The adsorption behavior of ion exchange resin D301 in the extraction of hexavalent molybdenum from high acidic leach solution was investigated. SEM, EDS and Raman spectra analyses were applied to studying the adsorpti...The adsorption behavior of ion exchange resin D301 in the extraction of hexavalent molybdenum from high acidic leach solution was investigated. SEM, EDS and Raman spectra analyses were applied to studying the adsorption capacity, reaction kinetics and possible adsorption mechanism in detail. Results showed that the adsorption capacity of D301 resin for molybdenum from high acidic leach solution was up to 463.63 mg/g. Results of the kinetic analysis indicated that the adsorption process was controlled by the particle diffusion with the activation energy 25.47 k J/mol（0.9-1.2 mm） and 20.38 k J/mol（0.6-0.9 mm）. Furthermore, the molybdenum loaded on the resin could be eluted by using 2 mol/L ammonia hydroxide solution. Besides, dynamic continuous column experiments verified direct extraction of molybdenum from acidic leach solutions by ion exchange resin D301 and the upstream flow improved dynamic continuous absorption.展开更多
AIM： To enrich hepatic progenitors using epithelial cell adhesion molecule （EpCAM） as a marker from human fetal liver and investigate the expression of human leukocyte antigen （HLA） and their markers associated w...AIM： To enrich hepatic progenitors using epithelial cell adhesion molecule （EpCAM） as a marker from human fetal liver and investigate the expression of human leukocyte antigen （HLA） and their markers associated with hepatic progenitor cells. METHODS： EpCAM ＋ve cells were isolated using magnetic cell sorting （MACS） from human fetuses （n = 10） at 15-25 wk gestation. Expression of markers for hepatic progenitors such as albumin, alpha-fetoprotein （AFP）, CD29 （integrin ~1）, CD49f （integrin c^6） and CD90 （Thy 1） was studied by using flow cytometry, immunocytochemistry and RT-PCR; HLA class Ⅰ （A, B, C） and class Ⅱ （DR） expression was studied by flow cytometry only. RESULTS： FACS analysis indicated that EpCAM ＋ve cells were positive for CD29, CD49f, CD90, CD34, HLA class I, albumin and AFP but negative for HLA class Ⅱ （DR） and CD45. RT PCR showed that EpCAM ＋ve cells expressed liver epithelial markers （CK18）, biliary specific marker （CK19） and hepatic markers （albumin, AFP）. On immunocytochemical staining, EpCAM ＋ve cells were shown positive signals for CK18 and albumin. CONCLUSION： Our study suggests that these EpCAM ＋ve cells can be used as hepatic progenitors for cell transplantation with a minimum risk of alloreactivity and these cells may serve as a potential source for enrichment of hepatic progenitor.展开更多
Objective To investigate the apoptosis of epididyrnis epithelium and the change of epididymissialic acid following torsed/detorsed testes. Methods Twenty four adult male Sprague-Dawley rats were subjected to unilatera...Objective To investigate the apoptosis of epididyrnis epithelium and the change of epididymissialic acid following torsed/detorsed testes. Methods Twenty four adult male Sprague-Dawley rats were subjected to unilateral 720 testicular torsion with the duration of 2h and 4h, then repaired. The ischemic epididymis were collected for detecting the content of sialic acid by using spectrophotometry and the apoptosis with TUNEL technique. Results There were no statistically significant difference in the apoptosis of epididymis epithelium [(9.51± 2.78)% vs (6.34±1. 98)%] and the content of epididymis sialic acid(23, 3851 ± 9. 2199mg/mgprot vs 19. 3661 6. 3373mg/mgprot) at 24h between following 2h-torsed/detorsed testes and those of sham group. There were statistically significant difference in the apopotosis of epididymis epithelium[ (46. 81 ±3. 55)% vs (6. 34±1. 98) % ] and the content of epididymis sialic acid (13. 7249±7. 8006mg/mgprot vs 19. 3661±6. 3373mg/mgprot) at 24h between following 4h-torsed/detorsed testes and those of sham group(P <0. 05). Conclusion The results suggest that the sialic acid-secreting-function of epididymis remain normal at 24h following 2h-torsed/detorsed testes, while the apoptosis index of epididymis epithelium do not increase. The epididymis would be injured at 24h following 4h-torsed/detorsed testes, while the apoptosis index increased.展开更多
