BACKGROUND CYP21A2 gene mutations may all cause reduction or loss of 21-hydroxylase activity,leading to development of congenital adrenal hyperplasia(CAH)with different clinical phenotypes.For families with CAH childr...BACKGROUND CYP21A2 gene mutations may all cause reduction or loss of 21-hydroxylase activity,leading to development of congenital adrenal hyperplasia(CAH)with different clinical phenotypes.For families with CAH children,genetic testing of the parents and genetic counseling are recommended to assess the risk of recurrence.CASE SUMMARY We report a case of CAH with a high suspicion before delivery.The risk of the child suffering from CAH during the pregnancy had been underestimated due to the deviation of genetic counseling and genetic testing results.Our report confirmed a CYP21A2 homozygous deletion in this case,CYP21A2 heterozygous deletion in the mother,and a rare 2+0 CYP21A2 deletion in the father.CONCLUSION It is important to analyze the distribution of CYP21A2 gene in the two alleles of parents of children with CAH.展开更多
AIM To study the correlationship between the changes of p53, Waf1p21 and the cell proliferation determined by PCNA at different stages of human esophageal carcinogenesis. METHODS Biopsied and resected esophageal ti...AIM To study the correlationship between the changes of p53, Waf1p21 and the cell proliferation determined by PCNA at different stages of human esophageal carcinogenesis. METHODS Biopsied and resected esophageal tissues from a high risk population for esophageal cancer in northern China were used in this study. All the specimens were fixed with 85% alcohol and further processed with routine histology. The avidin biotin peroxidase complex (ABC) method was used for the detection of p53, Waf1p21 and PCNA. RESULTS The strong nuclear staining for p53, Waf1p21 and PCNA was observed in the normal esophageal epithelium and the epithelia with different severities of lesions. As the lesions progressed to dysplasia (DYS) and to esophageal squamons cell carcinoma (SCC), the percentage of Waf1p21 immunoreactivity decreased. The number of Waf1p21 immunostaining positive cells increased slightly from normal to basal cell hyperplasia (BCH), but there was no further increase in DYS and in SCC. The total number of positive cells for Waf1p21 stain appeared to be lower than that of p53 in normal and BCH esophageal epithelia and much lower in DYS and SCC. The Waf1p21 positive immunostaining cells were located at the third and forth cell layers in half of the samples examined, which was 2~4 cell layers higher than that of PCNA and p53 in the same histological categories of normal, BCH and DYS. CLNCLUSION The low levels of Waf1p21 at the stage of DYS may be related to a functional loss of p53. Other mechanisms may also be responsible to the lack of Waf1p21 expression in DYS and SCC.展开更多
AIM: To investigate the association between the polymorphism of TBX21 gene and the risk of gastric cancer in a Chinese population. METHODS: The -1993 polymorphism located in TBX21 gene promoter region was identified b...AIM: To investigate the association between the polymorphism of TBX21 gene and the risk of gastric cancer in a Chinese population. METHODS: The -1993 polymorphism located in TBX21 gene promoter region was identified by polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) method. The risk between TBX21 gene genotype and gastric cancer was determined by multivariate logistic regression analysis in 220 gastric cancer patients and 262 cancer-free controls matched by age, sex and ethnicity. RESULTS: Compared with the TBX21 -1993TT genotype, the -1993CC genotype exhibited a significantly elevated risk for gastric cancer [Odds ratio (OR) = 3.42, 95% confidence interval (CI): 1.41-8.31]. The relation-ship between the -1993 polymorphic genotype and the invasive status such as lymph node and distant metastasis was found among the gastric cancer patients (OR = 4.02, 95% CI: 1.87-8.66; OR = 7.02, 95% CI: 3.44-14.34, respectively). CONCLUSION: TBX21 -1993 polymorphism might contribute to the risk of gastric cancer, especially to the distant metastasis.展开更多
Objective: To investigate the p21WAF1 /CIP1gene DNA sequence change and their relationship with the phenotype of human osteosarcoma. Methods: p21WAF1 /CIP1gene DNA of 36 osteosarcoma spec- ...Objective: To investigate the p21WAF1 /CIP1gene DNA sequence change and their relationship with the phenotype of human osteosarcoma. Methods: p21WAF1 /CIP1gene DNA of 36 osteosarcoma spec- imens was examined by using polymerase chain reaction-single strand conformation polymorphism (PCR- SSCP) method. The PCR products were sequenced directly. Results: In p21WAF1 /CIP1 gene exon3 of 36 cases of human osteosarcoma, the change of C→T in the p21WAF1 /CIP1gene CDNA sequence of position 609th occurred in 17 cases with the incidence being 44.4%. In 10 normal blood samples, DNA sequence analysis showed the change of C→T in the p21WAF1 /CIP1gene CDNA sequence of position 609th occurred in 8 cases with the incidence being 80%. Conclusion: The novel location of p21WAF1 /CIP1gene polymorphism of osteosarcoma, but not mutation was de?ned, and this location might provide the meaningful reference for the further research of p21WAF1/CIP1 gene.p2lWAF1/CIP1基因DNA序列分析及其与骨肉瘤表型的关系展开更多
