OBJECTIVETo investigate the role of CD40 ligand (CD40L) in dendritic cells (DC) of CEA transgenic mice and to evaluate the specific cellular immunity induced by activated DC.

METHODSBone marrow cells of the CEA transgenic mice were used to generate immature dendritic cells under the condition of GM-CSF and IL-4. CD40L was added to activate dendritic cells into mature phenotype. Dendritic cells cancer vaccine was pulsed with CEA526-533 peptide which made the vaccine specific for cancer immunity. The immunophenotype molecules were identified by flow cytometry. The cytokines produced by cells were determined by ELISA. T cells proliferation was measured by (3)H-thymidine essays.

RESULTSImmunophenotype molecules expressions of CD40L-activated dendritic cells were significantly higher than those in control group. IL-12 secretion by CD40L-activated dendritic cells was (937.81+/-51.99) pg/10(6) DC, significantly higher than that in control group [(83.06+/-8.58) pg/10(6) DC, P<0.01]. CD8(+) T cells proliferation induced by CD40 L-activated dendritic cells was stronger as compared to control group (P<0.05), and the secretion of IFN-gamma was(33.900+/-4.550) ng/L, significantly higher than that in control group [(5.226+/-0.460) ng/L, P<0.01]. Splenocytes proliferation induced by CD40 L-activated dendritic cells was stronger as compared to control group (P<0.01), and the secretion of IFN-gamma was (69.802+/-11.407) ng/L, significantly higher than that in control group [(2.912+/-0.562) ng/L, P<0.01].

CONCLUSIONThe method of using CD40L to stimulate bone marrow-delivered dendritic cells promotes the maturation and activation of dendritic cells, which enhances the cellular immunity in CEA transgenic mice.

Animals ; CD40 Ligand ; immunology ; physiology ; Dendritic Cells ; cytology ; immunology ; metabolism ; Immunity, Cellular ; immunology ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic

Objective To explore the short-term results of arthroscopic medial retinaculum placation (MRP) and lateral retinaculum release (LRR) in treatment of patients with traumatic patellar dislocation. Methods 17 cases (6 male, 11 female) with traumatic patellar instability from March 2012 to December 2015, with an average age of 16.8 years old (range from 14 to 37 years old). All of the patients had a clear history of trauma and experienced patellar dislocation for the first time, the patients experienced patellar dislocation 1 to 4 times preoperatively. The arthroscopic examination was undertaken before the repairing to observe the injured site of the medial retinaculum and the patellar track, as well as the dynamic patellofemoral congruence. All patients underwent arthroscopic MRP and LRR minimally invasive procedure. Results All patients were followed up for 9 to 28 months averaging (19.7 ± 1.3) months. The fear test was negative after operation. There was no redislocation during follow-up and their ranges of motion returned to normal. Postoperative CT images showed 15 cases regained normal anatomical relation of patellofemoral joint. 2 cases had mild semi-dislocation. Lysholm's score averaging (51.8 ± 4.5) points preoperatively and (92.4 ± 2.8) points postoperatively. According to Insall scale, the results were excellent in 11 knees, good in 5 knees, and fair in 1 knee at 1 year after operation with an excellent and good rate of 94.1％. Conclusions Arthroscopic MRP and LRR showed satisfactory results with limited morbidity in the short-term follow-up. This method can make the patients smaller wound,quicker recovered and lower recurrence rate.

Objective To explore the effects of Astragalus polysaccharides on liver injury induced by cadmium chloride. Methods The cadmium chloride solution was administrated i.v. to develope rat model of liver injury. Rats were randomized divided into three groups;control group, model group, Astragalus polysaccharides intervention group. After the 5 weeks, the rats were sacrificed and the livers were weighted, alanine aminotrans ferase(ALT), aspartate aminotransferase (AST) and lactate dehydrogenase(LDH), the contents of cadmium(Cd) was checked by plasma mass spectrometerthe(ICP-MS), the contents of interleukin-2(IL-2) and transforming growth factor-β1(TGF-β1) was measured by ELISA, the the protein expression of Bcl-2 and Bax was observed by immunohisto-chemistry staining method. Results The liver weight was increased in model group, the level of ALT, AST and LDH were also ascended in model group, the contents of Cd, TGF-β1 and the protein expression of Bax all increased in mode group(P＜0.05 or P＜0.01). The content of IL-2 and the protein expression of Bcl-2 were decreased in mode group (P＜0.05 or P＜0.01). Compared with the model group, the liver index was decreased in Astragalus polysaccharides intervention group, the levels of ALT, AST and LDH were also reduced in Astragalus poly-saccharides intervention group, the contents of Cd, TGF-β1 and the protein expression of Bax were all dropped in Astragalus polysaccharides intervention group(P＜0.05 or P＜0.01).The content of IL-2 and the protein expression of Bcl-2 increased in Astragalus polysaccharides intervention group(P＜0.05 or P＜0.01). Conclusions Astragalus polysaccharides may reduce the oxidative stress injury of rats liver cells indued by cadmium chloride, and its mechanism may be explained by regulation of Bcl- 2/Bax protein expression.

