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1.
Chinese Journal of Organ Transplantation ; (12): 152-159, 2023.
Article in Chinese | WPRIM | ID: wpr-994645

ABSTRACT

Objective:To explore the current status of surgery for portal hypertension to grasp current status and future development of surgery in China.Methods:This study is jointly sponsored by China Hepatobiliary & Pancreatic Specialist Alliance & Portal Hypertension Alliance in China (CHESS).Comprehensive surveying is conducted for basic domestic situations of surgery for portal hypertension, including case load, surgical approaches, management of postoperative complications, primary effects, existing confusion and obstacles, liver transplantation(LT), laparoscopic procedures and transjugular intrahepatic portosystemic shunt(TIPS), etc.Results:A total of 8 512 cases of portal hypertension surgery are performed at 378 hospitals nationwide in 2021.Splenectomy plus devascularization predominated(53.0%)and laparoscopy accounted for 76.1%.Primary goal is preventing rebleeding(67.0%) and 72.8% of hospitals used preventive anticoagulants after conventional surgery.And 80.7% of teams believe that the formation of postoperative portal vein thrombosis is a surgical dilemma and 65.3% of hospitals practiced both laparoscopy and TIPS.The major reasons for patients with portal hypertension not receiving LT are due to a lack of qualifications for LT(69.3%)and economic factors(69.0%).Conclusions:Surgery is an integral part of management of portal hypertension in China.However, it is imperative to further standardize the grasp of surgical indications, the handling of surgical operation and the management of postoperative complications.Moreover, prospective, multi-center randomized controlled clinical studies should be performed.

2.
Chinese Journal of Pancreatology ; (6): 61-65, 2022.
Article in Chinese | WPRIM | ID: wpr-931278

ABSTRACT

Objective:To compare the clinical effectiveness of laparoscopic cholecystectomy (LC)+ laparoscopic choledocholithotomy (LCBDE)+ one-stage suture and primary choledocholithotomy with endoscopic retrograde cholangiopancreatography (ERCP)+ endoscopic duodenal sphincterotomy (EST)+ nasobiliary drainage (ENBD)+ LC in the treatment of choledocholithiasis complicated with gallbladder stones.Methods:A total of 200 patients with choledocholithiasis complicated with gallbladder stones admitted to the General Surgery Department of Shanxi Bethune Hospital from June 2015 to February 2021 were collected, and patients were divided into LC+ LCBDE+ one-stage suture (one-stage suture group, n=130) and ERCP+ EST+ ENBD+ LC (endoscopic surgery group, n=70) according to different treatments. The amount of intraoperative blood loss, operation time, postoperative feeding time, postoperative incidence of pancreatitis, cholangitis and other complications (biliary leakage, abdominal bleeding, wound infection), hospitalization costs, postoperative hospital stay, etc were compared between two groups. Results:The postoperative incidence of pancreatitis in the one-stage suture group (0.7% vs 5.7%) and the hospitalization cost [(2.74±0.39) ten thousand yuan vs (3.86±0.63) ten thousand yuan] were significantly lower than those in the endoscopic surgery group. The operation time [(103.21±9.36) min vs (88.18±7.20)min] was significantly longer than that of the endoscopic surgery group, and postoperative feeding time [(3.3±0.3)d vs (2.2±0.8)d] were significantly later than the endoscopic surgery group ( P<0.05). The amount of intraoperative blood loss [(36.0±3.0)ml vs (37.3±2.7)ml], the incidence of postoperative cholangitis (1.5% vs 2.9%) and other complications [biliary leakage (2.3% vs 1.4%), abdominal bleeding (1.5% vs 4.3%), wound infection(0 vs 0)], postoperative hospital stay [(6.8±1.3)d vs (7.1)d] had no significant differences between the two group. Conclusions:The two minimally invasive methods for the treatment of choledocholithiasis complicated with gallbladder stones had good efficacy, but LC+ LCBDE+ one-stage suture can retain the sphincter function of Oddis, maintain the normal anatomy and physiology of the biliary tract, reduce the incidence of related complications, and contribute to the recovery of patients, with high safety, effectiveness and feasibility.

