BACKGROUND: The mortality rate of heavy type hepatitisis high. No special treatment is available except generaltreatment. This multicenter clinical study was designed toobserve the safety and efficacy of promoting hep...BACKGROUND: The mortality rate of heavy type hepatitisis high. No special treatment is available except generaltreatment. This multicenter clinical study was designed toobserve the safety and efficacy of promoting hepatic growthfactor (PHGF) in the treatment of heavy type hepatitis andsevere chronic hepatitis.METHODS: 347 patients with heavy type hepatitis and 324with severe chronic hepatitis were subjected to administra-tion of 120 μg of PHGF per day for 4 weeks on the basis ofgeneral treatment. Those who were being effectively treat-ed would last additional 2 to 4 weeks. Blood routine, urineroutine, blood urea nitrogen (BUN), blood creatinine(Cr), blood ammonia, alpha fetoprotein (AFP), electro-lyte, alanine transaminase (ALT), aspartate transaminase(AST), serum total bilirubin (TBIL), serum direct biliru-bin (DBIL), prothrombin time activity (PTA), total pro-tein (TP) and albumin (ALB) were detected in the pa-tients before treatment, 2 weeks after treatment, and at theend of the treatment. Any side-effect would be recorded.RESULTS: In the patients with severe chronic hepatitis, thetotal effective rate of the treatment was 88. 9% The levelsof ALT, AST and TBIL decreased significantly (P<0.001),whereas those of PTA and ALB increased significantly (P <0.001), and the level of AFP increased slightly. In patientswith heavy type hepatitis, the total effective rate of thistreatment was 78.4%, and patients at different stage showeddifferent results. The total effective rates of patients withearly, medium and terminal stage heavy type hepatitis were89.9%, 84.8% and 27.5%, respectively. No severe side-effect was shown.CONCLUSION: PHGF is effective and safe in the treat-ment of patients with heavy type hepatitis and severe chro-nic hepatitis. But it should be administered early in patientswith heavy type hepatitis so as to get better curative effects.展开更多
BACKGROUND Liver cancer has a high mortality and morbidity rate throughout the world.In clinical practice,the prognosis of liver cancer patients is poor,and the complex reasons contribute to treatment failures,includi...BACKGROUND Liver cancer has a high mortality and morbidity rate throughout the world.In clinical practice,the prognosis of liver cancer patients is poor,and the complex reasons contribute to treatment failures,including fibrosis,hepatitis viral infection,drug resistance and metastasis.Thus,screening novel prognostic biomarkers is of great importance for guiding liver cancer therapy.Orosomucoid genes(ORMs)encode acute phase plasma proteins,including orosomucoid 1(ORM1)and ORM2.Previous studies showed their upregulation upon inflammation,but the specific function of ORMs has not yet been determined,especially in the development of liver cancer.AIM To determine the expression of ORMs and their potential function in liver cancer.METHODS Analysis of the expression of ORMs in different human tissues was performed on data from the HPA RNA-seq normal tissues project.The expression ratio of ORMs was determined using the HCCDB database,including the ratio between liver cancer and other cancers,normal liver and other normal tissues,liver cancer and adjacent normal liver tissues.Analysis of ORM expression in different cancer types was performed using The Cancer Genome Atlas and TIMER database.The expression of ORMs in liver tumor tissues and adjacent normal tissues were further confirmed using Gene Expression Omnibus data,including GSE36376 and GSE14520.The 10-year overall survival(OS),progression-free survival(PFS)and relapse-free survival(RFS)rates between high and low ORM expression groups in liver cancer patients were determined using the Kaplan-Meier plotter tool.Gene Set Enrichment Analysis(GSEA)was employed to explore the ORM2-associated signaling network.Correlations between ORM2 expression and tumor purity or the infiltration level of macrophages in liver tumor tissues were determined using the TIMER database.The correlation between ORM2 gene levels,tumor-associated macrophage(TAM)markers(including CD68 and TGFβ1)and T cell immunosuppression(including CTLA4 and PD-1)in liver tumor tissues and liver GTEx was determined using the GEPIA database.RESULTS ORM1 and ORM2 were highly expressed in normal liver and liver tumor tissues.ORM1 and ORM2 expression was significantly decreased in liver tumor tissues compared with adjacent normal tissues,and similar results were also noted in cholangiocarcinoma,esophageal carcinoma,and lung squamous cell carcinoma.Further analysis of the Gene Expression Omnibus Database also confirmed the downregulation of ORM1 and ORM2 in liver tumors.Survival analysis showed that the high ORM2 group had better survival rates in OS,PFS and RFS.ORM1 only represented better performance in PFS,but not in OS or RFS.GSEA analysis of ORM2 from The Cancer Genome Atlas liver cancer data identified that ORM2 positively associated with the G2/M checkpoint,E2F target signaling,as well as Wnt/β-catenin and Hedgehog signaling.Moreover,apoptosis,IFN-αresponses,IFN-γresponses and humoral immune responses were upregulated in the ORM2 high group.ORM2 expression was negatively correlated with the macrophage infiltration level,CD68,TGFβ1,CTLA4 and PD-1 levels.CONCLUSION The results showed that ORM1 and ORM2 were highly expressed specifically in liver tissues,whereas ORM1 and ORM2 were downregulated in liver tumor tissues.ORM2 is a better prognostic factor for liver cancer.Furthermore,ORM2 is closely associated with cancer-promoting pathways.展开更多
This paper takes 30 hm2 wheat sprinkling irrigation land of Yunguo Family Farm in Yanzhou, Shandong Province, China as a sample, and draws a conclusion by comparing it with other households (including individual and l...This paper takes 30 hm2 wheat sprinkling irrigation land of Yunguo Family Farm in Yanzhou, Shandong Province, China as a sample, and draws a conclusion by comparing it with other households (including individual and large households). The conclusions are as follows: wheat sprinkling irrigation has a remarkable water-saving effect and comparative revenue. Water saving efficiency can reach 61.54% and a comprehensive income-increase rate can reach 38.67%. The main factors of increasing income and incentives of saving irrigation by sprinkling irrigation ranks as the following: saving land consolidation and water monitoring labors (accounts for 62.50%), saving land area of wheat bed to increase production and income (accounts for 23.44%), saving water bills (accounts for 14.06%). The incentive effect of water saving is not obvious mainly because the water price is low. The main obstacles to the promotion of sprinkling irrigation by individual household are the uneconomical scale and the barriers of coordination of proxy irrigation. Other large household’s (family farm) obstacles are mainly the instability of land tenure and mixed management. Suggestions on promoting sprinkling irrigation in wheat field: Accelerate land circulation and promote agricultural scale management to create basic scale conditions for spreading sprinkling irrigation;stabilizing farmland management rights as stabilizing farmland contractual rights, thus giving long-term business interests to all kinds of large household owners;guide the development of “scale + specialization” modern family farms;appropriate water saving subsidies should be given according to the positive externality of household water saving;confirm agricultural water rights to household and allow compensated transfer of “surplus water rights”.展开更多