Objective To investigate the effect of microRNA-205 reduction by antagomirs on adhesion ability of normal human corneal epithelial keratinocytes(NHCEKs).Methods Antagomir-205,complementary and inhibitory to microRNA-2...Objective To investigate the effect of microRNA-205 reduction by antagomirs on adhesion ability of normal human corneal epithelial keratinocytes(NHCEKs).Methods Antagomir-205,complementary and inhibitory to microRNA-205,was used to suppress endogenous microRNA-205 in NHCEKs.The adhesion ability of treated NHCEKs was then assessed by cell adhesion assay.Immunoblot and immunohistochemistry were conducted to determine the level of two focal adhesion-related proteins,focal adhesion kinase(FAK) and paxillin(Pax).Phalloidin staining was performed to measure the level of filamentous actin in antagomir-treated NHCEKs.Results Antagomir-205 markedly reduced the level of microRNA-205 in NHCEKs and significantly enhanced adhesion ability of NHCEKs(P<0.01).Further protein analysis validated that inhibition of mi-croRNA-205 increased the number of phosphorylated FAK and phosphorylated Pax,and decreased filamen-tous actin.Conclusion Our findings suggest that microRNA-205 has down-regulating effect on cell motility in NHCEKs.展开更多
Objective To discuss the application of semi-precision attachment in restoring a midfacial defect, including a nasal, upper lip, and anterior maxillary defect. Methods A splinted metal-ceramic crowns （from the upper ...Objective To discuss the application of semi-precision attachment in restoring a midfacial defect, including a nasal, upper lip, and anterior maxillary defect. Methods A splinted metal-ceramic crowns （from the upper right first molar to left molar） and a bar with stud attachments was done. A removable partial denture （RPD） containing the patrix portion of the attachment was then designed to restore the missing maxillary anterior teeth and alveolar ridge. Finally,, The facial prosthesis was joined to the denture utilizing two retentive metal posts on the superior aspect of the removable partial. Results A splinted metal-ceramic crowns （from the upper right frst molar to left molar） and a bar with stud attachments was done. A RPD containing the patrix portion of the attachment was then designed to restore the missing maxillary anterior teeth and alveolar ridge. Finally, The facial prosthesis was joined to the denture utilizing two retentive metal posts on the superior aspect of the removable partial. The prosthesis markedly improved the appearance of the patient and demonstrated good retention. Conclusion Using attachment in restoring rnaxillofacial defect may provide adequate retetion which lead to a sucessful treatment outcome.展开更多
To determine the role of corticotropin releasing factor receptor (CRF2) in epithelial permeability and enterocyte cell differentiation.METHODSFor this purpose, we used rat Sprague Dawley and various colon carcinoma ce...To determine the role of corticotropin releasing factor receptor (CRF2) in epithelial permeability and enterocyte cell differentiation.METHODSFor this purpose, we used rat Sprague Dawley and various colon carcinoma cell lines (SW620, HCT8R, HT-29 and Caco-2 cell lines). Expression of CRF2 protein was analyzed by fluorescent immunolabeling in normal rat colon and then by western blot in dissociated colonic epithelial cells and in the lysates of colon carcinoma cell lines or during the early differentiation of HT-29 cells (ten first days). To assess the impact of CRF2 signaling on colonic cell differentiation, HT-29 and Caco-2 cells were exposed to Urocortin 3 recombinant proteins (Ucn3, 100 nmol/L). In some experiments, cells were pre-exposed to the astressin 2b (A2b) a CRF2 antagonist in order to inhibit the action of Ucn3. Intestinal cell differentiation was first analyzed by functional assays: the trans-cellular permeability and the para-cellular permeability were determined by Dextran-FITC intake and measure of the transepithelial electrical resistance respectively. Morphological modifications associated to epithelial dysfunction were analyzed by confocal microscopy after fluorescent labeling of actin (phaloidin-TRITC) and intercellular adhesion proteins such as E-cadherin, p120ctn, occludin and ZO-1. The establishment of mature adherens junctions (AJ) was monitored by following the distribution of AJ proteins in lipid raft fractions, after separation of cell lysates on sucrose gradients. Finally, the mRNA and the protein expression levels of characteristic markers of intestinal epithelial cell (IEC) differentiation such as the transcriptional factor krüppel-like factor 4 (KLF4) or the dipeptidyl peptidase IV (DPPIV) were performed by RT-PCR and western blot respectively. The specific activities of DPPIV and alkaline phosphatase (AP) enzymes were determined by a colorimetric method.RESULTSCRF2 protein is preferentially expressed in undifferentiated epithelial cells from the crypts of colon and in human colon carcinoma cell lines. Furthermore, CRF2 expression is down regulated according to the kinetic of HT-29 cell differentiation. By performing functional assays, we found that Ucn3-induced CRF2 signaling alters both para- and trans-cellular permeability of differentiated HT-29 and Caco-2 cells. These effects are partly mediated by Ucn3-induced morphological changes associated with the disruption of mature AJ in HT-29 cells and tight junctions (TJ) in Caco-2 cells. Ucn3-mediated activation of CRF2 decreases mRNA and protein expression levels of KLF4 a transcription factor involved in IEC differentiation. This signaling is correlated to a down-regulation of key IEC markers such as DPPIV and AP, at both transcriptional and post-transcriptional levels.CONCLUSIONOur findings suggest that CRF2 signaling could modulate IEC differentiation. These mechanisms could be relevant to the stress induced epithelial alterations found in inflammatory bowel diseases.展开更多