3.
Chinese Journal of Digestive Surgery ; (12): 447-452, 2019.
Article in Chinese | WPRIM | ID: wpr-752962

ABSTRACT

Objective To investigate the application value of early and delayed laparoscopic cholecystectomy (LC) after percutaneous transhepatic gallbladder drainage (PTGD) in 65 years of age or older patients with severe acute cholecystitis.Methods The prospective study was conducted.The clinical data of 80 patients with severe acute cholecystitis who were admitted to Shanxi Dayi Hospital of Shanxi Academy of Medical Sciences from May 2016 to January 2018 were collected.All patients were divided into two groups by random number table,including patients undergoing LC 72 h later after extubation of PTGD in the PTGD + early LC group,and patients undergoing LC 5-14 days later after extubation of PTGD in the PTGD + delayed LC group.Observation indicators:(1) surgical situations;(2) analysis of liver function before and after LC in the two groups;(3) analysis of serum-related inflammatory factors before and after LC in the two groups;(4) follow-up situations.Patients were followed up by outpatient examination or telephone interview to detect the postoperative complications in the postoperative three months up to April 2018.Measurement data with normal distribution were represented as Mean ± SD,and comparison between groups was done using the paired t test.Count data were represented as absolute number,and comparison between groups was analyzed using the chi-square test or Fisher exact probability.Results Eighty patients were screened for eligibility,including 41 males and 39 females,aged from 65 to 70 years,with an average age of 67 years.There were 40 patients in the PTGD + early LC group and 40 in the PTGD + delayed LC group,respectively.(1) Surgical situations:the operation time,volume of intraoperative blood loss,and duration of postoperative hospital stay were (52± 15) minutes,(29± 11) mL,(18.9± 1.6) days in the PTGD + early LC group,and (88± 13)minutes,(69± 11)mL,(27.7±4.8)days in the PTGD + delayed LC group,respectively,showing significant differences in the above indicators between the two groups (t =11.668,16.219,11.000,P<0.05).(2) Analysis of liver function before and after LC in the two groups:the levels of aspartate transaminase (AST),alanine aminotransferase (ALT),gamma glutamyl transferase (GGT),and total bilirubin (TBil) of PTGD + early LC group were (53 ± 11) U/L,(203 ±40) U/L,(128± 22) U/L,(19± 6)U/L,(86±21)μmol/L before LC,and (26±5)U/L,(83±23)U/L,(29±3)U/L,(11±5)U/L,(27± 7) μmol/L at 24 hours after LC,showing significant differences in the above indicators before and after LC (t =12.562,16.448,28.199,6.478,16.857,P<0.05).The levels of AST,ALT,GGT,and TBil of PTGD + delayed LC group were (54± 12) U/L,(203±48) U/L,(130±24) U/L,(19±6) U/L,(85±20) μmol/L before LC,and (29±5) U/L,(151±36) U/L,(53±7)U/L,(17±3)U/L,(31±8)μmol/L at 24 hours after LC,showing significant differences in the above indicators before and after LC (t =13.622,5.481,2.169,1.988,15.855,P<0.05).There was no significant difference in the levels of AST,ALT,ALP,GGT,TBil before LC between the two groups (t=0.389,0.000,0.389,0.000,0.218,P>0.05),meanwhile,there were significant differences in the levels of AST,ALT,ALP,GGT,TBil after LC between the two groups (t =2.683,10.067,19.931,6.508,2.380,P<0.05).(3) Analysis of serum-related inflammatory factors before and after LC in the two groups:the levels of interleukin-1 (IL-1),interleukin-6 (IL-6),high-sensitivity C-reactive protein (CRP),interleukin-10 (IL-10),and tumor necrosis factor-α (TNF-α) of PTGD + early LC group were (71 ±9) ng/L,(82±9)ng/L,(137±16)ng/L,(75±6)ng/L,(67±9)μg,/L before LC,and (87±13)ng/L,(97±9)ng/L,(81± 19)ng/L,(145±6)ng/L,(85±6)μg/L at 24 hours after LC,showing significant differences in the above indicators before and after LC (t ==6.400,7.454,14.259,52.175,10.525,P<0.05).The levels of IL-1,IL-6,high-sensitivity CRP,IL-10,and TNF-α of PTGD + delayed LC group were (71±9) ng/L,(82± 10) ng/L,(145±28)ng/L,(75±6)ng/L,(67±10) μg/L before LC,and (145±7)ng/L,(135±16) ng/L,(101±1S)ng/L,(146±9) ng/L,(113±10)μg/L at 24 hours after LC,showing significant differences in the above indicators before and after LC (t =41.079,17.766,8.360,41.525,27.578,P < 0.05).There was no significant difference in the levels of IL-1,IL-6,high-sensitivity CRP,IL-10,and TNF-α before LC between the two groups (t =0.000,0.000,1.569,0.000,0.000,P>0.05),meanwhile,there were significant differences in the levels of IL-1,IL-6,high-sensitivity CRP,and TNF-α after LC between the two groups (t=24.844,13.092,4.833,15.185,P<0.05).(4) Follow-up situations:80 patients were followed up for 3 months.Two patients in the PTGD + early LC group had postoperative complications,including 1 of bile duct injury and 1 of incisional infection;9 patients of PTGD + delayed LC group had postoperative complications,including 3 of bile duct injury,3 of multiple organ failure,2 of incisional infection,1 of death.There was a significant difference in the postoperative complication between the two groups (x2 =5.165,P<0.05).Conclusion Early LC after PTGD can effectively shorten operation time,reduce volume of intraoperative blood loss,shorten duration of postoperative hospital stay,protect liver function,reduce the expression of serum inflammatory factors at 24 hours after surgery,and reduce postoperative complications.

