BACKGROUND: Excessive alcohol consumption can result in multiple organ injury, of which alcoholic liver disease (ALD) is the most common. With economic development and improvement of living standards, the incidence of...BACKGROUND: Excessive alcohol consumption can result in multiple organ injury, of which alcoholic liver disease (ALD) is the most common. With economic development and improvement of living standards, the incidence of diseases caused by alcohol abuse has been increasing in China, although its pathogenesis remains obscure. The aim of this study was to investigate the role of hypoxia in chronic ALD. METHODS: Twenty-eight male Sprague-Dawley rats were randomized into a control group (n=12) with a normal history and an experimental group (n=16) fed with 10 ml/ kg of 56% (vol/vol) ethanol once per day by gastric lavage for 24 weeks. At 24 weeks, blood samples were collected and then the rats were killed. Liver samples were frozen at -80 ℃ and used for RT-PCR; other liver samples were obtained for immunohistochemical staining. RESULTS: When the period of alcohol consumption increased, the positive rate of expression of hypoxia- inducible factor-1 alpha (HIF-1α) mRNA was more significantly elevated in the liver of the alcohol group than in the control group (P≤0.05). The HIF-1α protein located in the cytoplasm was seldom expressed in the control group, but significantly in the alcohol group (P≤0.01). CONCLUSION: HIF-1α mRNA expression was activated by ethanol-induced injury in this study, suggesting that hypoxia is involved in the underlying mechanism of ALD.展开更多
BACKGROUND:Alcohol abuse and dependence are major factors in the pathogenesis of alcoholic liver disease(ALD).Alcohol abuse is becoming an increasingly severe problem among the Han,Mongol,and Korean nationalities in n...BACKGROUND:Alcohol abuse and dependence are major factors in the pathogenesis of alcoholic liver disease(ALD).Alcohol abuse is becoming an increasingly severe problem among the Han,Mongol,and Korean nationalities in northeast China.This study aimed to investigate the relationship between ALD and the genetic polymorphism and expression levels of two enzymes,cytochrome P450ⅡE1(CYPⅡE1)and glutathione S-transferase P1(GSTP1)in patients of three nationalities.METHODS:Peripheral blood was collected from 353 Chinese patients with ALD,300 alcohol dependent patients without liver disease(alcoholic),and 360 healthy controls.Each group included patients from the Han,Mongol and Korean nationalities.Real-time polymerase chain reaction(PCR)and PCR-restriction fragment length polymorphism(PCR-RFLP)were used.RESULTS:Regardless of nationality,patients who carried the rare CYPⅡE1 C2 and GSTP1 Val alleles were at higher risk of ALD.The frequency of C2 and Val in patients with ALD was respectively 50.00%and 26.98%in the Han,31.36%and 22.87%in the Mongol,and 45.87%and 22.02% in the Korean nationality.No significant differences were seen in the frequency of either C2 or Val alleles in ALD patients among the three nationalities.In each nationality,the frequency of both C2 and Val alleles was significantly higher in ALD compared to alcoholic and healthy controls.Except for nationality,the average mRNA levels of CYPⅡ E1 in ALD patients and healthy controls were 10.05%and 2.21%,respectively.The average mRNA levels of GSTP1 in ALD patients and healthy controls were 0.53%and 2.12%,respectively.The mRNA level of CYPⅡE1 was higher,and that of GSTP1 was lower in patients with ALD compared to the controls.CONCLUSIONS:Except for nationality,patients with ALD in this series tended to have a higher mRNA expression of CYPⅡE1 and to carry the C2 allele,and tended to have a lower mRNA expression of GSTP1 and to carry the Val allele.There is a causal relationship between the polymorphic alleles,which leads to different mRNA levels and the development of ALD.展开更多
