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Objective:To preliminarily explore the association between single nucleotide polymorphisms (SNP) of five candidate genes (APH1B, PRNP, HMGCR, SIRT1, ApoE) and Alzheimer′s disease (AD), and to analyze the methylation levels of BAX and ApoE promoters on the pathogenesis of AD.Methods:Seventeen cases who were admitted to the Department of Geriatrics of the First Affiliated Hospital of Xinjiang Medical University from 2014 to 2015 and diagnosed as likely to be AD by geriatrician and neurologists according to the AD diagnostic criteria in 4th Revised Edition of the Diagnostic and Statistical Manual of Mental Disorders of the American Psychiatric Association served AD group, with an age of (75.65±5.86) years, and 34 non-AD patients with matching baseline data such as age, gender, ethnicity, and education status among patients hospitalized during the same period were selected as control group, with an age of (77.59±7.41) years. Sanger sequencing method was used for SNP typing of candidate genes. Methylation-specific polymerase chain reaction was used to determine the DNA methylation level.Results:The distribution of ApoE ε4 allele was statistically different between the AD group and the control group (χ 2=9.718, P=0.002). Candidate genes (SIRT1 rs7895833, APH1B rs1047552, PRNP rs1799990, HMGCR rs3846662) SNP locus genotypes and alleles had no statistically significant differences in the distribution between the AD group and the control group ( P>0.05). After stratification according to whether they carried ApoE ε4, no statistically significant difference was found between the two groups ( P>0.05). The BAX promoter methylation level of the AD group (0.045±0.025) was lower than that of the control group (0.061±0.028) ( t=-2.078, P=0.045). After gender stratification, the BAX methylation level of the female AD group (0.044±0.021) was lower than that of the control group (0.065±0.275) ( t=-2.230, P=0.045). There was no statistically significant difference in the methylation level of ApoE promoter between the AD group and the control group ( P>0.05). After stratification according to whether they carry ApoE ε4 or not, the methylation level of AD patients with ApoE ε4 allele (1.553±0.291) was higher than that of non-carriers (1.221±0.261) ( t=2.480, P=0.025). Conclusions:ApoE ε4 allele may be a risk factor for the onset of AD. BAX promoter hypomethylation contributes to AD in the elderly in Xinjiang, especially in female. ApoE ε4 allele may cause AD through the interaction with ApoE methylation.
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Objective To investigate the expression and clinicopathological significance of Bcl-2 and Bax genes in colorectal cancer (CRC) patients complicated with schistosomiasis. Methods The CRC patients receiving surgical treatment in the First Affiliated Hospital of Dali University from June 2016 to June 2020 were recruited as the study subjects, and 30 subjects were randomly sampled from the CRC patients complicated with schistosomiasis (CRC-S group) and 30 subjects were randomly sampled from the CRC patients without schistosomiasis (CRC group) using a random number table method. The cancer specimens were sampled from subjects in the CRC-S and CRC groups, and the peri-cancer specimens were sampled from subjects in the CRC group. The Bcl-2 and Bax expression was quantified in cancer and peri-cancer specimens using a real-time fluorescent quantitative PCR (qPCR) assay and immunohistochemistry at transcriptional and translational levels, and the cell apoptosis was detected in cancer specimens using HE staining. Results A total of 60 subjects were enrolled, including 30 cases in the CRC group and 30 cases in the CRC-S group. There were no significant differences between the two groups in terms of gender distribution (χ2 = 0.271, P > 0.05), mean age (t = -0.596, P > 0.05), tumor growth pattern (χ2 = 0.275, P > 0.05), tumor location (χ2 = 4.008, P > 0.05), tumor invasion depth (χ2 = 0.608, P > 0.05), degree of tumor differentiation (χ2 = 0.364, P > 0.05), or presence of vascular metastasis (χ2 = 1.111, P > 0.05), while significant differences were seen between the two groups in terms of histological type, presence of lymph node metastasis and TMN staging (χ2 = 5.963, 8.297 and 5.711, all P values < 0.05). qPCR assay and immunohistochemistry quantified significantly higher Bcl-2 and Bax expression in cancer specimens from the CRC and CRC-S groups than in the peri-cancer specimens from the CRC group at both translational and transcriptional levels (all P values < 0.05), and higher Bcl-2 and lower Bax expression were seen in the cancer specimens from the CSC-S group than that from the CRC group (all P values < 0.05). In addition, the cell apoptotic rate was significantly greater in the cancer specimens in the CRC group than in the CRC-S group (42.00% vs. 23.35%; χ2 = 41.500, P = 0.000). Conclusion Schistosomiasis may be involved in the development and progression of CRC through affecting Bcl-2 and Bax gene expression in the apoptosis signaling pathway.
