Objective To evaluate the effect of bone marrow mesenchymal stem cell (BMSC) on the expression of interleukin (IL)-10 and tumor necrosis factor (TNF)-α in mice with ischemia-reperfusion acute kidney injury (IR-AKI). Methods All mice were randomly divided into the sham operation group (control group), ischemia-reperfusion injury group (IRI group) and BMSC treatment group (BMSC group), with 6 mice in each group, respectively. The renal function and pathological changes of mice were detected. The cell apoptosis of renal tissues of mice was determined. The expression levels of serum IL-10 and TNF-α of mice were quantitatively measured. The mouse BMSC was randomly divided into the control and hypoxia-reoxygenation groups (IRI group), and the expression levels of IL-10 and TNF-α in cell supernatant were determined. Results The renal structure of mice was normal in the control group, severe damage was observed in the IRI group, and mild damage occurred in the BMSC group. Compared with the control group, the renal tissue injury scores were significantly higher in the IRI and BMSC groups (both P < 0.05). Compared with the IRI group, the renal tissue injury score was significantly lower in the BMSC group (P < 0.05). Compared with the control group, the levels of serum creatinine (Scr) and blood urea nitrogen (BUN) were remarkably up-regulated in the IRI group, and the level of BUN was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of Scr and BUN were significantly down-regulated in the BMSC group (both P < 0.05). In the IRI group, the quantity of apoptotic cells in the renal tissues was considerably higher than those in the BMSC and control groups, and the quantity of apoptotic cells in the BMSC group was significantly higher than that in the control group (all P < 0.05). Compared with the control group, the levels of serum IL-10 and TNF-α were significantly up-regulated in the IRI group, whereas the level of serum TNF-α was significantly down-regulated and the level of serum IL-10 was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of serum IL-10 and TNF-α were significantly down-regulated in the BMSC group (both P < 0.05). The levels of IL-10 and TNF-α in the cell supernatant did not significantly differ between the IRI and control groups (P=0.080、0.627). Conclusions BMSC infusion may reduce the incidence of renal IRI and inflammation, probably via the mechanism of down-regulating TNF-α expression rather than up-regulating IL-10 expression.

Objective To investigate the characteristics and manifestations of the different stages of BK virus infection in the recipients after renal transplantation.Methods A retrospective survey from January 2015 to December 2016 was done in our hospital.A total 135 recipients were included and accepted BK virus detection in 1,3,6,9,12,15 months respectively after renal transplantation.The prevalence of decoy cell,BK virus DNA load in urine and BK virus DNA load in blood was 56 cases (41.5％),9 cases (43.7％) and 30 cases (22.2％),5 cases of BK vims nephropathy confirmed by pathological biopsy (3.7％).At the same time,51 cases (37.8％) were combined with decoy cells and virus DNA load in urine.Positive decoy cells and negative BK virus DNA load in urine was 5 cases,and Positive BK virus DNA load in urine and negative decoy cells was 8 cases.The recipients were divided into positive group of urine decoy cell,positive group of urinary BK virus DNA load,and positive group of blood BK virus DNA load.Statistical correlation analysis was conducted on the laboratory test results of the 3 groups.Results The positive group of blood BK virus DNA load were detected the high level urine decoy cell count [median of 23/10HPF(2-48/10HPF)] and high level of urinary BK virus DNA load [4.52 × 106 copies/ml (6.51 × 103-7.89 × 109 copies/ml)],significantly higher than the positive group of decoy cells [8/10HPF(2-40/10HPF)] and the positive group of urine BK virus DNA load [4.56 × 105 copies/ml(5.62 × 103-7.89 ×109 copies/ml)] (P ＜ 0.05).The decoy cell count and urine DNA load has a significant linear correlation in viruria recipients,and the urinary BK DNA load and blood BK virus DNA load has the same significant 0.939 and 0.702 in 3 months,0.969 and 0.910 in 6 months,0.782 and 0.766 in 9 months,0.898 and 0.615 in 12 months after renal transplantation.Conclusions There is a linear correlation between decoy cell in urine,viruria and viremia,suggesting that the infection of BK virus in kidney transplant recipients is a continuous process.linear correlation in viremia recipients(P ＜ 0.05).The correlation coefficients at different time points were