HBV-associated acute-on-chronic liver failure is prevalent in China's Mainland. The prognosis of HBV-ACLF is poor. The mortality of HBV-ACLF is approximately 80%. Therefore, a prognostic indicator was needed in or...HBV-associated acute-on-chronic liver failure is prevalent in China's Mainland. The prognosis of HBV-ACLF is poor. The mortality of HBV-ACLF is approximately 80%. Therefore, a prognostic indicator was needed in order to allow us to intervene as soon as possible. The model for end-stage liver disease（MELD） scoring system is widely used to predict the prognosis of liver failure. However, the assessment is too complex to restrict its application. This study aimed to investigate the expression of IP-10 in peripheral blood mononuclear cells（PBMC）, in order to explore the relationship between the expression and prognosis of patients with HBV-ACLF. The mRNA level of IP-10 in PBMCs were analyzed in 80 patients with HBV-ACLF, 40 patients with chronic hepatitis B（CHB） and 40 healthy people by fluorescent quantitative PCR. IP-10 mRNA level was significantly higher in the HBV-ACLF group than in the other two groups（P〈0.01）. Group with MELD score below 30 had lower IP-10 mRNA level than group with MELD score over 30（P〈0.05）. The IP-10 mR NA level in PBMCs in positive group was higher than that in negative group（P〈0.01）. With a threshold of 0.925, the area under the receiver operating characteristic（ROC） curves was 0.815. These findings suggest that assessment of IP-10 mRNA level in the PBMCs would be helpful for evaluating the disease severity and prognosis in patients with HBV-ACLF.展开更多
Summary： In adult liver, CYP3A4 plays an important role in the metabolism of a wide range of en- dogenous and exogenous compounds. To investigate whether there is a single nucleotide polymorphism （SNP） of CYP3A4 in...Summary： In adult liver, CYP3A4 plays an important role in the metabolism of a wide range of en- dogenous and exogenous compounds. To investigate whether there is a single nucleotide polymorphism （SNP） of CYP3A4 intron 2 in the liver and its effects on the mRNA expression and enzymatic activity of CYP3A4, genomic DNA was extracted from 96 liver tissue samples obtained from patients who had undergone liver surgery. An SNP of CYP3A4 intron 2 was identified by polymerase chain reaction （PCR）-single-strand confirmation polymorphism and DNA sequencing. The mRNA expression of CYP3A4 was determined by the fluorescence quantitative PCR technique. The enzymatic activity of CYP3A4 was measured using erythromycin and testosterone as probe substrates. Twelve patients were found to have the SNP/T4127G CYP3A4 within intron 2. The mRNA levels of CYP3A4 in wild-type and SNP/T4127G samples were 2.62±1.09 and 2.79±1.63, respectively （P〉0.05）. Erythromycin N-demethylase activity in wild-type and SNP/T4127G samples were 121.2±32.8 and 124.7±61.6 nmol·mg^-1min^-1, respectively （P〉0.05）. The activity of testosterone 613-hydroxylase was significantly different between wild-type （648±173 pmol·mg^-1·min^-1） and SNP/T4127G samples （540-4-196 pmol.mg-l-minl; P〈0.05）. In conclusion, the SNP/T4127G of CYP3A4 intron 2 exists in the liver. This SNP does not affect the mRNA expression of CYP3A4 but significantly decreases the hepatic micro- somal testosterone 613-hydroxylase activity of CYP3A4. Furthermore, this study indicates that the ap- propriate selection of probe substrates is very important in studying the relationship between the geno- type and phenotype of CYP3A4.展开更多
Objective: Liver metastasis,which contributes substantially to high mortality,is the most common recurrent mode of colon carcinoma.Thus,it is necessary to identify genes implicated in metastatic colonization of the li...Objective: Liver metastasis,which contributes substantially to high mortality,is the most common recurrent mode of colon carcinoma.Thus,it is necessary to identify genes implicated in metastatic colonization of the liver in colon carcinoma.Methods: We compared mRNA profiling in 18 normal colon mucosa(N),20 primary tumors(T) and 19 liver metastases(M) samples from the dataset GSE49355 and GSE62321 of Gene Expression Omnibus(GEO) database.Gene ontology(GO) and pathways of the identified genes were analyzed.Co-expression network and proteinprotein interaction(PPI) network were employed to identify the interaction relationship.Survival analyses based on The Cancer Genome Atlas(TCGA) database were used to further screening.Then,the candidate genes were validated by our data.Results: We identified 22 specific genes related to liver metastasis and they were strongly associated with cell migration,adhesion,proliferation and immune response.Simultaneously,the results showed that C-X-C motif chemokine ligand 14(CXCL14) might be a favorable prediction factor for survival of patients with colon carcinoma.Importantly,our validated data further suggested that lower CXCL14 represented poorer outcome and contributed to metastasis.Gene set enrichment analysis(GSEA) showed that CXCL14 was negatively related to the regulation of stem cell proliferation and epithelial to mesenchymal transition(EMT).Conclusions: CXCL14 was identified as a crucial anti-metastasis regulator of colon carcinoma for the first time,and might provide novel therapeutic strategies for colon carcinoma patients to improve prognosis and prevent metastasis.展开更多