Transcription factors, which represent an important class of proteins that play key roles in controlling cellular proliferation and cell cycle modulation, are attractive targets for cancer therapy. Previous researches...Transcription factors, which represent an important class of proteins that play key roles in controlling cellular proliferation and cell cycle modulation, are attractive targets for cancer therapy. Previous researches have shown that the expression level of activating transcription factor 5 （ATF5） was frequently increased in glioma and its acetylation level was related to glioma. The purposes of this study were to explore the methylation level of ATF5 in clinical glioma tissues and to explore the effect of ATF5 methylation on the expression of ATF5 in glioma. Methylation of the promoter region of ATF5 was assayed by bisulflte-specific polymerase chain reaction （PCR） sequencing analysis in 35 cases of glioma and 5 normal tissues. Quantitative real-time PCR （qRT-PCR） was also performed to detect ATF5 mRNA expression in 35 cases of glioma and 5 normal tissues. Clinical data were collected from the patients and analyzed. The percentages of methylation of the ATF5 gene in the promoter region in healthy control, patients with well-differentiated glioma, and those with poorly differentiated glioma were 87.78%, 73.89%, and 47.70%, respectively. Analysis of the methylation status of the promoter region of the ATF5 gene showed a gradually de- creased methylation level in poorly differentiated glioma, well-differentiated glioma, and normal tissues （P〈0.05）. There was also a significant difference between well-differentiated glioma and poorly differentiated glioma （P〈0.05）. ATF5 mRNA expression in glioma was significantly higher than that in the normal tissues （P〈0.05）. This study provides the first evidence that the methylation level of ATF5 decreased, and its mRNA expression was evidently up-regulated in glioma.展开更多
Amh (anti-Müllerian hormone) is a single copy gene which is expressed strongly in Sertoli cells in the foetal testis and participates in the onset of sexual differentiation. Its promoter driving the expression of...Amh (anti-Müllerian hormone) is a single copy gene which is expressed strongly in Sertoli cells in the foetal testis and participates in the onset of sexual differentiation. Its promoter driving the expression of a reporter gene (d2EGFP) has been used to analyse the role of certain defined putative elements and a downstream enhancer element in gene expression. These experiments were carried out in vitro using a line of pre-pubertal mouse Sertoli cells, transienly transfected with circular DNA constructs with variously mutated promoter elements. A downstream enhancer element, situated immediately 3’ of the polyadenylation (PA) signal for Amh, has been inserted in an equivalent position in the d2EGFP construct. When the Amh promoter is unmodified, the downstream enhancer (DE) is positively associated with a large increase in EGFP expression. This is at least partly the consequence of an increased rate of expression by individual cells. Experiments using variously truncated Amh promoters indicate that an upstream region (-214 to -336) may play a minor role in facilitating enhancement. However mutation of the Wilms tumour factor-1 element, situated between the tata box and the start of translation, results in an almost complete suppression of enhancement.展开更多
Transcription factors（TFs）play critical roles in the development of the nervous system,but the transcriptional regulatory mechanisms of these genes are poorly understood.Here we analyzed 5-kb of the 5＇ flanking gen...Transcription factors（TFs）play critical roles in the development of the nervous system,but the transcriptional regulatory mechanisms of these genes are poorly understood.Here we analyzed 5-kb of the 5＇ flanking genomic DNA sequences of 41 TF genes involved in neuronal development.The results showed that the TF genes tend to have higher GC contents in the proximal region and most of the TF genes have at least one proximal GC-rich（GC content60%）promoter with a CpG island.The promoter distribution analysis showed that the GC-poor promoters were sporadically distributed within the 5-kb flanking genomic sequence（FGS）;however,more than half（37 of 70）of the GC-rich promoters were located in the proximal region between nucleotides—1 and—500.Luciferase assays showed that partial GC-rich promoters increased gene expression in SH-SY5Y cells and that CpG methylation repressed the promoter activity.This study suggests a potential general mechanism for regulation of TF expression.展开更多
The divergence and continuous evolution of plants and animals contribute to ecological diversity.Promoters and transcription factors(TFs) are key determinants of gene regulation and transcription throughoutlife.Howeve...The divergence and continuous evolution of plants and animals contribute to ecological diversity.Promoters and transcription factors(TFs) are key determinants of gene regulation and transcription throughoutlife.However,theevolutionary trajectories and relationships of promoters and TFs are still poorly understood. Here, we conducted extensive analysis of large-scale multi-omics sequences in 420 animal species and 223 plant species spanning nearly a billion years of evolutionary history. Results showed that promoter GC-contentandTFisoelectricpoints,as features/signatures that accompany long biological evolution, exhibited increasing growth in animal cells but a decreasing trend in plant cells. Furthermore, the evolutionary trajectories of promoter and TF signatures in the animal kingdom provided further evidence that Mammalia as well as Aves evolved directly from the ancestor Reptilia. The strong correlation between promoter and TF signatures indicates that promoters and TFs formed antagonistic coevolution in the animal kingdom, but mutualistic coevolution in the plant kingdom. The distinct coevolutionary patterns potentially drive the plant-animal divergence, divergent evolution and ecological diversity.展开更多
The present study examines the factorial structure and assesses the psychometric properties of the adapted multidimensional Health-Promoting Lifestyle Profile II Scale, considering a sample of Italian university stude...The present study examines the factorial structure and assesses the psychometric properties of the adapted multidimensional Health-Promoting Lifestyle Profile II Scale, considering a sample of Italian university students who participated to an online survey. The original 52-items Scale showed a high overall internal consistency. Four of the six subscales were associated with good values of the Cronbach’s α coefficient, whereas two subscales had lower values. Hence corrected item-total correlation was calculated and 26 items that decreased the scale’s reliability were deleted. The remaining 26 items were first subjected to Principal Component Analysis that suggested a conceptually meaningful five-factor model. This result was further supported by the first-order confirmatory factor analysis, in which all the factor loadings were statistically significant. The internal consistency and the composite reliability for the reduced version of the Scale and its subscales have shown a good reliability for the measurement models. The multidimensionality of the scale was also confirmed by a second-order factor model.展开更多
Objective:The purpose of this study was to determine the effectiveness of Mobile-Stroke Risk Scale and Life Style Guidance(M-SRSguide)in promoting a healthy lifestyle and reducing stroke risk factors in atrisk persons...Objective:The purpose of this study was to determine the effectiveness of Mobile-Stroke Risk Scale and Life Style Guidance(M-SRSguide)in promoting a healthy lifestyle and reducing stroke risk factors in atrisk persons.Methods:This research was an clinical trial with a pre-test and post-test control group design.The accessible population is persons at risk of stroke in the community(West and East Kalimantan Province,Indonesia).Thirty-two participants in the intervention group and 32 participants in the control group participated in this study.The sampling method was systematic random sampling.We allocate the sample into the intervention and control groups using a randomized block design.The intervention group used the M-SRSguide.The control group used manual book for a self-assessment of stroke risk.The measurement of a healthy lifestyle and the stroke risk factors was performed before and six months after the intervention.Results:There are no significant differences in healthy lifestyle and stroke risk factors between the two groups after the intervention(P>0.05).Analysis of healthy lifestyle behavior assessment items in the intervention group showed an increase in healthy diets,activity patterns,and stress control after the use of the M-SRSguide(P<0.01).Conclusion:The use of M-SRSguide is effective in promoting a healthy lifestyle.展开更多