4.
Chinese Journal of Hepatobiliary Surgery ; (12): 259-263, 2019.
Article in Chinese | WPRIM | ID: wpr-745374

ABSTRACT

Objective To analyze tumor immune microenvironment and related mechanisms in liver cancer.Methods We included 10 cases of hepatocellular carcinoma,hepatitis B patients and healthy volunteers from January 2015 to December 2017 in Shanxi Grand Hospital.We first detected the peripheral and local GM-CSF level in each group,detected myeloid-derived suppressor cells (MDSCs) GM-CSF and pathway-related protein expression.from liver cancer,tumor margin and normal liver tissue through flow cytometry and immunohistochemistry,Finally,we transfected the CCR4-NOT transcriptional complex subunit 7 (CNOT7) recombinant plasmid in the hepatoma cell line,and then detected the related protein expression.Results There was no significant difference for peripheral blood GM-CSF level between liver cancer group,hepatitis group and control group (P>0.05).The level of local GM-CSF was (32.2±8.9) ng/L,which was higher than that of hepatocellular carcinoma (9.7±2.7) ng/L and normal liver tissue (11.6±2.9) ng/L.The difference was statistically significant (P<0.05).The proportion of MDSCs at the edge of the tumor was (9.9 ±3.6) %,which was higher than that of liver cancer (4.0± 1.5) % and normal liver tissue (6.3±2.3) %,and the difference was statistically significant (P<0.05).Immunohistochemistrydata was consistent with previous data.Compared with normal liver tissue,CNOT7 and STAT3 were highly expressed in liver cancer tissues,while STAT1 was lowly expressed.HepG2 human hepatoma cells were selected for transfection.Compared with the empty plasmid group,CNOT7 expression was decreased in the knocking out group at the same time STAT1 expression was increased,STAT3 and GM-CSF expression was decreased.Conclusion In hepatocellular carcinoma,the secretion of GM-CSF increased and the number of MDSCs increased.Knocking out CNOT7 reduced GM-CSF secretion and activate the JAK/STAT signaling pathway.

5.
Journal of China Medical University ; (12): 225-229, 2019.
Article in Chinese | WPRIM | ID: wpr-744830

ABSTRACT

Objective To study the effect of human CCR4-NOT transcription complex subunit 7 (CNOT7) gene knockdown on the immune microenvironment of HepG2 cells and explore its significance. Methods We designed a cell transfection protocol and performed the experiment with three groups:CNOT7-targeted knockdown group, control group, and CNOT7 overexpression group. The transfection efficiency was assessed using inverted fluorescence microscopy, and the expression level of CNOT7, transforming growth factor-β1 (TGF-β1), and nuclear factor-kappa B (NF-κB) p65 proteins was determined by Western blotting. The concentration of TGF-β1 secreted in the cell culture supernatant was measured by ELISA. The sensitivity of tumor cells to the killing function of natural killer (NK) cells was detected by flow cytometry. Results Compared with the control group, the expression level of TGF-β1 and NF-κB p65 proteins was significantly decreased in the CNOT7-targeted knockdown group, and the TGF-β1 concentration in the culture supernatant was also significantly reduced. However, in the CNOT7 overexpression group, the expression level of the two proteins and TGF-β1 concentration were significantly increased. NK cells were co-cultured with tumor cells, and the apoptosis rate of HepG2 cells transfected with CNOT7-specific shRNA was significantly increased. However, in the CNOT7 overexpression group, the apoptosis rate was significantly decreased. Conclusion CNOT7 forms the immune microenvironment of hepatocellular carcinoma. Targeted knockdown of CNOT7 can reduce TGF-β1 secretion and enhance the killing function of NK cells toward HepG2 cells.