We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constr...We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed by sequentially inserting capsid precursor protein gene(P1) of FMDV and enhanced green fluorescent protein gene(EGFP) into pBABEpuro.The recombinant retroviral vector and the pVSV-G plasmid were co-transfected into packaging cells(GP2-293) by liposomemediated transduction to produce the pseudovirus.The pseudovirus was used to infect BHK-21 cells and resistant cells were screened with puromycin.Green fluorescent proteins were observed by fluorescence microscopy and expression of the capsid precursor protein gene of FMDV was detected by indirect immunofluorescence.The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed successfully.The capsid precursor protein of FMDV and green fluorescent protein were expressed in BHK-21 cells.The mammalian cell expression system for the capsid precursor protein of FMDV has been constructed successfully,which lays the foundation of development of a FMDV subunit vaccine.展开更多
INTRODUCTIONIn order to study the relationship between oncogeneexpression and HCC generation,we observed theprecancerous hepatic GGT loci,IGF-Ⅱ,p53 andp21 expression during hepatocarcinogenesis of treeshrew induced b...INTRODUCTIONIn order to study the relationship between oncogeneexpression and HCC generation,we observed theprecancerous hepatic GGT loci,IGF-Ⅱ,p53 andp21 expression during hepatocarcinogenesis of treeshrew induced by hepatitis B virus (HBV) and/oraflatoxin B1 (AFB1).展开更多
AIM: To construct the retroviral vector of p^125FAK specific ribozyme genes and to explore the feasibility of ribozyme in BGC-823 gene therapy in vitro. METHODS: A hammerhead ribozyme DNA targeting p^125FAK mRNA fro...AIM: To construct the retroviral vector of p^125FAK specific ribozyme genes and to explore the feasibility of ribozyme in BGC-823 gene therapy in vitro. METHODS: A hammerhead ribozyme DNA targeting p^125FAK mRNA from nt 1010 to nt 1032 was synthesized and recombinated into the retroviral vector pLXSN forming pLRZXSN recon. Using the lipofectin-mediated DNA transfection technique, pLRZXSN was introduced into BGC-823 cells. The effects of ribozyme on the growth of BGC-823 cells and apoptosis were studied by cell colony assay, flow cytometry (FCM), reverse transcriptasepolymerase chain reaction (RT-PCR), detection of DNA fragmentation and electron microscopy. RESULTS: The number of BGC-823 cell colonies was inhibited by 56% after the cells were treated for 48 h. The cell proliferation was inhibited effectively by p^125FAK ribozyme and the inhibitory effect depended on the concentration and the time of incubation. The expression of p^125FAK mRNA and protein p^125 decreased sharply in BGC-823 cells treated with p^125FAK ribozyme. The characteristics of apoptosis, namely sub-G1 peak, DNA fragmentation and morphological changes, were revealed in BGC-823 cells treated with p^125FAK ribozyme. CONCLUSION: p^125FAK ribozyme decreases p^125FAK gene expression and induces apoptosis of human gastric cancer cells in vitro.展开更多
Objective: To study the molecular mechanisms of TNF--a expression induced by lipopolysaccharide (LPS) for exploring novel methods to prevent or treat clinical patients with endotoxic shock. Methods:Protein kinase assa...Objective: To study the molecular mechanisms of TNF--a expression induced by lipopolysaccharide (LPS) for exploring novel methods to prevent or treat clinical patients with endotoxic shock. Methods:Protein kinase assay was used to detect the kinase activity stimulated by LPS; Con focal laser scan technique was used to show the translocation of p38 on the activation; RT PCR and reporter gene system were used to study the molecular mechanism of TNF -a gene transcription. Results: In RAW cells it was found that p38 was activated on the stimulation of LPS. and activated p38 moved into nucleus from cytosol; TNF--a mRNA increased on the stimulation of LPS and the increased promoter transactivity induced by LPS could be inhibited significantly by specific inhibitor for p38. Conclusion: p38 mitogen activated protein kinase (MAPK ) was activated by the stimulation of LPS,which brought about its entry to the nucleus to act on transcription factors to regulate cellular processes. p38 MAPK Is an important regulator of TNF--a gene expression induced by LPS.展开更多
BACKGROUND The single nucleotide polymorphisms of interleukin-21(IL-21)gene were confirmed to be related to various diseases,but no studies have examined the possible role of IL-21 single nucleotide polymorphisms(SNPs...BACKGROUND The single nucleotide polymorphisms of interleukin-21(IL-21)gene were confirmed to be related to various diseases,but no studies have examined the possible role of IL-21 single nucleotide polymorphisms(SNPs)(rs907715,rs2221903,and rs12508721)in gastric precancerous lesions.AIM To explore the associations between SNPs of IL-21 gene(rs907715,rs2221903,and rs12508721)and gastric precancerous lesions in a Chinese population.METHODS Three SNPs of IL-21 were genotyped using polymerase chain reaction–ligase detection reaction in 588 cases and 290 healthy controls from May 2013 to December 2016 in northwestern China.Gastric precancerous lesions were confirmed by endoscopic examination and categorized as non-atrophic gastritis,atrophic gastritis,and intestinal metaplasia.Descriptive statistic and logistic regression were used for data analyses.RESULTS IL-21 rs907715 genotype CC and C frequencies were higher in in patients with gastric precancerous lesions than in the controls(OR=1.59,95%CI:1.06-2.38,P=0.013;OR=1.28,95%CI:1.01-2.22,P=0.044,respectively)after adjusting for confounding factors.For SNP rs907715 in intestinal metaplasia patients,significant differences between cases and controls were observed in the frequencies of genotype CC and C(OR=1.92,95%CI:1.24-2.98,P=0.004;OR=1.53,95%CI:1.04-2.24,P=0.028,respectively);for non-atrophic gastritis and atrophic gastritis patients,the CC and C genotypes showed no significant association with risk in all models.No association between either rs2221903 or rs12508721 and gastric precancerous lesions was found in the present study.In the haplotype analysis,the TC haplotype(rs907715 and rs12508721)and TT haplotype(rs2221903 and rs907715)were more frequent in the case group than control group(P<0.05).CONCLUSION Our findings indicate that SNP rs907715 of IL-21 gene is associated with gastric precancerous lesions.The TC haplotype(rs907715 and rs12508721)and TT haplotype(rs2221903 and rs907715)increased the risk of gastric precancerous lesions.If confirmed,these findings will shed light on the etiology of precancerous lesions.展开更多