An Amh promoter driving expression of a reporter gene (d2EGFP) has been used to analyze the role of two specific promoter transcription factor binding elements. In addition a downstream (3’) enhancer (DE) was also in...An Amh promoter driving expression of a reporter gene (d2EGFP) has been used to analyze the role of two specific promoter transcription factor binding elements. In addition a downstream (3’) enhancer (DE) was also investigated. The transcription factors GATA-1 and GATA-4 had opposite effects, the former being incremental and the latter decremental. The quantitative balance between these two factors may provide a degree of control over the level of gene expression.展开更多
In order to elucidate the molecular mechanisms of globin gene expression during embryonic development, the nuclear extracts from mouse hematopoietic tissue at different stages of development have been prepared. By usi...In order to elucidate the molecular mechanisms of globin gene expression during embryonic development, the nuclear extracts from mouse hematopoietic tissue at different stages of development have been prepared. By using DNase I footprinting and gel mobility shift assays, the binding of protein factors in these extracts to the human βglobin promoter was analyzed. The differences in the binding patterns of protein factors during development were observed. An erythroid-specific and stage-specific nuclear protein in the nuclear extract from d 18 mouse fetal liver was identified, which can bind to the sequence (from -66bp to -90bp) of human β-globin promoter. We therefore speculate that the function of this cis-acting element may be similar to stage selector element (SSE) in chieken βA- promoter.展开更多
The DNasel hypersensitive site 2 (HS2) of human β-globin locus control region (LCR) is required fOr the high level expression of human d-globin genes. In the present study, a stage-specific protein factor (LPF-β) wa...The DNasel hypersensitive site 2 (HS2) of human β-globin locus control region (LCR) is required fOr the high level expression of human d-globin genes. In the present study, a stage-specific protein factor (LPF-β) was identified in the nuclear extract prepared from mouse fetal liver at d 18 of gestation, which could bind to the HS2 region of humanβ-globin LCRt We also found that the shift band of LPF-βfactor could be competed by humanβ-globin promoter. However, it couldn’t be competed by human E-globin promoter or by human Aβ-globin promoter. Furthermore, our data demonstrated that the binding-sequence of LPF-d factor is 5’CACACCCTA 3’,which is located at the HS2 region ofβ-LCR (from -10845 to -10853 bp) and humanβ-globin promoter (from -92 to -84 bp). We speculated that these regions containing the CACCC box in both the humallβ-globin promoter and HS2 might function as stage selector elements in the regulation of humanβd-globin switching and the LPF-βfactor might be a stage-specific protein factor involved in the regulation of humanβ-globin gene expression.展开更多
The promotion of energy efficiency(EE)helps address energy constraints and promote environmental sustainability.This study comprehensively explores the spatiotemporal variations,influencing factors,and configuration p...The promotion of energy efficiency(EE)helps address energy constraints and promote environmental sustainability.This study comprehensively explores the spatiotemporal variations,influencing factors,and configuration promotion paths of EE in 284 Chinese cities during 2003‒2019 using the global super-efficiency minimum distance to strong efficient frontier(G-S-MinDS),exploratory spatial data analysis(ESDA),multiscale geographically weighted regression(MGWR),and fuzzy set qualitative comparative analysis(fsQCA)methods.The findings are:①China’s cities have an annual average EE of 0.658 with a growth rate of 0.53%,showing considerable promotion potential.②Industrial structure optimization,population agglomeration,economic development,and increased green coverage contribute positively,while government intervention and openness hinder China’s urban EE.③Four configurational promotion paths for enhancing China’s urban EE are identified,where among those paths population density is a core condition,while government intervention is not.This study provides valuable insights into substantially improving urban EE,emphasizing the need for targeted policies to address energy and environmental crises in China.展开更多
Background: Although BCG is the most widely administered vaccine in the world, there have never been as many cases of TB as there are now. Globally, more than 8.8 million people developed active TB and 1.4 million—ma...Background: Although BCG is the most widely administered vaccine in the world, there have never been as many cases of TB as there are now. Globally, more than 8.8 million people developed active TB and 1.4 million—many of them—died in 2010. It is estimated that half of pulmonary TB cases arise from latent Mtb infection, making the study of latency and reactivation of utmost importance. Methods: Widely administered BCG vaccines and a gene modified recombinant BCG (rBCG) strain, AERAS-422, were used as models to investigate the growth promoting function of resuscitation-promoting factors (Rpfs) in different bacilli culture phases. Different supernatant fractions were prepared by ultrafiltration, and the promoting function of each fraction containing secreted Rpf(s) was evaluated by growth curve monitoring and colony counting on 7H10 agar plates. Results: The promoting effect of culture supernatants was mainly associated with the high molecular weight fraction (>30 kDa), which stimulated bacterial growth, but did not extend the exponential phase of stimulated culture. Anti-RpfB antibody showed significant growth restriction of the tested cultures. When comparing rBCG cultures containing 7H9 medium, the 10 - 30 kDa fraction, or the >30 kDa fraction, only the >30 kDa fraction was displayed with down-regulation of the secretion of RpfC, D and E. In colony counting tests, the plates containing the >30 kDa fraction had total countable colony numbers 2 to 3 fold higher than the plates with the 10 - 30 kDa fraction, and colonies appeared one to two weeks earlier than on the regular plates. The potential applications of the prepared supernatant fractions containing RpfA and RpfB are discussed, which may include accelerating diagnosis of Mtb infection and future TB vaccine development.展开更多
