Individualized Immunosuppressive Protocol of Liver Transplant Recipient Should be Made Based on Splenic Function Status / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Lymphocyte subsets play important roles in rejection in liver transplant recipients, and the effect of splenic function on these roles remains unknown. The aim of this study was to explore the feasibility to adjust immunosuppressive agents based on splenic function status through detecting the lymphocyte subsets in liver transplantBeijing recipients.</p><p><b>METHODS</b>The lymphocyte subsets of 49 liver transplant recipients were assessed in the 309th Hospital of Chinese People's Liberation Army between June 2014 and August 2015. The patients were divided into splenectomy group (n = 9), normal splenic function group (n = 24), and hypersplenism group (n = 16). The percentages and counts of CD4+ T, CD8+ T, natural killer (NK) cell, B-cell, regulatory B-cell (Breg), and regulatory T-cell (Treg) were detected by flow cytometer. In addition, the immunosuppressive agents, histories of rejection and infection, and postoperative time of the patients were compared among the three groups.</p><p><b>RESULTS</b>There was no significant difference of clinical characteristics among the three groups. The percentage of CD19+CD24+CD38+ Breg was significantly higher in hypersplenism group than normal splenic function group and splenectomy group (3.29 ± 0.97% vs. 2.12 ± 1.08% and 1.90 ± 0.99%, P = 0.001). The same result was found in CD4+CD25+FoxP3+ Treg percentage (0.97 ± 0.39% vs. 0.54 ± 0.31% and 0.56 ± 0.28%, P = 0.001). The counts of CD8+ T-cell, CD4+ T-cell, and NK cell were significantly lower in hypersplenism group than normal splenic function group (254.25 ± 149.08 vs. 476.96 ± 225.52, P= 0.002; 301.69 ± 154.39 vs. 532.50 ± 194.42, P= 0.000; and 88.56 ± 63.15 vs. 188.33 ± 134.51, P = 0.048). Moreover, the counts of CD4+ T-cell and NK cell were significantly lower in hypersplenism group than splenectomy group (301.69 ± 154.39 vs. 491.89 ± 132.31, P= 0.033; and 88.56 ± 63.15 vs. 226.00 ± 168.85, P = 0.032).</p><p><b>CONCLUSION</b>Splenic function status might affect the immunity of liver transplant recipients, that should be considered when we make immunosuppressive protocols.</p>

HLA-G expression in the peripheral blood of live kidney transplant recipients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The human leukocyte antigen-G (HLA-G) has been considered to be an important tolerogeneic molecule playing an essential role in maternal-fetal tolerance, upregulated in the context of transplantation, malignancy, and inflammation, and has been correlated with various clinical outcomes. The aim of this study was to investigate the clinical relevance of the expression of membrane HLA-G (mHLA-G), intracellular HLA-G (iHLA-G), and soluble HLA-G (sHLA-G) in the peripheral blood of live kidney transplant recipients.</p><p><b>METHODS</b>We compared the expression of the three HLA-G isoforms in three groups, healthy donors (n=20), recipients with acute rejection (n=19), and functioning transplants (n=30). Flow cytometry was used to detect the expression of mHLA-G and iHLA-G in the T lymphocytes of peripheral blood from subjects in the three groups. Enzyme-linked immunosorbent assays were used to detect sHLA-G in the plasma from the three groups.</p><p><b>RESULTS</b>There were no significant differences in mHLA-G and intracellular HLA-G among the three groups, but the sHLA-G plasma level was higher in the functioning group than in the acute rejection or healthy group. We found a subset of CD4(+)HLA-G(+) and CD8(+)HLA-G(+) T lymphocytes with low rates of mHLA-G expression in the peripheral blood of kidney transplantation recipients. Intracellular expression of HLA-G was detected in T lymphocytes. However, there was no correlation between acute rejection and the mHLA-G or intracellular HLA-G expression.</p><p><b>CONCLUSION</b>sHLA-G was the major isoform in the peripheral blood of live kidney transplant recipients and high sHLA-G levels were associated with allograft acceptance.</p>

