Accompanying rapid developments in hepatic surgery,the number of surgeries and identifications of histological types of primary hepatic space-occupying lesions (PHSOLs) have increased dramatically.This has led to many...Accompanying rapid developments in hepatic surgery,the number of surgeries and identifications of histological types of primary hepatic space-occupying lesions (PHSOLs) have increased dramatically.This has led to many changes in the surgicopathological spectrum of PHSOLs,and has contributed to a theoretical basis for modern hepatic surgery and oncological pathology.Between 1982 and 2009 at the Eastern Hepatobiliary Surgery Hospital (EHBH) in Shanghai,31 901 patients underwent surgery and were diagnosed as having a PHSOL.In this paper,we present an analysis of the PHSOL cases at the EHBH for this time period,along with results from a systematic literature review.We describe a surgicopathological spectrum comprising more than 100 types of PHSOLs that can be stratified into three types:tumor-like,benign,and malignant.We also stratified the PHSOLs into six subtypes derived from hepatocytes;cholangiocytes;vascular,lymphoid and hemopoietic tissues;muscular,fibrous and adipose tissues;neural and neuroendocrine tissues;and miscellaneous tissues.The present study provides a new classification system that can be used as a current reference for clinicians and pathologists to make correct diagnoses and differential diagnoses among various PHSOLs.展开更多
AIM:To evaluate the functions of a new bioartificial liver(BAL)system in vitro and in vitro.MEHTODS:The BAL system was configurated byinoculating porcine hepatocyte spheroids into the cellcircuit of a hollow fiber bio...AIM:To evaluate the functions of a new bioartificial liver(BAL)system in vitro and in vitro.MEHTODS:The BAL system was configurated byinoculating porcine hepatocyte spheroids into the cellcircuit of a hollow fiber bioreactor.In the experiments ofBAL in vitro,the levels of alanine aminotransferase(ALT),total bilirubin(TB),and albumin(ALB)in the circulatinghepatocyte suspension and RPMI-1640 medium weredetermined during 6 h of circulation in the BAL device.In the experiments of BAL in vitro,acute liver failure(ALF)model in canine was induced by an end-side portocavalshunt combined with common bile duct ligation andtransaction.Blood ALT,TB and ammonia levels ofALF in canines were determined before and after BALtreatment.RESULTS:During 6 h of circulation in vitro,therewas no significant change of ALT,whereas the TB andALB levels gradually increased with time both in thehepatocyte suspension and in RPMI-1640 medium.Inthe BAL treatment group,blood ALT,TB and ammonialevels of ALF in canines decreased significantly.CONCLUSION:The new BAL system has the ability toperform liver functions and can be used to treat ALF.展开更多
AIM: To evaluate the effects of survivin on cell proliferation and apoptosis in liver cancer. METHODS: MTT assay was used to generate and optimize phosphorothioate antisense oligonucleotides (ODNs)-LipofectamineTM2000...AIM: To evaluate the effects of survivin on cell proliferation and apoptosis in liver cancer. METHODS: MTT assay was used to generate and optimize phosphorothioate antisense oligonucleotides (ODNs)-LipofectamineTM2000 (LiP) compound by varying ODNs (μg): LiP (μL) ratios from 1:0.5 to 1:5. Then, liver cancer cells (HepG2) were transfected with the compound. By using RT-PCR and Western blot, the expression levels of survivin mRNA and proteins were detected in HepG2 cells treated with antisense compounds (ODNs:LiP=1:4), and compared with those treated with sense compounds (1:4) as control. MTT assay was applied to the determination of cell proliferation in HepG2 cells. Active caspase-3 was evaluated by flow cytometric analysis. The morphological changes were assessed by electron microscopy. Laser scanning confocal microscopy was performed to detect the subcellular localization of survivin proteins in treated and untreated cells. RESULTS: Antisense compounds (1:4) down-regulated survivin expression (mRNA and protein) in a dose-dependent manner with an IC50 of 250 nmol/L. Its maximum effect was achieved at a concentration of 500 nmol/L, at which mRNA and protein levels were down-regulated by 80%. The similar results were found in MTT assay. Antisense compound (l:4)-treated cells revealed increased caspase-3-like protease activity compared with untreated cells. Untreated cells as control were primarily negative for the presence of active-caspase-3. As shown by transmission electron microscopy, treated cells with antisense compounds (1:4) resulted in morphological changes such as blebbing and loss of microvilli, vacuolization in the cytoplasm, condensation of the cytoplasm and nuclei, and fragmented chromatin. Immunofluorescence analysis confirmed the presence of survivin protein pool inside the cytoplasm in untreated cells. Labeled-FITC immunofluorescence staining of survivin clearly showed that survivin was distributed mainly in a spotted form inside the cytoplasm. Whereas cells treated with antisense compounds were rare and weak inside the cytoplasm. CONCLUSION: Down-regulation of survivin expression induced by the antisense compounds reduces tumor growth potential, promotes apoptosis and affects the localization of survivin proteins in HepG2 cells. Furthermore, survivin protein is a key molecule associated with proliferation and apoptosis, and antisense oligonucleotides targeting survivin have a bright prospect in the therapy of liver cancer.展开更多
