Hepatitis B virus surface antigen （HBsAg）, a specific antigen on the membrane of Hepatitis B virus （HBV）-infected cells, provides a perfect target for therapeutic drugs. The development of reagents with high affin...Hepatitis B virus surface antigen （HBsAg）, a specific antigen on the membrane of Hepatitis B virus （HBV）-infected cells, provides a perfect target for therapeutic drugs. The development of reagents with high affinity and specificity to the HBsAg is of great significance to the early-stage diagnosis and treatment of HBV infection. Herein, we report the selection of RNA aptamers that can specifically bind to HBsAg protein and HBsAg-positive hepatocytes. One high affinity aptamer, HBs-A22, was isolated from an initial 115 met library of -1.1 ×10^15 random-sequence RNA molecules using the SELEX procedure. The selected aptamer HBs-A22 bound specifically to hepatoma cell line HepG2.2.15 that expresses HBsAg but did not bind to HBsAg-devoid HepG2 cells. This is the first reported RNA aptamer which could bind to a HBV specific antigen. This newly isolated aptamer could be modified to deliver imaging, diagnostic, and therapeutic agents targeted at HBV-infected cells.展开更多
AIM: To discover novel biomarkers for early diagnosis, prognosis or treatment of human colorectal cancer. METHODS: i TRAQ 2D LC-MS/MS analysis was used to identify differentially expressed proteins(DEPs) in the human ...AIM: To discover novel biomarkers for early diagnosis, prognosis or treatment of human colorectal cancer. METHODS: i TRAQ 2D LC-MS/MS analysis was used to identify differentially expressed proteins(DEPs) in the human colonic epithelial carcinogenic process using laser capture microdissection-purified colonic epithelial cells from normal colon, adenoma, carcinoma in situ and invasive carcinoma tissues. RESULTS: A total of 326 DEPs were identified, and four DEPs(DMBT1, S100A9, Galectin-10, and S100A8) with progressive alteration in the carcinogenic process were further validated by immunohistochemistry. The DEPs were involved in multiple biological processes including cell cycle, cell adhesion, translation, m RNA processing, and protein synthesis. Some of the DEPs involved in cellular process such as "translation" and "m RNA splicing" were progressively up-regulated, while some DEPs involved in other processes such as "metabolism" and "cell response to stress" was progressively downregulated. Other proteins with up- or down-regulation at certain stages of carcinogenesis may play various roles at different stages of the colorectal carcinogenic process. CONCLUSION: These findings give insights into our understanding of the mechanisms of colorectal carcinogenesis and provide clues for further investigation of carcinogenesis and identification of biomarkers.展开更多
This study investigated the expression profiles of IL-10 gene in three human hepatoma cell lines including Huh7, HepG2, and HepG2 transfected with a plasmid containing hepatitis B virus （HBV） named HepG2.2.15. RT-PC...This study investigated the expression profiles of IL-10 gene in three human hepatoma cell lines including Huh7, HepG2, and HepG2 transfected with a plasmid containing hepatitis B virus （HBV） named HepG2.2.15. RT-PCR analysis demonstrated that IL-10 message RNA was absent in HepG2 and Huh7 cells, whereas it was present in HepG2.2.15 cells, which was consistent with ELISA result. Furthermore, except for lamivudine other antiviral treatments did not significantly decrease the HBV DNA level in HepG2.2.15 cells, while they had different effects on the expression of IL-10 protein, although stimulation by LPS had no significant effect. In addition, except for poly（I：C）, the other treatments decreased the expression of IL-10 protein to different degrees, but had no sig-nificant effects on the expression of NF-κB and MyD88. Meanwhile, all treatments we used had effect on the expression of STAT1. In conclusion, IL-10 was expressed in HepG2.2.15 cells and STAT1 pathway might be involved in the regulation of IL-10 expression in HepG2.2.15 cells, but it was not the sole pathway, the exact mechanism warrants further study.展开更多
Objective： To prepare and identity monoclonal antibodies （McAbs） against the capsid proteins of adenovirus vector. Methods： BALB/c mice were immunized with a mixture of the purified adenovirus vector （Adv） and A...Objective： To prepare and identity monoclonal antibodies （McAbs） against the capsid proteins of adenovirus vector. Methods： BALB/c mice were immunized with a mixture of the purified adenovirus vector （Adv） and AI（OH）3. McAbs were produced using cell fusion technique in a conventional way. The sensitivity and specificity of monoclonal antibodies was identified by indirect enzyme linked immunosorbent assay （ELISA）, immunocytochemical staining and Western blotting. Results： Six strains of hybridoma cells （A4H11, A8C7, F1H5, G1D2, G4E3 and H2G8） that can stably secrete the IgG1 McAb against Adv were obtained. After 3 months subculture and low concentration of serum adapting culture, six strains retained their stability to secrete McAb. The ascites titers were between 1：10^6 and 1：10^8. Western blot analysis demonstrated that all the McAbs reacted with one protein （about 114 kDa） which is present in wild type 3 adenovirus （wtAd3）, wild type 5 adenovirus （wtAd5）, wild type 7 adenovirus （wtAd7） and adenovirus vector. Conclusion： Successfully prepared six strains of hybridoma cell secreted monoclonal antibodies against the hexon proteins of adenovirus vector, and provided the substantial foundation of preclinical research of adenovirus vectors.展开更多
