BACKGROUND Endoscopic ultrasonography(EUS)has become an established method in diagnostic and therapeutic procedures in gastroenterology;however,it has recently gained a growing role in hepatology.AIM To evaluate the r...BACKGROUND Endoscopic ultrasonography(EUS)has become an established method in diagnostic and therapeutic procedures in gastroenterology;however,it has recently gained a growing role in hepatology.AIM To evaluate the role of EUS features,strain elastography(SE),and EUS-tissue acquisition in diagnosing hepatic focal lesions(HFLs)that could affect further management.METHODS This cross-sectional study included 215 patients with pancreatic,biliary,or gastrointestinal malignancies referred for EUS examination.HFLs were identified in 43 patients(20%),and EUSguided tissue acquisition was performed from these lesions.RESULTS EUS features were highly sensitive(100%)but much less specific(57%)in diagnosing HFLs;the overall accuracy was 94%.Real-time elastography was also very sensitive(97%)but less specific(67%)in diagnosing HFLs;however,the overall accuracy was 92%.EUS tissue acquisition was extremely sensitive(100%)and specific(100%),with a 100%overall diagnostic accuracy.CONCLUSION The diagnostic utility of EUS-guided tissue acquisition was extremely accurate in diagnosing HFLs.EUS characteristics and real-time SE accurately predicted the histological diagnosis of both benign and malignant HFLs.展开更多
AIM:To evaluate the effect of nitric oxide(NO)on the development and degree of liver failure in an animal model of acute hepatic failure(AHF).METHODS:An experimental rat model of galactosamine-induced AHF was used.An ...AIM:To evaluate the effect of nitric oxide(NO)on the development and degree of liver failure in an animal model of acute hepatic failure(AHF).METHODS:An experimental rat model of galactosamine-induced AHF was used.An inhibitor of NO synthase,nitroarginine methyl ester,or an NO donor,arginine,were administered at various doses prior to or after the induction of AHF.RESULTS:All tested groups developed AHF.Following inhibition of the endogenous NO pathway,most liver parameters improved,regardless of the inhibitor dose before the induction of liver damage,and depending on the inhibitor dose after liver damage.Prophylactic administration of the inhibitor was more effective in improving liver function parameters than administration of the inhibitor after liver damage.An attempt to activate the endogenous NO pathway prior to the induction of liver damage did not change the observed liver function parameters.Stimulation of the endogenous NO pathway after liver damage,regardless of the NO donor dose used,improved most liver function parameters.CONCLUSION:The endogenous NO pathway plays an important role in the development of experimental galactosamine-induced AHF.展开更多
AIM:To explore the anti-inflammatory potential of adeno-associated virus-mediated delivery of Tregitope 167 in an experimental colitis model.METHODS:The trinitrobenzene sulfonate(TNBS) model of induced colitis was use...AIM:To explore the anti-inflammatory potential of adeno-associated virus-mediated delivery of Tregitope 167 in an experimental colitis model.METHODS:The trinitrobenzene sulfonate(TNBS) model of induced colitis was used in Balb/c mice.Subsequently after intravenous adeno-associated virusmediated regulatory T-cell epitopes(Tregitope) delivery,acute colitis was initiated by intra-rectal administration of 1.5 mg TNBS in 40% ethanol followed by a second treatment with TNBS(0.75 mg in 20% ethanol) 8 d later.Control groups included mice not treated with TNBS(healthy control group) and mice treated by TNBS only(diseased group).At the time of sacrifice colon weight,the disease activity index and histology damage score were determined.Immunohistochemical staining of the colonic tissues was performed to asses the cellular infiltrate and the presence of transcription factor forkhead Box-P3(Foxp3).Thymus,mesenteric lymph nodes,liver and spleen tissue were collected and the corresponding lymphocyte populations were further assessed by flow cytometry analysis for the expression of CD4+ T cell and regulatory T cell associated markers.RESULTS:The Tregitope 167 treated mice gained an average of 4% over their initial body weight at the time of sacrifice.In contrast,the mice treated with TNBS alone(no Tregitope) developed colitis,and lost 4% of their initial body weight at the time of sacrifice(P < 0.01).The body weight increase that had been observed in the mice pre-treated with Tregitope 167 was substantiated by a lower disease activity index and a decreased colon weight as compared to the diseased control group(P < 0.01 