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Objective To investigate the effect of hedyotic diffusa willd injection on osteosarcoma MG‐63 cells Bax gene expres‐sion .Methods MTT was be used to detect the inhibition rate of MG‐63 cells by hedyotic diffusa willd injection after 6 ,12 ,24 ,48 h certain concentration ,RT‐PCR was be used to detect the intracellular expression of Bax gene .Results Hedyotic diffusa willd injec‐tion can significantly inhibit the proliferation of MG‐63 cells ,as the concentration was 100 μL/mL ,Bax gene expression was signifi‐cantly increased over time(P<0 .05) .Conclusion Upregulating the expression of Bax gene by hedyotic diffusa willd injection can induced human osteosarcoma M G‐63 cells apoptosis to death .
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@#BACKGROUND: Many studies have showed that apoptosis of endothelial cells plays a curial role in the progress of sepsis. But the role of simvastatin in apoptosis of endothelial cells induced by sepsis is not clear. The present study aimed to investigate the role of simvastatin in apoptosis of endothelial cells induced by sepsis and its mechanism. METHODS: Human umbilical vein endothelial cells (HUVECs) were randomly divided into three groups: control group, sepsis serum intervention group (sepsis group) and simvastatin+sepsis serum intervention group (simvastatin group). After 24-hour incubation with corresponding culture medium, the relative growth rate of HUVECS in different groups was detected by MTT assay; the apoptosis of HUVECs was detected by Hoechst33258 assay and flow cytometry; and the expression of the Bcl-2 and Bax genes of HUVECs was detected by PCR. RESULTS: Compared with the sepsis group, HUVECs in the simvastatin group had a higher relative growth rate. Apoptotic HUVECs decreased significantly in the simvastatin group in comparison with the sepsis group. Expression of the Bcl-2 gene in HUVECs decreased obviously, but the expression of the Bax gene increased obviously after 24-hour incubation with sepsis serum;however, the expression of the Bcl-2 and Bax genes was just the opposite in the simvastatin group. CONCLUSIONS: Our study suggests that simvastatin can inhibit apoptosis of endothelial cells induced by sepsis through upregulating the expression of Bcl-2 and downregulating Bax. It may be one of the mechanisms for simvastatin to treat sepsis.
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Objective To study the function of Bak in mitochondrial signaling pathway and interaction between Bax and Bak during apoptosis.Methods Bax/Bak double knock out(Bax-/- and Bak-/-)MEF cells from mouse embryo fibroblasts(MEF) and Hela cells were divided into four groups according the cell different genotypes(wild type,Bax-/-,Bak-/- and double knock out) and treated with different chemical reagents after co-transfection with Bax,Bak,Mito-Red and empty pEGFP vector.Apoptosis,mitochondrial morphology and cytochrome C release were detected with confocal microscope,immunofluoresence and Western blotting techniques.Results There were correlations between the percentage of Hela cell apoptosis and mitochondrial fission(%) as well as cytochrome C(%)(P
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Objective To review the relationship between the expression levels of bcl-2,bax and bad gene and other biological factors of breast cancer in the growth and development of breast cancer.Methods Related literatures were summarized and reviewed.Results The expression level change of antiapoptosis gene bcl-2 was still under research and the expression levels of apoptosis gene bax and bad were down-regulated progressively in the evolution from benign breast tissue to breast cancer tissue.The expression level of bcl-2 had positive correlation with some positive factors in breast cancer such as estrogen receptor(ER) and progesterone receptor(PR),while it had negative correlation with some negative factors such as p53,EGFR,c-erbB-2 and lymph node metastasis.The levels of ER,PR and the expression level of p53 of breast cancer had no relationship with the expression level of bax.Up to now there was no report about the relationship between the expression level of bad and other biological factors of breast cancer.Conclusion The role of altered expression level of bcl-2,in the treatment and prognosis of breast cancer is still controversial,and the relationship between the expression of bad and the prognosis of breast cancer is still unknown,but expression level of bax is correlated positively with the prognosis of breast cancer.Research on these genes can provide us some new index to evaluate the prognosis of breast cancer and new ideas on treatment of breast cancer including gene therapy.