A rat liver epithelial cell line designated LW13 was established using a sequential sedimentation method. The cell line retained many normal properties of liver epithelial cells and showed some structural and function...A rat liver epithelial cell line designated LW13 was established using a sequential sedimentation method. The cell line retained many normal properties of liver epithelial cells and showed some structural and functional features resembling those of liver parenchymal cells. LW13 cells became malignant after the introduction of exogenous transforming EJ Ha ras gene. Tumors produced by inoculation of the transformed cells into baby rats .contained areas of poorly differentiated hepatocellular carcinoma. In situ hybridization analysis confirmed the random rather than specific integration of exogenous ras gene into host chromosomes. Furthermore , an at least tenfold increase in the expression of the endogenous c myc gene was detected among transformed cell lines, suggesting the involvement of the c myc proto oncogene in the in vitro transformation of rat liver epithelial cells by EJ Ha ras oncogene.展开更多
Objective To investigate the relationship between expression of somatostatin receptors(SSTRs) and activation of rat hepatic stellate cell (HSC). Methods HSCs were isolated from rats by in situ perfusion and single-ste...Objective To investigate the relationship between expression of somatostatin receptors(SSTRs) and activation of rat hepatic stellate cell (HSC). Methods HSCs were isolated from rats by in situ perfusion and single-step density gradient centrifugation, and then SSTR1-5 mRNA levels in the differentiated first passage HSCs were detected by means of reverse transcription polymerase chain reaction. On the other hand, hepatic fibrosis was induced in adult male Sprague-Dawley rats by carbon tetrachloride intoxication, and the expression of SSTR1-5 in normal as well as fibrotic liver was measured by immunohistochemical staining. Results SSTR mR-NA and SSTR could not be found in freshly isolated rat HSCs and normal rat liver. But SSTR1-3 mRNA appeared as HSCs became wholly activated, and SSTR1-3 could also be identified on the membrane of activated HSCs in the peri-sinusoid space, fibrous septa, etc. Conclusion The expression of SSTR1-3 in the rat HSC is closely related to its activation. This may reflect one of the main negative regulation mechanisms in the course of HSC activation.展开更多
AIM: To study genetic difference of mitochondrial DMA (mtDNA) between two hepatocarcinoma cell lines (Hca-F and Hca-P) with diverse metastatic characteristics and the relationship between mtDNA changes in cancer cells...AIM: To study genetic difference of mitochondrial DMA (mtDNA) between two hepatocarcinoma cell lines (Hca-F and Hca-P) with diverse metastatic characteristics and the relationship between mtDNA changes in cancer cells and ttieir oncogenic phenotype. METHODS: Mitochondrial DMA D-loop, tRNAMet+Glu+Ile and ND3 gene fragments from the hepatocarcinoma cell lines with 1100,1126 and 534 bp in length respectively were analysed by PCR amplification and restriction fragment length polymorphism techniques. The D-loop 3′ end sequence of the hepatocarcinoma cell lines was determined by sequencing. RESULTS: No amplification fragment length polymorphism and restriction fragment length polymorphism were observed in tRNAMet+Glu+Ile, ND3 and D-loop of mitochondrial DNA of the hepatocarcinoma cells. Sequence differences between Hca-F and Hca-P were found in mtDNA D-loop. CONCLUSION: Deletion mutations of mitochondrial DNA restriction fragment may not play a significant role in carcinogenesis. Genetic difference of mtDNA D-loop between Hca-F and Hca-P, which may reflect the environmental and genetic influences during tumor progression, could be linked to their tumorigenic phenotypes.展开更多
基金Supported by The National Nature Science Foundation of China,No.30872506 and No.81072026
