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.

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.

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 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 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 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 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 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 study the influential factors of tacrolimus'dosage and concentration differences between individuals in morning periods after renal transplantation.Methods The clinical data consisted of 118 receptors in morning periods after renal transplantation,whose immune suppressions were tacrolimus,mycophenolate and hormone.At 3,7,14 and 30 d after operation,all the receptors'weight,dosage of tacrolimus,dosage of hormone,diarrhea,blood fat,liver function,renal function,albumn and erythrocrit were recorded respectively,and at the same time their concentrations of tacrolimus and genetic polymorphisms of CYP3A5,MDRl 3435,MDR1 2677 and MDRl 1236 weredetected.Multiple linear regressions were performed.Results The fitting degrees of stepwise regression equations were low.At 3,7,14 and 30 d after operation,the adjusted R2was 0.284,0.267,0.417 and 0.324,respectively.From the aspect of pharmacogenomics,the main factors rela-ted to the differences of tacrolimus'dosage and concentration included MDR1 2677,MDRl 1236 and MDR13435,which varied intensively.Age,albumn,renal function,blood fat and liver function were important factors too.Conclusions The main reasons of the differences of tacrolimus'dosage and concentration between individuals in morning periods after renal transplantation are medicines and changes of internal environment after operation.The genetic polymorphisms of MDR1,age,albumn,renal runetion.blood fat and liver function are important factors too.

Objective To investigate a possible association of donor-recipient compatibility for ABO blood group alleles with acute rejection (AR) in renal transplantation. Methods A study comprising 87 pairs of donor and recipient was performed. The ABO genotype A1, A2, O1, O2, and B alleles of renal transplanted recipients and their respective donors were assessed by PCR amplification with sequence-specific primers (PCR-SSP). Accordingly, recipients were divided into donor-recipient ABO genotype matched and mismatched groups. Results The PCR-SSP based types of all cases showed total concordance with their serologically assigned ABO groups. Fifty pairs (57. 5%) were matched for ABO genotype among the 87 pairs of donor and recipient while 37 (42. 5%) were mismatched, including 1 allele mismatch in 31 pairs (83.8%), 2 alleles mismatches in 6 pairs (16. 2%).The incidence of AR was 12.0% (6 cases) and 29. 7% (11 cases) for ABO genotype matched and mismatched transplant patients, respectively ( P ＜ 0.05). After high dose methylprednisolone (MP)treatment, all cases exepienced reversion of AR except a A2O1 recipient receiving kidney from a A1O1enced 4 AR episodes within 3-10 months, and the period of AR was gradually shortened. After high dose MP was administered empirically, even though short-term improvement of renal function was observed, the serum creatinine continued to increase progressively with decreased efficacy of high dose MP. One year after operation the serum creatinine rose to 441 μmol/L. Conclusions Simultaneous definition of the ABO genotype and HLA is highly feasible. The A2 patient is suitable for receiving kidneys from blood group O donors. DNA mismatch for ABO genotype of renal transplant recipients and their respective donors is an independent risk factor for AR. Genotyping of ABO blood group is conducive to prevent AR.