Association of vitamin D receptor FokI and ApaI polymorphisms with human cytomegalovirus disease in the first three months following kidney transplantation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Untreated human cytomegalovirus (CMV) disease (CMVD) is an identified risk factor for reduced rates of patient (and graft) survival, death or retransplantation in kidney transplant recipients due to increased immunological tolerance after transplant. Vitamin D receptor (VDR) gene polymorphisms have an obvious relationship with autoimmune diseases but the relationship between VDR gene polymorphisms and CMVD are not well understood. This study investigated the relationship between VDR FokI and ApaI gene polymorphisms and CMVD, and their value for predicting risk of CMVD.</p><p><b>METHODS</b>Ninety-eight kidney transplantation recipients were randomly chosen for which peripheral blood samples and case histories for the first three months after kidney transplantation were obtained. Using polymerase chain reaction-restriction fragment length polymorphisms, 30 recipients were found to be homozygous for the FokI gene (FF), 47 heterozygous (Ff), and 21 were homozygous (ff). Likewise, similar analyses determined that 12 recipients were homozygous for the ApaI gene (AA), 36 heterozygous (Aa), and 50 homozygous (aa). Factors affecting the prognosis of the kidney transplantation were compared for all genotypes by statistical analysis before operation. Infection by CMV for all recipients was detected by immunofluorescence assay to diagnose CMVD.</p><p><b>RESULTS</b>No statistical significance was observed for the factors affecting the prognosis of the kidney transplantation between both genotypes; however, statistical differences in CMVD among the FokI genotypes were identified. It was determined that the risk of CMVD was significantly increased for recipients of the ff genotype than for other genotypes. There was no statistical significance observed for CMVD among ApaI genotypes.</p><p><b>CONCLUSIONS</b>The recessive f allelic gene of VDR can be regarded as a risk factor of CMVD while FF recipients have lower incidence of CMVD after kidney transplantation. ApaI genotypes showed no relationship with predisposition to CMVD.</p>

Programmed death 1 mRNA in peripheral blood as biomarker of acute renal allograft rejection / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Invasive kidney biopsy is a priority diagnostic method for the acute rejection after renal transplantation for the past decades. However, no effective and noninvasive assay for predicting the severity of acute rejection is in wide use at present. This study was designed to investigate the predictive value of programmed death 1 (PD-1) mRNA for acute rejection after renal transplantation with real-time reverse transcriptase polymerase chain reaction (RT-PCR). A noninvasive diagnostic method has been expected to replace the traditional kidney biopsy for the diagnosis of acute rejection and prediction of the outcome after kidney transplantation.</p><p><b>METHODS</b>The whole blood samples from 19 subjects with acute rejection, 20 subjects with delayed graft function (DGF) and 21 subjects with stable recipients after kidney transplantation in a single kidney transplantation center between 2006 and 2009 were collected. The messenger RNA (mRNA) of PD-1 was analyzed with real-time RT-PCR. The associations of PD-1 mRNA levels with acute rejection and disease severity were investigated.</p><p><b>RESULTS</b>The log-transformed ratio of PD-1 mRNA to GAPDH mRNA was higher in peripheral blood mononuclear cell (PBMC) from the group with acute rejection (4.52 ± 1.1) than that from the group with DGF (1.12 ± 0.6) or the group with normal biopsy results (0.7 ± 0.4) (P < 0.01, by the Kruskal-Wallis test). PD-1 mRNA levels were correlated with serum creatinine levels measured at the time of biopsy in the acute rejection group (Spearman's correlation coefficient, r = 0.81, P = 0.03), but not in the group with DGF or the group with normal biopsy results. PD-1 mRNA levels identified subjects at risk for graft failure within six months after the incident episode of acute rejection.</p><p><b>CONCLUSIONS</b>Our data suggest that PD-1 status may be a new predictor of acute rejection and the levels of PD-1 mRNA in whole blood cells may positively correlate with the severity of acute rejection after renal transplantation. Meanwhile, the data provide the rational for interfering into the PD-1/PD-L1 as a novel therapy against the acute rejection after renal transplantation in clinical settings.</p>

Immune regulation effect of rat dendritic cells phagocytosing photochemotherapy-treated allogeneic cells on syngeneic T cells / 中国实验血液学杂志

The aim of this study was to investigate the immune regulatory effect of dendritic cells phagocytosing photochemotherapy-treated allogeneic spleen lymphocytes on syngeneic T cells. DA rat spleen lymphocytes were treated with 8-methoxypsoralen plus UVA irradiation (PUVA). LEW rat bone marrow-derived DCs were co-cultured with PUVA-treated DA spleen lymphocytes (PUVA-SP), and the surface markers (MHC-II, CD86 and CD40) of treated DC were detected by flow cytometry. CFSE-labeled PUVA SP were incubated with LEW DCs and the phagocytosis of DCs on PUVA-SP was observed by using fluorescent microscope. The ability of DC phagocytosing allogeneic PUVA-SP (PUVA-SP DC) to stimulate the proliferation of LEW T cells was analyzed by mixed leukocyte reactions (MLR). The production of IL-4, IL-10, IL-2, IFN-gamma in MLR culture supernatant was determined by luminex method. The results indicated that the PUVA treatment effectively induced early apoptosis of DA rat spleen lymphocytes. After co-culture, DC efficiently phagocytosed allogeneic PUVA-SP and still maintained an immature phenotype with low levels of MHC II, CD40 and CD86. PUVA-SP DC induced LEW T cell hyporesponsiveness to DA rat antigen, and led to skewing of T cell cytokine expression toward Th2 (IL-10 and IL-4). It is concluded that the PUVA-SP DC effectively down-regulate T cell response to alloantigen and induce Th2 immune deviation in vitro.