BACKGROUND： The integrative analysis of microRNA and mRNA expression profiles can elucidate microRNA-targeted gene function. We used this technique to elucidate insights into the immunological pathology of non-alcoho...BACKGROUND： The integrative analysis of microRNA and mRNA expression profiles can elucidate microRNA-targeted gene function. We used this technique to elucidate insights into the immunological pathology of non-alcoholic fatty liver disease （NAFLD）. METHODS： We analyzed differentially expressed microRNA and mRNA expression profiles of CD4＋ T lymphocytes from the liver and mesenteric lymph nodes （MLNs） of mice with NAFLD using microarrays and RNA sequencing. Normal mice were used as controls. The target genes of microRNAs were predicted by TargetScan. Integrative analysis showed that the mRNAs were overlapped with microRNAs. Furthermore, the Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses were performed to predict the key genes and pathways. Then, 16 microRNAs and 10 mRNAs were validated by qRT-PCR. RESULTS： Microarray analysis suggested that 170 microRNAs were significantly de-regulated in CD4＋ T lymphocytes from the liver between the two groups. Eighty mRNAs corresponded with microRNA targeted genes. KEGG analysis indicated that the MAPK pathway was consistently augmented in the liver of NAFLD mice. miR-23b, let-7e, miR-128 and miR-130b possibly played significant parts in the MAPK pathways. Furthermore, between the two groups, 237 microRNAs were significantly deregulated in CD4＋ T lymphocytes from MLNs. 38 mRNAs coincided with microRNA target genes. The metabolic pathway was consistently enriched in the MLNs of NAFLD mice. miR- 206-3p, miR-181a-Sp, miR-29c-3p and miR-30d-5p likely play important roles in the regulation of metabolic pathways. CONCLUSION： The results of this study presented a new perspective on the application of integrative analysis to identify complex regulation means involved in the immunological pathogenesis of NAFLD.展开更多
Phenytoin(5,5-diphenylhydantoin;DPH) induces expression of cytochromes P450(CYPs). Interactions between DPH and tacrolimus suggested that the persistence of CYP induction after discontinuation of DPH is dependent on t...Phenytoin(5,5-diphenylhydantoin;DPH) induces expression of cytochromes P450(CYPs). Interactions between DPH and tacrolimus suggested that the persistence of CYP induction after discontinuation of DPH is dependent on the history of administration and dosing period of DPH. However, the relationship between the duration of DPH administration and expression of CYPs in the liver and small intestine of rats is not known. Alterations in levels of P-glycoprotein(P-gp;MDR1;ABCB1) as well as CYPs cause drug interactions in the small intestine. We examined the effects of the duration of DPH administration on expression of CYPs and P-gp in the liver and small intestine of rats. Rats were treated with DPH(100 mg/kg,peroral(p.o.) twice a day(b.d.)) for 2, 4, 8, and 16 d. mRNA levels of CYPs and P-gp were examined using the total RNA extracted from the liver and duodenum 2 h and 24 h after the final administration of DPH. CYP3 A activities were determined using microsomes. DPH administration for 2 d and 4 d markedly increased m RNA levels of CYPs such as CYP3 A1, CYP3 A2,CYP2 B1, and CYP2 B2 in the liver. A relatively long duration of DPH administration(8 d and16 d) resulted in abolition of the induction of hepatic CYP but increased CYP3 A activities were maintained. These results suggest that the duration of DPH administration could be an important determinant of hepatic CYP induction.展开更多