Previous studies proposed that the synergistic effect of fibroblast growth factor-21 (FGF-21) and insulin may be due to the improvement of insulin sensitivity by FGF-21. However, there is no experimental evidence to support this. This study was designed to elucidate the mechanism of synergistic effect of FGF-21 and insulin in the regulation of glucose metabolism. The synergistic effect of FGF-21 and insulin on regulating glucose metabolism was demonstrated by investigating the glucose absorption rate by insulin resistance HepG2 cell model and the blood glucose chances in type 2 diabetic db/db mice after treatments with different concentrations of FGF-21 or/and insulin; The synergistic metabolism was revealed through detecting GLUT1 and GLUT4 transcription levels in the liver by real-time PCR method. The experimental results showed that FGF-21 and insulin have a synergistic effect on the regulation of glucose metabolism. The results of real-time PCR showed that the effective dose of FGF-21 could up-regulate the transcription level of GLUT1 in a dose-dependent manner, but had no effect on the transcription level of GLUT4. Insulin (4 u) alone could up-regulate the transcription level of GLUT4, yet had no effect on that of GLUT1. Ineffective dose 0.1 mg kg(-1) FGF-21 alone could not change the transcription level of GLUT1 or GLUT4. However, when the ineffective dose 0.1 mg x kg(-1) FGF-21 was used in combination with insulin (4 u) significantly increased the transcription levels of both GLUT1 and GLUT4, the transcription level of GLUT1 was similar to that treated with 5 time concentration of FGF-21 alone; the transcription level of GLUT4 is higher than that treated with insulin (4 u) alone. In summary, in the presence of FGF-21, insulin increases the sensitivity of FGF-21 through enhancing GLUT1 transcription. Vice versa, FGF-21 increases the sensitivity of insulin by stimulating GLUT4 transcription in the presence of insulin. FGF-21 and insulin exert a synergistic effect on glucose metabolism through mutual sensitization.
Animals ; Blood Glucose ; Diabetes Mellitus, Experimental ; metabolism ; Drug Synergism ; Fibroblast Growth Factors ; pharmacology ; Glucose ; metabolism ; Glucose Transporter Type 1 ; metabolism ; Glucose Transporter Type 4 ; metabolism ; Hep G2 Cells ; Humans ; Insulin ; pharmacology ; Insulin Resistance ; Liver ; metabolism ; Mice

Objective To explore the role of Astragalus polysaccharidesin renal tissue under cadmium treatment in rats.Methods The SPFWistar rats were randomized divided into three groups:normal group,model group,experimental group.The normal group was intraperitoneallyinjected with 0.9％ saline solution,while the model group and theexperimental group were intraperitoneally injected daily with cadmiumchloride (CdC12) in a single dose of 1.5 mg· kg-1 body weight.At thesame time,the experimental group was given Astragalus polysaccharide20 mg · kg-1 once a day for five consecutive weeks.The model group andthe experimental group were given 0.9％ saline solution once a day for five consecutive weeks.After the last injection,all the rats were sacrificed by femoral artery sampling.The content of cadmium (Cd) was measured by atomic absorption spectrophotometry.The levels of superoxide dismutase (SOD),malondialdehyde (MDA),lactate dehydrogenase(LDH),glautathione peroxidase(GSH-Px) were checked by colorimetric analysis.The contents of interleukin -2(IL-2),transforming growth factor-3 (TGF-β) were determined by enzyme linked immunosorbent assay.The protein expressions of Bcl-2 and Bax were analyzed by immunohistochemical method.Results IL-2 level in normal group,model group,experimental group were (117.38 ± 10.89),(71.82 ± 14.56),(102.46 ±13.82)pg·mL-1,respectively;TGF-β1level in above groups were (4.88 ±0.38),(9.77 ±0.21),(6.35 ±0.41)pg·mL-1,respectively;GSH-Px level in above groups were (240.48 ± 18.24),(159.26 ± 21.06)(191.50 ± 23.82) U · g-1,respectively;LDH level in above groups were (1125.25 ± 37.91),(2089.46 ± 35.87)(1159.61 ± 28.86) U · g-1,respectively;Cd level in above groups were (0.02 ± 0.01),(19.68 ± 1.85)(12.99±2.11)mg · kg-1,respectively;SOD level in above groups were (71.55 ± 3.56),(39.30 ± 3.36)(67.25±2.68) U · mg-1,respectively;MDA level in above groups were (3.96 ± 0.79),(7.47 ± 0.78)(5.61 ±0.99) nmol · mg-1,respectively;Bcl-2 gray level in above groups were 37.54±3.23,68.43 ±5.2138.25 ± 6.17,respectively;Bax gray level in above groups were 57.87 ± 8.93,27.54 ± 5.45,48.23 ± 3.54.respectively.Compared with the normal group,all factors in the model group were significantly (all P ＜ 0.01).Compared with the model group,all factors in the experimental group were significandy (P ＜ 0.05 or P ＜0.01).Conclusion Astragalus polysaccharides pretreatment has a certain protective effect against cadmium-induced renal damage in rats and the protection is achieved by the regulation of Bcl-2/Bax protein expression.