6.
Chinese Journal of General Surgery ; (12): 596-600, 2018.
Article in Chinese | WPRIM | ID: wpr-710593

ABSTRACT

Objective To study the regulation of dendritic cells by recombinant glycated polylysine-coupled MIP-3α-FL double-gene targeting expression vector in liver cancer immune microenvironment.Methods H22 hepatocarcinoma cells were transfected with recombinant plasmid of MIP-3α-FL (shMIP-3α-FL) and injected into hepatoma model mice.The survival time,tumor size were compared.Flow cytometry was used to measure the number and phenotype of tumor infiltrating DCs.Results Western blot and ELISA demonstrated that the secretion of MIP-3α and FL in H22 cells was significantly increased after transfection with MIP-3α-FL.The survival time of the mice in the experimental group was significantly prolonged,the tumor size decreased.Flow cytometry showed that the number of tumor-infiltrating DCs in the experimental group was significantly higher than that in the control group;the expression of CD80 and CD86 in the infiltrating dendritic cells (TIDCs) was significantly higher than that of the control group.Conclusions The co-action of MIP-3α and FL can significantly promote DC accumulation,maturation,and conjugate glycosylated polylysine carriers increase the precision of targeting and enhance the antigenpresentation of the DCs.

7.
Chinese Journal of General Surgery ; (12): 57-60, 2017.
Article in Chinese | WPRIM | ID: wpr-620788

ABSTRACT

Objective To study the action of CNOT 7 (CCR4-NOT transcription complex subunit 7 human)gene and its mechanisms in the process of Vγ 9Vδ2T cell immunologic tolerance of HepG2 cells (Hepatoblastoma G2 Cell Line).Methods The shCNOT 7 (Recombinant plasmid of CNOT 7) and control vector shRNA were transfected into HepG2 cells.Vγ9Vδ2T cytokine stimulated each group before and after cell transfection,Cell apoptosis was detected by flow cytometry (FCM),CNOT 7 protein and STAT1,STAT3 expression level was detected by Western blot.CNOT 7,STAT1 and STAT3 protein expression levels of HepG2 liver cancer cell lines and L02 normal liver cell line was assayed by Western blot.Results When stimulated by Vγ9Vδ2T cytokine,the apoptosis rate of gene-knockdown group significantly improved from (7.55% ±2.63%) to (20.59% ±3.12%).Compared with L02 cells,the CNOT7 protein expression of HepG2 cells increased (F =28.76,P < 0.01),STAT3 protein expression increased (F =110.29,P < 0.01),while STAT1 protein expression was down-regulated (F =35.67,P < 0.01).CNOT 7 knockout could induce HepG2 cells STAT1 expression (t =6.69,P < 0.05).Conclusions CNOT 7 gene could induce HepG2 cells Vγ 9Vδ2T cellular immune tolerance.CNOT 7 knockout could reverse the Vγ 9Vδ2T cell immunologic tolerance of HCC.

8.
Chinese Journal of General Surgery ; (12): 549-552, 2016.
Article in Chinese | WPRIM | ID: wpr-497052

ABSTRACT

Objective To investigate the percentage,mature classification and Immune killing function of Vγ9Vδ2T cell in peripheral blood of HCC patients.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from HCC patients (n =25) and healthy donors (n =20) by discontinuous density gradient centrifugation.Proportion,mature and differentiate subtypes and IFN-γ and CD107a expressing of the delta 2 T cells were detect by using flow cytometry,δ2Tcell were selectively cultured with zoledronate and human IL-2.After 12-14 days cells were collected and tested for the second time.Results While the percentage of Vγ9Vδ2Tcell of total T cell in peripheral blood of HCC patients is lower than healthy people before culture (t =4.505,P < 0.001),after augmentation in vitro the proportion increased significantly (t =8.782,P < 0.001),to a level similar to healthy group (t =1.644,P =0.109).There was no statistically significant difference when differentiation subtypes of patient's Vγ9Vδ2Tcell were compared with healthy group before culturing (all P > 0.05),after culture the proportion of Tn,Tcm and Temra decreased [t(Tn) =2.081,t(Tcm) =2.478,t(Temra) =2.953,all P < 0.05],and the proportion of Tem,Tem+ Temra increased [t(Tem) =12.6,t (Tem + Temra) =9.843,all P < 0.001].Cell culture did not alter the proportion of IFN-γ and CD107a secreting Vγ9Vδ2T cells in the peripheral blood in both HCC patients and healthy people (all P > 0.05).Conclusions While the percentage of Vγ9Vδ2T cell of HCC patients in peripheral blood was lower than healthy people,its matured subtypes are similar to those of healthy people,and functions of expressing IFN-γ and CD107a are not different with healthy people.Applying ZOL + IL-2 can amplifyVγ9Vδ2T cells of patients with HCC.