and P < 0.001,respectively).Immunohistochemical staining of the colonic tissues for CD4+ showed that inflammatory cell infiltrates were present in TNBS treated mice with or without administration with tregitope 167 and that these cellular infiltrates consisted mainly of CD4+ cells.For both TNBS treated groups CD4+ T cell infiltrates were observed in the sub-epithelial layer and the lamina propria.CD4+ T cell infiltrates were also present in the muscularis mucosa layer of the diseased control mice,but were absent in the Tregitope 167 treated group.Numerous Foxp3 positive cells were detected in the lamina propria and sub-epithelium of the colon sections from mice treated with Tregitope 167.Furthermore,the Foxp3 and glycoprotein A repetitions predominant markers were significantly increased in the CD4+ T lymphocyte population in the thymus of the mice pre-treated with adeno-associated virus serotype 5(cytomegalovirus promoter-Tregitope 167),as cytomegalovirus promoter compared to lymphocyte populations in the thymus of diseased and the healthy control mice(P < 0.05 and P < 0.001,respectively).CONCLUSION:This study identifies adeno-associated virus-mediated delivery of regulatory T-cell epitope 167 as a novel anti-inflammatory approach with the capacity to decrease intestinal inflammation and induce longterm remission in inflammatory bowel disease.展开更多
AIM: p53-Inducible ribonucleotide reductase small subunit 2 (p53R2) encodes a 351-amino-acid peptide, which catalyzes conversion of ribonucleoside diphosphates to the corresponding deoxyribonucleotides required for DN...AIM: p53-Inducible ribonucleotide reductase small subunit 2 (p53R2) encodes a 351-amino-acid peptide, which catalyzes conversion of ribonucleoside diphosphates to the corresponding deoxyribonucleotides required for DNA replication and repair. A recent study reported that a point mutation (G/T) in the p53 binding sequence in a colon cancer cell line completely impaired p53R2 protein activity.METHODS: We screened the p53R2 gene coding regions and a regulatory region which contains a p53 binding sequence in 100 patients with colorectal adenoma and 100 control subjects using PCR, cold SSCP, and direct DNA sequencing.RESULTS: Although we did not identify genetic variation in all nine exons, four regulatory-region variants were found,of which three were single nucleotide polymorphisms (SNPs) (nt 1 789 C/G, nt 1 928 A/G, 1 933 T/C), and one was 20 bp insertion which replaced a ATTTT between nt 1 831 and 1 835. Additionally, we determined the frequency of these p53R2 variants in a recently concluded case-control study of incident sporadic colorectal adenomas (163 cases and 210 controls).CONCLUSION: Although more detailed functional characterizations of these polymorphisms remain to be undertaken, these polymorphic sites may be useful for identifying alleles associated with mis-splicing, additional transcript factors and, more generally, in cancer-susceptibility association studies.展开更多
BACKGROUND Hepatocellular carcinoma(HCC)is the fifth most common cancer worldwide.HCC patients suffer from a high mortality-to-incidence ratio and low cure rate since we still have no specific and effective treatment....BACKGROUND Hepatocellular carcinoma(HCC)is the fifth most common cancer worldwide.HCC patients suffer from a high mortality-to-incidence ratio and low cure rate since we still have no specific and effective treatment.Although tremendous advances have been made in the investigation of HCC,the specific mechanisms of the progression of this disease are still only partially established.Hence,more research is needed to elucidate the underlying potential mechanisms to develop effective strategies for HCC.AIM To determine the role of developing brain homeobox 2(Dbx2)gene in promoting the development of HCC.METHODS Dbx2 expression in clinical specimens and HCC cell lines was detected by Western blot(WB)and immunohistochemistry.Gain and loss of Dbx2 function assays were performed in vitro and in vivo.Cell viability assays were used to investigate cell growth,flow cytometry was employed to assess cell cycle and apoptosis,and trans-well assays were conducted to evaluate cell migration,invasion,and metastasis.The expression of key molecules in the sonic hedgehog(Shh)signaling was determined by WB.RESULTS Compared to matched adjacent non-tumorous tissues,Dbx2 was overexpressed in 5 HCC cell lines and 76 surgically resected HCC tissues.Dbx2 overexpression was correlated with large tumor size.Both gain and loss of function assays indicated that Dbx2 promoted HCC cell proliferation by facilitating the transition from G1 to S phase,attenuating apoptosis and promoted HCC proliferation,migration,and invasion in vitro and in vivo.Mechanistically,Dbx2 modulated Shh signaling by enhancing FTCH1 and GLi1 expression in HCC cells that overexpressed Dbx2,which was reversed in HCC cells with Dbx2 knockdown.CONCLUSION Our results indicate that Dbx2 is significantly upregulated in HCC tissues and plays significant roles in proliferation and metastasis of HCC cells by activating the Shh pathway.展开更多