AIM: To explore the role of mammalian target of rapamycin(m TOR) in the pathogenesis of cirrhotic cardiomyopathy and the potential of rapamycin to improve this pathologic condition.METHODS: Male albino Wistar rats wei...AIM: To explore the role of mammalian target of rapamycin(m TOR) in the pathogenesis of cirrhotic cardiomyopathy and the potential of rapamycin to improve this pathologic condition.METHODS: Male albino Wistar rats weighing 100-120 g were treated with tetrachloride carbon(CCl_4) for 8 wk to induce cirrhosis. Subsequently, animals were administered rapamycin(2 mg/kg per day). The QT_c intervals were calculated in a 5-min electrocardiogram. Then, the left ventricular papillary muscles wereisolated to examine inotropic responsiveness to β-adrenergic stimulation using a standard organ bath equipped by Powerlab system. Phosphorylated-m TOR localization in left ventricles was immunohistochemically assessed, and ventricular tumor necrosis factor(TNF)-α was measured. Western blot was used to measure levels of ventricular phosphorylated-m TOR protein.RESULTS: Cirrhosis was confirmed by hematoxylin and eosin staining of liver tissues, visual observation of lethargy, weight loss, jaundice, brown urine, ascites, liver stiffness, and a significant increase of spleen weight(P < 0.001). A significant prolongation in QTc intervals occurred in cirrhotic rats exposed to CCl_4(P < 0.001), while this prolongation was decreased with rapamycin treatment(P < 0.01). CCl_4-induced cirrhosis caused a significant decrease of contractile responsiveness to isoproterenol stimulation and a significant increase in cardiac TNF-α. These findings were correlated with data from western blot and immunohistochemical studies on phosphorylated-m TOR expression in left ventricles. Phosphorylated-m TOR was significantly enhanced in cirrhotic rats, especially in the endothelium, compared to controls. Rapamycin treatment significantly increased contractile force and myocardial localization of phosphorylated-m TOR and decreased cardiac TNF-α concentration compared to cirrhotic rats with no treatment. CONCLUSION: In this study, we demonstrated a potential role for cardiac m TOR in the pathophysiology of cirrhotic cardiomyopathy. Rapamycin normalized the inotropic effect and altered phosphorylated-m TOR expression and myocardial localization in cirrhotic rats.展开更多
Objective: The significance of isolated high-grade prostatic intraepithelial neoplasia in initial biopsy as an predic-tor for prostate cancer has been extensively research, and the true relationship remnant is no clea...Objective: The significance of isolated high-grade prostatic intraepithelial neoplasia in initial biopsy as an predic-tor for prostate cancer has been extensively research, and the true relationship remnant is no clear till now. The aim of this study is to evaluate prediction value of cancer on repeat biopsy in patients with high-grade prostatic intraepithelial neoplasia, using multivariate analysis. Methods: Thirty-eight men with a diagnosis of isolated high-grade prostatic intraepithelial neo-plasia in initial needle biopsy were studies, in the Fist Affiliated Hospital of Medical School of Xi'an Jiaotong University, from January 2003 to March 2009. These samples were using immunostaining of p63 and 34βE12 and P504s, with a median fol-low-up of 525 (range, 7 to 1650) days, and to researched the incidence of subsequent prostate cancer, and to predicted the risk of prostate cancer in clinicopathological parameters of isolated high-grade prostatic intraepithelial neoplasia on repeat biopsies by logistic regression analysis. Results: There were 10 of 38 (26.3%) men with prostate cancer on repeat biopsies after diagnosis isolated high-grade prostatic intraepithelial neoplasia in initial biopsy, of the rates of prostate cancer were 80% for micropapillary and 75% for cribriform high-grade prostatic intraepithelial neoplasia (P < 0.05), respectively. The positive cores of isolated high-grade prostatic intraepithelial neoplasia was the important for the risk of prostate cancer using Multi-factor logistic regression analysis. The time range in 30 to 690 days was stronger risk for prostate cancer detection after diagnosis isolated HGPIN in initial biopsy. p63 and 34βE12 were disrupted positive expression, and P504S was weak posi-tive expression in the 61% isolated high-grade prostatic intraepithelial neoplasia. Conclusion: Isolated high-grade prostatic intraepithelial neoplasia on repeat biopsy conferred a 26.3% risk of prostate cancer, and this risk level is lower than the previ-ously reported risk of 24% to 58%. The number of positive cores and the histopathological pattern with high-grade prostatic intraepithelial neoplasia on initial biopsy was significantly associated with the risk of cancer.展开更多