9.
Chinese Journal of Hepatobiliary Surgery ; (12): 373-376, 2015.
Article in Chinese | WPRIM | ID: wpr-466289

ABSTRACT

Objective To analyze the differentially expressed genes between the NCAM + c-Kit +RBE and NCAM-c-Kit-RBE of intrahepatic cholangiocarcinoma (ICC) cell lines,and to screen out the differentially expressed genes that are related to the stem cell signaling pathways.Methods Magnetic activated cell sorting was used to isolate the NCAM + c-Kit +/NCAM-c-Kit-subset cells,and then Agilent Whole Human Genome Microarray Kit was used to test the difference in gene expressions between the NCAM + cKit + and NCAM-c-Kit-subset cells.The difference in gene expressions related to the stem cell signaling pathways was analyzed by the SAS system.The result of the microarray was further confirmed by RT-PCR.Results The total differentially expressed genes which could be found through gene microarray were 7270 [foldchange(fc) ≥2 or fc ≤0.5].Compared with the NCAM-c-Kit-RBE,3572 genes were upregulated while 3698 genes were downregulated.The differences in gene expressions related to the stem cell signaling pathways were 421 (fc ≥2 or fc ≤ 0.5),among which 231 genes were upregulated while 190 genes were downregulated.Conclusions High-flux microarray could be used to screen out lots of differentially expressed genes between the NCAM + c-Kit + and NCAM-c-Kit-RBE cells.The differences in gene expression in the stem cell signaling pathways could also be further analyzed using the SAS system.

10.
Chinese Journal of General Surgery ; (12): 382-384, 2014.
Article in Chinese | WPRIM | ID: wpr-447049

ABSTRACT

Objective To study the relationship between Treg cell numbers and level of TGF-β1,IL-10 in the immune microenvironment of rat liver cancer.Method 40 male Wistar rats were randomly divided into three groups,liver cancer model group (n =20),saline control group (n =10) and blank control group (n =10).Liver cancer model was established by intraperitoneal injection of DEN (100 mg/kg).Percentage of Treg cells in cancer tissues was detected by fiow cytometry and the expression level of TGF-β1 and IL-10 by immunohistochemical SABC.Results The ratio of Treg cells in CD4 + T cells was 14.32% ± 4.84% in liver cancer tissues,significantly higher than that in the blank control group 5.64% ±6.10% and the saline control group 7.95% ±3.55%.The difference was statistically significant (F =211.279,P < 0.05).The expression level of TGF-β1 and IL-10 in cancer tissues was significantly higher than the blank control group and the saline control group (FTGF-β1 =250.740,FIL-10 =152.744,all P <0.05).The number of Treg cells was positively correlated with TGF-β1,IL-10 level (P < 0.05).Conclusions TGF-β1,IL-10 and Treg cells significantly increased in rat experimental liver cancer tissues,and there was positive correlation between Treg cells,and TGF-β1 and IL-10 in liver cancer tissues.

11.
Cancer Research and Clinic ; (6): 191-193, 2011.
Article in Chinese | WPRIM | ID: wpr-413252

ABSTRACT

Objective To investigate the expression of NF-κB and epithelial-mesenchymal transition (EMT) related markers E-cadherin and Vimentin proteins in human pancreatic cancer tissues and its relation with the malignant features. Methods The expression of NF-κB、E-cadherin and Vimentin proteins in 62 cases of pancreatic cancer tissues were detected by using immunohistochemistry and compared with the clinicopathological data of pancreatic cancer. Results The positive expression rate of NF-κB was 81% (50/62), Vimentin protein increased of expression was 61% (38/62), and E-cadherin protein loss of expression was 55 % (34/62) in pancreatic cancer. The positive expression rate of NF-κB was significantly related with the lymph node metastasis (x2=11.761, P<0.05), distant metastasis (x2=9.225, P<0.05), the absent expression of epithelial marker E-cadherin protein (r =0.352, P <0.05) and the positive expression of mesenchymal marker Vimentin protein (r=0.343, P <0.05), but there was no relation with the patients gender,age, tumor location, tumor type and tumor differentiation (P >0.05). In addition, the significant correlation of E-cadherin expression loss and Vimentin expression with tumor lymph node metastasis and distant metastasis was found (x 2= 6.914, 4.984, 7.753, 5.144, P <0.05). Conclusion The overexpression of NF-κB in pancreatic cancer may accelerate invasion and metastasis of pancreatic cancer through inducing EMT.