Human primary epithelial cells of renal pelvis was established to investigate the adherence of uropathogenic Escherichia coli （UPEC） to this cell line, in which the primary cell culture was performed by using cultiv...Human primary epithelial cells of renal pelvis was established to investigate the adherence of uropathogenic Escherichia coli （UPEC） to this cell line, in which the primary cell culture was performed by using cultivation of the normal epithelium of renal pelvis in keratinocyte serum free medium （K-SFM） with epidermal growth factor （EGF） and bovine pituitary extract （BPE）. Both UPEC132 obtained from urine specimen of patients with pyelonephritis and the pilus-free representative strain E. coli K-12p678- 54 were used to study the adherence of these strains on human primary epithelial cells of renal pelvis. The UPEC adherence was performed with observation on the morphological changes of the adhered cells, while the adhesion rates and indices were calculated in different times of experiment. In addition, the virulence genes hly and cnfl of UPEC132 were detected by multiplex PCR assay. In this study, the human primary epithelial cells of renal pelvis was found to exhibit the character of the transitional epithelial cells. Compared with the control group, the adhesion rates and indices began to increase from 15 min of the experiment time and reached its peak in 120 min. The adhesion rate and index of UPEC132 to human primary epithelial cells of renal pelvis were 74.4% and 34.0 respectively. Many microscopic changes in the primary cells adhered with UPEC132 could be detected, such as rounding or irregularity in shape, unevenness in staining and the cytoplasmic and nuclear changes. It suggests that human primary epithelial cells of renal pelvis can be used for the experiment on UPEC adhesion, thus providing a basis for the further study on the pathogenesis of UPEC.展开更多
基金Projects(21376251,21406233) supported by the National Natural Science Foundation of China
文摘The adsorption behavior of ion exchange resin D301 in the extraction of hexavalent molybdenum from high acidic leach solution was investigated. SEM, EDS and Raman spectra analyses were applied to studying the adsorption capacity, reaction kinetics and possible adsorption mechanism in detail. Results showed that the adsorption capacity of D301 resin for molybdenum from high acidic leach solution was up to 463.63 mg/g. Results of the kinetic analysis indicated that the adsorption process was controlled by the particle diffusion with the activation energy 25.47 k J/mol（0.9-1.2 mm） and 20.38 k J/mol（0.6-0.9 mm）. Furthermore, the molybdenum loaded on the resin could be eluted by using 2 mol/L ammonia hydroxide solution. Besides, dynamic continuous column experiments verified direct extraction of molybdenum from acidic leach solutions by ion exchange resin D301 and the upstream flow improved dynamic continuous absorption.
基金Council of Scientific and Industrial Research Network Grant CMM002ICMR Grant (GAP 0215)
文摘AIM： To enrich hepatic progenitors using epithelial cell adhesion molecule （EpCAM） as a marker from human fetal liver and investigate the expression of human leukocyte antigen （HLA） and their markers associated with hepatic progenitor cells. METHODS： EpCAM ＋ve cells were isolated using magnetic cell sorting （MACS） from human fetuses （n = 10） at 15-25 wk gestation. Expression of markers for hepatic progenitors such as albumin, alpha-fetoprotein （AFP）, CD29 （integrin ~1）, CD49f （integrin c^6） and CD90 （Thy 1） was studied by using flow cytometry, immunocytochemistry and RT-PCR; HLA class Ⅰ （A, B, C） and class Ⅱ （DR） expression was studied by flow cytometry only. RESULTS： FACS analysis indicated that EpCAM ＋ve cells were positive for CD29, CD49f, CD90, CD34, HLA class I, albumin and AFP but negative for HLA class Ⅱ （DR） and CD45. RT PCR showed that EpCAM ＋ve cells expressed liver epithelial markers （CK18）, biliary specific marker （CK19） and hepatic markers （albumin, AFP）. On immunocytochemical staining, EpCAM ＋ve cells were shown positive signals for CK18 and albumin. CONCLUSION： Our study suggests that these EpCAM ＋ve cells can be used as hepatic progenitors for cell transplantation with a minimum risk of alloreactivity and these cells may serve as a potential source for enrichment of hepatic progenitor.
