Objective:To explore the changes and possible mechanisms of circulating Tc 17 cells in patients with colorectal cancer ( CRC) following disease progression .Methods:The peripheral blood were collected from 54 patients with colorectal cancer . the proportions of Tc17 cells in peripheral blood mononuclear cells (PBMCs) and cultured PBMCs treated by IL-1β, IL-6 and TGF-βin different concentrations were determined by flow cytometry;the levels of IL-1β, IL-17A, IL-23 and IL-6 in sera were measured by ELISA.Results:The proportions of Tc17 cells and the levels of IL-1β, IL-17A, IL-23 and IL-6 were significantly higher than those in healthy controls (P<0.05).Compared with early CRC, the proportions of Tc17 cells and the levels of IL-1β, IL-17A in advanced CRC were significantly decreased, but the level of IL-6 was significantly increased (P<0.05).The difference of IL-23 level between early and late groups was not significant (P>0.05).In vitro experiments confirmed that IL-1βor high concentrations of IL-6 and TGF-βcould significantly increase the number of Tc 17 cells in PBMCs.Conclusion: The changes of circulating Tc 17 cells in the progression of colorectal cancer are possibly modulated by IL-1β, IL-23, IL-6 and TGF-β.

Objective To investigate the effect of high volume hemofiltration (HVHF) on expression of miRNA-146 in peripheral blood mononuclear cells in posttraumatic sepsis patients and its therapeutic mechanism.Methods Twenty-five cases of posttraumatic sepsis were included as HVHF group.Another 25 age-and gender-matched traumatic sepsis patients with similar APACHE-Ⅱ who received no HVHF treatment for some reasons were used as controls.Therapeutic measurements were the same of the two groups except for HVHF.At 0-,6-,12-,24-and 48-hour time points,the peripheral blood mononuclear cells were isolated from patients of both groups to detect level of miRNA-146.Peripheral blood mononuclear cells isolated at 24 hours were incubated with lipopolysaccharide (LPS) in vitro.At hours 4,8,12,24 and 48 after incubation,level of miRNA-146 in mononuclear cells was determined and levels of TNF-α,IL-6,and IL-10 in the substrate was detected by ELISA.Results (1) Level of miRNA-146 in HVHF group was decreased significantly over time as compared with that in sepsis group;(2) Before incubation and at 4-and 8-hour after incubation,miRNA-146 level was lowered significantly in HVHF group as compared with that in sepsis group.After LPS stimulation,mononuclear cell also presented a stronger inflammatory response in HVHF group than in sepsis group.Conclusions HVHF provides a definite effect on immune function recovery and a significant improvement in prognosis.Moreover,HVHF may attenuate the impact of miRNA-146 on mononuclear cell inflammatory factor release and enhance the cell ability to respond to external stimuli again via down-regulating miRNA-146,as may be one of the therapeutic mechanisms of HVHF for posttraumatic sepsis.

Objective To investigate the expression of soluble triggering receptors expressed on myeloid cells-1 ( sTREM-1 ) in intraperitoneal drainage fluid of patients with abdorminal trauma and its predictive value for post-traumatic sepsis. Methods A total of 80 abdominal trauma patients were served as the trauma group and 25 patients treated with subtotal gastrectomy as the control group.Intraperitoneal drainage fluid sTREM-1,serum sTREM-1,procalcitonin (PCT) and C-reactive protein (CRP)at 0,24,48,72 hours after admission were determined in two groups for assessing their value in early prediction of post-traumatic sepsis. Results The levels of drainage fluid sTREM-1,serum sTREM-1,PCT and CRP in the trauma group were significantly higher than those in the control group (P ＜ 0.05 ).Drainage fluid sTREM-1 showed the area under the receiver operating characteristic (ROC) curve,sensitivity and specificity for 0.84,77％,and 83％ in the prediction of post-traumatic sepsis,which was superior to the serum sTREM-1,PCT and CRP. Conclusion Intraperitoneal drainage fluid sTREM-1 has high accuracy in predicting the sepsis in abdominal trauma patients.