Objective: We aimed to analysis the HER2 gene status and its relationship with p21 protein expression in gastric carcinoma. Methods: Fluorescence in situ hybridisation (FISH) and immunohistochemistry (IHC) techn...Objective: We aimed to analysis the HER2 gene status and its relationship with p21 protein expression in gastric carcinoma. Methods: Fluorescence in situ hybridisation (FISH) and immunohistochemistry (IHC) techniques were used to detect HER2 gene status and p53 protein in 59 cases of gastric cancer. Results: FISH detection of HER2 gene amplification rate was 16.9% (10/59), HER2 gene amplification in 49 cases without copy number gain and gene amplification were a total of 49.2% (29/59). HER2 protein expression was 42.4% (25/59), HER2 gene amplification rates in patients with +++, ++ HER2 protein expression were 3/3 and 5/8, while in patients with + HER2 protein expression, it was 2/14, there was significant difference (P 0.05). p21 protein expression rate was 49.2% (29/59), HER2 gene amplification rates and p21 protein expression had significant difference in tumor invasion depth, lymph node metastasis (P 0.05); had no statistical significance in histological type, age, gender differences (P 0.05). Conclusion: HER2 gene amplification rate and gene copy number had positively correlation with p21 protein expression, HER2 gene status and expression of p21 protein combined detection can provide a reference value in gastric cancer metastasis, patient’s condition development and prognosis, it also can guide clinical development of individual treatment.展开更多
The quantitative resistance gene pi21 from Sensho was introgressed to an indica breeding line IR63307-4B-13-2,a pyramiding line IRBB4/5/13/21,and a tropical japonica line Kinandang Patong by marker-assisted backcrossi...The quantitative resistance gene pi21 from Sensho was introgressed to an indica breeding line IR63307-4B-13-2,a pyramiding line IRBB4/5/13/21,and a tropical japonica line Kinandang Patong by marker-assisted backcrossing.A total of 192 improved lines at the BC4F3 and BC4F4 generations were developed and confirmed to have the gene introgression via genotyping using a pi21-specific InDel marker.Thirteen randomly selected improved lines,representing all the three genetic backgrounds,demonstrated resistance against leaf blast composites in the field and a broader spectrum resistance against individual isolates compared to the recurrent parents in the glasshouse.Specifically,the tested lines exhibited pi21-acquired resistance against 11 leaf blast isolates that elicited susceptible reactions from the recurrent parents.All the tested lines maintained a comparative heading date,and similar or improved panicle length,number of primary branches per panicle and number of total grains per panicle relative to the recurrent parents.The physical grain characteristics of the recurrent parents were also maintained in the 13 lines tested,although variability in the amylose content and chalkiness degree was observed.The successful marker-assisted introgression of pi21 in diverse genetic backgrounds and the resulting broader spectrum resistance of improved lines against leaf blast indicate the potential of pi21 for deployment in cultivars grown across other rice growing regions in Asia.展开更多
Objective To study Chinese children with congenital adrenal hyperplasia (CAH) in the types ofpoint mutation in steroid 21 - hydroxylase gene (CYP21). Methods By using PCR-ASO hybridization analysiswith amplified segme...Objective To study Chinese children with congenital adrenal hyperplasia (CAH) in the types ofpoint mutation in steroid 21 - hydroxylase gene (CYP21). Methods By using PCR-ASO hybridization analysiswith amplified segments involving exon 3-4 and exon 6-8 of the gene to investigate for the type ofmutations. Results The results showed among 5 point mutations detected positive findings being in 28/66 (42%)of CAH chromosomes, or 17/33 (52%) of the CAH cases examined. The only 1 non - classic form CAH case wasfound as homonsous for val - 281→len mutation. Three classic cases were heterozygous for compound mutations asnitron 2 A, C→G associated with lie-172→Asn or lie - 172→Asn with Gin-318→stop. The other point mutationsall revealed as homozygous alleies with the most freguent matations as nitron 2A, C→G. No PCR product wereprovided by 3 cases who had been verilied by Southern blotting with CYP21 B gene deletion (not shown). NO pointmutations were illustrated in normal controls. Conclusion This report presentS data in 17/33 (52K) of Chinesechildren with CAH in CYP 21 B gene had point mutations documenting the type and location of mutation indiagnosis of CAH.展开更多