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Objective To investigate the proliferation rate of HepG2 cell after multiple thermotherapy and the possible reasons related to it. Methods After HepG2 cell were treaded by ten repeated cycles of heat exposure at 43 ℃ for 80 minutes twice a day, the doubling time of cell was analyzed, and the cell cycle, bcl-2 mRNA and bax mRNA were detected. Results The proliferation rate of HepG2 cell which treated with heat speeded up, the percentage of G2 and S in cell cycle increased, the expression of bcl-2 mRNA strengthened and the rate of bcl-2/bax increased. Conclusion The speeded proliferation of HepG2 cell after multiple thermotherapy is related to its high percentage of DNA duplicated and dividing cell, strengthened expression of bcl-2 mRNA and increased rate of bcl-2/bax.
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Objective To explore the relationship between mitochondrial DNA microsatellite instability(mtMSI) and expression of Bcl 2 and Bax mRNA in gastric cancer and its precancerous lesions. Methods mtMSI and expressions of Bcl 2 and Bax mRNA were detected with PCR SSCP and RT PCR. Results Expressions of Bcl 2 mRNA in intestinal metaplasia (IM,53 3%) and dysplasia (Dys, 70%) were significantly higher than that in normal control(10%, P
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Objective To explore the role of 1, 4, 5-trisphosphate inositol ( IP3) and bax gene expression in apoptosis of HepG2 cells induced by Quercetin.Methods HepG2 cells were cultured for 72h as control. HepG2 cells were treated with different concentrations including 20,40.60,80?mol/L Quercetin for 72h, and treated with 60?mol/ L Quercetin for 12h, 24h, 48h and 72h. IP3, bax mRNA, bax protein and apoptosis rate were assayed by IP3-[3H] Birtrak assay,RT-PCR, Western blotting and flow cytometry, respectively.Results In HepG2 cells incubated with each of the concentrations of Quercetin for 72 h,IP3continent was lower than that of control(P
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Apoptosis is involved in a variety of physiological and path ol ogical conditions. Bax is proapoptotic member in signal pathway of apoptosis. It will contribute to the survival of cells if reducing its expression and activit y. Advances in factors influencing Bax, biological effects induce d by Bax knock-out, and antisense nucleotides targeted to Bax-mRNA were r eviewed in this article during recent years.
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Radiation therapy is one of the most important treatment modalities following surgery of the primary malignant or metastatic brain tumors. But radiation can be harmful to normal healthy brain tissues around the tumor. There have been numerous reports of radiation induced damage such as delayed necrosis to human brain after therapeutic exposure. Apoptosis is a form of cell death with morphological and biochemical features that differ from those of necrosis. The aim of this study is to evaluate the apoptosis in normal rat brain after irradiation. Twenty one Sprague-Dawley rats were given a single dose of 10 Gy using high dose rate Ir-192 over 5 minutes at the right frontal region. Apoptosis was evaluated by the TUNEL method(In-situ end labelling technique) and mutant p53 protein, bc1-2 and bax genes were evaluated by immunohistochemical stain. Apoptosis was assessed at 1 week(group A, n=5), 2 week(group B, n=), 4 week(group C, n=), 6 week(group D, n=), 8 week(group E, n=) after irradiation. Apoptosis was noted with 20% of cases(1/5) in group A, 40% of cases(2/5) in group B, 60% of cases(3/5) in group C, 67% of cases(2/3) in group D and 100% of cases(3/3) in group E. Overall apoptosis positive rate was 52.4%(11/21). Apoptosis was most prominently found in external granular and external pyramidal layer(82%, 9/11) and found one case in internal pyramidal layer and the other one case in corticowhite matter junction. There were no positive stainning for mutant p53 protein, bc1-2 and bax gene in all cases pertaining to the phenomenon of apoptosis. In conclusion, apoptosis was evident in the rat brain after irradiation and the incidence of apoptosis was increased with time after irradiation. But the genes related to apoptosis after irradiation were not apparent in this study. Further evaluation including biochemical and clonogenic study needs to clarify the mechanism of apoptosis in normal brain after irradiation.