·AIM: To investigate the impact of cirrhosis on retinal morphology and to evaluate the role of endogenous opioids as a mediator in cirrhosis induced retinal change.·METHODS: Thirty-six male rats were divided...·AIM: To investigate the impact of cirrhosis on retinal morphology and to evaluate the role of endogenous opioids as a mediator in cirrhosis induced retinal change.·METHODS: Thirty-six male rats were divided into 3main groups; the common bile duct ligated(BDL) group,the sham-operated(Sham) group and the unoperated(Unop) group. Then each of these three main groups was divided into two subgroups; the first subgroup received daily injection of naltrexone hydrochloride(NTX) and the second group was injected with normal saline(Saline)daily. After 28 d, rats were anesthetized and their right eyes were enucleated and assessed for histological changes. The thickness of the rod and cons layer, outer nuclear layer, outer plexiform layer, inner nuclear layer,inner plexiform layer and ganglion cell layer for each eye were measured in micrometers by light microscope.· RESULTS: Ganglion cell layer showed significant increase in thickness in the BDL group(P 【0.05). This increase was eliminated in the group where BDL rats received daily intraperitoneal injection of naltrexone hydrochloride(20 mg/kg). No other histological changes were detected in the other 5 layers we measured·CONCLUSION: The morphological change we detected in the retina of cirrhotic rats is probably due to opioids increased tone in cirrhosis since the increase in thickness in the ganglion cell layer was almost eliminated when naltrexone hydrochloride was injected.These results suggest a possible role for endogenous opioids in the morphological retinal changes detected in cirrhotic rats.展开更多
Cellular metabolism-induced epigenetic regulation is essential for the maintenance of cellular homeostasis.Nicotinamide N-methyltransferase(NNMT)is emerging as a key point of intersection between cellular metabolism a...Cellular metabolism-induced epigenetic regulation is essential for the maintenance of cellular homeostasis.Nicotinamide N-methyltransferase(NNMT)is emerging as a key point of intersection between cellular metabolism and epigenetic regulation and has a central role in various physiological and pathological processes.NNMT catalyzes the methylation of nicotinamide(NAM)using the universal methyl donor S-adenosyl methionine(SAM)to yield S-adeno-syl-L-homocysteine(SAH)and N1-methylnicotinamide(MNAM),directly linking methylation balance with nicotinamide adenosine dinucleotide(NAD+)contents.NNMT acts on either the SAM-methylation balance or both NAD+metabolism,depending on the tissue involved or pathological settings where metabolic demand is increased.Under physiological conditions,the liver act as an essential metabolic organ with abundant NNMT expression,while NNMT hepatic function is not mediated by its methyltransferase activity due to other major methyltransferases such as glycine N-methyltransferase(GNMT)in the liver.However,hepatic NNMT,as well as its metabolite is improperly regulated and linked to the worse pathological states in liver diseases,including alcoholic liver disease,non-alcoholic fatty liver disease(NAFLD),liver cirrhosis,and hepatocellular carcinoma(HCC),suggesting a potential role in the process of liver diseases.In this review,we summarize how NNMT regulates cell methylation balance and NAD metabolism,and extensively outline the current knowledge concerning the functions of NNMT in hepatic metabolism including glucose,lipid and energy,with a specific focus on the contribution of NNMT to the pathophysiology of liver-related diseases.NNMT is involved in the development and progression of liver diseases.Understanding the complex NNMT regulatory network and its effects on pathogenesis could provide new therapeutic strategies in the context of liver diseases.展开更多
Mesenchymal stem cell(MSC)-mediated immunomodulation has been harnessed for the treatment of human diseases,but its underlying mechanism has not been fully understood.Dead cells,including apoptotic cells have immunomo...Mesenchymal stem cell(MSC)-mediated immunomodulation has been harnessed for the treatment of human diseases,but its underlying mechanism has not been fully understood.Dead cells,including apoptotic cells have immunomodulatory properties.It has been repeatedly reported that the proportion of nonviable MSCs in a MSC therapeutic preparation varied from 5-50%in the ongoing clinical trials.It is conceivable that the nonviable cells in a MSC therapeutic preparation may play a role in the therapeutic effects of MSCs.We found that the MSC therapeutic preparation in the present study had about 5%dead MSCs(DMSCs),characterized by apoptotic cells.Namely,1×10^(6) MSCs in the preparation contained about 5×10^(4) DMSCs.We found that the treatment with even 5×10^(4) DMSCs alone had the equal therapeutic effects as with 1×10^(6) MSCs.This protective effect of the dead MSCs alone was confirmed in four mouse models,including concanavalin A(ConA)-and carbon tetrachloride(CCI4)-induced acute liver injury,LPS-induced lung injury and spinal cord injury.We also found that the infused MSCs died by apoptosis in vivo.Furthermore,the therapeutic effect was attributed to the elevated level of phosphatidylserine(PS)upon the injection of MSCs or DMSCs.The direct administration of PS liposomes(PSLs)mimic apoptotic cell fragments also exerted the protective effects as MSCs and DMSCs.The Mer tyrosine kinase(MerTK)deficiency or the knockout of chemokine receptor C-C motif chemokine receptor 2(CCR2)reversed these protective effects of MSCs or DMSCs.These results revealed that DMSCs alone in the therapeutic stem cell preparation or the apoptotic cells induced in vivo may exert the same immunomodulatory property as the'living MSCs preparation"through releasing PS,which was further recognized by MerTK and participated in modulating immune cells.展开更多
