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Objective To observe the effects of exogenous adrenomedullin(ADM)on expressions of IL-4 and IFN-γ following mechanical renal trauma in the rats. Methods A total of 104 healthy adult plain grade Wistar rats were,randomly divided into four groups,ie,normal control group(eight rats),simple trauma group(32 rats),prevention group(32 rats,injected with ADM before trauma)and treatment group(32 rats,injected with ADM after trauma).The experimental model of rat with mechanical renal trauma was prepared by striking the ridge of the left rib area with free dropping forrous hammer in all groups except for the control group.Ten minutes before and after mechanical renal trauma,ADM ly.All rats were sacrificed by draining out all the blood in their hearts at 1,6,12 and 24 hours after mechanical renal trauma.Then,the renal tissues were removed and fixed with 10% formalin for observing the positive expressions of IL-4 and IFN-γ by means of SABC staining. Results Compared with the simple trauma group,the positive expression of IL-4 in the prevention group Wag observed at 1 hour,gradually increased at 6 and 12 hours and reached the peak at 24 hours after mechanical renal trauma,with statistical difference(P<0.05).The positive expression of IL-4 in the treatment group WaS increased at one hour and reached the peak at 6 hours after mechanical renal trauma,with statistical difference(P<0.05).Compared with the simple trauma group,the IFN-γ expression in the prevention group was,increased at 6 and 24 hours but decreased at 12 hour,while that in the treatment group was decreased significantly at 1,12 and 24 hours.Compared with the normal control group,the IFN-γ expression in the treatment group was significantly decreased at 1 and 12 hours.The IFN-γ expression in the treatment group was lower than that in the prevention group at every time points,with statistical difference(P<0.05). Conclusions Preventive and therapeutic administration of exogenous ADM can enhance the expression of IL-4.IL-4 and IFN-γ play an antagonistic role in repair ofthe renal injury.The primary role of exogenous ADM is the dynamic regulation of IFN-γ expression.
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Objective To study the expressions of intercellular adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1) and E-selectin on the aorta of artherosclerosis and explore the relationship between these molecules and artherosclerosis(AS).Methods Human arterial specimens were collected from 18 cadavers,including 5 with normal arterial tunica intima(control) and 13 with pathologically identified AS(AS group).Analysis were carried out in consecutive sections by immunohistochemical staining for ICAM-1,VCAM-1 and E-selectin.Results ICAM-1,VCAM-1 and E-selectin positive cells were mainly located in the tunica intima and adventitia of AS aorta.More positive cells were observed obviously in AS group than in control(P
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Objective To investigate the efficiency of antitumor immune responses induced by a controlled live dendritic cell(DC)vaccine Methods DC precursors were isolated from Fischer 344 rat bone marrow and cultured with granulocyte macrophage colony stimulating factor and interleukin 4 The rat ovarian tumor cell line NuTu 19 was genetically modified by retroviral mediated suicide gene(HSV 1 TK), and the positive clones were selected using G418 Live DC vaccine was then fused with DC and NuTu 19/TK cell by polyethylene glycol The characteristics of live DC vaccine were assayed with flow cytometry and confocal laser scanning microscopy The specific expression of HSV 1 TK gene in live DC vaccine was evaluated by RT PCR and western blot The sensitivity of live DC vaccine to ganciclovir (GCV) was evaluated by methylthiazoletetrazolium assay In vivo, rats vaccinated twice with live DC vaccine were compared to those vaccinated with killed DC vaccine, unfused DC and NuTu 19/TK cell or phosphate buffered saline Seven days following the last immunization, the rats were sacrificed to test the specific cytotoxic T lymphocyte (CTL) activity by lactate dehydrogenase release assay, or challenged with NuTu 19 and tumor incidence was observed Results The fusion efficiency was approximately (23?14) Live DC vaccine displayed an up regulated expression of major histocompatibility complex (MHC) IIOX6 (87 6?3 4)%, costimulatory molecule B 1 2 (71 1?9 3)%, integrin OX 62 (68 0?7 4)%, and adhesion ICAM 1 (77 1?2 0)%, and specifically expressed HSV 1 TK gene. Our data showed that spleen T lymphocytes from rats vaccinated with live vaccine displayed enhanced CTL aetivity (61 8?8 3)% contrast to that of rats vaccinated with killed vaccines (26 0?3 8)% ( P
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Objective The present study was designed to investigate whether pulsing DCs with tumor-derived extracts is an ef- fective way to induce CTL. and antitumor immunity, Methods DCs were propagated from bone marrow (BM)of C57BL/6J(H-2Kb. I- Ab)mice in vitro with GM-CSF + IL-4tumor associated antigen (TAA) extracted from actively growing Hepa 1-6 cells was used to activate DCs. The phenotypes of DCs were detected by FACS, the cytotoxicity of CTL was as- sayed by 3H-TdR labbel assay. Result and Conclusion The TAA extract pulsed DCs exhibited much more and longer cell processes and increased expression of MHC- Ⅰ, MHC-Ⅱ, CD80 (B7-1 ) 、 CD86 (B7-2 ). This experiment has shown that DCs pulsed with TAA extracts of C57B/6J cells could stimulate effectively the responsiveness of syngenic splenic T cells to induce specific CTL against C57BL/6J cells.
