Objective: To determine the ultrastructural findings on Rectal Mucosa (RM) of patients with HIV/AIDS and anorectal pathologies (ARP), at micrometric and nanometric scales. Materials and methods: 5 patients were evalua...Objective: To determine the ultrastructural findings on Rectal Mucosa (RM) of patients with HIV/AIDS and anorectal pathologies (ARP), at micrometric and nanometric scales. Materials and methods: 5 patients were evaluated, 18 - 55 years old, with ARP (HIV co-infection with HPV, n = 4, and HIV-negative patient with HPV infection) (control n = 1), who were referred to the Coloproctology Unit of the HUC, and subjected to rectoscopy and biopsy. RM samples were identified, placed in a sterile plastic bottle with 1 mL of 2% glutaraldehyde and immediately transported for routine processing of fine cut (60 - 90 nm) to be evaluated via Transmission Electron Microscopy (TEM). They were fixed with Karnovsky solution with Millonig phosphate buffer (pH 7.4 and 320 mOsm) and post-fixed with OsO4 under the same conditions of pH and osmolarity. Results: Ultrastructural findings, at 10−6 scale: 1) Intestinal mucosa: vacuoles of mucus of different sizes that seem to be fused. 2) Smooth muscle cells: loss of definition of contractile myofilaments mass. 3) Unmyelinated axons and terminals of Schwann cells (SC): Edema and loss of their plasma membranes in some areas of association with axon terminals as well as abundant collagen fibers associated with SC. Ultrastructural findings, at 10−9 scale: 1) Smooth muscle cells: folded wrapper cores and edema of mitochondria and rough endoplasmic reticulum cisterns (RER). 2) Myelinated axon terminals: Loss of synaptic vesicles. 3) Fibroblasts: One observes mitochondria and cisterns of RER with alterations. All these alterations can generate intestinal and anorectal dysfunction in these patients. Conclusions: The HIV causes changes in rectal and muscular mucosa despite HAART treatment with undetectable viral load.展开更多
AIM: To evaluate antihepatoma effect of antisense phosphorothioate oligodeoxyribonucleotides (S-ODNs) targeted to alpha-fetoprotein (AFP) genes in vitro and in nude mice. METHODS: AFP gene expression was examined by i...AIM: To evaluate antihepatoma effect of antisense phosphorothioate oligodeoxyribonucleotides (S-ODNs) targeted to alpha-fetoprotein (AFP) genes in vitro and in nude mice. METHODS: AFP gene expression was examined by immunocytochemical method or enzyme-linked immunosorbent assay. Effect of S-ODNs on SMMC-7721 human hepatoma cell growth in vitro was determined using microculture tetrazolium assay. In vitro antitumor activities of S-ODNs were monitored by measuring tumor weight differences in treated and control mice bearing SMMC-7721 xenografts. Induction of cell apoptosis was evaluated by fluorescence-activated cell sorter (FACS) analysis. RESULTS: Antisense S-ODN treatment led to reduced AFP gene expression. Specific antisense S-ODNs, but not control S-ODNs, inhibited the growth of hepatoma cells in vitro. In vitro, only antisense S-ODNs exhibited obvious antitumor activities. FACS analysis revealed that the growth inhibition by antisense S-ODNs was associated with their cell apoptosis induction. CONCLUSION: Antisense S-ODNs targeted to AFP genes inhibit the growth of human hepatoma cells and solid hepatoma, which is related to their cell apoptosis induction.展开更多
AIM: To investigate the specific mechanisms of intrinsic and acquired resistance to taxotere (TXT) in pancreatic adenocarcinoma (PAC). METHODS: MTT assay was used to detect the sensitivity of PAC cell line SUIT-2 and ...AIM: To investigate the specific mechanisms of intrinsic and acquired resistance to taxotere (TXT) in pancreatic adenocarcinoma (PAC). METHODS: MTT assay was used to detect the sensitivity of PAC cell line SUIT-2 and its sublines (S-007, S-013, S-020, S-028 and TXT selected SUIT-2 cell line, S2/TXT) to TXT. Mdr1 (P-gp), multidrug resistance associated protein (MRP), lung resistance protein (LRP) and beta-tubulin isotype gene expressions were detected by RT-PCR. The functionality of P-gp and MRP was tested using their specific blocker verapamil (Ver) and indomethacin (IMC), respectively. The transporter activity of P-gp was also confirmed by Rhodamine 123 accumulation assay. RESULTS: S-020 and S2/TXT were found to be significantly resistant to TXT(19 and 9.5-fold to their parental cell line SUIT-2, respectively). RT-PCR demonstrated strong expression of Mdr1 in these two cell lines, but weaker expression or no expression in other cells lines. MRP and LRP expressions were found in most of these cell lines. The TXT-resistance in S2-020 and S2/TXT could be reversed almost completely by Ver, but not by IMC. Flow cytometry showed that Ver increased the accumulation of Rhodamine-123 in these two cell lines. Compared with S-020 and SUIT-2, the levels of beta-tubulin isotype II, III expressions in S-2/TXT were increased remarkably. CONCLUSION: The both intrinsic and acquired TXT-related drug resistance in these PAC cell lines is mainly mediated by P-gp, but had no relationship to MRP and LRP expressions. The increases of beta-tubulin isotype II, III might be collateral changes that occur when the SUIT-2 cells are treated with TXT.展开更多