AIM: To investigate the inhibitory effect of tumor suppressor p33ING1b and its synergy with p53 gene in hepatocellular carcinoma (HCC).METHODS: Recombinant sense and antisense p33ING1b plasmids were transfected into h...AIM: To investigate the inhibitory effect of tumor suppressor p33ING1b and its synergy with p53 gene in hepatocellular carcinoma (HCC).METHODS: Recombinant sense and antisense p33ING1b plasmids were transfected into hepatoma cell line HepG2 with lipofectamine. Apoptosis, G0/G1 arrest, cell growth rate and cloning efficiency in soft agar of HepG2 were analyzed after transfection. In three hepatoma cell lineswith different endogenous p53 gene expressions, the synergistic effect of p33ING1b with p53 was analyzed by flow cytometry and luciferase assay was performed to detect the activation of p53 downstream gene p21WAF1/CIP1. In addition, the expression and mutation rates of p33ING1b in HCC tissues were measured by immunohistochemistry and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP).RESULTS: Overexpression of p33ING1b inhibited cell growth of HepG2, induced more apoptosis and protected cells from growth in soft agar. Combined transfer of p33ING1b and p53 gene promoted hepatoma cell apoptosis, G0/G1 arrest and elevated expression of p21WAF1/CIP1. Immunostaining results showed co-localized P33ING1b with P53 protein in HCC tissues and there was a significant relation between protein expression rates of these two genes (P<0.01).Among 28 HCC samples, p33ING1b presented a low gene mutation rate (7.1%).CONCLUSION: p33ING1b collaborates with p53 in cell growth inhibition, cell cycle arrest and apoptosis in HCC. Loss or inactivation of p33ING1b normal function may be an important mechanism for the development of HCC retaining wildtype p53.展开更多
Formation of apoptotic bodies is a typical character ofaPoptotic cell death, but how the processes are controlledis not known. In this study, we compared two apoptosisinducing systems in vascular endothelial cells (VE...Formation of apoptotic bodies is a typical character ofaPoptotic cell death, but how the processes are controlledis not known. In this study, we compared two apoptosisinducing systems in vascular endothelial cells (VEC). Wefound that the formation of aPoptotic bodies during apop-tosis induced by rattlesnake venom, which is an unique andspecific aPoptosis inducer to vascular endotheliaI cells, wasmuch faster than that induced by deprivation of survivalfactors (aFGF and serum). When we blocked the synthesisof mRNAs in cells treated with rattlesnake venom by DRB(5, 6- dichloro- 1 -β- D- rib ofur anosylb enzimidazole ), an in-hibitor of transcription, the formation of aPoptotic bodieswas dramatically inhibited. We examined the expressionof Psa gene and found that its expression was much higherin apoptosis induced by rattlesnake venom than that inaPoptosis induced by deprivation of aFGF and serum. Ourresults suggest that gene expression is important and P53gene may play a major role in inducing the formation ofapoptotic bodies in VEC.展开更多
By southern hybridization with 1. 8 kb cDNA probe,a high frequency (40. 5 % ) of structural abnor- mality of p53 gene was observed in primary nasopharyngeal carcinoma (NPC) biopsies. The regions of ex- ons 1 to 4 ...By southern hybridization with 1. 8 kb cDNA probe,a high frequency (40. 5 % ) of structural abnor- mality of p53 gene was observed in primary nasopharyngeal carcinoma (NPC) biopsies. The regions of ex- ons 1 to 4 of the gene were examined by polymerase chain reaction-single strand con formation polymor- phism,no point mutation was found. Because very low rate of point mutation had been reported in exons 5 to 8, we considered that structural abnormality in the region of e-cons 1 to 8 of the gene might be uncom- mon in NPC. The spectrophotometer scanning analysis of autoradiograms and rehybridization investigation of nitrocellulose filter with exon 11 probe indicated that most of structure aberrations we observed might be rearrangement occurring in exon 11.展开更多
Objective The radiation sensitive gene rad 21 of Schizosaccharomyces pombe is involved in the repair of double-stranded breaks in DNA and is essential for mitotic growth. The hHR21 sp g...Objective The radiation sensitive gene rad 21 of Schizosaccharomyces pombe is involved in the repair of double-stranded breaks in DNA and is essential for mitotic growth. The hHR21 sp gene is its human homologue. In an attempt to investigate the role of hHR21 sp in DNA repair, we studied the effects of UV and γ-ray irradiation on hHR21 sp gene expression in normal human peripheral blood cells, and non-iradiated peripheral and bone marrow cells from Fanconi anemia (FA) patients who have shown DNA repair deficiency.Methods Total steady state RNA was extracted from peripheral blood cells and bone marrow. RNA transcripts were quantified after RT-PCR and Southern blot, phosphoimmage and autoradiogram analysis. The results were compared with control groups. Results hHR21 sp expression was significantly increased from 3?h to 9?h after UV irradiation in peripheral blood cells from normal subjects at doses of 40-80?j/m 2 (P<0.05). hHR21 sp was also up-regulated by γ-ray irradiation at 6?h to 9?h at dose of 1 to 5?Gy (P<0.01), which was more significant than the UV irradiation. In the non-irradiated FA patient group, hHR21 sp expression was decreased in bone marrow hematopoietic cells (P<0.05). After activation by PHA and IL-2, there was still a significant depression in expression by the FA patients peripheral blood cells compared with control groups (P<0.05). Conclusion hHR21 sp was up-regulated at doses and times irradiated at the range tested in normal peripheral blood cells, and is more affected by γ-ray irradiation than UV irradiation. FA patient bone marrow hematopoietic cells and peripheral blood mononuclear cells showed down-regulation of hHR21 sp expression. The results imply that defects in DNA repair via hHR21 sp expression may play an important role in the pathogenesis of FA syndrome.展开更多
基金Supported by the Fundamental Research Funds of Health Commission of Sichuan Province,No.17ZD035.