Objective:This study aimed to examine the influence of behavioral lifestyle factors on recent episodic memory retention capacity among young-old adults(aged 60-69 years)in China.The findings provide scientific evidenc...Objective:This study aimed to examine the influence of behavioral lifestyle factors on recent episodic memory retention capacity among young-old adults(aged 60-69 years)in China.The findings provide scientific evidence to inform proactive strategies to mitigate cognitive decline risk within China’s rapidly aging population.Methods:Utilizing data from the 2022 wave of the China Family Panel Studies(CFPS),a total of 2,772 adults aged 60-69 were included in the analytical sample.Recent episodic memory retention capacity(scored 0-5 points,based on self-reported assessment)served as the dependent variable.Six categories of behavioral lifestyle indicators(including exercise frequency,sleep quality,dietary patterns,etc.)were analyzed as independent variables.Associations were assessed using multivariate ordinal logistic regression models,controlling for relevant covariates.Results:Self-reported potential impairment in recent episodic memory was identified by 47.19%of respondents.Multivariate analysis revealed significant associations between behavioral lifestyle factors and memory retention capacity.Regular exercise(OR=1.297,95%CI:1.118-1.504),meat consumption(OR=1.765,95%CI:1.393-2.237),regular reading habits(OR=1.599,95%CI:1.283-1.992),and internet use(OR=1.413,95%CI:1.217-1.641)emerged as significant protective factors.Abnormal sleep duration was detrimentally associated with retention capacity(too short:OR=0.728,95%CI:0.591-0.897;too long:OR=0.810,95%CI:0.670-0.980).Significant associations were also observed for control variables:urban residence(OR=1.270,95%CI:1.100-1.467),high school education or above(OR=1.543,95%CI:1.293-1.841),and better self-rated health status(OR=1.156,95%CI:1.089-1.227)were positively correlated with better memory retention.Conclusions:Optimal sleep duration,regular physical exercise,meat intake,habitual reading,and internet engagement positively predict self-assessed recent episodic memory retention capacity in Chinese young-old adults.These findings underscore the potential for multi-faceted lifestyle interventions to enhance cog-nitive health in aging populations.Specifically,strategies should encompass community-based sleep hygiene management,tailored nutritional interventions(especially promoting adequate protein sources like meat),enhanced digital literacy and internet accessibility programs,and the promotion of age-appropriate physical activity initiatives.Furthermore,implementing culturally responsive strategies adapted to urban-rural contexts-such as deploying“mobile cognitive health units”in rural areas and fostering digital reading platforms in urban settings-is recommended to optimize intervention effectiveness.展开更多
AIM:To examine how the expression of caudal type homebox transcription factor 2(Cdx2) is regulated in the development of malignancy in Barrett's esophagus.METHODS:Cdx2,mucin(MUC) series(MUC2,MUC5AC and MUC6),p53 a...AIM:To examine how the expression of caudal type homebox transcription factor 2(Cdx2) is regulated in the development of malignancy in Barrett's esophagus.METHODS:Cdx2,mucin(MUC) series(MUC2,MUC5AC and MUC6),p53 and E-cadherin expression in Barrett's esophagus and adenocarcinoma specimens were examined by immunostaining.Isolated clusters of cells from(1) MUC2 and Cdx2-positive intestinal metaplastic mucosa;(2) MUC5AC and MUC6-positive,and MUC2 and Cdx2-negative high-grade dysplasia(HD),or intramucosal adenocarcinoma(IMC);and(3) MUC5AC,MUC6 and Cdx2-positive poorly-differentiated invasive adenocarcinoma(PDA) were analyzed by methylationspecific polymerase chain reaction using sets of primers for detecting methylation status of the Cdx2 gene.RESULTS:Most of the non-neoplastic Barrett's esophageal mucosa showing intestinal-type metaplasia with or without low-grade dysplasia was positive for E-cadherin,MUC series and Cdx2,but negative for p53.A portion of the low-grade to HD was positive for E-cadherin,MUC5AC,MUC6 and p53,but negative for MUC2 and Cdx2.The definite IMC area was strongly positive for MUC5AC,MUC6 and p53,but negative for MUC2 and Cdx2.Methylation of the Cdx2 promoter was not observed in intestinal metaplasia,while hypermethylation of part of its promoter was observed in hot dipped and IMC.Hypermethylation of a large fraction of the Cdx2 promoter was observed in PDA.CONCLUSION:Cdx2 expression is restored irrespective of the methylation status of its promoter.Apparent positive immunohistochemical results can be a molecular mark for gene silencing memory.展开更多
Myostatin(MSTN) is one of the key factors regulating myogenesis. Because of its role as a negative regulator of muscle mass deposition, much interest has been given to its protein and, in recent years, several studies...Myostatin(MSTN) is one of the key factors regulating myogenesis. Because of its role as a negative regulator of muscle mass deposition, much interest has been given to its protein and, in recent years, several studies have analysed MSTN gene regulation. This review discusses the MSTN gene promoter, focusing on its structure in several animal species, both vertebrate and invertebrate. We report the important binding sites considering their degree of phylogenetic conservation and roles they play in the promoter activity. Finally, we discuss recent studies focusing on MSTN gene regulation via promoter manipulation and the potential applications they have both in medicine and agriculture.展开更多
AIM: To test the hypothesis to block VEGF expression of SMMC-7721 hepatoma cells may inhibit tumor growth using the rat hepatoma model. METHODS: Amplify the 200 VEGF cDNA fragment and insert it into human U6 gene cass...AIM: To test the hypothesis to block VEGF expression of SMMC-7721 hepatoma cells may inhibit tumor growth using the rat hepatoma model. METHODS: Amplify the 200 VEGF cDNA fragment and insert it into human U6 gene cassette in the reverse orientation transcribing small antisense RNA which could specifically interact with VEGF165, and VEGF121 mRNA. Construct the retroviral vector containing this antisense VEGF U6 cassette and package the replication-deficient recombinant retrovirus. SMMC-7721 cells were transduced with these virus and positive clones were selected with G418. PCR and Southern blot analysis were performed to determine if U6 cassette integrated into the genomic DNA of positive clone. Transfected tumor cells were evaluated for RNA expression by ribonuclease protection assays. The VEGF protein in the supernatant of parental tumor cells and genetically modified tumor cells was determined with ELISA. In vitro and in vivo growth properties of antisense VEGF cell clone in nude mice were analyzed. RESULTS: Restriction enzyme digestion and PCR sequencing verified that the antisense VEGF RNA retroviral vector was successfully constructed.After G418 selection, resistant SMMC-7721 cell clone was picked up. PCR and Southern blot analysis suggested that U6 cassette was integrated into the cell genomic DNA. Stable SMMC-7721 cell clone transduced with U6 antisense RNA cassette could express 200 bp small antisense VEGF RNA and secrete reduced levels of VEGF in culture condition. Production of VEGF by antisense transgene-expressing cells was 65+/-10 ng/L per 10(6) cells, 42045 ng/L per 10(6) cells in sense group and 485+/-30 ng/L per 10(6) cells in the negative control group, (P【 0.05). The antisense-VEGF cell clone appeared phenotypically indistinguishable from SMMC-7721 cells and SMMC-7721 cells transfected sense VEGF. The growth rate of the antisense-VEGF cell clone was the same as the control cells. When S.C. was implanted into nude mice, growth of antisense-VEGF cell lines was greatly inhibited compared with control cells. CONCLUSION: Expression of antisense VEGF RNA in SMMC-7721 cells could decrease the tumorigenicity, and antisense-VEGF gene therapy may be an adjuvant treatment for hepatoma.展开更多
BACKGROUND Aberrant expression of stanniocalcin 2(STC2)is implicated in colon adenocarcinoma(COAD).A previous study identified that STC2 functions as a tumor promoter to drive development of some cancers,but the role ...BACKGROUND Aberrant expression of stanniocalcin 2(STC2)is implicated in colon adenocarcinoma(COAD).A previous study identified that STC2 functions as a tumor promoter to drive development of some cancers,but the role of its overexpression in the development of COAD remains unclear.AIM To evaluate the regulation mechanism of STC2 overexpression in COAD.METHODS The expression of STC2 in COAD was assessed by TCGA COAD database and GEO(GSE50760).Methylation level of the STC2 promoter was evaluated with beta value in UALCAN platform,and the correlation between STC2 expression and survival rate was investigated with TCGA COAD.Transcription binding site prediction was conducted by TRANSFAC and LASAGNA,and a luciferase reporter system was used to identify STC2 promoter activity in several cell lines,including HEK293T,NCM460,HT29,SW480,and HCT116.Western blotting was performed to evaluate the role of Sp1 on the expression of STC2.RESULTS The central finding of this work is that STC2 is overexpressed in COAD tissues and positively correlated with poor prognosis.Importantly,the binding site of the transcription factor Sp1 is widely located in the promoter region of STC2.A luciferase reporter system was successfully constructed to analyze the transcription activity of STC2,and knocking down the expression of Sp1 significantly inhibited the transcription activity of STC2.Furthermore,inhibition of Sp1 remarkably decreased protein levels of STC2.CONCLUSION Our data provide evidence that the transcription factor Sp1 is essential for the overexpression of STC2 in COAD through activation of promoter activity.Taken together,our finding provides new insights into the mechanism of oncogenic function of COAD by STC2.展开更多