OBJECTIVETo evaluate the clinical results and perioperative management of primary total knee arthroplasty (TKA) in hemophilic patients.

METHODSFrom February 1997 to February 2006, the data of 6 total knee arthroplasty performed in 4 hemophilic patients was reviewed retrospectively. The values of coagulation factor were maintained at suitable level by monitoring the activity of the factors and their inhibitors during perioperative period. The mean follow-up time was 4.4 years, knee society score and the last postoperative radiographs were recorded.

RESULTSAfter TKA, the hemophilic patients felt pain of knee relieved, the knee function was improved, but the range of motion increased limitedly. At the early post-operative stage, 3 knees in 2 patients with hemarthrosis or muscle bleeding, 1 of the 2 patients complicated with formation of inhibitor of factor VIII and healing problem in 1 knee after TKA, 1 patient with transient paralysis of the common peroneal nerve, 1 patient with venous circulation insufficiency crisis, but no compartment syndrome. In the late stage after TKA, 1 patient with hemarthrosis of both elbows, but no late infection, loosening, displacement and fracture of the prosthesis in the 6 knees.

CONCLUSIONSTotal knee arthroplasty could alleviate knee pain and improve joint function in advanced severe hemophilic arthritic patients. It is important to monitor the activity and inhibitors of coagulation factor VIII or IX, which could decrease the early and late postoperative complications.

Adult ; Aged ; Arthritis ; etiology ; surgery ; Arthroplasty, Replacement, Knee ; Follow-Up Studies ; Hemophilia A ; complications ; Humans ; Male ; Middle Aged ; Perioperative Care ; Retrospective Studies ; Treatment Outcome

BACKGROUNDRenal transplantation in sensitized candidates remains a highly significant challenge worldwide. The production of panel reactive antibody (PRA) against human leukocyte antigen (HLA) is a major risk factor in presensitized recipients. The aim of this study was to evaluate the impact of HLA matching and recipients' PRA on two-year outcome in presensitized renal allograft recipients.

METHODSWe determined the percentage of panel reactivity and specificity of anti-HLA immunoglobulin (Ig) G antibodies in 73 presensitized renal allograft recipients compared with 81 unsensitized recipients (control group). HLA genotyping of both recipients and corresponding donors was performed by PCR with sequence-specific primers (PCR-SSP). We analyzed the factors influencing the early graft outcome (two-year rejection rates and survival rates of the grafts), including HLA mismatching, class and degree of panel reactivity, and target antigen of donors.

RESULTSPresensitized recipients had a worse two-year outcome than unsensitized recipients (P = 0.019 for rejection rate, P = 0.01 for survival rate). The difference in number of HLA-mismatched alleles with either 6-antigen matching (Ag M) standard or amino acid residue matching (Res M) standard was not significant between the rejection and non-rejection groups of presensitized recipients or between the graft survival group and graft loss group. Compared with the control group, recipients with both PRA-I and PRA-II antibodies had a significantly worse two-year outcome (P = 0.001 for rejection rate, P = 0.002 for survival rate). The two-year outcomes of the peak PRA >/= 50% group and its subgroup, at-transplant PRA > or = 50% group, were significantly worse compared with the control group (P = 0.025 and P = 0.001 for rejection rate, P = 0.043 and P = 0.024 for survival rate). The rejection rates of the at-transplant target antigen positive group and its subgroup, HLA-I target antigen positive group, were significantly higher than the control group (P = 0.001 and P = 0.001), target antigen negative group (P = 0.003 and P = 0.001), and peak target antigen positive with negative at-transplant target antigen group (P = 0.024 and P = 0.002). Two-year graft survival rates of the target antigen positive group and HLA-I target antigen positive group were significantly lower than the control group (P = 0.012 and P = 0.001). The two-year outcome of target antigen unknown group was similar to that of the target antigen positive group. Presensitized recipients with pre-transplant plasmapheresis or immunoadsorption (PRA prepared group) had a better but non-significant two-year outcome than the control group. However, the PRA unprepared presensitized recipients were different to the control group (P = 0.004 for rejection rate and P = 0.005 for survival rate). Hyperacute rejection (HR) occurred in three recipients with positive HLA-I target antigen and without mismatch according to Res M and in one case with positive PRA-II (for an unknown target antigen). No HR occurred in eight cases with positive HLA-II target antigens.

