The effects of dexamethasone(Dex)on the 3H-Dex specific binding were studied in the human heptatocellular carcinoma cell line(SMMC-7721) by using counting vial culture assay.Cells were cultivated in the counting vials, as th; grew well and the monolayer cells adhered to the wall of vials.The cells were cultivated in the mdium containing 10-3,10-7, and 10-6mol/L Dex for 6, 12, 24 and 48 h, and then the specific binding of 3H-Dex at 30nmol/L of the ligand was determined after elimination of the effect of the occupancy of GR.It was found that 3H-Dex specific binding was not changed within 6 h, but decreased to 70% at 12 h and maintained at this level from 12 to 48 h in the Dex pretreated cells.The specific binding was recovered approximately to the control level 24 h after removal of Dex.These results indicate that glucocorticoids may down-regulate glucocorticoid receptor in vitro.

Objective To investigate the effeet of eyclosporine A (CsA) on CD4+ Foxp3+ T cells in C57BL/6 mice and the underlying mechanism.Methods C57BL/6 mice, aging 8～ 10 weeks, were administered intraperitoneally with CsA at a dose of 20 mg· kg-1·d-1 for 2 weeks, while the control mice given sterile PBS.After 2 weeks, peripheral blood was collected, and splenoeytes and thymoeytes were prepared for the detection of CD4 + Foxp3 + T cells by using flow cytometry.Results The proportion of CD4+ Foxp3+ T cells in peripheral blood, spleen and thymus of CsA-treated mice was (0.581 ± 0.089) %, (2.189 ± 0.046) % and (0.472 ± 0.049) %, while that in PBS-treated mice was (1.751 ± 0.227) %, (3.684 ± 0.169) % and (1.412 ± 0.188) %, respectively (P < 0.001).Conclusion The reduction of CD4 + Foxp3+ T cells in CsA-treated mice suggests that CsA can inhibit the development of regulatory T cells.

Objective To investigate the effects of FTY720 on cardiac allograft survival in rats.Methods Wistar rats underwent abdominal heterotopic heart transplantation from SD rats and randomly divided into control group, CsA group, FTY720 group, MP group, FTY720+CsA group and FTY720+CsA+MP group.Results The number of peripheral blood lymphocytes in FTY720 group, FTY720+CsA group and FTY720+CsA+MP group was decreased at 3 h after administration, and recovered at 4th week. The median survival time in control group, CsA group, MP group and FTY720+CsA+MP group was 7.8 days, 16 days, 27.6 days and 16.8 days, but that in FTY720 group and FTY720+CsA group was respectively more than 150 days and 124 days.Conclusion FTY720 induces long-term survival of cardiac allograft in rats.

BACKGROUND: In the field of organ transplantation, patients often take immunosuppressants after organ transplantation, such as CsA, FK506, DEX and MPA. However, their mechanisms of immunosuppression are different. The effect of immunosuppressive drugs on monocyte chemoattractant protein-1 (MCP-1) remains poorly understood. OBJECTIVE: To investigate the effects of different immunosuppressants on the secretions of MCP-1 in whole blood. METHODS: The whole blood of healthy volunteers was mixed with different immunosuppressants for 6 hours, such as CsA, FK506, DEX and MPA, which included low, middle and high concentrations, followed by PMA and IONO stimulation for 6 hours. MCP-1 levels in whole blood samples were compared. The whole blood cultured alone served as control. RESULTS AND CONCLUSION: MCP-1 secretion was inhibited by DEX (1, 10 mg/L) and CsA (0.25,1.25 mg/L)- However, FK and MPA exhibited no such effect. Therefore, DEX and CsA may inhibit the function of monocytes and macrophages in immune system by diminishing the secretion of MCP-1. The combination of FK (5 μg/L), MPA (10 mg/L) and DEX (1 mg/L) or CsA (0.25 mg/L), MPA (10 mg/L) and DEX (1 mg/L) can inhibit the secretion of MCP-1, but only DEX among all the immunosuppressants mentioned above exhibited significant effect on inhibiting the secretion of MCP-1 when using alone.