Northern blot analysis of glutathione S-transferase (GST) Yb1 mRNA in different tissues of male and female rats revealed that its tissue-specific transcription patterns were highly sex hormone related. Although the GS...Northern blot analysis of glutathione S-transferase (GST) Yb1 mRNA in different tissues of male and female rats revealed that its tissue-specific transcription patterns were highly sex hormone related. Although the GST Yb1 mRNA could be detected in most of the tissues examined at various levels, the highest abundance was observed in the ventral prostate, uterus and liver, which were the main target tissue for androgen, estrogen and glucocorticoid respectively The effect of androgen on the transcription of GST Yb1 was also tissue-specific. Since androgen withdrawal by castration caused the up-regulation of GST Yb1 mRNA in the ventral prostate but down-regulation in the liver and no effect in the brain, evaluation of this system for studying the regulation mechanisms of gene expression by which androgen exerts its differential effects has been discussed.展开更多
文摘BACKGROUND: Excessive alcohol consumption can result in multiple organ injury, of which alcoholic liver disease (ALD) is the most common. With economic development and improvement of living standards, the incidence of diseases caused by alcohol abuse has been increasing in China, although its pathogenesis remains obscure. The aim of this study was to investigate the role of hypoxia in chronic ALD. METHODS: Twenty-eight male Sprague-Dawley rats were randomized into a control group (n=12) with a normal history and an experimental group (n=16) fed with 10 ml/ kg of 56% (vol/vol) ethanol once per day by gastric lavage for 24 weeks. At 24 weeks, blood samples were collected and then the rats were killed. Liver samples were frozen at -80 ℃ and used for RT-PCR; other liver samples were obtained for immunohistochemical staining. RESULTS: When the period of alcohol consumption increased, the positive rate of expression of hypoxia- inducible factor-1 alpha (HIF-1α) mRNA was more significantly elevated in the liver of the alcohol group than in the control group (P≤0.05). The HIF-1α protein located in the cytoplasm was seldom expressed in the control group, but significantly in the alcohol group (P≤0.01). CONCLUSION: HIF-1α mRNA expression was activated by ethanol-induced injury in this study, suggesting that hypoxia is involved in the underlying mechanism of ALD.
基金supported by a grant from the Youth Science Fund of Heilongjiang Province(No.QC08C77)
文摘BACKGROUND:Alcohol abuse and dependence are major factors in the pathogenesis of alcoholic liver disease(ALD).Alcohol abuse is becoming an increasingly severe problem among the Han,Mongol,and Korean nationalities in northeast China.This study aimed to investigate the relationship between ALD and the genetic polymorphism and expression levels of two enzymes,cytochrome P450ⅡE1(CYPⅡE1)and glutathione S-transferase P1(GSTP1)in patients of three nationalities.METHODS:Peripheral blood was collected from 353 Chinese patients with ALD,300 alcohol dependent patients without liver disease(alcoholic),and 360 healthy controls.Each group included patients from the Han,Mongol and Korean nationalities.Real-time polymerase chain reaction(PCR)and PCR-restriction fragment length polymorphism(PCR-RFLP)were used.RESULTS:Regardless of nationality,patients who carried the rare CYPⅡE1 C2 and GSTP1 Val alleles were at higher risk of ALD.The frequency of C2 and Val in patients with ALD was respectively 50.00%and 26.98%in the Han,31.36%and 22.87%in the Mongol,and 45.87%and 22.02% in the Korean nationality.No significant differences were seen in the frequency of either C2 or Val alleles in ALD patients among the three nationalities.In each nationality,the frequency of both C2 and Val alleles was significantly higher in ALD compared to alcoholic and healthy controls.Except for nationality,the average mRNA levels of CYPⅡ E1 in ALD patients and healthy controls were 10.05%and 2.21%,respectively.The average mRNA levels of GSTP1 in ALD patients and healthy controls were 0.53%and 2.12%,respectively.The mRNA level of CYPⅡE1 was higher,and that of GSTP1 was lower in patients with ALD compared to the controls.CONCLUSIONS:Except for nationality,patients with ALD in this series tended to have a higher mRNA expression of CYPⅡE1 and to carry the C2 allele,and tended to have a lower mRNA expression of GSTP1 and to carry the Val allele.There is a causal relationship between the polymorphic alleles,which leads to different mRNA levels and the development of ALD.