CONCLUSIONSPre-transplant PRA preparations might improve the access of presensitized patients to renal donors. Avoiding antigen-positive donors remains a fundamental measure in preventing HR and early rejections.

Adult ; Enzyme-Linked Immunosorbent Assay ; Female ; Graft Rejection ; immunology ; Graft Survival ; immunology ; HLA Antigens ; immunology ; Histocompatibility Testing ; Humans ; Isoantibodies ; blood ; Kidney Transplantation ; adverse effects ; immunology ; mortality ; Male ; Middle Aged ; Transplantation, Homologous ; immunology ; Treatment Outcome

OBJECTIVETo compare the levels of STAT4 tyrosine phosphorylation in peripheral T-lymphocytes induced by IL-12 in rheumatoid arthritis (RA) and osteoarthritis (OA).

METHODSFrom May 2007 to August 2009, peripheral blood mononuclear cells (PBMCs) were isolated from RA patients [RA group, all the cases were female, the age was from 28 to 55 years with an average of (45.0 +/- 13.0) years] and OA patients [OA group, all the cases also were female; the age was from 55 to 75 years with an average of (67.0 +/- 9.6) years]. The purity of T-lymphocytes from PBMCs was accredited by flow cytometry. The IL-12 of 50 ng/ml added in T-lymphocytes, the levels of STAT4 tyrosine phosphorylation were detected by western blot after different time intervals (0, 10, 30, 60 min).

RESULTSThe purity of T-lymphocytes were above 91% through diremption and depuration for peripheral blood monouclear cells. The levels of STAT4 tyrosine phosphorylation in T-lymphocytes from RA induced by IL-12 were higher than that from OA in the different times (10, 30, 60 min); after 30 min, its levels from RA and OA achieved to crest value.

CONCLUSIONSTAT4 in peripheral T-lymphocytes of rheumatoid arthritis was more easily to be activated than osteoarthritis.

Adult ; Aged ; Arthritis, Rheumatoid ; immunology ; Female ; Humans ; Interleukin-12 ; pharmacology ; Middle Aged ; Osteoarthritis ; immunology ; Phosphorylation ; Polymorphism, Single Nucleotide ; STAT4 Transcription Factor ; genetics ; metabolism ; T-Lymphocytes ; drug effects ; metabolism ; Tyrosine ; metabolism

BACKGROUNDImmunosuppression for immunologically high-risk kidney transplant patients usually involves antithymocyte globulin induction with triple drug maintenance therapy. Alemtuzumab, a humanized anti-CD52 antibody, was expected to be a promising induction therapy agent for kidney transplantation. However, currently no consensus is available about its efficacy and safety. This study aimed to evaluate the efficacy and safety of alemtuzumab as immune induction therapy in highly sensitized kidney transplant recipients.

METHODSIn this prospective, open-label, randomized, controlled trial, we enrolled 23 highly immunological risk patients (panel reactive antibody > 20%). They were divided into two groups: alemtuzumab group (trial group) and anti-thymocyte globulin (ATG) group (control group). Patients in the alemtuzumab group received intravenous alemtuzumab (15 mg) as a single dose before reperfusion. At the 24th hour post-operation, another dosage of alemtuzumab (15 mg) was given. The control group received a bolus of rabbit ATG (9 mg/kg), which was given 2 hours before kidney transplantation and lasted until the removal of vascular clamps when the anastomoses were completed. Maintenance immunosuppression in both groups comprised standard triple therapy consisting of tacrolimus, prednisone, and mycophenolate mofetil (MMF). Acute rejection (AR) and infection episodes were recorded, and kidney function was monitored during a 2-year follow-up. χ(2) test, t test and Kaplan-Meier analysis were performed with SPSS17.0 software.