文摘Accompanying rapid developments in hepatic surgery,the number of surgeries and identifications of histological types of primary hepatic space-occupying lesions (PHSOLs) have increased dramatically.This has led to many changes in the surgicopathological spectrum of PHSOLs,and has contributed to a theoretical basis for modern hepatic surgery and oncological pathology.Between 1982 and 2009 at the Eastern Hepatobiliary Surgery Hospital (EHBH) in Shanghai,31 901 patients underwent surgery and were diagnosed as having a PHSOL.In this paper,we present an analysis of the PHSOL cases at the EHBH for this time period,along with results from a systematic literature review.We describe a surgicopathological spectrum comprising more than 100 types of PHSOLs that can be stratified into three types:tumor-like,benign,and malignant.We also stratified the PHSOLs into six subtypes derived from hepatocytes;cholangiocytes;vascular,lymphoid and hemopoietic tissues;muscular,fibrous and adipose tissues;neural and neuroendocrine tissues;and miscellaneous tissues.The present study provides a new classification system that can be used as a current reference for clinicians and pathologists to make correct diagnoses and differential diagnoses among various PHSOLs.
基金Supported by the Key Project Foundation of the Health Department of Jiangsu Province,China,No.BQ2000202002 Qinglan Project Foundation of the Education Department of Jiangsu Province,China
文摘AIM:To evaluate the functions of a new bioartificial liver(BAL)system in vitro and in vitro.MEHTODS:The BAL system was configurated byinoculating porcine hepatocyte spheroids into the cellcircuit of a hollow fiber bioreactor.In the experiments ofBAL in vitro,the levels of alanine aminotransferase(ALT),total bilirubin(TB),and albumin(ALB)in the circulatinghepatocyte suspension and RPMI-1640 medium weredetermined during 6 h of circulation in the BAL device.In the experiments of BAL in vitro,acute liver failure(ALF)model in canine was induced by an end-side portocavalshunt combined with common bile duct ligation andtransaction.Blood ALT,TB and ammonia levels ofALF in canines were determined before and after BALtreatment.RESULTS:During 6 h of circulation in vitro,therewas no significant change of ALT,whereas the TB andALB levels gradually increased with time both in thehepatocyte suspension and in RPMI-1640 medium.Inthe BAL treatment group,blood ALT,TB and ammonialevels of ALF in canines decreased significantly.CONCLUSION:The new BAL system has the ability toperform liver functions and can be used to treat ALF.
基金Supported by the National Natural Science Foundation of China, No.30171059
文摘AIM: To evaluate the effects of survivin on cell proliferation and apoptosis in liver cancer. METHODS: MTT assay was used to generate and optimize phosphorothioate antisense oligonucleotides (ODNs)-LipofectamineTM2000 (LiP) compound by varying ODNs (μg): LiP (μL) ratios from 1:0.5 to 1:5. Then, liver cancer cells (HepG2) were transfected with the compound. By using RT-PCR and Western blot, the expression levels of survivin mRNA and proteins were detected in HepG2 cells treated with antisense compounds (ODNs:LiP=1:4), and compared with those treated with sense compounds (1:4) as control. MTT assay was applied to the determination of cell proliferation in HepG2 cells. Active caspase-3 was evaluated by flow cytometric analysis. The morphological changes were assessed by electron microscopy. Laser scanning confocal microscopy was performed to detect the subcellular localization of survivin proteins in treated and untreated cells. RESULTS: Antisense compounds (1:4) down-regulated survivin expression (mRNA and protein) in a dose-dependent manner with an IC50 of 250 nmol/L. Its maximum effect was achieved at a concentration of 500 nmol/L, at which mRNA and protein levels were down-regulated by 80%. The similar results were found in MTT assay. Antisense compound (l:4)-treated cells revealed increased caspase-3-like protease activity compared with untreated cells. Untreated cells as control were primarily negative for the presence of active-caspase-3. As shown by transmission electron microscopy, treated cells with antisense compounds (1:4) resulted in morphological changes such as blebbing and loss of microvilli, vacuolization in the cytoplasm, condensation of the cytoplasm and nuclei, and fragmented chromatin. Immunofluorescence analysis confirmed the presence of survivin protein pool inside the cytoplasm in untreated cells. Labeled-FITC immunofluorescence staining of survivin clearly showed that survivin was distributed mainly in a spotted form inside the cytoplasm. Whereas cells treated with antisense compounds were rare and weak inside the cytoplasm. CONCLUSION: Down-regulation of survivin expression induced by the antisense compounds reduces tumor growth potential, promotes apoptosis and affects the localization of survivin proteins in HepG2 cells. Furthermore, survivin protein is a key molecule associated with proliferation and apoptosis, and antisense oligonucleotides targeting survivin have a bright prospect in the therapy of liver cancer.
