Objective To observe the effect of exercise on the expression of TNF-α in the adipose tissue of insulin resistant rats fed a high fat diet. Methods Thirty healthy male rats were randomly divided into a high fat di- et group and a normal chow group. Eighteen weeks later, the high fat group was randomly divided into a resting group fed with the high fat diet only, and an exercise group fed the high fat diet, but receiving swimming training for 6 weeks. Changes in their metabolism of glucose and lipids were observed, and the insulin sensitivity index was calcu-lated. Meanwhile, the level of TNF-α mRNA in their adipose tissue was detected with a real-time fluorescence quan-titative polymerase chain reaction (PCR), and protein in the adipose tissue was measured using Western blotting. Results After 18 weeks of high fat diet feeding, the insulin sensitivity index of the high fat diet group decreased sig-nificantly as compared to the normal chow group, suggesting that insulin resistance had been acquired in the high fat diet group. 24 weeks later, the insulin sensitivity index of the resting group had decreased further, again significantly when compared to the normal chow group. Compared to the resting group, the insulin sensitivity index of exercise group was significantly higher, and the expression of TNF-α mRNA and protein in their adipose tissue was significant- ly increased. Conclusion Insulin resistance can be induced by high fat diet feeding. Exercise can improve insulin resistance by increasing the expression of TNF-α in adipose tissue.

To further analyze immunoglobulin (Ig) properties of panel reactive antibodies (PRA) and their potential harmful effects on allogeneic renal transplants. Methods Since dithiothreitol (DTT) is able to crack disulphuric bond of Ig and to depoly-merize IgM with heavy molecular weight, PRA study was performed with treatment of DTT. Thus, a DTT-ERA method was established, which can identify immunoglobulin class in sera of high PRA patients. Results Among 701 recipients, whose sera were positive by the standard PRA, the positive rate of DTT-PRA was 33.2% . Besides, positive PRA patients could be divided into three groups according to their sera's sensitivity to DTT: Group 1 consisted of patients whose sera contained exclusively IgM antibodies; Group 2 consisted of patients whose sera contained of IgG antibodies only; Group 3 consisted of patients whose sera were found to contain both IgM class antibodies and IgG antibodies. Conclusion IgM class antibodies are not associated with posttrans-plant rejection, while IgG antibodies alone and mixture of IgG and IgM antibodies may mediate acute, or even hyperacute rejection.

Objective To investigate the changes of panel reaction antibody (PRA) and the effects of HLA sensitized paths in patients waiting for renal transplantation.Methods PRA of 10 555 samples from 8926 renal transplant recipients in 51 transplant centers was analyzed.In 1991-1998 group,PRA was by using NIH-CDC technique,and in 1999-2010 group,PRA was detected by using ELISA.The effects of blood transfusion,pregnancy and transplantation on the PRA positive rate were analyzed.Results There were 1324 recipients positive for PRA in 8926 (14.83 ％ ).The average PRA positive rate in 1991 1998 group and 1999-2010 group was 18.17％ (513/2823) and 13.29％ (811/6103) repectively (P＜0.01).Among 1324 PRA positive recipients,the number of recipients with PRA of 1％-30％ and PRA of ≥30％ respectively accounted for 71.83％ and 28.17％.There were statistically significant differences in the PRA positive rate between the recipients receiving blood transfusion and those without blood transfusion (31.77％ vs 1.21％,P ＜ 0.01 ),between the recipients having pregnancy history and those without pregnancy history (24.64％ vs 7.19％,P＜ 0.01 ),and between primary transplant and re-transplant recipients (13.35 ％ vs 40.62％,P＜0.01).Conclusion In last 20 years, PRA in majority of PRA positive recipients was ＜ 30％ (low sensitized).Renal transplantation and blood transfusion were the important influencing factors that led to positive PRA,and pregnancy history was a related factor.