文摘BACKGROUND: The mortality rate of heavy type hepatitisis high. No special treatment is available except generaltreatment. This multicenter clinical study was designed toobserve the safety and efficacy of promoting hepatic growthfactor (PHGF) in the treatment of heavy type hepatitis andsevere chronic hepatitis.METHODS: 347 patients with heavy type hepatitis and 324with severe chronic hepatitis were subjected to administra-tion of 120 μg of PHGF per day for 4 weeks on the basis ofgeneral treatment. Those who were being effectively treat-ed would last additional 2 to 4 weeks. Blood routine, urineroutine, blood urea nitrogen (BUN), blood creatinine(Cr), blood ammonia, alpha fetoprotein (AFP), electro-lyte, alanine transaminase (ALT), aspartate transaminase(AST), serum total bilirubin (TBIL), serum direct biliru-bin (DBIL), prothrombin time activity (PTA), total pro-tein (TP) and albumin (ALB) were detected in the pa-tients before treatment, 2 weeks after treatment, and at theend of the treatment. Any side-effect would be recorded.RESULTS: In the patients with severe chronic hepatitis, thetotal effective rate of the treatment was 88. 9% The levelsof ALT, AST and TBIL decreased significantly (P<0.001),whereas those of PTA and ALB increased significantly (P <0.001), and the level of AFP increased slightly. In patientswith heavy type hepatitis, the total effective rate of thistreatment was 78.4%, and patients at different stage showeddifferent results. The total effective rates of patients withearly, medium and terminal stage heavy type hepatitis were89.9%, 84.8% and 27.5%, respectively. No severe side-effect was shown.CONCLUSION: PHGF is effective and safe in the treat-ment of patients with heavy type hepatitis and severe chro-nic hepatitis. But it should be administered early in patientswith heavy type hepatitis so as to get better curative effects.
基金Supported by Medicine and Health Science and Technology Plan Projects of Zhejiang Province,No.2018KY569Zhejiang Provincial Natural Science Foundation of China,No.LY17H030002
文摘BACKGROUND Liver cancer has a high mortality and morbidity rate throughout the world.In clinical practice,the prognosis of liver cancer patients is poor,and the complex reasons contribute to treatment failures,including fibrosis,hepatitis viral infection,drug resistance and metastasis.Thus,screening novel prognostic biomarkers is of great importance for guiding liver cancer therapy.Orosomucoid genes(ORMs)encode acute phase plasma proteins,including orosomucoid 1(ORM1)and ORM2.Previous studies showed their upregulation upon inflammation,but the specific function of ORMs has not yet been determined,especially in the development of liver cancer.AIM To determine the expression of ORMs and their potential function in liver cancer.METHODS Analysis of the expression of ORMs in different human tissues was performed on data from the HPA RNA-seq normal tissues project.The expression ratio of ORMs was determined using the HCCDB database,including the ratio between liver cancer and other cancers,normal liver and other normal tissues,liver cancer and adjacent normal liver tissues.Analysis of ORM expression in different cancer types was performed using The Cancer Genome Atlas and TIMER database.The expression of ORMs in liver tumor tissues and adjacent normal tissues were further confirmed using Gene Expression Omnibus data,including GSE36376 and GSE14520.The 10-year overall survival(OS),progression-free survival(PFS)and relapse-free survival(RFS)rates between high and low ORM expression groups in liver cancer patients were determined using the Kaplan-Meier plotter tool.Gene Set Enrichment Analysis(GSEA)was employed to explore the ORM2-associated signaling network.Correlations between ORM2 expression and tumor purity or the infiltration level of macrophages in liver tumor tissues were determined using the TIMER database.The correlation between ORM2 gene levels,tumor-associated macrophage(TAM)markers(including CD68 and TGFβ1)and T cell immunosuppression(including CTLA4 and PD-1)in liver tumor tissues and liver GTEx was determined using the GEPIA database.RESULTS ORM1 and ORM2 were highly expressed in normal liver and liver tumor tissues.ORM1 and ORM2 expression was significantly decreased in liver tumor tissues compared with adjacent normal tissues,and similar results were also noted in cholangiocarcinoma,esophageal carcinoma,and lung squamous cell carcinoma.Further analysis of the Gene Expression Omnibus Database also confirmed the downregulation of ORM1 and ORM2 in liver tumors.Survival analysis showed that the high ORM2 group had better survival rates in OS,PFS and RFS.ORM1 only represented better performance in PFS,but not in OS or RFS.GSEA analysis of ORM2 from The Cancer Genome Atlas liver cancer data identified that ORM2 positively associated with the G2/M checkpoint,E2F target signaling,as well as Wnt/β-catenin and Hedgehog signaling.Moreover,apoptosis,IFN-αresponses,IFN-γresponses and humoral immune responses were upregulated in the ORM2 high group.ORM2 expression was negatively correlated with the macrophage infiltration level,CD68,TGFβ1,CTLA4 and PD-1 levels.CONCLUSION The results showed that ORM1 and ORM2 were highly expressed specifically in liver tissues,whereas ORM1 and ORM2 were downregulated in liver tumor tissues.ORM2 is a better prognostic factor for liver cancer.Furthermore,ORM2 is closely associated with cancer-promoting pathways.
文摘This paper takes 30 hm2 wheat sprinkling irrigation land of Yunguo Family Farm in Yanzhou, Shandong Province, China as a sample, and draws a conclusion by comparing it with other households (including individual and large households). The conclusions are as follows: wheat sprinkling irrigation has a remarkable water-saving effect and comparative revenue. Water saving efficiency can reach 61.54% and a comprehensive income-increase rate can reach 38.67%. The main factors of increasing income and incentives of saving irrigation by sprinkling irrigation ranks as the following: saving land consolidation and water monitoring labors (accounts for 62.50%), saving land area of wheat bed to increase production and income (accounts for 23.44%), saving water bills (accounts for 14.06%). The incentive effect of water saving is not obvious mainly because the water price is low. The main obstacles to the promotion of sprinkling irrigation by individual household are the uneconomical scale and the barriers of coordination of proxy irrigation. Other large household’s (family farm) obstacles are mainly the instability of land tenure and mixed management. Suggestions on promoting sprinkling irrigation in wheat field: Accelerate land circulation and promote agricultural scale management to create basic scale conditions for spreading sprinkling irrigation;stabilizing farmland management rights as stabilizing farmland contractual rights, thus giving long-term business interests to all kinds of large household owners;guide the development of “scale + specialization” modern family farms;appropriate water saving subsidies should be given according to the positive externality of household water saving;confirm agricultural water rights to household and allow compensated transfer of “surplus water rights”.