BACKGROUND: Panel reactive antibodies (PRA) easily appear in the peripheral blood of organ transplant recipients sensitized by allogeneic human leukocyte antigen (HLA).How to enhance the success rate of renal transplantation.and long-term survival rate of renal allografts in sensitized recipients should be further studied.OBJECTIVE: This study was to detecthuman leukocyte antigen immunoglobulin G(HLA-IgG) antibody level and its specificity in renal transplant recipients,evaluate humoral immunity sensitization,and investigate the relationship of the acceptable mismatching of HLA cross-reactive group and survival rate of renal allograft.DESIGN: A clinical observation.SETTING: Zhujiang Hospital Affiliated to Southern Medical University.PARTICIPANTS: A total of 1297 patients,824 males and 473 females,averaging (42±16) years of age,received renal transplantation in the Department of Organ Transplantation,Zhujiang Hospital,Southern Medical University between January 1998 and December 2005,were recruited for this study.Among these patients,165 were HLA-IgG antibody-positive recipients,1132 were HLA-IgG antibody-negative ones,1217 received renal transplantation for the first time,77 received renal transplantation twice,2 three times,and 1 four times.Written informed consent was obtained from each subject for related laboratory measurements and treatment.The protocol was approved by the Hospital's Ethics Committee.Reagents:Lamhda antigen tray (LAT),Lambda antigen tray mixed (LATM),Special Monocloneal Tray-Asian HLA Class Ⅰ,and Micro SSP? Generic HLA Class Ⅱ were purchased from One Lambda Company,USA.Taq polymerase was purchased from PE Company,USA. DNA extract reagent was from Qiagen Company,Germany.Anti-human complement 4d (C4d) polyclonal antibody and chrornogenic substrate DAB were purchased from Biomedica Company,Austria.METHODS: Prior to operation,serum HLA-IgG antibody in the recipients was determined by an enzyme linked immunosorbent assay (ELISA).HLA-IgG antibody-positive serum was further detected by antigen tray (LAT1240 and LATIHDS) for antibody-positive rate and specificity.HLA genotyping was performed by a sequence specific primer polymerase chain reaction (PCR-SSP).For 40 recipients who had elevated serum creatinine (Scr),anti-HLA antibody detection and renal transplant needle biopsy were conducted.At the same time,C4d deposition on the capillary wall around the renal tubule was observed by immunohistochemical staining.Survival rate of renal allografts in recipients 1,3,and 5 years after transplantation,and relationships of gender and renal transplantation and antibody-positive rate were investigated.Survival rate of renal allograft in recipients that received different mismatch of HLA cross-reactive group was analyzed.MAIN OUTCOME MEASURES: Prior to and after renal transplantation,HLA-IgG antibody-positive rate and HLA genotyping in renal transplant recipients.Characterization of C4d deposition on the capillary wall around the renal tubule in the renal transplant biopsy tissue.Difference of survival rate of renal allograft.RESULTS: All 1297 recipients were included in the final analysis.Among them,1132 were HLA-IgG antibody-negative recipients,165 were HLA-IgG antibody-positive ones,126 were anti-HLA class Ⅰ IgG antibody-positive ones,90 were anti-HLA class Ⅱ IgG antibody-pesitive ones,51 were anti-HLA class Ⅰ and Ⅱ IgG antibody-positive ones,and 94 were highly sensitized ones (antibody-positive rate ＞50%).Among 40 recipients with needle biopsy,C4d deposition was found in the 13 recipients,but not found in the 27 recipients.Ten out of thirteen C4d-positive recipients presented with anti-HLA antibody-positive in the peripheral circulation.The incidence for delayed graft function (DGF) was significantly higher in recipients with HLA-IgG antibody-positive than in recipients with HLA-IgG antibody-negative (P ＜ 0.01).There was no significant difference in the survival rates of renal allografts between recipients with HLA-IgG antibody-positive and with HLA-IgG antibody-negative 1 ,3,and 5 years after renal transplantation (P ＞ 0.05).Antibody-positive rate was significantly higher in female recipients than in male recipients (P ＜ 0.01).Antibody-positive rate was significantly higher in recipients that received renal transplantation for the second time than in recipients that received renal transplantation for the first time (P ＜ 0.01).With HLA cross-reactive group mismatching increasing,survival rate of renal allograft presented a tendency of decline.One,three and five years after renal transplantation,the survival rate of renal allograft was respectively 97%,94%,and 92% for recipients with no mismatching,and 91%,82%,and 77% for recipients with two mismatches,which was respectively decreased by 6%,12%,and 15% compared to recipients that received no mismatching.For recipients with three mismatches,the survival rate of renal allograft was respectively decreased by 9%,15%,and 24% compared to recipients with no mismatching.CONCLUSION: C4d deposition on the capillary wall around the renal tubule can be detected as an indicator of antibody-mediated humoral rejection.A good HLA matching can noticeably decrease the incidence of rejection and improve the survival of renal allograft.