文摘BACKGROUND CYP21A2 gene mutations may all cause reduction or loss of 21-hydroxylase activity,leading to development of congenital adrenal hyperplasia(CAH)with different clinical phenotypes.For families with CAH children,genetic testing of the parents and genetic counseling are recommended to assess the risk of recurrence.CASE SUMMARY We report a case of CAH with a high suspicion before delivery.The risk of the child suffering from CAH during the pregnancy had been underestimated due to the deviation of genetic counseling and genetic testing results.Our report confirmed a CYP21A2 homozygous deletion in this case,CYP21A2 heterozygous deletion in the mother,and a rare 2+0 CYP21A2 deletion in the father.CONCLUSION It is important to analyze the distribution of CYP21A2 gene in the two alleles of parents of children with CAH.
文摘AIM To study the correlationship between the changes of p53, Waf1p21 and the cell proliferation determined by PCNA at different stages of human esophageal carcinogenesis. METHODS Biopsied and resected esophageal tissues from a high risk population for esophageal cancer in northern China were used in this study. All the specimens were fixed with 85% alcohol and further processed with routine histology. The avidin biotin peroxidase complex (ABC) method was used for the detection of p53, Waf1p21 and PCNA. RESULTS The strong nuclear staining for p53, Waf1p21 and PCNA was observed in the normal esophageal epithelium and the epithelia with different severities of lesions. As the lesions progressed to dysplasia (DYS) and to esophageal squamons cell carcinoma (SCC), the percentage of Waf1p21 immunoreactivity decreased. The number of Waf1p21 immunostaining positive cells increased slightly from normal to basal cell hyperplasia (BCH), but there was no further increase in DYS and in SCC. The total number of positive cells for Waf1p21 stain appeared to be lower than that of p53 in normal and BCH esophageal epithelia and much lower in DYS and SCC. The Waf1p21 positive immunostaining cells were located at the third and forth cell layers in half of the samples examined, which was 2~4 cell layers higher than that of PCNA and p53 in the same histological categories of normal, BCH and DYS. CLNCLUSION The low levels of Waf1p21 at the stage of DYS may be related to a functional loss of p53. Other mechanisms may also be responsible to the lack of Waf1p21 expression in DYS and SCC.
基金Supported by National Natural Science Foundation, No.81000187
文摘AIM: To investigate the association between the polymorphism of TBX21 gene and the risk of gastric cancer in a Chinese population. METHODS: The -1993 polymorphism located in TBX21 gene promoter region was identified by polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) method. The risk between TBX21 gene genotype and gastric cancer was determined by multivariate logistic regression analysis in 220 gastric cancer patients and 262 cancer-free controls matched by age, sex and ethnicity. RESULTS: Compared with the TBX21 -1993TT genotype, the -1993CC genotype exhibited a significantly elevated risk for gastric cancer [Odds ratio (OR) = 3.42, 95% confidence interval (CI): 1.41-8.31]. The relation-ship between the -1993 polymorphic genotype and the invasive status such as lymph node and distant metastasis was found among the gastric cancer patients (OR = 4.02, 95% CI: 1.87-8.66; OR = 7.02, 95% CI: 3.44-14.34, respectively). CONCLUSION: TBX21 -1993 polymorphism might contribute to the risk of gastric cancer, especially to the distant metastasis.