Neuropeptide Y（NPY） is widely expressed in the central nervous system and influences many physiological processes.It is located within the rat quantitative trait locus（QTL） for alcohol preference on chromosome 4.A...Neuropeptide Y（NPY） is widely expressed in the central nervous system and influences many physiological processes.It is located within the rat quantitative trait locus（QTL） for alcohol preference on chromosome 4.Alcohol-nonpreferring（NP） rats consume very little alcohol,but have significantly higher NPY expression in the brain than alcohol-preferring（P） rats.We capitalized on this phenotypic difference by creating an Npy knockout（KO） rat using the inbred NP background to evaluate NPY effects on alcohol consumption.Zinc finger nuclease（ZNF） technology was applied,resulting in a 26-bp deletion in the Npy gene.RT-PCR,Western blotting and immunohistochemistry confirmed the absence of Npy mRNA and protein in KO rats.Alcohol consumption was increased in Npy＋/- but not Npy-/- rats,while Npy-/- rats displayed significantly lower body weight when compared to Npy^（＋/＋） rats.In whole brain tissue,expression levels of Npy-related and other alcohol-associated genes,Npy1 r,Npy2r,Npy5 r,Agrp,Mc3 r,Mc4r,Crh and CrMr,were significantly greater in Npy-/- rats,whereas Pome and Crhr2 expressions were highest in Npy＋/- rats.These findings suggest that the NPY-system works in close coordination with the melanocortin（MC） and corticotropin-releasing hormone（CRH） systems to modulate alcohol intake and body weight.展开更多
Background:In vivo cell tracking after transplantation in regenerative medicine remains an unmet challenge and limits current understanding of the wound healing mechanism through cell-based therapies.This study invest...Background:In vivo cell tracking after transplantation in regenerative medicine remains an unmet challenge and limits current understanding of the wound healing mechanism through cell-based therapies.This study investigated tracking of human Wharton’s jelly stem cells(hWJSCs)seeded onto an acellular dermal matrix(ADM)and labeled with superparamagnetic iron oxide nanoparti-cles(SPIONs)by magnetic resonance imaging(MRI)in burn injury.Method:The hWJSCs were characterized and assessed for growth kinetics.A total of 30 rats were enrolled in three equal groups.Group 1 underwent scald burn injury left without treatment,the group 2 was treated by an ADM that was prepared from cosmetic surgery skin samples and the group 3 received hWJSCs labeled with SPIONs seeded onto an ADM.Tensile strength was evaluated before and after interventions,real time PCR assessed apoptosis,and Prussian blue staining,scanning electron microscopy(SEM)and MRI were used for the tracking of labeled cells.Results:The hWJSCs exhibited mesenchymal stem cell properties.Population doubling time was 40.1 hours.SPIONs did not show any toxic effect.The hWJSCs seeded onto an ADM decreased Bax and increased Bcl-2 gene expression.Internalization of SPIONs within hWJSCs was confirmed by Prussian blue staining,SEM and MRI until day 21.There was a significant difference between the Young’s moduli of normal skin and the group receiving hWJSCs seeded onto an ADM.Histological observations and SEM imaging confirmed that MRI is an accurate method to track SPION-labeled hWJSCs in vivo.Conclusions:This study showed that SPION labeling coupled with MRI can be used to further understand the fate of stem cells after transplantation in a burn model.展开更多
Peroxisome proliferator-activated receptorγ(PPARγ)is a transcriptional coactivator that binds to a diverse range of transcription factors.PPARγcoactivator 1(PGC-1)coactivators possess an extensive range of biologic...Peroxisome proliferator-activated receptorγ(PPARγ)is a transcriptional coactivator that binds to a diverse range of transcription factors.PPARγcoactivator 1(PGC-1)coactivators possess an extensive range of biological effects in different tissues,and play a key part in the regulation of the oxidative metabolism,consequently modulating the production of reactive oxygen species,autophagy,and mitochondrial biogenesis.Owing to these findings,a large body of studies,aiming to establish the role of PGC-1 in the neuromuscular system,has shown that PGC-1 could be a promising target for therapies targeting neuromuscular diseases.Among these,some evidence has shown that various signaling pathways linked to PGC-1αare deregulated in muscular dystrophy,leading to a reduced capacity for mitochondrial oxidative phosphorylation and increased reactive oxygen species(ROS)production.In the light of these results,any intervention aimed at activating PGC-1 could contribute towards ameliorating the progression of muscular dystrophies.PGC-1αis influenced by different patho-physiological/pharmacological stimuli.Natural products have been reported to display modulatory effects on PPARγactivation with fewer side effects in comparison to synthetic drugs.Taken together,this review summarizes the current knowledge on Duchenne muscular dystrophy,focusing on the potential effects of natural compounds,acting as regulators of PGC-1α.展开更多