基金supported by the National Natural Science Foundation(Nos.81471958 and 31401258)the Natural Science Foundation of Shandong Province(No.ZR2012BM006),China
文摘Transcription factors, which represent an important class of proteins that play key roles in controlling cellular proliferation and cell cycle modulation, are attractive targets for cancer therapy. Previous researches have shown that the expression level of activating transcription factor 5 （ATF5） was frequently increased in glioma and its acetylation level was related to glioma. The purposes of this study were to explore the methylation level of ATF5 in clinical glioma tissues and to explore the effect of ATF5 methylation on the expression of ATF5 in glioma. Methylation of the promoter region of ATF5 was assayed by bisulflte-specific polymerase chain reaction （PCR） sequencing analysis in 35 cases of glioma and 5 normal tissues. Quantitative real-time PCR （qRT-PCR） was also performed to detect ATF5 mRNA expression in 35 cases of glioma and 5 normal tissues. Clinical data were collected from the patients and analyzed. The percentages of methylation of the ATF5 gene in the promoter region in healthy control, patients with well-differentiated glioma, and those with poorly differentiated glioma were 87.78%, 73.89%, and 47.70%, respectively. Analysis of the methylation status of the promoter region of the ATF5 gene showed a gradually de- creased methylation level in poorly differentiated glioma, well-differentiated glioma, and normal tissues （P〈0.05）. There was also a significant difference between well-differentiated glioma and poorly differentiated glioma （P〈0.05）. ATF5 mRNA expression in glioma was significantly higher than that in the normal tissues （P〈0.05）. This study provides the first evidence that the methylation level of ATF5 decreased, and its mRNA expression was evidently up-regulated in glioma.
文摘Amh (anti-Müllerian hormone) is a single copy gene which is expressed strongly in Sertoli cells in the foetal testis and participates in the onset of sexual differentiation. Its promoter driving the expression of a reporter gene (d2EGFP) has been used to analyse the role of certain defined putative elements and a downstream enhancer element in gene expression. These experiments were carried out in vitro using a line of pre-pubertal mouse Sertoli cells, transienly transfected with circular DNA constructs with variously mutated promoter elements. A downstream enhancer element, situated immediately 3’ of the polyadenylation (PA) signal for Amh, has been inserted in an equivalent position in the d2EGFP construct. When the Amh promoter is unmodified, the downstream enhancer (DE) is positively associated with a large increase in EGFP expression. This is at least partly the consequence of an increased rate of expression by individual cells. Experiments using variously truncated Amh promoters indicate that an upstream region (-214 to -336) may play a minor role in facilitating enhancement. However mutation of the Wilms tumour factor-1 element, situated between the tata box and the start of translation, results in an almost complete suppression of enhancement.
基金supported by the National Natural Science Foundation of China (Nos. 31070928, 30600198, 30870876 and 81000558)the Guangzhou Scholar Project (Nos. 10A011G and 10A012G)the Scientific Research of Guangzhou Municipal Colleges and Universities (No. 10A211).
文摘Transcription factors（TFs）play critical roles in the development of the nervous system,but the transcriptional regulatory mechanisms of these genes are poorly understood.Here we analyzed 5-kb of the 5＇ flanking genomic DNA sequences of 41 TF genes involved in neuronal development.The results showed that the TF genes tend to have higher GC contents in the proximal region and most of the TF genes have at least one proximal GC-rich（GC content60%）promoter with a CpG island.The promoter distribution analysis showed that the GC-poor promoters were sporadically distributed within the 5-kb flanking genomic sequence（FGS）;however,more than half（37 of 70）of the GC-rich promoters were located in the proximal region between nucleotides—1 and—500.Luciferase assays showed that partial GC-rich promoters increased gene expression in SH-SY5Y cells and that CpG methylation repressed the promoter activity.This study suggests a potential general mechanism for regulation of TF expression.
基金supported by the National Key Research and Development Program of China (2017YFA0505500 to L.N.C., 2017YFC0909502 to J.S.Z.)Strategic Priority Research Program of the Chinese Academy of Sciences (XDB38040400 to L.N.C., XDB13000000 to W.W.)+3 种基金National Science Foundation of China (12131020 and 31930022 to L.N.C, 61602460 to J.S.Z.)Major Key Project of PCL (PCL2021A12 to L.N.C.)Special Fund for Science and Technology Innovation Strategy of Guangdong Province(2021B0909050004 and 2021B0909060002 to L.N.C.)Fundamental Research Funds for the Central Universities(3102019JC007 to W.W.)。
文摘The divergence and continuous evolution of plants and animals contribute to ecological diversity.Promoters and transcription factors(TFs) are key determinants of gene regulation and transcription throughoutlife.However,theevolutionary trajectories and relationships of promoters and TFs are still poorly understood. Here, we conducted extensive analysis of large-scale multi-omics sequences in 420 animal species and 223 plant species spanning nearly a billion years of evolutionary history. Results showed that promoter GC-contentandTFisoelectricpoints,as features/signatures that accompany long biological evolution, exhibited increasing growth in animal cells but a decreasing trend in plant cells. Furthermore, the evolutionary trajectories of promoter and TF signatures in the animal kingdom provided further evidence that Mammalia as well as Aves evolved directly from the ancestor Reptilia. The strong correlation between promoter and TF signatures indicates that promoters and TFs formed antagonistic coevolution in the animal kingdom, but mutualistic coevolution in the plant kingdom. The distinct coevolutionary patterns potentially drive the plant-animal divergence, divergent evolution and ecological diversity.
文摘The present study examines the factorial structure and assesses the psychometric properties of the adapted multidimensional Health-Promoting Lifestyle Profile II Scale, considering a sample of Italian university students who participated to an online survey. The original 52-items Scale showed a high overall internal consistency. Four of the six subscales were associated with good values of the Cronbach’s α coefficient, whereas two subscales had lower values. Hence corrected item-total correlation was calculated and 26 items that decreased the scale’s reliability were deleted. The remaining 26 items were first subjected to Principal Component Analysis that suggested a conceptually meaningful five-factor model. This result was further supported by the first-order confirmatory factor analysis, in which all the factor loadings were statistically significant. The internal consistency and the composite reliability for the reduced version of the Scale and its subscales have shown a good reliability for the measurement models. The multidimensionality of the scale was also confirmed by a second-order factor model.
基金We thank the Politeknik Kesehatan Kementerian Kesehatan Indonesia,LB.01.01/I.1/2657/2019 for funding this study,the respondents for participating in this study,and Marshall Godwin for granting permission to use the Simple Lifestyle Indicator Questionnaire(SLIQ).