Background Wilms tumor (WT) is the most common childhood kidney cancer worldwide, yet its incidence and clinical behavior vary according to race and access to adequate healthcare resources. To guide and streamline the...Background Wilms tumor (WT) is the most common childhood kidney cancer worldwide, yet its incidence and clinical behavior vary according to race and access to adequate healthcare resources. To guide and streamline therapy in the war-torn and resource-constrained city of Baghdad, Iraq, we conducted a first-ever molecular analysis of 20 WT specimens to characterize the biological features of this lethal disease within this challenged population. Methods Next-generation sequencing of ten target genes associated with WT development and treatment resistance (WT1, CTNNB1, WTX, IGF2, CITED1, SIX2, p53, N-MYC, CRABP2, and TOP2A) was completed. Immunohistochemistry was performed for 6 marker proteins of WT (WT1, CTNNB1, NCAM, CITED1, SIX2, and p53). Patient outcomes were compiled. Results Mutations were detected in previously described WT 'hot spots' (e.g., WT1 and CTNNB1) as well as novel loci that may be unique to the Iraqi population. Immunohistochemistry showed expression domains most typical of blastemal-predominant WT. Remarkably, despite the challenges facing families and care providers, only one child, with combined WT1 and CTNNB1 mutations, was confirmed dead from disease. Median clinical follow-up was 40.5 months (range 6–78 months). Conclusions These data suggest that WT biology within a population of Iraqi children manifests features both similar to and unique from disease variants in other regions of the world. These observations will help to risk stratify WT patients living in this difficult environment to more or less intensive therapies and to focus treatment on cell-specific targets.展开更多
Androgen deficiency is strongly associated with erectile dysfunction (ED). Inadequate penile arterial blood flow is one of the major causes of ED. The blood flow to the corpus cavernosum is mainly derived from the i...Androgen deficiency is strongly associated with erectile dysfunction (ED). Inadequate penile arterial blood flow is one of the major causes of ED. The blood flow to the corpus cavernosum is mainly derived from the internal pudendal arteries (IPAs); however, no study has evaluated the effects of androgen deprivation on IPA's function. We hypothesized that castration impairs IPAs reactivity and structure, contributing to ED. In our study, Wistar male rats, 8-week-old, were castrated and studied 30 days after orchiectomy. Functional and structural properties of rat IPAs were determined using wire and pressure myograph systems, respectively. Protein expression was determined by Western blot and immunohistochemistry. Plasma testosterone levels were determined using the IMMULITE 1000 Immunoassay System. Castrated rats exhibited impaired erectile function, represented by decreased intracavernosal pressure/mean arterial pressure ratio. IPAs from castrated rats exhibited decreased phenylephrine- and electrical field stimulation (EFS)-induced contraction and decreased acetylcholine- and EFS-induced vasodilatation. IPAs from castrated rats exhibited decreased internal diameter, external diameter, thickness of the arterial wall, and cross-sectional area. Castration decreased nNOS and α-actin expression and increased collagen expression, p38 (Thr180/ryr182) phosphorylation, as well as caspase 3 cleavage. In conclusion, androgen deficiency is associated with impairment of IPA reactivity and structure and increased apoptosis signaling markers. Our findings suggest that androgen deficiency-induced vascular dysfunction is an event involving hypotrophic vascular remodeling of IPAs.展开更多
Mutant KRAs is a common driver in lung cancer.Nevertheless,the occurrence of human lung cancer and its subsequent development after activation of oncogenic KRAS in epithelial cells remain poorly understood.Organoids a...Mutant KRAs is a common driver in lung cancer.Nevertheless,the occurrence of human lung cancer and its subsequent development after activation of oncogenic KRAS in epithelial cells remain poorly understood.Organoids are embedded in three-dimensional Matrigel in which tissue-derived human adult stem cells can efficiently grow for a long time,maintaining genetic and phenotypic stability."But until now,the use of organoids to simulate human non-small cell lung cancer has not been reported.展开更多