基金National Mega Research Program of China(2008ZX10002-011)National Natural Science Foundation of China(30700701)National High Tech-nology Research and Development program of China(2006AA02Z128)
文摘Hepatitis B virus surface antigen （HBsAg）, a specific antigen on the membrane of Hepatitis B virus （HBV）-infected cells, provides a perfect target for therapeutic drugs. The development of reagents with high affinity and specificity to the HBsAg is of great significance to the early-stage diagnosis and treatment of HBV infection. Herein, we report the selection of RNA aptamers that can specifically bind to HBsAg protein and HBsAg-positive hepatocytes. One high affinity aptamer, HBs-A22, was isolated from an initial 115 met library of -1.1 ×10^15 random-sequence RNA molecules using the SELEX procedure. The selected aptamer HBs-A22 bound specifically to hepatoma cell line HepG2.2.15 that expresses HBsAg but did not bind to HBsAg-devoid HepG2 cells. This is the first reported RNA aptamer which could bind to a HBV specific antigen. This newly isolated aptamer could be modified to deliver imaging, diagnostic, and therapeutic agents targeted at HBV-infected cells.
基金Supported by the National “973” Project of China,No.2011CB910704National Natural Science Foundation of China,No.81372904,No.81570537 and No.81272971
文摘AIM: To discover novel biomarkers for early diagnosis, prognosis or treatment of human colorectal cancer. METHODS: i TRAQ 2D LC-MS/MS analysis was used to identify differentially expressed proteins(DEPs) in the human colonic epithelial carcinogenic process using laser capture microdissection-purified colonic epithelial cells from normal colon, adenoma, carcinoma in situ and invasive carcinoma tissues. RESULTS: A total of 326 DEPs were identified, and four DEPs(DMBT1, S100A9, Galectin-10, and S100A8) with progressive alteration in the carcinogenic process were further validated by immunohistochemistry. The DEPs were involved in multiple biological processes including cell cycle, cell adhesion, translation, m RNA processing, and protein synthesis. Some of the DEPs involved in cellular process such as "translation" and "m RNA splicing" were progressively up-regulated, while some DEPs involved in other processes such as "metabolism" and "cell response to stress" was progressively downregulated. Other proteins with up- or down-regulation at certain stages of carcinogenesis may play various roles at different stages of the colorectal carcinogenic process. CONCLUSION: These findings give insights into our understanding of the mechanisms of colorectal carcinogenesis and provide clues for further investigation of carcinogenesis and identification of biomarkers.
基金supported by grants from the National Major Science and Technology Special Project for Infectious Diseases of China (No. 2008ZX10002-011)
文摘This study investigated the expression profiles of IL-10 gene in three human hepatoma cell lines including Huh7, HepG2, and HepG2 transfected with a plasmid containing hepatitis B virus （HBV） named HepG2.2.15. RT-PCR analysis demonstrated that IL-10 message RNA was absent in HepG2 and Huh7 cells, whereas it was present in HepG2.2.15 cells, which was consistent with ELISA result. Furthermore, except for lamivudine other antiviral treatments did not significantly decrease the HBV DNA level in HepG2.2.15 cells, while they had different effects on the expression of IL-10 protein, although stimulation by LPS had no significant effect. In addition, except for poly（I：C）, the other treatments decreased the expression of IL-10 protein to different degrees, but had no sig-nificant effects on the expression of NF-κB and MyD88. Meanwhile, all treatments we used had effect on the expression of STAT1. In conclusion, IL-10 was expressed in HepG2.2.15 cells and STAT1 pathway might be involved in the regulation of IL-10 expression in HepG2.2.15 cells, but it was not the sole pathway, the exact mechanism warrants further study.
文摘Objective： To prepare and identity monoclonal antibodies （McAbs） against the capsid proteins of adenovirus vector. Methods： BALB/c mice were immunized with a mixture of the purified adenovirus vector （Adv） and AI（OH）3. McAbs were produced using cell fusion technique in a conventional way. The sensitivity and specificity of monoclonal antibodies was identified by indirect enzyme linked immunosorbent assay （ELISA）, immunocytochemical staining and Western blotting. Results： Six strains of hybridoma cells （A4H11, A8C7, F1H5, G1D2, G4E3 and H2G8） that can stably secrete the IgG1 McAb against Adv were obtained. After 3 months subculture and low concentration of serum adapting culture, six strains retained their stability to secrete McAb. The ascites titers were between 1：10^6 and 1：10^8. Western blot analysis demonstrated that all the McAbs reacted with one protein （about 114 kDa） which is present in wild type 3 adenovirus （wtAd3）, wild type 5 adenovirus （wtAd5）, wild type 7 adenovirus （wtAd7） and adenovirus vector. Conclusion： Successfully prepared six strains of hybridoma cell secreted monoclonal antibodies against the hexon proteins of adenovirus vector, and provided the substantial foundation of preclinical research of adenovirus vectors.