12.
Cancer Research and Clinic ; (6): 307-309,313, 2011.
Article in Chinese | WPRIM | ID: wpr-597880

ABSTRACT

Objective To investigate the significance of CerbB-2 oncogene in the occurrence and development of gastric adenocarcinoma. Methods By using immunohistochemistry and fluorescence in situ hybridization, 60 cases of gastric adenocarcinoma tissue, 60 cases of normal margin of mucosal tissue and 30 cases of normal gastric mucosa tissue were detected for the expression and amplification of CerbB-2. Results The positive rate of CerbB-2 expression in gastric adenocarcinoma tissues, normal margin of mucosal tissues and normal gastric mucosa tissues were 31.7 %, 8.33 % and 3.33 %; The positive rate of CerbB-2 amplification of these were 28.3 %, 5 % and 0. There were significant differences between gastric adenocarcinoma tissues and the others in the expression and amplification of CerbB-2. The expression and amplification of CerbB-2 in gastric adenocarcinoma tissues had no correlation with age,gender,tumor size or tumor differentiation degree of patients' (P >0.05), but were correlated with the extent of tumor invasion, lymph node metastasis, distant metastasis and clinical stage(P <0.05). Conclusion CerbB-2 is closely related to the occurrence and development of gastric adenocarcinoma, and can be considered as an important index for determining the biological behavior of gastric cancer.

13.
Cancer Research and Clinic ; (6): 371-373, 2009.
Article in Chinese | WPRIM | ID: wpr-380746

ABSTRACT

Objective To investigate the inhibitory effect of immunosuppressive agent FRY720 on hepatocellular carcinoma Hepal-6. Methods Hepal-6 cells were cultured, and divided into 4 groups, namely control group and 0.1 μg/ml, 10 μg/ml, 100 μg/ml quality concentration groups. The cells were treated by the drugs for 24 to 48 hours respectively. The inhibitory rate of the cells was measured by MTT assay, and cell cycle and cell apoptotic rate were detected by flow cytometry (FCM). Results The ability of tumor cell growth were inhibited by FTY720 after 48 h. The maximal inhibition rate was 62.10 %, The apoptosis ratio was increased when FTY720 was 0.1-100 μg/ml, and it was 4.07 %, 8.16 %, 19.84 % respectively. FTY720 significantly prolonged cell G1 phase. Conclusion FTY720 could inhibit the growth of hepatocellular carcinoma, arrest the cell in G1 phase, and increase apoptosis.

14.
Chinese Journal of Pathophysiology ; (12)2000.
Article in Chinese | WPRIM | ID: wpr-529770

ABSTRACT

AIM:To study the influence of stimulation by LPS and CD40 ligandization in vitro on the TLR4-MD2 expression and IL-12 production in dendritic cells (DCs) modified by sCD40 gene and provide the experimental clues of inducing dornor-specific immune tolerance.METHODS: Plasmid pEGFP-N1/sCD40 and pEGFP-N1 was transfected into DC2.4 cell line with lipofectamine. After 6 h of treatment with LPS and anti-CD40mAb, the expression of TLR4-MD2 on DCs was determined with flow cytometry (FCM) and RT-PCR. IL-12p70 protein was detected by ELISA.RESULTS: LPS treatment of DCs down-regulated surface expression of TLR4-MD2, LPS treatment along with anti-CD40mAb significantly up-regulated TLR4-MD2 surface expression. CD40 ligandization did not affect TLR4-MD2 mRNA expression in DCs but partly increased its low level induced by LPS and markly enhanced IL-12p70 secretion after LPS stimulation. DCs modified by sCD40 gene inhibited the above effect.CONCLUSION: After treatment with LPS and anti-CD40mAb, DCs modified by sCD40 gene down-regulate surface expression of TLR4-MD2 and IL-12p70 secretion decreases significantly, which might be linked with the interruption of TLR4-MD2 transportation from cytoplasm.

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