文摘Objective: To investigate the p21WAF1 /CIP1gene DNA sequence change and their relationship with the phenotype of human osteosarcoma. Methods: p21WAF1 /CIP1gene DNA of 36 osteosarcoma spec- imens was examined by using polymerase chain reaction-single strand conformation polymorphism (PCR- SSCP) method. The PCR products were sequenced directly. Results: In p21WAF1 /CIP1 gene exon3 of 36 cases of human osteosarcoma, the change of C→T in the p21WAF1 /CIP1gene CDNA sequence of position 609th occurred in 17 cases with the incidence being 44.4%. In 10 normal blood samples, DNA sequence analysis showed the change of C→T in the p21WAF1 /CIP1gene CDNA sequence of position 609th occurred in 8 cases with the incidence being 80%. Conclusion: The novel location of p21WAF1 /CIP1gene polymorphism of osteosarcoma, but not mutation was de?ned, and this location might provide the meaningful reference for the further research of p21WAF1/CIP1 gene.p2lWAF1/CIP1基因DNA序列分析及其与骨肉瘤表型的关系
基金supported by the National Programs for High Technology Research and Development of China (2006AA10A204)the Gansu Key Technologies R&D Program(ZGS-052-A41-0006-03)the Programs for Director Fund of Lanzhou Veterinary Research Institute
文摘We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed by sequentially inserting capsid precursor protein gene(P1) of FMDV and enhanced green fluorescent protein gene(EGFP) into pBABEpuro.The recombinant retroviral vector and the pVSV-G plasmid were co-transfected into packaging cells(GP2-293) by liposomemediated transduction to produce the pseudovirus.The pseudovirus was used to infect BHK-21 cells and resistant cells were screened with puromycin.Green fluorescent proteins were observed by fluorescence microscopy and expression of the capsid precursor protein gene of FMDV was detected by indirect immunofluorescence.The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed successfully.The capsid precursor protein of FMDV and green fluorescent protein were expressed in BHK-21 cells.The mammalian cell expression system for the capsid precursor protein of FMDV has been constructed successfully,which lays the foundation of development of a FMDV subunit vaccine.
基金the National Natural Science Foundation of China,No.39260033.
文摘INTRODUCTIONIn order to study the relationship between oncogeneexpression and HCC generation,we observed theprecancerous hepatic GGT loci,IGF-Ⅱ,p53 andp21 expression during hepatocarcinogenesis of treeshrew induced by hepatitis B virus (HBV) and/oraflatoxin B1 (AFB1).
基金Supported by the Natural Science Foundation of Fujian Province,No.C0010015
文摘AIM: To construct the retroviral vector of p^125FAK specific ribozyme genes and to explore the feasibility of ribozyme in BGC-823 gene therapy in vitro. METHODS: A hammerhead ribozyme DNA targeting p^125FAK mRNA from nt 1010 to nt 1032 was synthesized and recombinated into the retroviral vector pLXSN forming pLRZXSN recon. Using the lipofectin-mediated DNA transfection technique, pLRZXSN was introduced into BGC-823 cells. The effects of ribozyme on the growth of BGC-823 cells and apoptosis were studied by cell colony assay, flow cytometry (FCM), reverse transcriptasepolymerase chain reaction (RT-PCR), detection of DNA fragmentation and electron microscopy. RESULTS: The number of BGC-823 cell colonies was inhibited by 56% after the cells were treated for 48 h. The cell proliferation was inhibited effectively by p^125FAK ribozyme and the inhibitory effect depended on the concentration and the time of incubation. The expression of p^125FAK mRNA and protein p^125 decreased sharply in BGC-823 cells treated with p^125FAK ribozyme. The characteristics of apoptosis, namely sub-G1 peak, DNA fragmentation and morphological changes, were revealed in BGC-823 cells treated with p^125FAK ribozyme. CONCLUSION: p^125FAK ribozyme decreases p^125FAK gene expression and induces apoptosis of human gastric cancer cells in vitro.
文摘Objective: To study the molecular mechanisms of TNF--a expression induced by lipopolysaccharide (LPS) for exploring novel methods to prevent or treat clinical patients with endotoxic shock. Methods:Protein kinase assay was used to detect the kinase activity stimulated by LPS; Con focal laser scan technique was used to show the translocation of p38 on the activation; RT PCR and reporter gene system were used to study the molecular mechanism of TNF -a gene transcription. Results: In RAW cells it was found that p38 was activated on the stimulation of LPS. and activated p38 moved into nucleus from cytosol; TNF--a mRNA increased on the stimulation of LPS and the increased promoter transactivity induced by LPS could be inhibited significantly by specific inhibitor for p38. Conclusion: p38 mitogen activated protein kinase (MAPK ) was activated by the stimulation of LPS,which brought about its entry to the nucleus to act on transcription factors to regulate cellular processes. p38 MAPK Is an important regulator of TNF--a gene expression induced by LPS.