文摘Objective: To determine the ultrastructural findings on Rectal Mucosa (RM) of patients with HIV/AIDS and anorectal pathologies (ARP), at micrometric and nanometric scales. Materials and methods: 5 patients were evaluated, 18 - 55 years old, with ARP (HIV co-infection with HPV, n = 4, and HIV-negative patient with HPV infection) (control n = 1), who were referred to the Coloproctology Unit of the HUC, and subjected to rectoscopy and biopsy. RM samples were identified, placed in a sterile plastic bottle with 1 mL of 2% glutaraldehyde and immediately transported for routine processing of fine cut (60 - 90 nm) to be evaluated via Transmission Electron Microscopy (TEM). They were fixed with Karnovsky solution with Millonig phosphate buffer (pH 7.4 and 320 mOsm) and post-fixed with OsO4 under the same conditions of pH and osmolarity. Results: Ultrastructural findings, at 10−6 scale: 1) Intestinal mucosa: vacuoles of mucus of different sizes that seem to be fused. 2) Smooth muscle cells: loss of definition of contractile myofilaments mass. 3) Unmyelinated axons and terminals of Schwann cells (SC): Edema and loss of their plasma membranes in some areas of association with axon terminals as well as abundant collagen fibers associated with SC. Ultrastructural findings, at 10−9 scale: 1) Smooth muscle cells: folded wrapper cores and edema of mitochondria and rough endoplasmic reticulum cisterns (RER). 2) Myelinated axon terminals: Loss of synaptic vesicles. 3) Fibroblasts: One observes mitochondria and cisterns of RER with alterations. All these alterations can generate intestinal and anorectal dysfunction in these patients. Conclusions: The HIV causes changes in rectal and muscular mucosa despite HAART treatment with undetectable viral load.
基金Supported by the National Postdoctoral Science Foundation of China,No.199711.
文摘AIM: To evaluate antihepatoma effect of antisense phosphorothioate oligodeoxyribonucleotides (S-ODNs) targeted to alpha-fetoprotein (AFP) genes in vitro and in nude mice. METHODS: AFP gene expression was examined by immunocytochemical method or enzyme-linked immunosorbent assay. Effect of S-ODNs on SMMC-7721 human hepatoma cell growth in vitro was determined using microculture tetrazolium assay. In vitro antitumor activities of S-ODNs were monitored by measuring tumor weight differences in treated and control mice bearing SMMC-7721 xenografts. Induction of cell apoptosis was evaluated by fluorescence-activated cell sorter (FACS) analysis. RESULTS: Antisense S-ODN treatment led to reduced AFP gene expression. Specific antisense S-ODNs, but not control S-ODNs, inhibited the growth of hepatoma cells in vitro. In vitro, only antisense S-ODNs exhibited obvious antitumor activities. FACS analysis revealed that the growth inhibition by antisense S-ODNs was associated with their cell apoptosis induction. CONCLUSION: Antisense S-ODNs targeted to AFP genes inhibit the growth of human hepatoma cells and solid hepatoma, which is related to their cell apoptosis induction.
基金Supported in part by Phne-Poulenc Rorer Pharmaceuticals INC.
文摘AIM: To investigate the specific mechanisms of intrinsic and acquired resistance to taxotere (TXT) in pancreatic adenocarcinoma (PAC). METHODS: MTT assay was used to detect the sensitivity of PAC cell line SUIT-2 and its sublines (S-007, S-013, S-020, S-028 and TXT selected SUIT-2 cell line, S2/TXT) to TXT. Mdr1 (P-gp), multidrug resistance associated protein (MRP), lung resistance protein (LRP) and beta-tubulin isotype gene expressions were detected by RT-PCR. The functionality of P-gp and MRP was tested using their specific blocker verapamil (Ver) and indomethacin (IMC), respectively. The transporter activity of P-gp was also confirmed by Rhodamine 123 accumulation assay. RESULTS: S-020 and S2/TXT were found to be significantly resistant to TXT(19 and 9.5-fold to their parental cell line SUIT-2, respectively). RT-PCR demonstrated strong expression of Mdr1 in these two cell lines, but weaker expression or no expression in other cells lines. MRP and LRP expressions were found in most of these cell lines. The TXT-resistance in S2-020 and S2/TXT could be reversed almost completely by Ver, but not by IMC. Flow cytometry showed that Ver increased the accumulation of Rhodamine-123 in these two cell lines. Compared with S-020 and SUIT-2, the levels of beta-tubulin isotype II, III expressions in S-2/TXT were increased remarkably. CONCLUSION: The both intrinsic and acquired TXT-related drug resistance in these PAC cell lines is mainly mediated by P-gp, but had no relationship to MRP and LRP expressions. The increases of beta-tubulin isotype II, III might be collateral changes that occur when the SUIT-2 cells are treated with TXT.