基金Supported by Tehran University of Medical Sciences and Health Services grant,No.92033024196
文摘AIM: To explore the role of mammalian target of rapamycin(m TOR) in the pathogenesis of cirrhotic cardiomyopathy and the potential of rapamycin to improve this pathologic condition.METHODS: Male albino Wistar rats weighing 100-120 g were treated with tetrachloride carbon(CCl_4) for 8 wk to induce cirrhosis. Subsequently, animals were administered rapamycin(2 mg/kg per day). The QT_c intervals were calculated in a 5-min electrocardiogram. Then, the left ventricular papillary muscles wereisolated to examine inotropic responsiveness to β-adrenergic stimulation using a standard organ bath equipped by Powerlab system. Phosphorylated-m TOR localization in left ventricles was immunohistochemically assessed, and ventricular tumor necrosis factor(TNF)-α was measured. Western blot was used to measure levels of ventricular phosphorylated-m TOR protein.RESULTS: Cirrhosis was confirmed by hematoxylin and eosin staining of liver tissues, visual observation of lethargy, weight loss, jaundice, brown urine, ascites, liver stiffness, and a significant increase of spleen weight(P < 0.001). A significant prolongation in QTc intervals occurred in cirrhotic rats exposed to CCl_4(P < 0.001), while this prolongation was decreased with rapamycin treatment(P < 0.01). CCl_4-induced cirrhosis caused a significant decrease of contractile responsiveness to isoproterenol stimulation and a significant increase in cardiac TNF-α. These findings were correlated with data from western blot and immunohistochemical studies on phosphorylated-m TOR expression in left ventricles. Phosphorylated-m TOR was significantly enhanced in cirrhotic rats, especially in the endothelium, compared to controls. Rapamycin treatment significantly increased contractile force and myocardial localization of phosphorylated-m TOR and decreased cardiac TNF-α concentration compared to cirrhotic rats with no treatment. CONCLUSION: In this study, we demonstrated a potential role for cardiac m TOR in the pathophysiology of cirrhotic cardiomyopathy. Rapamycin normalized the inotropic effect and altered phosphorylated-m TOR expression and myocardial localization in cirrhotic rats.
基金Supported by a grant from the Key Sci-tech Research Project of Shanxi Province, China (No. 2003K10-G38)
文摘Objective: The significance of isolated high-grade prostatic intraepithelial neoplasia in initial biopsy as an predic-tor for prostate cancer has been extensively research, and the true relationship remnant is no clear till now. The aim of this study is to evaluate prediction value of cancer on repeat biopsy in patients with high-grade prostatic intraepithelial neoplasia, using multivariate analysis. Methods: Thirty-eight men with a diagnosis of isolated high-grade prostatic intraepithelial neo-plasia in initial needle biopsy were studies, in the Fist Affiliated Hospital of Medical School of Xi'an Jiaotong University, from January 2003 to March 2009. These samples were using immunostaining of p63 and 34βE12 and P504s, with a median fol-low-up of 525 (range, 7 to 1650) days, and to researched the incidence of subsequent prostate cancer, and to predicted the risk of prostate cancer in clinicopathological parameters of isolated high-grade prostatic intraepithelial neoplasia on repeat biopsies by logistic regression analysis. Results: There were 10 of 38 (26.3%) men with prostate cancer on repeat biopsies after diagnosis isolated high-grade prostatic intraepithelial neoplasia in initial biopsy, of the rates of prostate cancer were 80% for micropapillary and 75% for cribriform high-grade prostatic intraepithelial neoplasia (P < 0.05), respectively. The positive cores of isolated high-grade prostatic intraepithelial neoplasia was the important for the risk of prostate cancer using Multi-factor logistic regression analysis. The time range in 30 to 690 days was stronger risk for prostate cancer detection after diagnosis isolated HGPIN in initial biopsy. p63 and 34βE12 were disrupted positive expression, and P504S was weak posi-tive expression in the 61% isolated high-grade prostatic intraepithelial neoplasia. Conclusion: Isolated high-grade prostatic intraepithelial neoplasia on repeat biopsy conferred a 26.3% risk of prostate cancer, and this risk level is lower than the previ-ously reported risk of 24% to 58%. The number of positive cores and the histopathological pattern with high-grade prostatic intraepithelial neoplasia on initial biopsy was significantly associated with the risk of cancer.