基金supported by Natural Science Foundation of Hubei Province(No.2013CFB385)
文摘HBV-associated acute-on-chronic liver failure is prevalent in China's Mainland. The prognosis of HBV-ACLF is poor. The mortality of HBV-ACLF is approximately 80%. Therefore, a prognostic indicator was needed in order to allow us to intervene as soon as possible. The model for end-stage liver disease（MELD） scoring system is widely used to predict the prognosis of liver failure. However, the assessment is too complex to restrict its application. This study aimed to investigate the expression of IP-10 in peripheral blood mononuclear cells（PBMC）, in order to explore the relationship between the expression and prognosis of patients with HBV-ACLF. The mRNA level of IP-10 in PBMCs were analyzed in 80 patients with HBV-ACLF, 40 patients with chronic hepatitis B（CHB） and 40 healthy people by fluorescent quantitative PCR. IP-10 mRNA level was significantly higher in the HBV-ACLF group than in the other two groups（P〈0.01）. Group with MELD score below 30 had lower IP-10 mRNA level than group with MELD score over 30（P〈0.05）. The IP-10 mR NA level in PBMCs in positive group was higher than that in negative group（P〈0.01）. With a threshold of 0.925, the area under the receiver operating characteristic（ROC） curves was 0.815. These findings suggest that assessment of IP-10 mRNA level in the PBMCs would be helpful for evaluating the disease severity and prognosis in patients with HBV-ACLF.
基金supported by grants from the Scientific Research Foundation for Returned Scholars of the Ministry of Education of ChinaPre-research Foundation of Wuhan University,China(No.301270050)
文摘Summary： In adult liver, CYP3A4 plays an important role in the metabolism of a wide range of en- dogenous and exogenous compounds. To investigate whether there is a single nucleotide polymorphism （SNP） of CYP3A4 intron 2 in the liver and its effects on the mRNA expression and enzymatic activity of CYP3A4, genomic DNA was extracted from 96 liver tissue samples obtained from patients who had undergone liver surgery. An SNP of CYP3A4 intron 2 was identified by polymerase chain reaction （PCR）-single-strand confirmation polymorphism and DNA sequencing. The mRNA expression of CYP3A4 was determined by the fluorescence quantitative PCR technique. The enzymatic activity of CYP3A4 was measured using erythromycin and testosterone as probe substrates. Twelve patients were found to have the SNP/T4127G CYP3A4 within intron 2. The mRNA levels of CYP3A4 in wild-type and SNP/T4127G samples were 2.62±1.09 and 2.79±1.63, respectively （P〉0.05）. Erythromycin N-demethylase activity in wild-type and SNP/T4127G samples were 121.2±32.8 and 124.7±61.6 nmol·mg^-1min^-1, respectively （P〉0.05）. The activity of testosterone 613-hydroxylase was significantly different between wild-type （648±173 pmol·mg^-1·min^-1） and SNP/T4127G samples （540-4-196 pmol.mg-l-minl; P〈0.05）. In conclusion, the SNP/T4127G of CYP3A4 intron 2 exists in the liver. This SNP does not affect the mRNA expression of CYP3A4 but significantly decreases the hepatic micro- somal testosterone 613-hydroxylase activity of CYP3A4. Furthermore, this study indicates that the ap- propriate selection of probe substrates is very important in studying the relationship between the geno- type and phenotype of CYP3A4.
基金supported by grants from the National Natural Science Foundation of China(No.8177061284)
文摘Objective: Liver metastasis,which contributes substantially to high mortality,is the most common recurrent mode of colon carcinoma.Thus,it is necessary to identify genes implicated in metastatic colonization of the liver in colon carcinoma.Methods: We compared mRNA profiling in 18 normal colon mucosa(N),20 primary tumors(T) and 19 liver metastases(M) samples from the dataset GSE49355 and GSE62321 of Gene Expression Omnibus(GEO) database.Gene ontology(GO) and pathways of the identified genes were analyzed.Co-expression network and proteinprotein interaction(PPI) network were employed to identify the interaction relationship.Survival analyses based on The Cancer Genome Atlas(TCGA) database were used to further screening.Then,the candidate genes were validated by our data.Results: We identified 22 specific genes related to liver metastasis and they were strongly associated with cell migration,adhesion,proliferation and immune response.Simultaneously,the results showed that C-X-C motif chemokine ligand 14(CXCL14) might be a favorable prediction factor for survival of patients with colon carcinoma.Importantly,our validated data further suggested that lower CXCL14 represented poorer outcome and contributed to metastasis.Gene set enrichment analysis(GSEA) showed that CXCL14 was negatively related to the regulation of stem cell proliferation and epithelial to mesenchymal transition(EMT).Conclusions: CXCL14 was identified as a crucial anti-metastasis regulator of colon carcinoma for the first time,and might provide novel therapeutic strategies for colon carcinoma patients to improve prognosis and prevent metastasis.
