.2%(6/37), 22.2%(8/36)and 25.0%(3/12)in the age group of 1 month ~1 year, ~3 years, ~6 years respectively, and there were significant differences among these groups( χ2= 27.742, P< 0.05). After the epidemic, the positive detection rates of the patients in the age group of 1 month ~1 year, ~3 years, ~6 years and>6 years were 5.9%(4/68), 4.0%(3/75), 5.7%(6/106)and 10.8%(4/37), with no statistical significance( χ2=2.235, P>0.05); Among them, the positive rates of IVB were 25.0%(1/4), 33.3%(1/3)and 66.7%(4/6)in the age group of 1 month ~1 year, ~3 years, ~6 years respectively, and in the age group>6 years was not detected, and there were significant differences among these groups( χ2= 96.022, P< 0.05). The detection rates of mixed infection of pathogens before and after the epidemic were 5.6%(16/286)and 0.3%(1/286)respectively, with no statistical significance( χ2= 2.314, P>0.05). Conclusion:The distribution of common pathogens of acute upper respiratory tract infection among children in Lanzhou was different before and after COVID-19 epidemic.
10.Construction of Sox9 Gene Eukaryotic Expression Vector and Its Inductive Effects on Directed Differentiation of Bone Marrow Stromal Cells into Precartilaginous Stem Cells in Rats
HU WEIHUA ; GUO FENGJING ; LI FENG ; HUANG HUI ; ZHANG WEIKAI ; CHEN ANMIN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(3):291-295
-tiated into the precartilaginons stem cells.