基金Supported by the Fundamental Research Funds for the Central Universities,No.zdyf2017007Project of Shaanxi Health and Culture Research Center,No.JKWH2019-Z02
文摘BACKGROUND The single nucleotide polymorphisms of interleukin-21(IL-21)gene were confirmed to be related to various diseases,but no studies have examined the possible role of IL-21 single nucleotide polymorphisms(SNPs)(rs907715,rs2221903,and rs12508721)in gastric precancerous lesions.AIM To explore the associations between SNPs of IL-21 gene(rs907715,rs2221903,and rs12508721)and gastric precancerous lesions in a Chinese population.METHODS Three SNPs of IL-21 were genotyped using polymerase chain reaction–ligase detection reaction in 588 cases and 290 healthy controls from May 2013 to December 2016 in northwestern China.Gastric precancerous lesions were confirmed by endoscopic examination and categorized as non-atrophic gastritis,atrophic gastritis,and intestinal metaplasia.Descriptive statistic and logistic regression were used for data analyses.RESULTS IL-21 rs907715 genotype CC and C frequencies were higher in in patients with gastric precancerous lesions than in the controls(OR=1.59,95%CI:1.06-2.38,P=0.013;OR=1.28,95%CI:1.01-2.22,P=0.044,respectively)after adjusting for confounding factors.For SNP rs907715 in intestinal metaplasia patients,significant differences between cases and controls were observed in the frequencies of genotype CC and C(OR=1.92,95%CI:1.24-2.98,P=0.004;OR=1.53,95%CI:1.04-2.24,P=0.028,respectively);for non-atrophic gastritis and atrophic gastritis patients,the CC and C genotypes showed no significant association with risk in all models.No association between either rs2221903 or rs12508721 and gastric precancerous lesions was found in the present study.In the haplotype analysis,the TC haplotype(rs907715 and rs12508721)and TT haplotype(rs2221903 and rs907715)were more frequent in the case group than control group(P<0.05).CONCLUSION Our findings indicate that SNP rs907715 of IL-21 gene is associated with gastric precancerous lesions.The TC haplotype(rs907715 and rs12508721)and TT haplotype(rs2221903 and rs907715)increased the risk of gastric precancerous lesions.If confirmed,these findings will shed light on the etiology of precancerous lesions.
文摘Objective: We aimed to analysis the HER2 gene status and its relationship with p21 protein expression in gastric carcinoma. Methods: Fluorescence in situ hybridisation (FISH) and immunohistochemistry (IHC) techniques were used to detect HER2 gene status and p53 protein in 59 cases of gastric cancer. Results: FISH detection of HER2 gene amplification rate was 16.9% (10/59), HER2 gene amplification in 49 cases without copy number gain and gene amplification were a total of 49.2% (29/59). HER2 protein expression was 42.4% (25/59), HER2 gene amplification rates in patients with +++, ++ HER2 protein expression were 3/3 and 5/8, while in patients with + HER2 protein expression, it was 2/14, there was significant difference (P 0.05). p21 protein expression rate was 49.2% (29/59), HER2 gene amplification rates and p21 protein expression had significant difference in tumor invasion depth, lymph node metastasis (P 0.05); had no statistical significance in histological type, age, gender differences (P 0.05). Conclusion: HER2 gene amplification rate and gene copy number had positively correlation with p21 protein expression, HER2 gene status and expression of p21 protein combined detection can provide a reference value in gastric cancer metastasis, patient’s condition development and prognosis, it also can guide clinical development of individual treatment.
基金supported by the Japan International Cooperation Agency as part of the Wonder Rice Initiative for Food Security and Health (WISH) Projectin part by the Canon Foundation and the SATREPS project entitled ‘Improvement in Productivity and Yield Stability of Rice under Kenya’s Biotic and Abiotic Stress Conditions Through Tailor-made Breeding and Development of Cultivation Methods’.
文摘The quantitative resistance gene pi21 from Sensho was introgressed to an indica breeding line IR63307-4B-13-2,a pyramiding line IRBB4/5/13/21,and a tropical japonica line Kinandang Patong by marker-assisted backcrossing.A total of 192 improved lines at the BC4F3 and BC4F4 generations were developed and confirmed to have the gene introgression via genotyping using a pi21-specific InDel marker.Thirteen randomly selected improved lines,representing all the three genetic backgrounds,demonstrated resistance against leaf blast composites in the field and a broader spectrum resistance against individual isolates compared to the recurrent parents in the glasshouse.Specifically,the tested lines exhibited pi21-acquired resistance against 11 leaf blast isolates that elicited susceptible reactions from the recurrent parents.All the tested lines maintained a comparative heading date,and similar or improved panicle length,number of primary branches per panicle and number of total grains per panicle relative to the recurrent parents.The physical grain characteristics of the recurrent parents were also maintained in the 13 lines tested,although variability in the amylose content and chalkiness degree was observed.The successful marker-assisted introgression of pi21 in diverse genetic backgrounds and the resulting broader spectrum resistance of improved lines against leaf blast indicate the potential of pi21 for deployment in cultivars grown across other rice growing regions in Asia.
文摘Objective To study Chinese children with congenital adrenal hyperplasia (CAH) in the types ofpoint mutation in steroid 21 - hydroxylase gene (CYP21). Methods By using PCR-ASO hybridization analysiswith amplified segments involving exon 3-4 and exon 6-8 of the gene to investigate for the type ofmutations. Results The results showed among 5 point mutations detected positive findings being in 28/66 (42%)of CAH chromosomes, or 17/33 (52%) of the CAH cases examined. The only 1 non - classic form CAH case wasfound as homonsous for val - 281→len mutation. Three classic cases were heterozygous for compound mutations asnitron 2 A, C→G associated with lie-172→Asn or lie - 172→Asn with Gin-318→stop. The other point mutationsall revealed as homozygous alleies with the most freguent matations as nitron 2A, C→G. No PCR product wereprovided by 3 cases who had been verilied by Southern blotting with CYP21 B gene deletion (not shown). NO pointmutations were illustrated in normal controls. Conclusion This report presentS data in 17/33 (52K) of Chinesechildren with CAH in CYP 21 B gene had point mutations documenting the type and location of mutation indiagnosis of CAH.