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Animales , Humanos , Ratas , Apoptosis , Neoplasias Encefálicas , Encéfalo , Muerte Celular , Etiquetado Corte-Fin in Situ , Incidencia , Necrosis , Ratas Sprague-DawleyRESUMEN
Objective To detect the protective mechanism of trehalose in tracheal cryopreservation.Methods Inbred male Sprague-Dawley (SD) rats were sacrificed with intraperitoneal injection of ketamine(150mg?kg -1).The tracheas were removed and immersed immediately in the freezing medium of low potassium dextran (LPD) solution only(Group Ⅰ) ,containing with 10% dimethylsulfoxide(DMSO)(Group Ⅱ), containing with 0.15mol?L -1 trehalose (Group Ⅲ),and containing with 10% DMSO and 0.15mol?L -1 trehalose (Group Ⅳ) respectively. A sterile plastic tube containing a 1-cm-long trachea was filled with the freezing medium,sealed,and frozen to -80℃ at rate of -1℃ per minute in a programmable freezer.Then the tube was stored in liquid nitrogen(-196℃) for 20 days. Then the specimen was thawed in a 37℃ water bath and rinsed with physiologic saline solution 10 times.Histologic changes before cryopreservation and after thawing were examined in each group. After the specimens were embedded in paraffin,5-(m-thick sections were stained with hematoxylin and eosin.The epithelium and cartilage was assessed. We also observed Bcl-2 and Bax gene expression by immunohistochemistry. At last, some tracheas(SD) after cryopreservation were thawed and transplanted into the abdominal cavity of Wistar rats. The transplanted tracheas were retrieved and assessed histologically.Results Microscopic findings of the tracheas in Group Ⅲ and Group Ⅳ showed their structure were intact and Bax gene expression was lower in cartilage after cryopreservation(20d) compared with other groups,especially in Group Ⅳ.The tracheas in Group Ⅲ and Group Ⅳ grew well after they were transplanted into cavity of Wistar rats heterotopically,too.There were no significant differences among 4 groups in Bcl-2 gene expression.Conclusion In tracheal cryopreservation the trehalose can protect the trachea by protecting the tracheal cartilage.It is one of the protective mechanism that the trehalose inhibit the Bax gene expression of cartilage cells.The concomitant use of trehalose and DMSO has a synergistic effect.
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Objective To explore Bcl-2 and Bax gene expression in hippocampus region after cerebral ischemia in rats and the modulation of expression by Nimodipine.Methods The cerebral ischemic model of rat was made by occluding left middle cerebral artery according to Nagasawy and Zea Longa improvement method. The rats in one group were pre-treated with Nimodipine. The expression of Bcl-2 and Bax mRNA were measured by RT-PCR method.Results Both Bcl-2 and Bax mRNA were induced in the hippocampus regions after middle cerebral artery occlusion. The Bcl-2 mRNA level was continuously high. However,the level of Bax mRNA increased gradually at first,reached a peak at 24 h,then decreased slowly.For the rats pretreated with Nimodipine Bcl-2 mRNA was up-regulated and Bax mRNA was down-regulated in the hippocampus at 6 and 24 h after ischemia.Conclusion Calcium antagonist can regulate Bcl-2 and Bax genes expression in the hippocampus region after cerebral ischemia.This study indicates that pharmacological modulation of Bcl-2 family member expression may become a new strategy to interfere with neuronal damage.
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To explore the role of apoptosis, apoptosis regulating genes in the pathogenesis and development of smoke inhalation injury. With smoke inhalation injury rat model, the changes int the expression of Bcl 2, Bax, Fas, FasL genes mRNA and protein contents and their relationship with apoptosis of lung tissue cells at different time points after the injury were observed with TUNEL, immunohistochemistry and RT PCR techniques. The results showed that: ①apoptosis indet of lung colls after smoke inhalation injury increased, ②expressions of Bcl 2, Bax, Fas, FasL genes were obviously up regulated in injury group, peaking at the 12th hour, whereas the peak of protein expression was at the 24th hour. Furthermore, a significant correlation was found between the expression of Fas, Fasl, Bax gene and apoptosis indices in lung cells. The results suggested that apoptosis participated in the early pathological process of smoke inhalation injury, and apoptosis regulating genes foot part in the regulation of apoptosis in smoke inhalation injury.