文摘BACKGROUND Endoscopic ultrasonography(EUS)has become an established method in diagnostic and therapeutic procedures in gastroenterology;however,it has recently gained a growing role in hepatology.AIM To evaluate the role of EUS features,strain elastography(SE),and EUS-tissue acquisition in diagnosing hepatic focal lesions(HFLs)that could affect further management.METHODS This cross-sectional study included 215 patients with pancreatic,biliary,or gastrointestinal malignancies referred for EUS examination.HFLs were identified in 43 patients(20%),and EUSguided tissue acquisition was performed from these lesions.RESULTS EUS features were highly sensitive(100%)but much less specific(57%)in diagnosing HFLs;the overall accuracy was 94%.Real-time elastography was also very sensitive(97%)but less specific(67%)in diagnosing HFLs;however,the overall accuracy was 92%.EUS tissue acquisition was extremely sensitive(100%)and specific(100%),with a 100%overall diagnostic accuracy.CONCLUSION The diagnostic utility of EUS-guided tissue acquisition was extremely accurate in diagnosing HFLs.EUS characteristics and real-time SE accurately predicted the histological diagnosis of both benign and malignant HFLs.
基金Supported by A grant from the Ministry of Science and Higher Education,No.216/KBL/12
文摘AIM:To evaluate the effect of nitric oxide(NO)on the development and degree of liver failure in an animal model of acute hepatic failure(AHF).METHODS:An experimental rat model of galactosamine-induced AHF was used.An inhibitor of NO synthase,nitroarginine methyl ester,or an NO donor,arginine,were administered at various doses prior to or after the induction of AHF.RESULTS:All tested groups developed AHF.Following inhibition of the endogenous NO pathway,most liver parameters improved,regardless of the inhibitor dose before the induction of liver damage,and depending on the inhibitor dose after liver damage.Prophylactic administration of the inhibitor was more effective in improving liver function parameters than administration of the inhibitor after liver damage.An attempt to activate the endogenous NO pathway prior to the induction of liver damage did not change the observed liver function parameters.Stimulation of the endogenous NO pathway after liver damage,regardless of the NO donor dose used,improved most liver function parameters.CONCLUSION:The endogenous NO pathway plays an important role in the development of experimental galactosamine-induced AHF.
基金Supported by Grant from the Broad Medical Research Program of The Broad Foundation,No. IBD-029 5R
文摘AIM:To explore the anti-inflammatory potential of adeno-associated virus-mediated delivery of Tregitope 167 in an experimental colitis model.METHODS:The trinitrobenzene sulfonate(TNBS) model of induced colitis was used in Balb/c mice.Subsequently after intravenous adeno-associated virusmediated regulatory T-cell epitopes(Tregitope) delivery,acute colitis was initiated by intra-rectal administration of 1.5 mg TNBS in 40% ethanol followed by a second treatment with TNBS(0.75 mg in 20% ethanol) 8 d later.Control groups included mice not treated with TNBS(healthy control group) and mice treated by TNBS only(diseased group).At the time of sacrifice colon weight,the disease activity index and histology damage score were determined.Immunohistochemical staining of the colonic tissues was performed to asses the cellular infiltrate and the presence of transcription factor forkhead Box-P3(Foxp3).Thymus,mesenteric lymph nodes,liver and spleen tissue were collected and the corresponding lymphocyte populations were further assessed by flow cytometry analysis for the expression of CD4+ T cell and regulatory T cell associated markers.RESULTS:The Tregitope 167 treated mice gained an average of 4% over their initial body weight at the time of sacrifice.In contrast,the mice treated with TNBS alone(no Tregitope) developed colitis,and lost 4% of their initial body weight at the time of sacrifice(P < 0.01).The body weight increase that had been observed in the mice pre-treated with Tregitope 167 was substantiated by a lower disease activity index and a decreased colon weight as compared to the diseased control group(P < 0.01 and P < 0.001,respectively).Immunohistochemical staining of the colonic tissues for CD4+ showed that inflammatory cell infiltrates were present in TNBS treated mice with or without administration with tregitope 167 and that these cellular infiltrates