OBJECTIVETo evaluate the myocardial viability with (201)Tl/(18)F-FDG DISA-SPECT technique in patients with acute myocardial infarction underwent emergent intracoronary autologous bone marrow mononuclear cells (BM-MNC) transplantation.

METHODSPatients with first acute myocardial infarction underwent emergent percutaneous coronary intervention (PCI) were randomized in a 1:1 ratio to either intracoronary transplantation of autologous BM-MNC (n = 20) or to sodium chloride concluding heparin (control, n = 20) via a micro infusion catheter group immediately after PCI. Change in global left ventricular function (LVEF measured by echocardiography) and the myocardial viability detected by (201)Tl/(18)F-FDG DISA-SPECT from baseline and 6-months post transplantation were analyzed.

RESULTSLeft ventricular ejection fraction (LVEF) was improved in both groups and the absolute increase (DeltaLVEF) in BM-MNC group was significantly higher than that in control group (7.6% +/- 2.8% vs. 3.0% +/- 2.8%, P < 0.001). In addition, the absolute decrease of myocardial infusion defect detected by (201)Tl SPECT was more significant in BM-MNC group than that in control group (6.7% +/- 3.0% vs. 2.6% +/- 2.6%, P < 0.001) and the number of mismatched segments (indicating viable myocardium) detected by (18)F-FDG SPECT in border zone was also significantly higher in BM-MNC group than that in control group.

CONCLUSIONImproved myocardial viability and reduced myocardial infusion defect post emergent intracoronary transplantation of autologous BM-MNC in patients with acute myocardial infarction could be detected by (201)Tl/(18)F-FDG DISA-SPECT technique.

Aged ; Bone Marrow Transplantation ; Cell Survival ; Female ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Myocardial Infarction ; diagnostic imaging ; therapy ; Myocytes, Cardiac ; diagnostic imaging ; Tomography, Emission-Computed, Single-Photon ; Ventricular Function, Left

OBJECTIVES: To observe the effects of Yougui pill (Traditional Chinese Medicine) on the related factors of Wnt signal pathway of rats with knee osteoarthritis (KOA), and explore its protective mechanism. METHODS: Sixty SPF SD rats were randomly divided into the sham-operative group, model group, glucosamine sulfate group, high-dose, middle-dose, low-dose of Yougui pill treated group (=10). KOA model was established by modified Hulth method for six weeks. The rats in the high, middle and low-dose of Yougui pill group were treated with Yougui pills at the doses of 20,10 and 5 g/kg respectively by gastrogavage once a day for 8 weeks, while equal volume of normal saline was given to those in the sham and model control group and an equal volume of glucosamine sulfate (1.7 g/kg·d) was given to those in glucosamine sulfate group for 8 weeks. The knee joint was removed after the last dose of drug. The pathological changes of cartilaginous tissues were observed under a microscope. The mRNA levels of Dickkopf homolog 1(DKK1), Wnt induced secreted protein 1(WISP1), Wnt1, low density lipoprotein receptor related protein 5(LRP5) and beta -catenin in rats cartilaginous tissues were analyzed by using RT-PCR method, and the protein contents of DKK1, WISP1, Wnt1, LRP5 and beta-catenin in cartilaginous tissues were detected by Western blot. RESULTS: Compared with the sham group, the articular cartilage was severely damaged, the Mankin score was increased significantly (<0. 05), the mRNA and protein expression levels of DKK1 in cartilaginous tissue were markedly decreased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were increased significantly in model group(<0.05). Compared with model group, the articular cartilage lesions was light (<0.05), the Mankin Score was decreased significantly(<0.05), and the mRNA and protein levels of DKK1 in cartilaginous tissue were increased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were decreased in Yougui pill high-dose group and glucosamine sulfate group (<0.05). CONCLUSIONS Yougui pill has protective effects on the KOA by inhibiting the expressions of WISP, Wnt1, LRP5, beta-catenin and increasing the expression of DKK1 cytokine in the Wnt signaling pathway.
Animals ; CCN Intercellular Signaling Proteins ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Glucosamine ; pharmacology ; Intercellular Signaling Peptides and Proteins ; metabolism ; Osteoarthritis, Knee ; drug therapy ; Proto-Oncogene Proteins ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Wnt Signaling Pathway ; Wnt1 Protein ; metabolism ; beta Catenin ; metabolism