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Objective To study the expression of intercellular adhesion molecule-1 (ICAM-1 ) in the rat cardiac heterotopic allo- grafts and the effect of cyclosporin A on prevention of allograft rejection. Methods Heart transplantated animals were divided into three groups: Group Ⅰ(control), Group Ⅱ(CsA 7. 5mg/kg B W, daily) and Group Ⅲ(CsA l5mg/kg B W, daily). Acute cardiac rejection grade was valued by the standrd of ISHLT(1990 ). Immunohistochemistry was performed to analyze the expression of ICAM-1 in heart grafts and donor aorta segments. Results After heart transplantation, it was found that from day 1 to 3, there was sligtly inflammatory infiltratiation, the rejection was graded 1A of 1B, But from day 11 to 12, there were disseminated inflammation and cardiac necrosis with serous hyperemia, exutation, edema, acute vasculitis and myocarditis. The rejection grade was 3B or 4, and could be reduced 1to 2. 5 grades by administration of CsA. It was also found that both in heart graft and donor aorta segments the expression of ICAM-1 on the endothelium cells, infiltrated lymphocytes was clearly increased. It was time-dependent and could be down-regulated by administra- tion of CsA. On Day 1 and Day 3 the suppressing function of CsA on expressing of ICAM-1 showed singificant dosage-dependent. But from Day 7 to Day 11, it appeared dosage- independent. Conclusiou Trea tment with CsA is an effect ive methed to down- regulate I - CAM-1 expression and could reduce the lympphocyte migration and filtration. These results may explain, in part, the mechanism of CsA reducing acute rejection in a rat cardiac transplantation medel.
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Objective:The present study was designed to investigate whether transfecting DC with tumor derived total RNA is an effective way to induce CTL and antitumor immunity.Methods:DC were propagated from bone marrow(BM) of C57BL/6J(H 2k b?I A b)mice in vitro with GM CSF+IL 4.Tumor derived total RNA extracted from actively growing Hepa 1 6 cells was used to transfected DC.The phenotypes of DC were detected by FACS,the cytotoxicity of CTL was assayed by MTT method.Results:The tumor derived total RNA transfected DC exhibited much more and longer plasm membrane processes and increased expression of MHC Ⅰ?MHC Ⅱ?CD80(B7 1)?CD86(B7 2).Conclusion:This experiment has shown that DC transfected with tumor derived total RNA of C57BL/6J cells could stimulate effectively the responsiveness of syngenic splenic T cells to induce specific CTL against C57BL/6J cells.
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Thirty-six male adult rats were divided into 12 groups. The rats, except the control group, were partially (68％) hepatectomized and then killed at intervals between 12-120h after operation. Isolated hepatocytes were prepared and flow cytometry was used to study the proliferative cycle. Mitotic index and binuclear cell count have been performed in liver sections and smears of isolated hepatocytes separately. Tetraploid cells acounted for 76% of all hepatocytes in normal rats and they also constituted the main proliferative population in regenerating liver. During 12-20h after operation, some of the tetraploid cells that remained in G_2 phase entered into mitosis. At 24h after operation, peaks of S phase in tetraploid cells and of octoploid cells occurred. At 36h after operation, mitotic index reached a maxium value and thereafter the percentage of binuclear cells were reduced rapidly. At 48-72h after operation, second peak of DNA synthesis occurred, but showed wide individual variations in time and cell proportion.
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Male Wistar rats were performed 70% partial hepatectomy (PH) or 1.5mg/g body weight D-galactosamine injection (Gal) intraperitoneally. From the 1st to the 7th day after treatment, the serum concentration of alpha-fetoprotein (AFP) and albumin (ALB) was measured and the liver sections were stained with routine and immunohistochemical methods and observed with light and electron microscopy. The mitotic index (MI) reached a maximum level (1.5%) on the 2nd day after PH, while the Gal group showed a lower (0.8%) and delayed (on the 3rd day) MI peak. On the 2nd and 3rd day after PH, AFP positive hepatocytes were predominantly observed in the periportal area and ALB positive cells mainly distributed around the central veins. In Gal group, AFP was detected both in hepatocytes near the necrotic areas and particularly in some special small cells which had not been seen in PH group. These small cells seemed to be able to transform into typical hepatocytes. Intracellular localization of AFP and ALB was observed in the perinuclear space, RER, and Golgi complex. Serum AFP and ALB levels in both groups presented reciprocal changes from the 1st to the 7th day after treatment.