文摘Objective:The purpose of this study was to determine the effectiveness of Mobile-Stroke Risk Scale and Life Style Guidance(M-SRSguide)in promoting a healthy lifestyle and reducing stroke risk factors in atrisk persons.Methods:This research was an clinical trial with a pre-test and post-test control group design.The accessible population is persons at risk of stroke in the community(West and East Kalimantan Province,Indonesia).Thirty-two participants in the intervention group and 32 participants in the control group participated in this study.The sampling method was systematic random sampling.We allocate the sample into the intervention and control groups using a randomized block design.The intervention group used the M-SRSguide.The control group used manual book for a self-assessment of stroke risk.The measurement of a healthy lifestyle and the stroke risk factors was performed before and six months after the intervention.Results:There are no significant differences in healthy lifestyle and stroke risk factors between the two groups after the intervention(P>0.05).Analysis of healthy lifestyle behavior assessment items in the intervention group showed an increase in healthy diets,activity patterns,and stress control after the use of the M-SRSguide(P<0.01).Conclusion:The use of M-SRSguide is effective in promoting a healthy lifestyle.
文摘An Amh promoter driving expression of a reporter gene (d2EGFP) has been used to analyze the role of two specific promoter transcription factor binding elements. In addition a downstream (3’) enhancer (DE) was also investigated. The transcription factors GATA-1 and GATA-4 had opposite effects, the former being incremental and the latter decremental. The quantitative balance between these two factors may provide a degree of control over the level of gene expression.
文摘In order to elucidate the molecular mechanisms of globin gene expression during embryonic development, the nuclear extracts from mouse hematopoietic tissue at different stages of development have been prepared. By using DNase I footprinting and gel mobility shift assays, the binding of protein factors in these extracts to the human βglobin promoter was analyzed. The differences in the binding patterns of protein factors during development were observed. An erythroid-specific and stage-specific nuclear protein in the nuclear extract from d 18 mouse fetal liver was identified, which can bind to the sequence (from -66bp to -90bp) of human β-globin promoter. We therefore speculate that the function of this cis-acting element may be similar to stage selector element (SSE) in chieken βA- promoter.
文摘The DNasel hypersensitive site 2 (HS2) of human β-globin locus control region (LCR) is required fOr the high level expression of human d-globin genes. In the present study, a stage-specific protein factor (LPF-β) was identified in the nuclear extract prepared from mouse fetal liver at d 18 of gestation, which could bind to the HS2 region of humanβ-globin LCRt We also found that the shift band of LPF-βfactor could be competed by humanβ-globin promoter. However, it couldn’t be competed by human E-globin promoter or by human Aβ-globin promoter. Furthermore, our data demonstrated that the binding-sequence of LPF-d factor is 5’CACACCCTA 3’,which is located at the HS2 region ofβ-LCR (from -10845 to -10853 bp) and humanβ-globin promoter (from -92 to -84 bp). We speculated that these regions containing the CACCC box in both the humallβ-globin promoter and HS2 might function as stage selector elements in the regulation of humanβd-globin switching and the LPF-βfactor might be a stage-specific protein factor involved in the regulation of humanβ-globin gene expression.
基金the financial support provided by the National Natural Science Foundation of China[Grant No.72373138 and 71973131]Major Project of National Social Science Foundation of China[Grant No.19VHQ002].
文摘The promotion of energy efficiency(EE)helps address energy constraints and promote environmental sustainability.This study comprehensively explores the spatiotemporal variations,influencing factors,and configuration promotion paths of EE in 284 Chinese cities during 2003‒2019 using the global super-efficiency minimum distance to strong efficient frontier(G-S-MinDS),exploratory spatial data analysis(ESDA),multiscale geographically weighted regression(MGWR),and fuzzy set qualitative comparative analysis(fsQCA)methods.The findings are:①China’s cities have an annual average EE of 0.658 with a growth rate of 0.53%,showing considerable promotion potential.②Industrial structure optimization,population agglomeration,economic development,and increased green coverage contribute positively,while government intervention and openness hinder China’s urban EE.③Four configurational promotion paths for enhancing China’s urban EE are identified,where among those paths population density is a core condition,while government intervention is not.This study provides valuable insights into substantially improving urban EE,emphasizing the need for targeted policies to address energy and environmental crises in China.
文摘Background: Although BCG is the most widely administered vaccine in the world, there have never been as many cases of TB as there are now. Globally, more than 8.8 million people developed active TB and 1.4 million—many of them—died in 2010. It is estimated that half of pulmonary TB cases arise from latent Mtb infection, making the study of latency and reactivation of utmost importance. Methods: Widely administered BCG vaccines and a gene modified recombinant BCG (rBCG) strain, AERAS-422, were used as models to investigate the growth promoting function of resuscitation-promoting factors (Rpfs) in different bacilli culture phases. Different supernatant fractions were prepared by ultrafiltration, and the promoting function of each fraction containing secreted Rpf(s) was evaluated by growth curve monitoring and colony counting on 7H10 agar plates. Results: The promoting effect of culture supernatants was mainly associated with the high molecular weight fraction (>30 kDa), which stimulated bacterial growth, but did not extend the exponential phase of stimulated culture. Anti-RpfB antibody showed significant growth restriction of the tested cultures. When comparing rBCG cultures containing 7H9 medium, the 10 - 30 kDa fraction, or the >30 kDa fraction, only the >30 kDa fraction was displayed with down-regulation of the secretion of RpfC, D and E. In colony counting tests, the plates containing the >30 kDa fraction had total countable colony numbers 2 to 3 fold higher than the plates with the 10 - 30 kDa fraction, and colonies appeared one to two weeks earlier than on the regular plates. The potential applications of the prepared supernatant fractions containing RpfA and RpfB are discussed, which may include accelerating diagnosis of Mtb infection and future TB vaccine development.
文摘Objective:This study aimed to examine the influence of behavioral lifestyle factors on recent episodic memory retention capacity among young-old adults(aged 60-69 years)in China.The findings provide scientific evidence to inform proactive strategies to mitigate cognitive decline risk within China’s rapidly aging population.Methods:Utilizing data from the 2022 wave of the China Family Panel Studies(CFPS),a total of 2,772 adults aged 60-69 were included in the analytical sample.Recent episodic memory retention capacity(scored 0-5 points,based on self-reported assessment)served as the dependent variable.Six categories of behavioral lifestyle indicators(including exercise frequency,sleep quality,dietary patterns,etc.)were analyzed as independent variables.Associations were assessed using multivariate ordinal logistic regression models,controlling for relevant covariates.Results:Self-reported potential impairment in recent episodic memory was identified by 47.19%of respondents.Multivariate analysis revealed significant associations between behavioral lifestyle factors and memory retention capacity.Regular exercise(OR=1.297,95%CI:1.118-1.504),meat consumption(OR=1.765,95%CI:1.393-2.237),regular reading habits(OR=1.599,95%CI:1.283-1.992),and internet use(OR=1.413,95%CI:1.217-1.641)emerged as significant protective factors.Abnormal sleep duration was detrimentally associated with retention capacity(too short:OR=0.728,95%CI:0.591-0.897;too long:OR=0.810,95%CI:0.670-0.980).Significant associations were also observed for control variables:urban residence(OR=1.270,95%CI:1.100-1.467),high school education or above(OR=1.543,95%CI:1.293-1.841),and better self-rated health status(OR=1.156,95%CI:1.089-1.227)were positively correlated with better memory retention.Conclusions:Optimal sleep duration,regular physical exercise,meat intake,habitual reading,and internet engagement positively predict self-assessed recent episodic memory retention capacity in Chinese young-old adults.These findings underscore the potential for multi-faceted lifestyle interventions to enhance cog-nitive health in aging populations.Specifically,strategies should encompass community-based sleep hygiene management,tailored nutritional interventions(especially promoting adequate protein sources like meat),enhanced digital literacy and internet accessibility programs,and the promotion of age-appropriate physical activity initiatives.Furthermore,implementing culturally responsive strategies adapted to urban-rural contexts-such as deploying“mobile cognitive health units”in rural areas and fostering digital reading platforms in urban settings-is recommended to optimize intervention effectiveness.