RESULTSMedian follow-up was 338 days. In both the alemtuzumab group and ATG group, creatinine and blood urea nitrogen values in surviving recipients were similar (P > 0.05). White blood cell counts were significantly reduced in the alemtuzumab group for the most time points up to 6 months (P < 0.05). One patient receiving alemtuzumab died for acute myocardial infarction at the 65th day post-operation. Two ATG patients died for severe pulmonary infection or cardiac and pulmonary failure. Cumulative 2-year graft survival rate was 90.9% in the alemtuzumab group and 81.8% in ATG group (P > 0.05) respectively. There was one graft failure in the alemtuzumab group and two graft failures in ATG group, with all graft failures at tributed to rejection episodes. The alemtuzumab group had a 2-year cumulative freedom from rejection rate of 81.8%, compared with 72.7% for the ATG group (P > 0.05).

CONCLUSIONAlemtuzumab induction therapy for highly sensitized kidney transplant recipients is an effective and safe protocol yielding an acceptable acute rejection rate.

Adult ; Aged ; Alemtuzumab ; Antibodies, Monoclonal ; therapeutic use ; Antibodies, Monoclonal, Humanized ; Antibodies, Neoplasm ; therapeutic use ; Antilymphocyte Serum ; therapeutic use ; Female ; Graft Rejection ; immunology ; Graft Survival ; immunology ; Humans ; Immunosuppressive Agents ; therapeutic use ; Kidney Transplantation ; immunology ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the effects of naloxone on the expression of c-kit receptor (c-kit R) and its ligand stem cell factor (SCF) in human embryo neuronal hypoxic injury.

METHODSSerum-free cerebral cortical cultures prepared from embryonic human brains were deprived of both oxygen and glucose which would set up an environment more likely with that of in vivo ischemic injury. Neurons in 24-well culture plates were randomly divided into four groups: control group, hypoxia group, naloxone 0.5 microg/ml group and naloxone 10 microg/ml group. MTT assay and biological analysis were performed to study the cell death and the changes of extracellular concentrations of lactate dehydrogenase (LDH) after combined oxygen-glucose deprivation. Neurons in 25 ml culture flasks were also randomly allocated into four groups as previously described. Intracellular total RNA were extracted at different time points: pre-hypoxia, immediately after hypoxia, and 3, 6, 12, and 24 hours after reoxygenation. The changes of SCF/c-kit R mRNA expression in hypoxic neurons treated with different concentrations of naloxone pre and post oxygen-glucose deprivation were determined with RT-PCR.

RESULTSThe cell vitality detected by MTT assay decreased significantly in hypoxia group and naloxone 0.5 microg/ml group when compared with control group (P<0.01), while no significant difference was found between naloxone 0.5 microg/ml group and hypoxia group or between naloxone 10 microg/ml group and control group. Extracellular concentration of LDH significantly increased in hypoxia group (P<0.05), while no difference was found between naloxone 0.5 microg/ml group and control group, between naloxone 0.5 microg/ml and hypoxia group, or between naloxone 10 microg/ml and control group (all P>0.05). Immediately after oxygen-glucose deprivation, the expression of SCF/c-kit R mRNA increased significantly (P<0.01). Among those the expression of SCF presented a distribution of double-peak value within 24 hours. After treated with different concentrations of naloxone, the peak value of each group were delayed to appear and went down with the increasing of naloxone concentration. The peak values in all treated groups were significantly different from that in control group (P<0.01).

CONCLUSIONSThe expression of SCF/c-kit R mRNA increases at the early stage after combined oxygen-glucose deprivation. Naloxone 0.5 microg/ml can attenuate cell injuries and regulate the expression of SCF/c-kit R. Naloxone may protect neurons by modulating the expressions of some cytokines.

Cell Hypoxia ; drug effects ; physiology ; Cells, Cultured ; Cerebral Cortex ; cytology ; Humans ; Naloxone ; pharmacology ; Neurons ; drug effects ; metabolism ; pathology ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; RNA, Messenger ; genetics ; Stem Cell Factor ; genetics ; metabolism

OBJECTIVETo discuss the outcomes and complications of total knee arthroplasty (TKA) for extension ankylosing deformity of the knee.