基金supported by a grant from the National Natural Science Foundation of China(81170375)
文摘BACKGROUND： The integrative analysis of microRNA and mRNA expression profiles can elucidate microRNA-targeted gene function. We used this technique to elucidate insights into the immunological pathology of non-alcoholic fatty liver disease （NAFLD）. METHODS： We analyzed differentially expressed microRNA and mRNA expression profiles of CD4＋ T lymphocytes from the liver and mesenteric lymph nodes （MLNs） of mice with NAFLD using microarrays and RNA sequencing. Normal mice were used as controls. The target genes of microRNAs were predicted by TargetScan. Integrative analysis showed that the mRNAs were overlapped with microRNAs. Furthermore, the Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses were performed to predict the key genes and pathways. Then, 16 microRNAs and 10 mRNAs were validated by qRT-PCR. RESULTS： Microarray analysis suggested that 170 microRNAs were significantly de-regulated in CD4＋ T lymphocytes from the liver between the two groups. Eighty mRNAs corresponded with microRNA targeted genes. KEGG analysis indicated that the MAPK pathway was consistently augmented in the liver of NAFLD mice. miR-23b, let-7e, miR-128 and miR-130b possibly played significant parts in the MAPK pathways. Furthermore, between the two groups, 237 microRNAs were significantly deregulated in CD4＋ T lymphocytes from MLNs. 38 mRNAs coincided with microRNA target genes. The metabolic pathway was consistently enriched in the MLNs of NAFLD mice. miR- 206-3p, miR-181a-Sp, miR-29c-3p and miR-30d-5p likely play important roles in the regulation of metabolic pathways. CONCLUSION： The results of this study presented a new perspective on the application of integrative analysis to identify complex regulation means involved in the immunological pathogenesis of NAFLD.
文摘Phenytoin(5,5-diphenylhydantoin;DPH) induces expression of cytochromes P450(CYPs). Interactions between DPH and tacrolimus suggested that the persistence of CYP induction after discontinuation of DPH is dependent on the history of administration and dosing period of DPH. However, the relationship between the duration of DPH administration and expression of CYPs in the liver and small intestine of rats is not known. Alterations in levels of P-glycoprotein(P-gp;MDR1;ABCB1) as well as CYPs cause drug interactions in the small intestine. We examined the effects of the duration of DPH administration on expression of CYPs and P-gp in the liver and small intestine of rats. Rats were treated with DPH(100 mg/kg,peroral(p.o.) twice a day(b.d.)) for 2, 4, 8, and 16 d. mRNA levels of CYPs and P-gp were examined using the total RNA extracted from the liver and duodenum 2 h and 24 h after the final administration of DPH. CYP3 A activities were determined using microsomes. DPH administration for 2 d and 4 d markedly increased m RNA levels of CYPs such as CYP3 A1, CYP3 A2,CYP2 B1, and CYP2 B2 in the liver. A relatively long duration of DPH administration(8 d and16 d) resulted in abolition of the induction of hepatic CYP but increased CYP3 A activities were maintained. These results suggest that the duration of DPH administration could be an important determinant of hepatic CYP induction.
文摘Northern blot analysis of glutathione S-transferase (GST) Yb1 mRNA in different tissues of male and female rats revealed that its tissue-specific transcription patterns were highly sex hormone related. Although the GST Yb1 mRNA could be detected in most of the tissues examined at various levels, the highest abundance was observed in the ventral prostate, uterus and liver, which were the main target tissue for androgen, estrogen and glucocorticoid respectively The effect of androgen on the transcription of GST Yb1 was also tissue-specific. Since androgen withdrawal by castration caused the up-regulation of GST Yb1 mRNA in the ventral prostate but down-regulation in the liver and no effect in the brain, evaluation of this system for studying the regulation mechanisms of gene expression by which androgen exerts its differential effects has been discussed.