Objective To study the preoperative treatment and prognosis observation? in sensitized recipients of kidney transplantation.Method Forty-one recipients positive for preoperative PRA accepted renal allograft transplantation from January 2007 to July 2012.All recipients were given immunosuppressant or immune induction by anti-CD25rnAb in advance,and plasma exchange,immunoadsorption and intravenous high-dose immune globulin were administered.Meanwhile,donor HLA antigens had to avoid all stored HLA antibodies of the recipient,and lymphocyte cytotoxicity cross test (CDC) had to be negative.Anti-human lymphocyte globulin (ATG) was used to strengthen the immune induction,and tacrolimus + mycophenolate mofetil (MMF) + corticosteroids triple immunosuppressive regimen was adopted after transplantation.Then intravenous micafungin would be given after transplanted kidney function was normal,and ganciclovir and sulfamethoxazole were taken orally to prevent infection.Result In 41 recipients positive for preoperative PRA,13 cases were positive for only HLA class Ⅰ antibodies,15 cases for only HLA class Ⅱ antibodies,and there existed 13 cases of both HLA class Ⅰ and class Ⅱ antibodies also with PRA≥50％.Fifteen patients achieved normal serum creatinine in one week,and no hyperacute rejection and accelerated rejection occurred.Fourteen recipients experienced an episode of acute rejection (34.15％,14/41): 12 recovered by steroids bolus therapy,and the other two reversed in 3-5 days by cyclophosphamide or ATG treatment.One case died of mycotic pneumonia in 4 months later.One-year recipient/kidney survival rate was 97.6％ (40/41).Conclusion The recipients positive for preoperative PRA only can accept renal allograft transplant while the donor's HLA antigens had to avoid all stored HLA antibodies of recipients themselves and CDC test was negative.After that the combination of desensitization therapy,immune induction therapy and postoperative potent immunosuppressant can prevent acute rejection effectively and increase postoperative recipient/kidney survival rate.

Objective To observe the effects of exercise on serum adiponectin and adiponectin receptor (AdipoR) level in skeletal muscle of insulin-resistant rats. Methods A total of 30 healthy male rats were randomly divided into a control group ( NC, n = 8) and a high-fat group ( HF, n = 22), fed with normal chow and high fat diet, respectively. Eighteen weeks later, the high-fat group was randomly divided into a high-fat diet control group (HC, n = 10) and an exercise group (HE, n = 12). The HC and HE group were continually fed with high fat diet, while the HE group was administered with swimming training for 6 weeks in addition at the same time. After 24 weeks, the insulin sensitivity index was calculated, and serum adiponectin level was detected by using ELISA. The expressions of AdipoR mRNA in skeletal muscle were detected with real-time fluorescence quantitative polymerase chain reaction (PCR). Results After 18 weeks, compared to NC group, the insulin sensitivity index of HF group decreased significantly. It suggested that insulin resistance appeared in HF group. Twenty-four weeks later, compared to NC group, the ISI of HC group was significantly decreased, meanwhile the level of serum adiponectin, expression of AdipoR1 and AdipoR2 mRNA in skeletal muscle of HC group were 71.9% , 59.9% and 69.2% of those of the NC group, respectively; compared to HC group, the ISI was increased significantly by exercise, meanwhile the expression of AdipoR1 mRNA in skeletal muscle was significantly increased by 1.33 times, however the level of serum adiponectin and the expression of AdipoR2 mRNA in skeletal muscle were not altered in HE group. Conclusion Six weeks of exercise improves insulin sensitivity through increasing the expression of AdipoRI mRNA in skeletal muscle.

Objective To investigate the correlation between serum C-reactive protein (CRP) level and carotid atherosclerotic plaque calcification in patients with ischemic stroke or transient ischemic attack (TIA).Methods The patients with non-cardiogenic ischemic stroke or TIA in anterior circulation performed head and neck vascular CTA at 1-6 months from the time of onset were enrolled prospectively.The demographic and clinical data were collected and serum CRP levels were detected.Univariate and multivariate logistic regression analyses were used to determine the correlation between the serum CRP level and the carotid atherosclerotic plaque calcification.Results A total of 165 patients were enrolled.Their age was 62.4± 10.6years,male patients accotnted for 66.7％;113 patients (68.5％)had carotid atherosclerotic plaque calcification (calcification group),52 (31.5％) did not have carotid atherosclerotic plaque calcification (non-calcification group).The age of the calcification group (median,interquartlle;66 [58-73] years vs.58 [51-66] years;Z=-3.738,P＜0.001) and CRP levels (1.9 [0.5-3.8] mg/L vs.0.0 [0.0-2.2] mg/L;Z =-4.126,P ＜ 0.001) were significantly higher than those of the non-calcification group.There were no significant differences in other baseline clinical data between the two groups.Multivariate logistic regression analysis showed that age (odds ratio 1.063,95％ confidence interval 1.024-1.104;P =0.001) and CRP levels (odds ratio 1.209,95％ confidence interval 1.030-1.419;P=0.020) were still significantly correlated with the plaque calcification after adjusting for other confounding factors.Conclucions Carotid plaque calcification was correlated with older age and increased serum CRP level in patients with ischemic stroke or TIA.