基金Supported by Grant-in Aid from Ministry of Education,Sports and Culture (GP Program for Basic Science),Japan
文摘AIM:To examine how the expression of caudal type homebox transcription factor 2(Cdx2) is regulated in the development of malignancy in Barrett's esophagus.METHODS:Cdx2,mucin(MUC) series(MUC2,MUC5AC and MUC6),p53 and E-cadherin expression in Barrett's esophagus and adenocarcinoma specimens were examined by immunostaining.Isolated clusters of cells from(1) MUC2 and Cdx2-positive intestinal metaplastic mucosa;(2) MUC5AC and MUC6-positive,and MUC2 and Cdx2-negative high-grade dysplasia(HD),or intramucosal adenocarcinoma(IMC);and(3) MUC5AC,MUC6 and Cdx2-positive poorly-differentiated invasive adenocarcinoma(PDA) were analyzed by methylationspecific polymerase chain reaction using sets of primers for detecting methylation status of the Cdx2 gene.RESULTS:Most of the non-neoplastic Barrett's esophageal mucosa showing intestinal-type metaplasia with or without low-grade dysplasia was positive for E-cadherin,MUC series and Cdx2,but negative for p53.A portion of the low-grade to HD was positive for E-cadherin,MUC5AC,MUC6 and p53,but negative for MUC2 and Cdx2.The definite IMC area was strongly positive for MUC5AC,MUC6 and p53,but negative for MUC2 and Cdx2.Methylation of the Cdx2 promoter was not observed in intestinal metaplasia,while hypermethylation of part of its promoter was observed in hot dipped and IMC.Hypermethylation of a large fraction of the Cdx2 promoter was observed in PDA.CONCLUSION:Cdx2 expression is restored irrespective of the methylation status of its promoter.Apparent positive immunohistochemical results can be a molecular mark for gene silencing memory.
文摘Myostatin(MSTN) is one of the key factors regulating myogenesis. Because of its role as a negative regulator of muscle mass deposition, much interest has been given to its protein and, in recent years, several studies have analysed MSTN gene regulation. This review discusses the MSTN gene promoter, focusing on its structure in several animal species, both vertebrate and invertebrate. We report the important binding sites considering their degree of phylogenetic conservation and roles they play in the promoter activity. Finally, we discuss recent studies focusing on MSTN gene regulation via promoter manipulation and the potential applications they have both in medicine and agriculture.
基金Project supported by National Natural Science Foundation of China,No.863 Z2001-04
文摘AIM: To test the hypothesis to block VEGF expression of SMMC-7721 hepatoma cells may inhibit tumor growth using the rat hepatoma model. METHODS: Amplify the 200 VEGF cDNA fragment and insert it into human U6 gene cassette in the reverse orientation transcribing small antisense RNA which could specifically interact with VEGF165, and VEGF121 mRNA. Construct the retroviral vector containing this antisense VEGF U6 cassette and package the replication-deficient recombinant retrovirus. SMMC-7721 cells were transduced with these virus and positive clones were selected with G418. PCR and Southern blot analysis were performed to determine if U6 cassette integrated into the genomic DNA of positive clone. Transfected tumor cells were evaluated for RNA expression by ribonuclease protection assays. The VEGF protein in the supernatant of parental tumor cells and genetically modified tumor cells was determined with ELISA. In vitro and in vivo growth properties of antisense VEGF cell clone in nude mice were analyzed. RESULTS: Restriction enzyme digestion and PCR sequencing verified that the antisense VEGF RNA retroviral vector was successfully constructed.After G418 selection, resistant SMMC-7721 cell clone was picked up. PCR and Southern blot analysis suggested that U6 cassette was integrated into the cell genomic DNA. Stable SMMC-7721 cell clone transduced with U6 antisense RNA cassette could express 200 bp small antisense VEGF RNA and secrete reduced levels of VEGF in culture condition. Production of VEGF by antisense transgene-expressing cells was 65+/-10 ng/L per 10(6) cells, 42045 ng/L per 10(6) cells in sense group and 485+/-30 ng/L per 10(6) cells in the negative control group, (P【 0.05). The antisense-VEGF cell clone appeared phenotypically indistinguishable from SMMC-7721 cells and SMMC-7721 cells transfected sense VEGF. The growth rate of the antisense-VEGF cell clone was the same as the control cells. When S.C. was implanted into nude mice, growth of antisense-VEGF cell lines was greatly inhibited compared with control cells. CONCLUSION: Expression of antisense VEGF RNA in SMMC-7721 cells could decrease the tumorigenicity, and antisense-VEGF gene therapy may be an adjuvant treatment for hepatoma.
基金the Natural Science Foundation of Liaoning Province,China,No.20180550769
文摘BACKGROUND Aberrant expression of stanniocalcin 2(STC2)is implicated in colon adenocarcinoma(COAD).A previous study identified that STC2 functions as a tumor promoter to drive development of some cancers,but the role of its overexpression in the development of COAD remains unclear.AIM To evaluate the regulation mechanism of STC2 overexpression in COAD.METHODS The expression of STC2 in COAD was assessed by TCGA COAD database and GEO(GSE50760).Methylation level of the STC2 promoter was evaluated with beta value in UALCAN platform,and the correlation between STC2 expression and survival rate was investigated with TCGA COAD.Transcription binding site prediction was conducted by TRANSFAC and LASAGNA,and a luciferase reporter system was used to identify STC2 promoter activity in several cell lines,including HEK293T,NCM460,HT29,SW480,and HCT116.Western blotting was performed to evaluate the role of Sp1 on the expression of STC2.RESULTS The central finding of this work is that STC2 is overexpressed in COAD tissues and positively correlated with poor prognosis.Importantly,the binding site of the transcription factor Sp1 is widely located in the promoter region of STC2.A luciferase reporter system was successfully constructed to analyze the transcription activity of STC2,and knocking down the expression of Sp1 significantly inhibited the transcription activity of STC2.Furthermore,inhibition of Sp1 remarkably decreased protein levels of STC2.CONCLUSION Our data provide evidence that the transcription factor Sp1 is essential for the overexpression of STC2 in COAD through activation of promoter activity.Taken together,our finding provides new insights into the mechanism of oncogenic function of COAD by STC2.