文摘A rat liver epithelial cell line designated LW13 was established using a sequential sedimentation method. The cell line retained many normal properties of liver epithelial cells and showed some structural and functional features resembling those of liver parenchymal cells. LW13 cells became malignant after the introduction of exogenous transforming EJ Ha ras gene. Tumors produced by inoculation of the transformed cells into baby rats .contained areas of poorly differentiated hepatocellular carcinoma. In situ hybridization analysis confirmed the random rather than specific integration of exogenous ras gene into host chromosomes. Furthermore , an at least tenfold increase in the expression of the endogenous c myc gene was detected among transformed cell lines, suggesting the involvement of the c myc proto oncogene in the in vitro transformation of rat liver epithelial cells by EJ Ha ras oncogene.
基金Supported by the Scientific Development Programs of Science and Technology Commission Foundation of Shanghai (004119047).
文摘Objective To investigate the relationship between expression of somatostatin receptors(SSTRs) and activation of rat hepatic stellate cell (HSC). Methods HSCs were isolated from rats by in situ perfusion and single-step density gradient centrifugation, and then SSTR1-5 mRNA levels in the differentiated first passage HSCs were detected by means of reverse transcription polymerase chain reaction. On the other hand, hepatic fibrosis was induced in adult male Sprague-Dawley rats by carbon tetrachloride intoxication, and the expression of SSTR1-5 in normal as well as fibrotic liver was measured by immunohistochemical staining. Results SSTR mR-NA and SSTR could not be found in freshly isolated rat HSCs and normal rat liver. But SSTR1-3 mRNA appeared as HSCs became wholly activated, and SSTR1-3 could also be identified on the membrane of activated HSCs in the peri-sinusoid space, fibrous septa, etc. Conclusion The expression of SSTR1-3 in the rat HSC is closely related to its activation. This may reflect one of the main negative regulation mechanisms in the course of HSC activation.
基金Supported by the National Natural Science Foundation of China, No. 39900173
文摘AIM: To study genetic difference of mitochondrial DMA (mtDNA) between two hepatocarcinoma cell lines (Hca-F and Hca-P) with diverse metastatic characteristics and the relationship between mtDNA changes in cancer cells and ttieir oncogenic phenotype. METHODS: Mitochondrial DMA D-loop, tRNAMet+Glu+Ile and ND3 gene fragments from the hepatocarcinoma cell lines with 1100,1126 and 534 bp in length respectively were analysed by PCR amplification and restriction fragment length polymorphism techniques. The D-loop 3′ end sequence of the hepatocarcinoma cell lines was determined by sequencing. RESULTS: No amplification fragment length polymorphism and restriction fragment length polymorphism were observed in tRNAMet+Glu+Ile, ND3 and D-loop of mitochondrial DNA of the hepatocarcinoma cells. Sequence differences between Hca-F and Hca-P were found in mtDNA D-loop. CONCLUSION: Deletion mutations of mitochondrial DNA restriction fragment may not play a significant role in carcinogenesis. Genetic difference of mtDNA D-loop between Hca-F and Hca-P, which may reflect the environmental and genetic influences during tumor progression, could be linked to their tumorigenic phenotypes.