consisted mainly of CD4+ cells.For both TNBS treated groups CD4+ T cell infiltrates were observed in the sub-epithelial layer and the lamina propria.CD4+ T cell infiltrates were also present in the muscularis mucosa layer of the diseased control mice,but were absent in the Tregitope 167 treated group.Numerous Foxp3 positive cells were detected in the lamina propria and sub-epithelium of the colon sections from mice treated with Tregitope 167.Furthermore,the Foxp3 and glycoprotein A repetitions predominant markers were significantly increased in the CD4+ T lymphocyte population in the thymus of the mice pre-treated with adeno-associated virus serotype 5(cytomegalovirus promoter-Tregitope 167),as cytomegalovirus promoter compared to lymphocyte populations in the thymus of diseased and the healthy control mice(P < 0.05 and P < 0.001,respectively).CONCLUSION:This study identifies adeno-associated virus-mediated delivery of regulatory T-cell epitope 167 as a novel anti-inflammatory approach with the capacity to decrease intestinal inflammation and induce longterm remission in inflammatory bowel disease.
基金Supported by the No. R03 CA92773-01A1 Grant to DX No. R01 CA66539 Grant to RMB from the National Cancer Institute from National Institutes of Health, Department of Health and Human Services
文摘AIM: p53-Inducible ribonucleotide reductase small subunit 2 (p53R2) encodes a 351-amino-acid peptide, which catalyzes conversion of ribonucleoside diphosphates to the corresponding deoxyribonucleotides required for DNA replication and repair. A recent study reported that a point mutation (G/T) in the p53 binding sequence in a colon cancer cell line completely impaired p53R2 protein activity.METHODS: We screened the p53R2 gene coding regions and a regulatory region which contains a p53 binding sequence in 100 patients with colorectal adenoma and 100 control subjects using PCR, cold SSCP, and direct DNA sequencing.RESULTS: Although we did not identify genetic variation in all nine exons, four regulatory-region variants were found,of which three were single nucleotide polymorphisms (SNPs) (nt 1 789 C/G, nt 1 928 A/G, 1 933 T/C), and one was 20 bp insertion which replaced a ATTTT between nt 1 831 and 1 835. Additionally, we determined the frequency of these p53R2 variants in a recently concluded case-control study of incident sporadic colorectal adenomas (163 cases and 210 controls).CONCLUSION: Although more detailed functional characterizations of these polymorphisms remain to be undertaken, these polymorphic sites may be useful for identifying alleles associated with mis-splicing, additional transcript factors and, more generally, in cancer-susceptibility association studies.
基金Supported by the National Natural Science Foundation of China,No.81571781
文摘BACKGROUND Hepatocellular carcinoma(HCC)is the fifth most common cancer worldwide.HCC patients suffer from a high mortality-to-incidence ratio and low cure rate since we still have no specific and effective treatment.Although tremendous advances have been made in the investigation of HCC,the specific mechanisms of the progression of this disease are still only partially established.Hence,more research is needed to elucidate the underlying potential mechanisms to develop effective strategies for HCC.AIM To determine the role of developing brain homeobox 2(Dbx2)gene in promoting the development of HCC.METHODS Dbx2 expression in clinical specimens and HCC cell lines was detected by Western blot(WB)and immunohistochemistry.Gain and loss of Dbx2 function assays were performed in vitro and in vivo.Cell viability assays were used to investigate cell growth,flow cytometry was employed to assess cell cycle and apoptosis,and trans-well assays were conducted to evaluate cell migration,invasion,and metastasis.The expression of key molecules in the sonic hedgehog(Shh)signaling was determined by WB.RESULTS Compared to matched adjacent non-tumorous tissues,Dbx2 was overexpressed in 5 HCC cell lines and 76 surgically resected HCC tissues.Dbx2 overexpression was correlated with large tumor size.Both gain and loss of function assays indicated that Dbx2 promoted HCC cell proliferation by facilitating the transition from G1 to S phase,attenuating apoptosis and promoted HCC proliferation,migration,and invasion in vitro and in vivo.Mechanistically,Dbx2 modulated Shh signaling by enhancing FTCH1 and GLi1 expression in HCC cells that overexpressed Dbx2,which was reversed in HCC cells with Dbx2 knockdown.CONCLUSION Our results indicate that Dbx2 is significantly upregulated in HCC tissues and plays significant roles in proliferation and metastasis of HCC cells by activating the Shh pathway.