文摘Objective To investigate the apoptosis of epididyrnis epithelium and the change of epididymissialic acid following torsed/detorsed testes. Methods Twenty four adult male Sprague-Dawley rats were subjected to unilateral 720 testicular torsion with the duration of 2h and 4h, then repaired. The ischemic epididymis were collected for detecting the content of sialic acid by using spectrophotometry and the apoptosis with TUNEL technique. Results There were no statistically significant difference in the apoptosis of epididymis epithelium [(9.51± 2.78)% vs (6.34±1. 98)%] and the content of epididymis sialic acid(23, 3851 ± 9. 2199mg/mgprot vs 19. 3661 6. 3373mg/mgprot) at 24h between following 2h-torsed/detorsed testes and those of sham group. There were statistically significant difference in the apopotosis of epididymis epithelium[ (46. 81 ±3. 55)% vs (6. 34±1. 98) % ] and the content of epididymis sialic acid (13. 7249±7. 8006mg/mgprot vs 19. 3661±6. 3373mg/mgprot) at 24h between following 4h-torsed/detorsed testes and those of sham group(P <0. 05). Conclusion The results suggest that the sialic acid-secreting-function of epididymis remain normal at 24h following 2h-torsed/detorsed testes, while the apoptosis index of epididymis epithelium do not increase. The epididymis would be injured at 24h following 4h-torsed/detorsed testes, while the apoptosis index increased.
基金Supported by Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences Grant (2009RC03)National Institutes of Health Grants (EY017536,EY019463)
文摘Objective To investigate the effect of microRNA-205 reduction by antagomirs on adhesion ability of normal human corneal epithelial keratinocytes(NHCEKs).Methods Antagomir-205,complementary and inhibitory to microRNA-205,was used to suppress endogenous microRNA-205 in NHCEKs.The adhesion ability of treated NHCEKs was then assessed by cell adhesion assay.Immunoblot and immunohistochemistry were conducted to determine the level of two focal adhesion-related proteins,focal adhesion kinase(FAK) and paxillin(Pax).Phalloidin staining was performed to measure the level of filamentous actin in antagomir-treated NHCEKs.Results Antagomir-205 markedly reduced the level of microRNA-205 in NHCEKs and significantly enhanced adhesion ability of NHCEKs(P<0.01).Further protein analysis validated that inhibition of mi-croRNA-205 increased the number of phosphorylated FAK and phosphorylated Pax,and decreased filamen-tous actin.Conclusion Our findings suggest that microRNA-205 has down-regulating effect on cell motility in NHCEKs.
基金grant of Shanghai Leading Academic Discipline Fund(T0202)
文摘Objective To discuss the application of semi-precision attachment in restoring a midfacial defect, including a nasal, upper lip, and anterior maxillary defect. Methods A splinted metal-ceramic crowns （from the upper right first molar to left molar） and a bar with stud attachments was done. A removable partial denture （RPD） containing the patrix portion of the attachment was then designed to restore the missing maxillary anterior teeth and alveolar ridge. Finally,, The facial prosthesis was joined to the denture utilizing two retentive metal posts on the superior aspect of the removable partial. Results A splinted metal-ceramic crowns （from the upper right frst molar to left molar） and a bar with stud attachments was done. A RPD containing the patrix portion of the attachment was then designed to restore the missing maxillary anterior teeth and alveolar ridge. Finally, The facial prosthesis was joined to the denture utilizing two retentive metal posts on the superior aspect of the removable partial. The prosthesis markedly improved the appearance of the patient and demonstrated good retention. Conclusion Using attachment in restoring rnaxillofacial defect may provide adequate retetion which lead to a sucessful treatment outcome.