Objective To compare the effects of cyclosporine A (CsA) and tacrolimus (FK506) on BK virus infection after renal transplantation by retrospective clinical study.Methods The data of calcineurin inhibitor (CNI)-based immunosuppression and virus infection were collected in allograft renal transplantation recipients (n =135) from Jan.2014 to Dec.2015.According to the severity of the virus infection the recipients were divided into three groups:viruria,viremia and virus nephropathy.The difference in BK virus infection between FK506 and CsA was compared.Results A total of 135 cases of transplant recipients,postoperative were enrolled.The number of viruria recipients given FK506 and CsA was 41 cases (69.5％) and 18 cases (30.5％),and that of viremia recipients was 26 cases (86.7 ％) and 4 cases (13.3 ％).Statistical analysis showed that CNI immunosuppressive agents had a significant correlation with viremia only (P＜0.05).There was a positive correlation between FK506 and viremia (r =0.423,P =0.018),and CsA showed a negative correlation yet (r =-0.336,P =0.022).Conclusion Tacrolimus is independent risk factors for early BK viremia after kidney transplantation,and CsA may inhibit the progression of BK viremia.

Objective To analyze the impairment of renal allograft function in renal transplant recipients caused by BK virus infection after renal transplantation. Methods Clinical data of 210 recipients who underwent allogenic renal transplantation and received BK virus monitoring regularly were analyzed retrospectively. The incidence of BK viruria, viremia and BK virus nephropathy (BKVN) after renal transplantation was summarized. The effect of BK virus infection on renal allograft function and prognosis of renal allograft function after the removement of BK virus were analyzed. Results Among the 210 recipients, there were 46 cases with pure viruria, 46 cases with viremia complicated with viruria and 7 cases with BKVN confirmed by pathological biopsy. The level of serum creatinine (Scr) in the recipients with viremia after renal transplantation was linearly related to BK viral load in urine and blood (r=0.594, 0.672, both P<0.01). The level of Scr increased significantly when BK viral load in blood of the recipients with viremia was found positive for the first time, and increased continuously after viremia sustained. And the level of Scr decreased slightly when blood viral load turned to negative after treatment, but still significantly higher than before virus infection. All the above differences were statistically significant (all P<0.05). Compared with the basic level, there was no significant difference in the level of Scr of recipients with pure viruria during positive viruria (all P>0.05). Conclusions It will impair the renal allograft function when BK viremia occurs after renal transplantation, and it is necessary to monitor viral infection regularly. Once the blood BK virus is found positive, it shall be implemented immediately to reduce the intensity of immunosuppression as the preferred clinical intervention.

Objective To evaluate the effect of γ-aminobutyric acid (GABA) and its receptors upon the proliferation of CD8+T cells.Methods The splenic CD8+T cells of Balb/c mice were obtained by CD8+f cell magnetic bead sorting kit.Under the effect of CD3/CD28-activated magnetic bead,CD8+T cells were stimulated by different concentrations of GABA.5-bromo-2-deoxyuridine (BrdU) labeling and flow cytometry were performed to detect the proliferation of CD8+T cells.The expression levels of GABA-A and GABA-B receptor before and after CD8+T cell activation were compared by fluorescent quantitative real-time polymerase chain reaction (PCR).Result Flow cytometry result revealed that GABA could inhibit the proliferation of activated CD8+T cells,manifested as significant decrease in the quantity of CD152+CD8+T cells.Fluorescent quantitative real-time PCR demonstrated that GABA-A receptor subtypes α2,α6 and GABA-B receptor subtype 1a were expressed only when the CD8+T cells were activated.After CD8+T cell activation,the quantity of GABA-A receptor subtypes α3,αs,β2,β3,γ1,γ2 and θ were significantly increased,whereas the quantity of GABA-B2R and GABA-B1b did not significantly differ before and after CD8+T cell activation.Conclusions GABA can suppress the proliferation of activated CD8+T cells.The activation of CD8+T cells is regulated by GABA receptors.However,the underlying mechanism remains to be elucidated.

Objective To analyze and discuss the dynamics of cellular immune response to persistent infection with BK virus after renal transplantation.Methods The recipients of renal transplantation in our center were selected and BK virus load in urine and blood was regularly observed.The victims of persistent infection with BK virus (defined as two successive positive results of BK virus load in urine or blood) were followed up and peripheral blood mononuclear cells (PBMCs) were collected for mixed cultivation with overlapping peptide pool,which contained peptide fragments (VP1,VP2,VP3,LT-Ag and st-Ag) extracted from BK virus.Flow cytometry was used to examine the in vitro proliferation of IFN-γ/IL-2/TNF-ininduced T cells and analyze the dynamics of cellular immune response to BK virus.Results A total of 46 recipients of renal transplantation were enrolled and 6 victims of persistent viruria were identified.Of the 6 victims,3 were complicated with persistent viremia,and 2 were diagnosed as BK virus nephropathy by biopsy,presenting with persistent viruria and viremia.The victims of persistent BK viremia after renal transplantation showed a significantly decreasing trend in cellular immune response to 5 BKV-specific proteins,according to the proliferation of TNF-γ/IL-2/TNF-α-induced T cells.However,this trend was not observed in the victims of persistent BK viruria.Conclusion At the stage of viremia,the victims of BKV infection after renal transplantation have seriously inhibited specific immune response to BKV.Thus,if the antiviral mechanisms are not restored in time,these recipients suffering persistent viremia are prone to virus nephropathy (BKVN),delayed graft function,and even graft loss.