基金Supported by the National Natural Science Foundation of China (Grants No.30070344 and No.30070839)
文摘AIM: To investigate the inhibitory effect of tumor suppressor p33ING1b and its synergy with p53 gene in hepatocellular carcinoma (HCC).METHODS: Recombinant sense and antisense p33ING1b plasmids were transfected into hepatoma cell line HepG2 with lipofectamine. Apoptosis, G0/G1 arrest, cell growth rate and cloning efficiency in soft agar of HepG2 were analyzed after transfection. In three hepatoma cell lineswith different endogenous p53 gene expressions, the synergistic effect of p33ING1b with p53 was analyzed by flow cytometry and luciferase assay was performed to detect the activation of p53 downstream gene p21WAF1/CIP1. In addition, the expression and mutation rates of p33ING1b in HCC tissues were measured by immunohistochemistry and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP).RESULTS: Overexpression of p33ING1b inhibited cell growth of HepG2, induced more apoptosis and protected cells from growth in soft agar. Combined transfer of p33ING1b and p53 gene promoted hepatoma cell apoptosis, G0/G1 arrest and elevated expression of p21WAF1/CIP1. Immunostaining results showed co-localized P33ING1b with P53 protein in HCC tissues and there was a significant relation between protein expression rates of these two genes (P<0.01).Among 28 HCC samples, p33ING1b presented a low gene mutation rate (7.1%).CONCLUSION: p33ING1b collaborates with p53 in cell growth inhibition, cell cycle arrest and apoptosis in HCC. Loss or inactivation of p33ING1b normal function may be an important mechanism for the development of HCC retaining wildtype p53.
文摘Formation of apoptotic bodies is a typical character ofaPoptotic cell death, but how the processes are controlledis not known. In this study, we compared two apoptosisinducing systems in vascular endothelial cells (VEC). Wefound that the formation of aPoptotic bodies during apop-tosis induced by rattlesnake venom, which is an unique andspecific aPoptosis inducer to vascular endotheliaI cells, wasmuch faster than that induced by deprivation of survivalfactors (aFGF and serum). When we blocked the synthesisof mRNAs in cells treated with rattlesnake venom by DRB(5, 6- dichloro- 1 -β- D- rib ofur anosylb enzimidazole ), an in-hibitor of transcription, the formation of aPoptotic bodieswas dramatically inhibited. We examined the expressionof Psa gene and found that its expression was much higherin apoptosis induced by rattlesnake venom than that inaPoptosis induced by deprivation of aFGF and serum. Ourresults suggest that gene expression is important and P53gene may play a major role in inducing the formation ofapoptotic bodies in VEC.
文摘By southern hybridization with 1. 8 kb cDNA probe,a high frequency (40. 5 % ) of structural abnor- mality of p53 gene was observed in primary nasopharyngeal carcinoma (NPC) biopsies. The regions of ex- ons 1 to 4 of the gene were examined by polymerase chain reaction-single strand con formation polymor- phism,no point mutation was found. Because very low rate of point mutation had been reported in exons 5 to 8, we considered that structural abnormality in the region of e-cons 1 to 8 of the gene might be uncom- mon in NPC. The spectrophotometer scanning analysis of autoradiograms and rehybridization investigation of nitrocellulose filter with exon 11 probe indicated that most of structure aberrations we observed might be rearrangement occurring in exon 11.
文摘Objective The radiation sensitive gene rad 21 of Schizosaccharomyces pombe is involved in the repair of double-stranded breaks in DNA and is essential for mitotic growth. The hHR21 sp gene is its human homologue. In an attempt to investigate the role of hHR21 sp in DNA repair, we studied the effects of UV and γ-ray irradiation on hHR21 sp gene expression in normal human peripheral blood cells, and non-iradiated peripheral and bone marrow cells from Fanconi anemia (FA) patients who have shown DNA repair deficiency.Methods Total steady state RNA was extracted from peripheral blood cells and bone marrow. RNA transcripts were quantified after RT-PCR and Southern blot, phosphoimmage and autoradiogram analysis. The results were compared with control groups. Results hHR21 sp expression was significantly increased from 3?h to 9?h after UV irradiation in peripheral blood cells from normal subjects at doses of 40-80?j/m 2 (P<0.05). hHR21 sp was also up-regulated by γ-ray irradiation at 6?h to 9?h at dose of 1 to 5?Gy (P<0.01), which was more significant than the UV irradiation. In the non-irradiated FA patient group, hHR21 sp expression was decreased in bone marrow hematopoietic cells (P<0.05). After activation by PHA and IL-2, there was still a significant depression in expression by the FA patients peripheral blood cells compared with control groups (P<0.05). Conclusion hHR21 sp was up-regulated at doses and times irradiated at the range tested in normal peripheral blood cells, and is more affected by γ-ray irradiation than UV irradiation. FA patient bone marrow hematopoietic cells and peripheral blood mononuclear cells showed down-regulation of hHR21 sp expression. The results imply that defects in DNA repair via hHR21 sp expression may play an important role in the pathogenesis of FA syndrome.