文摘Background Wilms tumor (WT) is the most common childhood kidney cancer worldwide, yet its incidence and clinical behavior vary according to race and access to adequate healthcare resources. To guide and streamline therapy in the war-torn and resource-constrained city of Baghdad, Iraq, we conducted a first-ever molecular analysis of 20 WT specimens to characterize the biological features of this lethal disease within this challenged population. Methods Next-generation sequencing of ten target genes associated with WT development and treatment resistance (WT1, CTNNB1, WTX, IGF2, CITED1, SIX2, p53, N-MYC, CRABP2, and TOP2A) was completed. Immunohistochemistry was performed for 6 marker proteins of WT (WT1, CTNNB1, NCAM, CITED1, SIX2, and p53). Patient outcomes were compiled. Results Mutations were detected in previously described WT 'hot spots' (e.g., WT1 and CTNNB1) as well as novel loci that may be unique to the Iraqi population. Immunohistochemistry showed expression domains most typical of blastemal-predominant WT. Remarkably, despite the challenges facing families and care providers, only one child, with combined WT1 and CTNNB1 mutations, was confirmed dead from disease. Median clinical follow-up was 40.5 months (range 6–78 months). Conclusions These data suggest that WT biology within a population of Iraqi children manifests features both similar to and unique from disease variants in other regions of the world. These observations will help to risk stratify WT patients living in this difficult environment to more or less intensive therapies and to focus treatment on cell-specific targets.
文摘Androgen deficiency is strongly associated with erectile dysfunction (ED). Inadequate penile arterial blood flow is one of the major causes of ED. The blood flow to the corpus cavernosum is mainly derived from the internal pudendal arteries (IPAs); however, no study has evaluated the effects of androgen deprivation on IPA's function. We hypothesized that castration impairs IPAs reactivity and structure, contributing to ED. In our study, Wistar male rats, 8-week-old, were castrated and studied 30 days after orchiectomy. Functional and structural properties of rat IPAs were determined using wire and pressure myograph systems, respectively. Protein expression was determined by Western blot and immunohistochemistry. Plasma testosterone levels were determined using the IMMULITE 1000 Immunoassay System. Castrated rats exhibited impaired erectile function, represented by decreased intracavernosal pressure/mean arterial pressure ratio. IPAs from castrated rats exhibited decreased phenylephrine- and electrical field stimulation (EFS)-induced contraction and decreased acetylcholine- and EFS-induced vasodilatation. IPAs from castrated rats exhibited decreased internal diameter, external diameter, thickness of the arterial wall, and cross-sectional area. Castration decreased nNOS and α-actin expression and increased collagen expression, p38 (Thr180/ryr182) phosphorylation, as well as caspase 3 cleavage. In conclusion, androgen deficiency is associated with impairment of IPA reactivity and structure and increased apoptosis signaling markers. Our findings suggest that androgen deficiency-induced vascular dysfunction is an event involving hypotrophic vascular remodeling of IPAs.
基金This work was supported by the National Natural Science Foundation of China(No.82070002,82072329 and 81872511)Frontier Research Program of Bioland Laboratory(Guangzhou Regenerative Medicine and Health GuangdongLaboratory,Guangdong,China)(No.2018GZR110105005).
文摘Mutant KRAs is a common driver in lung cancer.Nevertheless,the occurrence of human lung cancer and its subsequent development after activation of oncogenic KRAS in epithelial cells remain poorly understood.Organoids are embedded in three-dimensional Matrigel in which tissue-derived human adult stem cells can efficiently grow for a long time,maintaining genetic and phenotypic stability."But until now,the use of organoids to simulate human non-small cell lung cancer has not been reported.