Objective To investigate the correlation between human leukocyte antigens-A,-B,-DRB1 (HLA-A,-B,-DRB1) high resolution alleles and chronic renal failure (CRF) caused by immunoglobulin-a nephropathy (IgAN).Method The polymerase chain reaction-sequence-based typing (PCR-SBT) method was used to investigate the genotypes of HLA-A,-B and-DRB1 high-resolution alleles in 191 cases of CRF caused by IgAN (experimental group) and 503 healthy blood donors (control group).The alleles frequencies between two groups were compared and the association between CRF caused by IgAN and the polyrnorphism of HLA was analyzed.Result (1) There were 25 alleles at A locus,48 alleles at B locus and 32 alleles at DRB1 locus in experimental group.(2) The genetic frequency of HLAA * 2901 [Pc =0.033,OR =10.738,95％ CI (1.193,96.691)],HLA DRB1 * 1106 [Pc =0.0001,OR =0.969,95％ CI (0.944,0.994)],HLA-DRB1 * 1202[Pc =0.002,OR =1.859,95％ CI (1.259,2.745)],HLA-DRB1 * 1401 [Pc =0.021,OR =0.984,95％ CI (0.967,0.998)],HLA-DRB1 * 1602[Pc=0.015,OR=1.915,95％ CI (1.157,3.17)] in experimental group was higher than in control group (P＜0.05).Conclusion There is susceptibility association of HLA-A * 2901,HLA-DRB1 * 1106,HLA-DRE * 1202,HLA-DRB1 * 1401,HLA-DRB1 * 1602 with CRF caused by IgAN.It is concluded that there is a close genetic and immunological correlation between HLA alleles and the pathogenesis of CRF caused by IgAN.

Objective To establish a novel flow cytometric crossmatch assay allowing detection of complement-fixing donor specific anti-HLA IgG alloantibodies (Flow cytometry complement-dependent crossmatch,Flow-CDC). Methods One hundred pretransplant crossmatchings were performed using Flow-CDC and NIH-CDC between 62 patients awaiting renal transplantation and 33 donor cells.These crossmatchings were divided into two groups according to PRA.Group 1 consisted of 25 sera with negative PRA,and group 2 consisted of 75 sera with positive PRA.All of the sera were pretreated with DTT to inactivate IgM. Lymphocytes were isolated from peripheral blood (or,in a few instances,from the spleen) of the cadaveric donors. The correlation between different techniques for detection of donor specific anti-HLA antibodies was evaluated.The effect of both methods on clinical transplantation outcome was observed. Results In group 1,NIH-CDC and Flow-CDC were negative for all 25 sera.In group 2,24 (32.0%) had a positive NIH-CDC,31 (41.3%) had a positive Flow-CDC.There was a significant difference between two methods (?2=5.14, P= 0.016 ).Overall concordance between both tests was 93% with 69 concordant negatives and 24 concordant positives. The correlation coefficient (r) was 0.80.In group 2,5 patients received transplantation. One of them with negative NIH-CDC and positive Flow-CDC suffered from acute rejection after transplantation and lost the graft,and the other patients with negative NIH-CDC and Flow-CDC had good outcome. Conclusions Flow-CDC can detect specifically complement-fixing IgG alloantibodies against donor HLA and is more sensitive than NIH-CDC.Additionally,the computer printouts represent a permanent record of the crossmatch for retrospective review.Flow-CDC may become the standard crossmatch method as a possible alternative to conventional NIH-CDC testing.