基金Supported by grants from Association pour la Recherche sur le Cancer,Ligue Nationale contre le Cancer,No.GEFLUC and No.ESPOIR
文摘To determine the role of corticotropin releasing factor receptor (CRF2) in epithelial permeability and enterocyte cell differentiation.METHODSFor this purpose, we used rat Sprague Dawley and various colon carcinoma cell lines (SW620, HCT8R, HT-29 and Caco-2 cell lines). Expression of CRF2 protein was analyzed by fluorescent immunolabeling in normal rat colon and then by western blot in dissociated colonic epithelial cells and in the lysates of colon carcinoma cell lines or during the early differentiation of HT-29 cells (ten first days). To assess the impact of CRF2 signaling on colonic cell differentiation, HT-29 and Caco-2 cells were exposed to Urocortin 3 recombinant proteins (Ucn3, 100 nmol/L). In some experiments, cells were pre-exposed to the astressin 2b (A2b) a CRF2 antagonist in order to inhibit the action of Ucn3. Intestinal cell differentiation was first analyzed by functional assays: the trans-cellular permeability and the para-cellular permeability were determined by Dextran-FITC intake and measure of the transepithelial electrical resistance respectively. Morphological modifications associated to epithelial dysfunction were analyzed by confocal microscopy after fluorescent labeling of actin (phaloidin-TRITC) and intercellular adhesion proteins such as E-cadherin, p120ctn, occludin and ZO-1. The establishment of mature adherens junctions (AJ) was monitored by following the distribution of AJ proteins in lipid raft fractions, after separation of cell lysates on sucrose gradients. Finally, the mRNA and the protein expression levels of characteristic markers of intestinal epithelial cell (IEC) differentiation such as the transcriptional factor krüppel-like factor 4 (KLF4) or the dipeptidyl peptidase IV (DPPIV) were performed by RT-PCR and western blot respectively. The specific activities of DPPIV and alkaline phosphatase (AP) enzymes were determined by a colorimetric method.RESULTSCRF2 protein is preferentially expressed in undifferentiated epithelial cells from the crypts of colon and in human colon carcinoma cell lines. Furthermore, CRF2 expression is down regulated according to the kinetic of HT-29 cell differentiation. By performing functional assays, we found that Ucn3-induced CRF2 signaling alters both para- and trans-cellular permeability of differentiated HT-29 and Caco-2 cells. These effects are partly mediated by Ucn3-induced morphological changes associated with the disruption of mature AJ in HT-29 cells and tight junctions (TJ) in Caco-2 cells. Ucn3-mediated activation of CRF2 decreases mRNA and protein expression levels of KLF4 a transcription factor involved in IEC differentiation. This signaling is correlated to a down-regulation of key IEC markers such as DPPIV and AP, at both transcriptional and post-transcriptional levels.CONCLUSIONOur findings suggest that CRF2 signaling could modulate IEC differentiation. These mechanisms could be relevant to the stress induced epithelial alterations found in inflammatory bowel diseases.
基金This work was supported by the grants from the National Natural Science Foundation of China （30470096）.
文摘Human primary epithelial cells of renal pelvis was established to investigate the adherence of uropathogenic Escherichia coli （UPEC） to this cell line, in which the primary cell culture was performed by using cultivation of the normal epithelium of renal pelvis in keratinocyte serum free medium （K-SFM） with epidermal growth factor （EGF） and bovine pituitary extract （BPE）. Both UPEC132 obtained from urine specimen of patients with pyelonephritis and the pilus-free representative strain E. coli K-12p678- 54 were used to study the adherence of these strains on human primary epithelial cells of renal pelvis. The UPEC adherence was performed with observation on the morphological changes of the adhered cells, while the adhesion rates and indices were calculated in different times of experiment. In addition, the virulence genes hly and cnfl of UPEC132 were detected by multiplex PCR assay. In this study, the human primary epithelial cells of renal pelvis was found to exhibit the character of the transitional epithelial cells. Compared with the control group, the adhesion rates and indices began to increase from 15 min of the experiment time and reached its peak in 120 min. The adhesion rate and index of UPEC132 to human primary epithelial cells of renal pelvis were 74.4% and 34.0 respectively. Many microscopic changes in the primary cells adhered with UPEC132 could be detected, such as rounding or irregularity in shape, unevenness in staining and the cytoplasmic and nuclear changes. It suggests that human primary epithelial cells of renal pelvis can be used for the experiment on UPEC adhesion, thus providing a basis for the further study on the pathogenesis of UPEC.