基金Supported by Lorestan University of Medical Sciences
文摘·AIM: To investigate the impact of cirrhosis on retinal morphology and to evaluate the role of endogenous opioids as a mediator in cirrhosis induced retinal change.·METHODS: Thirty-six male rats were divided into 3main groups; the common bile duct ligated(BDL) group,the sham-operated(Sham) group and the unoperated(Unop) group. Then each of these three main groups was divided into two subgroups; the first subgroup received daily injection of naltrexone hydrochloride(NTX) and the second group was injected with normal saline(Saline)daily. After 28 d, rats were anesthetized and their right eyes were enucleated and assessed for histological changes. The thickness of the rod and cons layer, outer nuclear layer, outer plexiform layer, inner nuclear layer,inner plexiform layer and ganglion cell layer for each eye were measured in micrometers by light microscope.· RESULTS: Ganglion cell layer showed significant increase in thickness in the BDL group(P 【0.05). This increase was eliminated in the group where BDL rats received daily intraperitoneal injection of naltrexone hydrochloride(20 mg/kg). No other histological changes were detected in the other 5 layers we measured·CONCLUSION: The morphological change we detected in the retina of cirrhotic rats is probably due to opioids increased tone in cirrhosis since the increase in thickness in the ganglion cell layer was almost eliminated when naltrexone hydrochloride was injected.These results suggest a possible role for endogenous opioids in the morphological retinal changes detected in cirrhotic rats.
基金supported by grants from the National Natural Science Fund of China(NSFC)(No.82071590).
文摘Cellular metabolism-induced epigenetic regulation is essential for the maintenance of cellular homeostasis.Nicotinamide N-methyltransferase(NNMT)is emerging as a key point of intersection between cellular metabolism and epigenetic regulation and has a central role in various physiological and pathological processes.NNMT catalyzes the methylation of nicotinamide(NAM)using the universal methyl donor S-adenosyl methionine(SAM)to yield S-adeno-syl-L-homocysteine(SAH)and N1-methylnicotinamide(MNAM),directly linking methylation balance with nicotinamide adenosine dinucleotide(NAD+)contents.NNMT acts on either the SAM-methylation balance or both NAD+metabolism,depending on the tissue involved or pathological settings where metabolic demand is increased.Under physiological conditions,the liver act as an essential metabolic organ with abundant NNMT expression,while NNMT hepatic function is not mediated by its methyltransferase activity due to other major methyltransferases such as glycine N-methyltransferase(GNMT)in the liver.However,hepatic NNMT,as well as its metabolite is improperly regulated and linked to the worse pathological states in liver diseases,including alcoholic liver disease,non-alcoholic fatty liver disease(NAFLD),liver cirrhosis,and hepatocellular carcinoma(HCC),suggesting a potential role in the process of liver diseases.In this review,we summarize how NNMT regulates cell methylation balance and NAD metabolism,and extensively outline the current knowledge concerning the functions of NNMT in hepatic metabolism including glucose,lipid and energy,with a specific focus on the contribution of NNMT to the pathophysiology of liver-related diseases.NNMT is involved in the development and progression of liver diseases.Understanding the complex NNMT regulatory network and its effects on pathogenesis could provide new therapeutic strategies in the context of liver diseases.
基金This work was supported by the National Natural Science Foundation Regional Innovation and Development(number U19A2003)National Major Scientific and Technological Special Project for"Significant New Drugs Development”(number 2018ZX09733001)+3 种基金Excellent Youth Foundation of the Sichuan Scientific Committee Grant in China(number 2019JDJQ008)Development Program of China(number 2016YFA0201402)the National Natural Science Foundation of China(number 81800421)the National Natural Science Foundation of China(number 81821002).
文摘Mesenchymal stem cell(MSC)-mediated immunomodulation has been harnessed for the treatment of human diseases,but its underlying mechanism has not been fully understood.Dead cells,including apoptotic cells have immunomodulatory properties.It has been repeatedly reported that the proportion of nonviable MSCs in a MSC therapeutic preparation varied from 5-50%in the ongoing clinical trials.It is conceivable that the nonviable cells in a MSC therapeutic preparation may play a role in the therapeutic effects of MSCs.We found that the MSC therapeutic preparation in the present study had about 5%dead MSCs(DMSCs),characterized by apoptotic cells.Namely,1×10^(6) MSCs in the preparation contained about 5×10^(4) DMSCs.We found that the treatment with even 5×10^(4) DMSCs alone had the equal therapeutic effects as with 1×10^(6) MSCs.This protective effect of the dead MSCs alone was confirmed in four mouse models,including concanavalin A(ConA)-and carbon tetrachloride(CCI4)-induced acute liver injury,LPS-induced lung injury and spinal cord injury.We also found that the infused MSCs died by apoptosis in vivo.Furthermore,the therapeutic effect was attributed to the elevated level of phosphatidylserine(PS)upon the injection of MSCs or DMSCs.The direct administration of PS liposomes(PSLs)mimic apoptotic cell fragments also exerted the protective effects as MSCs and DMSCs.The Mer tyrosine kinase(MerTK)deficiency or the knockout of chemokine receptor C-C motif chemokine receptor 2(CCR2)reversed these protective effects of MSCs or DMSCs.These results revealed that DMSCs alone in the therapeutic stem cell preparation or the apoptotic cells induced in vivo may exert the same immunomodulatory property as the'living MSCs preparation"through releasing PS,which was further recognized by MerTK and participated in modulating immune cells.