Objective To investigate the production path of major histocompatibility complex class Ⅰ chain-related gene A(MICA) antibodies and the impact on the therapeutic efficacy after acute rejection in renal transplantation recipients.Methods Luminex flow cytometry was used to detect antiMICA antibodies and the antibody specificity in 157 pre-transplant kidney transplant recipients randomly selected.The clinical data were collected,anti-MICA antibody production pathway and immunoglobulin types were analyzed,and the impact of IgM anti-MICA antibody and IgM&IgG complex anti-MICA antibodies on acute rejection (AR) incidence and therapeutic efficacy after renal transplantation.Results Of the total 157 recipients,19 recipients were positive for anti-MICA antibodies before renal transplantation in 68 recipients who had history of blood transfusion,pregnancy and transplant sensitized experience (27.9％ ); In 89 recipients having no sensitized experience,MICA antibodies were positive in 26 recipients (29.2％ ) (P＞0.05).In 45 anti-MICA antibody-positive recipients,the anti-MICA antibodies type was IgM in 26 cases having no sensitized experience; and that was IgG and IgM complex in 19 cases having sensitized experience.In 38 antiMICA antibody-positive recipients undergoing kidney transplantation,7 out of 22 IgM anti-MICA antibodies recipients had AR (31.8％) that was reversed by methylprednisolone pulse therapy,and 7out of 16 IgM&IgG complex anti-MICA antibodies recipients had AR (43.8％) and treated with methylprednisolone pulse therapy:reversion in 3 recipients (42.9％),and the graft function loss in 4 recipients.The AR incidence was not associated with the two immunoglobulin types of MICA antibodies(P＞0.05),but there was significant difference in the reversal rate of AR (P＜0.05).Conclusion For non-allergenic history recipients,there exists the classic “natural antibodies” pathway in the production of the anti-MICA antibodies whose immunoglobulin type was IgM.In addition,the reversal effect of AR in recipients with IgM anti-MICA antibodies was much better.We need to attach importance to IgM&IgG complex anti-MICA antibodies for the pre-transplant anti-MICA antibodies in renal transplant recipients,because their AR treatment outcome is poor.

ObjectivesTo evaluate the effect of combination of liraglutide,a glucagon-like peptide-1 analogue and pioglitazone,an insulin sensitizer,on diabetic db/db mice.MethodsThirty-five 8-week old male db/db mice were divided into control group (n = 8 ),pioglitazone group (n =9 ),liraglutide group (n =9) and combined therapeutic group (n =9),which was given normal saline 0.1 ml,2/d,pioglitazone 24 mg· kg-1 · d-1 (feed contained 0.02％ pioglitazone) + normal saline 0.1 ml,2/d,liraglutide 300 mg/kg,2/d,and pioglitazone 20 mg · kg-1 · d -1 ( feed contained 0.02％ pioglitazone) +liraglutide 300 mg/kg,2/d,respectively.Liraglutide were given at 8:00 and 16:00 via subcutaneous injection after having been diluted with sterilized normal saline.Effect on glucose,lipid metabolism and islet β-cell preservation were assessed after 4 weeks.Oneway ANOVA was adopted for statistical analysis.Results Combination therapy displayed promising anti-hyperglycemic[glycosylated hemoglobin Alc: (4.5 ± 0.6)％vs.(7.3 ±0.4)％,P ＜ 0.001].Glucose tolerance were improved assessed by area under curve(AUC) of glucose by intraperitoneal glucose tolerance test (IPGTT)[(1814 ±91 ) mmol · min · L-1 vs.(4042 ±183) mmol · min · L-1,P ＜0.001];insulin release response to glucose were also preserved as AUC of insulin by IPGTT was higher[( 1639 ±372) μg · min · L-1 vs.(834 ±201 )μg · min · L-1].Combination therapy also reduced circulated free fatty acids and TG[( 202.0 ± 20.4 ) μmol/L vs.( 272.5 ± 21.7 )μmol/L,(0.81 ± 0.28) mmol/L vs.( 1.35 ± 0.21 ) mmol/L],and increased plasma adiponectin [(16.7±2.0)mg/L vs.(10.2±1.8)mg/L].All P value ＜0.05.Islet immunohistochemistry showed that combination therapy significantly increased insulin positive area were[( 59.5 ± 1.5 ) ％ vs.( 22.4 ±1.5) ％]and ratio of Brdu positive β-cells was three folds than vehicle-treated mice[( 2.4 ± 0.5 ) ％ vs.(0.8 ±0.3)％],both greater than each single treatment.Combined therapy significantly improved islet β cell/α cell distribution,which led to islet recovery.ConclusionsCombined therapy improves glucose and lipid metabolism,preserves islet β-cell function and stimulates β-cell proliferation,greater than either liraglutide or pioglitazone treatment alone.