Objective To compare the value of assessment with DWI and contrast-enhanced MRI (CE-MRI) in activity of sacroiliitis of patients with ankylosing spondylitis(AS).Methods Ninety-six patients conforming to modified New York criteria were prospectively collectedas the AS group, and twenty-one healthy volunteers were enrolled into the control group. According to the Bath AS disease activity index (BASDAI), erythrocyte sedimentation rate and C-reaction protein, AS patients were divided into the active AS group (n=60) and the chronic AS group (n=36) . All subjects were performed with conventional MRI, DWI and CE-MRI of bilateral sacroiliac joints. The MRI manifestations were reviewed and the ADC values and signal intensity enhancement rate (ΔSI) were measured.ANOVA was performed for the comparison ofΔSI and ADC values among active AS group, chronic AS group and control group with BASDAI and lab test results as the gold standards. ROC was analyzed with ΔSI and ADC values for activity of AS and paired
samples t test was obtained to comparethe areas under the ROC ofΔSI and ADC values.Results Among 96 cases of AS patients, MRI of sacroiliac jointsshowed that 62 cases had subchondral bone edema (57 cases of active group, 5 cases of chronic group), that 11 cases had bone surface erosion(4 cases of active group, 7 cases of chronic group), that 15 cases had bone sclerosis(6 cases of active group, 9 cases of chronic group) and that 58 cases had fat deposition on the sacroiliac joints (27 cases of active group, 31 cases of chronic group). The ΔSI values of the active group, the chronic group and control group were respectively (2.51 ± 1.69)%,(1.19 ± 0.67)%and(0.75 ± 0.21)%, and the ADCvalues were(1.33 ± 0.33)× 10-3,(1.00 ± 0.43)× 10-3 and(0.38±0.13)×10-3mm2/s. There were significant differences forΔSI and ADC values among three groups (F=18.375, 16.366. P<0.01), and statistical significance ofΔSI and ADC values were found between every two groups of three(P< 0.05).The area under the ROC between ΔSI and ADC to determine activity of AS patients were respectively 0.814 and 0.730, which had nostatistical significance(t=1.632, P=0.103). The sensitivity and specificity to determine activity of AS patients byΔSI=1.44%were 81.67%and 80.00%.The sensitivity and specificity to determine activity of AS patients by ADC=1.15 × 10-3/mm2 were 76.67% and 71.43%.Conclusion DWI and CE-MRI performed equally in detecting activity of AS patients.

BACKGROUND: Panel reactive antibody (PRA) can mediate hyperacute rejection, and lead to decrease in success rate of transplantation and survival rate of renal graft in highly sensitized recipients compared to non-sensitized recipients.OBJECTIVE: According to human leucocyte antigen (HLA) cross-matching standards to select suitable donors for sensitized recipients and to evaluate the incidence of acute rejection and survival rate of renal allografts.DESIGN: Case observation.SETTING: Zhujiang Hospital of Southern Medical University.PARTICIPANTS: 136 sensitized recipients with positive PRA underwent renal transplantation in Department of Organ Transplantation, Zhujiang Hospital of Southern Medical University between January 1997 and December 2003 were selected, including 41 males and 95 females, aged (45±9) years. Recipients of first, second, third, and fourth transplant were 115, 18, 2 and 1 case, respectively. The informed consent was obtained from all patients. The protocol was approved by Hospital Ethics Committee. Lambda antigen tray (LAT) and LAT-Mix were purchased from One Lambda, Inc, USA. Special monoclonal tray -Asian HLA class Ⅰ (SMT72R) and Micro SSP Generic HLA Class Ⅱ (DRB/DQB) were also purchased from One Lambda, Inc, USA.METHODS: Pre-operative PRA levels and specificity of recipients were detected by ELISA test with Lambda antigen tray (LAT). Donor and recipient HLA class Ⅰ typing was performed with special tray - Asian HLA class Ⅰ (SMT72R), and HLA class Ⅱ gene typing with Micro SSP Generic HLA Class Ⅱ (DRB/DQB) (Micro-SSP). HLA-matching between donor and recipient was performed according to HLA cross-reactive group (CREG) standards by UNOS and class Ⅱ antigen permissible mismatch. The incidence of acute rejection and survival rate of renal allografts were evaluated within 1, 3 and 5 years.MAIN OUTCOME MEASURES: ①PRA levels and specificity of sensitized recipients before and after transplantation; ②HLA-matching between donor and recipient; ③Incidence of acute rejection and survival rate of renal allografts after transplantation.RESULTS: 136 PRA positive sensitized recipients were all included in final analysis. ① There were 104 recipients with anti-HLA class Ⅰ IgG antibody, 76 with anti-HLA class Ⅱ IgG antibody, and 44 with both anti-HLA class Ⅰ and Ⅱ IgG antibodies in 136 recipients. ②The number of cases of 0, 1, 2, 3, and 4 mismatch (MM) was 7, 26, 47, 39 and 17, respectively by the standard of conventional HLA antigen matching; However, the number of the recipients with 0, 1, 2, 3, and 4MM was 31, 53, 36, 16, and 0, respectively according to the principle of HLA CREG matching. ③By the principle of HLA CREG matching, rates of acute rejection in sensitized recipients with 2MM and 3MM HLA-CREG were significantly higher than those with 0MM (P < 0.05). Renal allograft survival rate in sensitized recipients with 0MM was significantly higher than those with 2MM and 3MM (P < 0.05).CONCLUSION: ①HLA CREG matching can significantly improve the ratio of well-matched. ② Good HLA matching can reduce the incidence of acute rejection in sensitized recipients and increase the survival rate of renal grafts.