Objective To explore the clinical application experience of leflunomide in rescuing therapy of BK virus nephropathy (BKVN ) after renal transplantation in the case of ineffective treatment with reduction of immunosuppressant.Methods Four recipients with BKVN after renal transplantation were diagnosed at 1 35 th-737 th day after operation,with the pathological staging as following:2 cases in stage A1 ,1 case in stage B1 and 1 case in stage B2.For all recipients, leflunomide was used for rescuing therapy due to ineffective treatment with reduction of immunosuppressant for 0.5-3.0 months.Initially,50 mg/d of leflunomide was given continuously for 3 days,so as to reach therapeutic serum concentration,and then 20 mg/d of leflunomide was given for maintaining.The efficacy and safety were observed.Results After a follow-up for an average of 6 months (5-7 months),3 recipients with development of BKVN were controlled effectively,1 recipient (stage B2)with ineffective treatment.No obvious adverse reactions occurred during medication.Conclusions It is possible to slow down the development of BKVN and reduce the incidence of renal allograft loss by using leflunomide to conduct rescuing therapy of BKVN after renal transplantation in the case of ineffective treatment with reduction of immunosuppressant.Better effect can be achieved if early detection and diagnosis of BKVN are conducted as well as effective measures are taken timely in the early pathological stage.

Objective To investigate the change rules and its significance of erythrocytes surface molecule CD35 , CD58 and CD59 expression in recipients infected with cytomegalovirus (CMV)after renal transplantation. Methods Eighty-two recipients undergoing allogeneic renal transplantation were selected and divided into the negative (n=21 )and positive CMV groups (n=61 )based on the qualitative detection of CMV-pp65 antigen in peripheral blood. According to the results of CMV-pp65 (+)leucocyte count,all 61 patients in positive CMV group were further divided into low (n=55)and high active infection subgroups (n =6 ). Healthy adults were recruited into the normal control group (n =30 ). The expression levels of CMV-pp65 antigen,erythrocytes surface molecule CD35,CD58 and CD59 were measured by flow cytometry. Results Compared with normal control group,the expression levels of erythrocytes surface molecule CD35 , CD58 and CD59 in the positive CMV group were significantly down-regulated,and the CD35 and CD59 expression in the negative CMV group were considerably down-regulated (all P<0. 05 ). Compared with negative CMV group,the expression levels of CD58 and CD59 in the positive CMV group were significantly down-regulated (both P<0. 05 ). The expression levels of CD35 and CD59 in the high active infection subgroup were significantly lower than those in the low active infection subgroup (both P<0. 05 ). Conclusions The more severe active CMV infection after renal transplantation,the lower expression of erythrocytes surface molecule CD35,CD58 and CD59,hinting that red cell immune dysfunction is probably involved with active CMV infection.

Objective To explore the characteristics of diffusion weighted imaging (DWI) of lumbar sacral nerve roots (LSNR)in normal and degenerative lumbosacral vertebrae. Methods The research recruited 20 normal volunteers and 31 patients with spinal stenosis on conventional MRI and DWI scans in lumbosacral spine. We measured the areas from lumbar 3 to sacral 1 at the intervertebral spaces and reconstructed the 3D maximum intensity projection (MIP) and counted the apparent diffusion coefficient (ADC)of LSNR and ganglions. Results In the control group, 196 (98%) LSNR ran symmetrically and lateroinferiorly and 200 ganglions were well defined on MIP of DWI. In the patients group, 74 LSNR showed changes of compression on both T1WI and T2WI, in which DWI appeared thin and distorted in 59 (80%). The ADC value of LSNR were(1.70 ± 0.40)× 10-3 mm2/s and(1.98 ± 0.57) × 10-3 mm2/s separately in normal volunteers and patients (P=0.000), while the ADC values of ganglions were(1.42 ± 0.21)× 10-3 mm2/s and (1.54 ± 0.53)× 10-3 mm2/s respectively in normal volunteers and patients (P=0.000). Conclusion DWI can display the pattern and course of LSNR and ganglions, which indicate that ADC values of compressed LSNR and ganglions are higher than normal ones.