基金supported by grants from the National Basic Research Program(No.2013CB945001)the National Science Foundation of China(No.81272273)+1 种基金the State High-Tech Program(No.2012AA022403),PUMC Youth Fundthe National Institutes of Health(Nos.NIAAA P60AA007611,U01AA013522 and R24AA015512)
文摘Neuropeptide Y（NPY） is widely expressed in the central nervous system and influences many physiological processes.It is located within the rat quantitative trait locus（QTL） for alcohol preference on chromosome 4.Alcohol-nonpreferring（NP） rats consume very little alcohol,but have significantly higher NPY expression in the brain than alcohol-preferring（P） rats.We capitalized on this phenotypic difference by creating an Npy knockout（KO） rat using the inbred NP background to evaluate NPY effects on alcohol consumption.Zinc finger nuclease（ZNF） technology was applied,resulting in a 26-bp deletion in the Npy gene.RT-PCR,Western blotting and immunohistochemistry confirmed the absence of Npy mRNA and protein in KO rats.Alcohol consumption was increased in Npy＋/- but not Npy-/- rats,while Npy-/- rats displayed significantly lower body weight when compared to Npy^（＋/＋） rats.In whole brain tissue,expression levels of Npy-related and other alcohol-associated genes,Npy1 r,Npy2r,Npy5 r,Agrp,Mc3 r,Mc4r,Crh and CrMr,were significantly greater in Npy-/- rats,whereas Pome and Crhr2 expressions were highest in Npy＋/- rats.These findings suggest that the NPY-system works in close coordination with the melanocortin（MC） and corticotropin-releasing hormone（CRH） systems to modulate alcohol intake and body weight.
文摘Background:In vivo cell tracking after transplantation in regenerative medicine remains an unmet challenge and limits current understanding of the wound healing mechanism through cell-based therapies.This study investigated tracking of human Wharton’s jelly stem cells(hWJSCs)seeded onto an acellular dermal matrix(ADM)and labeled with superparamagnetic iron oxide nanoparti-cles(SPIONs)by magnetic resonance imaging(MRI)in burn injury.Method:The hWJSCs were characterized and assessed for growth kinetics.A total of 30 rats were enrolled in three equal groups.Group 1 underwent scald burn injury left without treatment,the group 2 was treated by an ADM that was prepared from cosmetic surgery skin samples and the group 3 received hWJSCs labeled with SPIONs seeded onto an ADM.Tensile strength was evaluated before and after interventions,real time PCR assessed apoptosis,and Prussian blue staining,scanning electron microscopy(SEM)and MRI were used for the tracking of labeled cells.Results:The hWJSCs exhibited mesenchymal stem cell properties.Population doubling time was 40.1 hours.SPIONs did not show any toxic effect.The hWJSCs seeded onto an ADM decreased Bax and increased Bcl-2 gene expression.Internalization of SPIONs within hWJSCs was confirmed by Prussian blue staining,SEM and MRI until day 21.There was a significant difference between the Young’s moduli of normal skin and the group receiving hWJSCs seeded onto an ADM.Histological observations and SEM imaging confirmed that MRI is an accurate method to track SPION-labeled hWJSCs in vivo.Conclusions:This study showed that SPION labeling coupled with MRI can be used to further understand the fate of stem cells after transplantation in a burn model.
基金supported by the crowd funding#Sport4Therapy to Giuseppe D’Antona(Italy)supported by Instituto de Salud CarlosⅢ,Grant Number:CIBEROBN CB12/03/30038
文摘Peroxisome proliferator-activated receptorγ(PPARγ)is a transcriptional coactivator that binds to a diverse range of transcription factors.PPARγcoactivator 1(PGC-1)coactivators possess an extensive range of biological effects in different tissues,and play a key part in the regulation of the oxidative metabolism,consequently modulating the production of reactive oxygen species,autophagy,and mitochondrial biogenesis.Owing to these findings,a large body of studies,aiming to establish the role of PGC-1 in the neuromuscular system,has shown that PGC-1 could be a promising target for therapies targeting neuromuscular diseases.Among these,some evidence has shown that various signaling pathways linked to PGC-1αare deregulated in muscular dystrophy,leading to a reduced capacity for mitochondrial oxidative phosphorylation and increased reactive oxygen species(ROS)production.In the light of these results,any intervention aimed at activating PGC-1 could contribute towards ameliorating the progression of muscular dystrophies.PGC-1αis influenced by different patho-physiological/pharmacological stimuli.Natural products have been reported to display modulatory effects on PPARγactivation with fewer side effects in comparison to synthetic drugs.Taken together,this review summarizes the current knowledge on Duchenne muscular dystrophy,focusing on the potential effects of natural compounds,acting as regulators of PGC-1α.