METHODSFrom January 1996 to June 2006, total knee arthroplasty was performed on 8 patients (9 knees) with extension ankylosing deformity. The preoperative ROM of all patients was 0 degrees . Preoperative knee and function score of KSS were 44 points (from 10 to 68) and 17 points (from -10 to 55) respectively.

RESULTSThe complications of all TKAs included patellar tendon avulsion in 1 knee, partial fracture of inferior patella in 1 knee, hematoma in 1, superficial infection in 1. All patients were followed up for an average of 40.4 months (from 7.0 to 120.0). The average postoperative ROM was 89 degrees (from 50 degrees to 120 degrees ). Postoperative knee and function score of KSS were 81 points (from 55 to 93) and 79 points (from 50 to 90) respectively. Extension lag occurred in 2 knees, one was 10 degrees and the other was 25 degrees . One knee had undergone re-revision of changing the thicker tibial spacer for the reason of instability of joint 1 year after revision.

CONCLUSIONSTKA performed in extension ankylosing deformity can get less satisfactory clinical results comparing with fixed flexion deformity. Exposure of the knee joint and separation of the fused bones, providing a mobile joint space plays crucial procedure for the next step of surgery. Preservation of sufficient bone stock of patella, protection of patellar tendon and blood supply of the knee and proper soft tissue balance are the key to TKA for extension ankylosing deformity.

Adult ; Aged ; Ankylosis ; physiopathology ; surgery ; Arthroplasty, Replacement, Knee ; adverse effects ; methods ; Female ; Humans ; Joint Deformities, Acquired ; physiopathology ; surgery ; Knee Joint ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; Range of Motion, Articular ; Retrospective Studies ; Treatment Outcome

Gerbil forebrain ischemia/reperfusion(I/R) injury model was used to study the effects of D(1) and D(2) receptor agonists and antagonists on neuronal apoptosis of hippocampal CA1 area. All animals were tested for habituation deficits in an open field test on the 1st, 3rd and 7th days after reperfusion. The animals were then killed, and brains underwent paraffin embedding for hematoxylin-eosin staining, in situ terminal deoxynucleotidyl transferase-mediated deoxy-UTP nick end labeling (TUNEL) staining and immunohistochemistry (bax, bcl-2). The result of open field test showed that the I/R group was significantly impaired (higher activity scores) when compared with the control group. Pretreatment with pergolide significantly reduced this habituation impairment. Forebrain ischemia for 5 min resulted in extensive CA1 apoptosis on the 3rd and 7th days after I/R injury. About 95% neurons in hippocampal CA1 area entered apoptosis and only 2%-7% pyramidal neurons stayed alive due to an inhibition of bcl-2 expression and an increase in bax expression. Pretreatment of pergolide attenuated neuronal damage caused by transient ischemia. Infusion of pergolide could induce the expression of bcl-2 and reduce the expression of bax. Pretreatment with SKF38393, SCH23390 and spiperone had no effects on these changes in this transient I/R injury model. All these results indicate that pergolide plays an important role in the protection of hippocampal neurons from apotosis through upregulating the expression of bcl-2 protein and reducing the expression of bax protein.
2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine ; pharmacology ; Animals ; Apoptosis ; Brain ; physiopathology ; Brain Ischemia ; physiopathology ; Dopamine Agonists ; pharmacology ; Dopamine Antagonists ; pharmacology ; Gerbillinae ; Hippocampus ; physiopathology ; Ischemic Attack, Transient ; physiopathology ; Male ; Neurons ; physiology ; Neuroprotective Agents ; pharmacology ; Pergolide ; pharmacology ; Prosencephalon ; physiopathology ; Proto-Oncogene Proteins ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Receptors, Dopamine D1 ; Receptors, Dopamine D2 ; Reperfusion Injury ; physiopathology ; bcl-2-Associated X Protein