Objective To investigate the clinical application value of fluorescence laparoscopy in radical resection of hepatocellular carcinoma (HCC).Methods Data of totally 113 patients with HCC in Henan Provincial People's Hospital between June 2016 to June 2018 were retrospectively analyzed.Among the 113 patients,46 patients underwent laparoscopic hepatectomy (LLR),and 67 patients underwent fluorescence guided laparoscopic hepatectomy (FLLR).Results No significant differences were observed between LLR group and FLLR group in terms of age,male proportion,liver function classification,surgical resection methods,and operation time (P＞0.05).The positive ratio of specimen surgical margin in LLR group was significantly higher than that in FLLR group,13.0％ vs.3.0％,and the difference was statistically significant (P＜0.05).In the FLLR group,22 patients received fluorescence guided anatomic hepatectomy with indocyanine green (ICG),10 with positive staining and 12 with negative staining,and fluorescence imaging was observed 2 minutes after ICG injection.There was no significant difference in operation time,hospitalization cost and length of stay between positive and negative staining (P＞ 0.05).Conclusion Fluorescence laparoscopy has certain advantages in hepatectomy,and can display the boundary of hepatocellular carcinoma in real time to ensure the safe margin of tumor resection.

Metastasis is a multistep process involving modification of morphology to suit migration, reduction of tumor cell adhesion to the extracellular matrix, increase of cell mobility, tumor cell resistance to anoikis, and other steps. MicroRNAs are well-suited to regulate tumor metastasis due to their capacity to repress numerous target genes in a coordinated manner, thereby enabling their intervention at multiple steps of the invasion-metastasis cascade. In this study, we identified a microRNA exemplifying these attributes, miR-124, whose expression was reduced in aggressive MDA-MB-231 and SK-3rd breast cancer cells. Down-regulation of miR-124 expression in highly aggressive breast cancer cells contributed in part to DNA hypermethylation around the promoters of the three genes encoding miR-124. Ectopic expression of miR-124 in MDA-MB-231 cells suppressed metastasis-related traits including formation of spindle-like morphology, migratory capacity, adhesion to fibronectin, and anoikis. These findings indicate that miR-124 suppresses multiple steps of metastasis by diverse mechanisms in breast cancer cells and suggest a potential application of miR-124 in breast cancer treatment.
Anoikis ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Adhesion ; Cell Line, Tumor ; Cell Movement ; Connective Tissue Growth Factor ; metabolism ; DNA Methylation ; Down-Regulation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; MicroRNAs ; genetics ; metabolism ; Neoplasm Metastasis ; rho GTP-Binding Proteins ; metabolism ; rho-Associated Kinases ; metabolism

OBJECTIVETo investigate the effect of transfusion of necrotic cells on regulatory T (Treg) and Th17 cell balance in septic mice.

METHODSThirty-four C57BL/6 mice were randomized into PBS group (n=5), sham-operated group (n=5), sepsis group (n=12), and necrotic cell transfusion group (n=12) and subjected to intraperitoneal PBS injection, sham operation by separating the cecum only, cecal ligation and puncture (CLP), and injection of 2 × 10⁷ necrotic cells 5 days before CLP, respectively. All the mice were sacrificed 2 weeks after CLP for analyzing the proportion of CD4⁺Foxp3⁺Treg cells and CD4⁺IL17A⁺Th17 cells in the peripheral blood, spleen and thymus by flow cytometry.

RESULTSThe percentage of Th17 cells and Treg/Th17 ratio in the spleen was significantly higher in CLP group than in the sham-operated group and PBS group (P<0.01). The percentage of Treg cells in the thymus was significantly lower in CLP group than in the sham-operated group (P<0.01). Pre-infusion of necrotic cells redressed the abnormality of Treg and Th17 cell percentages and Treg/Th17 imbalance in mice following CLP (P<0.05).

CONCLUSIONPre-infusion of necrotic cells can reverse Treg/Th17 imbalance in septic mice.

Animals ; Cecum ; Flow Cytometry ; Interleukin-17 ; Ligation ; Mice ; Mice, Inbred C57BL ; Necrosis ; Sepsis ; therapy ; Spleen ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology ; Thymus Gland