Risk factors of long-term cardiovascular diseases and cerebrovascular diseases in kidney allograft recipients / 中国医学科学院学报

<p><b>OBJECTIVE</b>To identify the risk factors of cardiovascular diseases and cerebrovascular diseases (CVD) events in kidney allograft recipients.</p><p><b>METHODS</b>We followed up 361 renal transplant recipients who had undergone renal transplantation in our center from January 2000 to December 2003 and evaluated the cumulative incidences and mortalities of CVD complications at baseline and post-transplantation 1, 3, 6, 12, 24, 36, 48, and 60 months. Kaplan-Meier plot was used to assess the incidence and Cox's proportional hazards model to determine the risk factors for cardiovascular complications.</p><p><b>RESULTS</b>The cumulative incidences of CVD were 3.1%, 5.4%, 9.9%, 13.0%, 18.0%, 21.1%, and 24.1%, 1, 6, 12, 36, 48, and 60 months after transplantation, respectively. History of diabetes mellitus (RR 2.19, 95% CI 1.32-3.97, P = 0.009) and CVD (RR 6.34, 95% CI 3.76-14.60, P = 0.002) as well as the post-transplantation hypertension (RR 1.18, 95% CI 1.02-1.34, P = 0.04), diabetes mellitus (RR 2.82, 95% CI 1.33-7.26, P = 0.002), hyperlipidemia (RR 2.04, 95% CI 1.26-5.17, P = 0.008) and abnormal creatinine (> 200 micromol/L, RR 1.81, 95% CI 1.08-3.21, P = 0.03), and proteinuria (> 0.3 g/d , RR 1.56, 95% CI 1.12-3.54, P = 0.05) were independently correlated with the development of cardiovascular events.</p><p><b>CONCLUSION</b>History of diabetes mellitus and CVD, post-transplant hypertension, diabetes mellitus, hyperlipidemia, abnormal creatinine and proteinuria are the independent risk factors of the development of CVD events.</p>

Pathogenesis of post-transplantation diabetes mellitus in 40 renal transplantation recipients / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore pathogenesis of post-transplantation diabetes mellitus (PTDM) in renal transplantation recipients.</p><p><b>METHODS</b>A total of 40 renal transplantation recipients were divided into three groups based on oral glucose tolerance test results: normal glucose tolerance (NGT) group (n = 10), impaired fasting glycaemia + impaired glucose tolerance (IFG + IGT) group (n = 16), and PTDM group (n = 14). Insulin resistance (IR) and beta cell function were assessed by homeostasis model.</p><p><b>RESULTS</b>The differences of the immunosuppressive agents used in these groups were not statistically significant (P > 0.05). Compared with NGT group, insulin area under curve and homeostasis model assessment-insulin resistance index were significantly higher in IGT + IFG group and PTDM group (P < 0.05). Compared with NGT group and IGT + IPG group, insulin secretion index at 30 min and homeostasis model assessment-insulin secretion index were significantly lower in PTDM group (P < 0.05).</p><p><b>CONCLUSION</b>Insulin resistance and beta-cell dysfunction may play a key role in the pathogenesis of PTDM.</p>

Role of IL-6/STAT3 in rat cholangiocyte proliferation induced by lipopolysaccharide / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate whether lipopolysaccharide (LPS) stimulates cholangiocyte proliferation via the IL-6/STAT3 pathway in vivo.</p><p><b>METHODS</b>Rats were randomized into three groups: LPS group (injected intravenously with LPS 2.5 mg/kg), anti-IL-6 group (injected intravenously with anti-IL-6 0.5 mg/kg 1hr after LPS injection), and control group. At 6, 12, 24, 48 and 72 h after LPS injection, LPS concentration in plasma was detected by kinetic turbidimetric limulus test. IL-6 concentrations in liver homogenate was determinded by ELISA, cholangiocyte proliferation was checked by immunohistochemistry, expression of IL-6 mRNA was quantified by real-time RT-PCR, the level of phophorylated-STAT3 (P-STAT3) protein was analyzed by western blotting.</p><p><b>RESULTS</b>Cholangiocytes responded to LPS by a marked increase in cell proliferation, IL-6 secretion and P-STAT3 expression. Anti-IL-6 neutralizing antibody inhibited LPS-induced cholangiocytes proliferation, and decreased levels of IL-6 and p-STAT3.</p><p><b>CONCLUSIONS</b>LPS promotes cholangiocyte proliferation through the IL-6/STAT3 pathway.</p>

Role of the activation of interleukin-6/STAT3 in cholangiocyte proliferation induced by cold ischemia reperfusion and injury / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the role of IL-6/STAT3 pathway in the proliferation of cholangiocyte induced by cold ischemia and reperfusion injury.</p><p><b>METHODS</b>Rats were randomized into CP 1 h and CP 12 h groups (supplied livers were preserved for 1 or 12 h), anti-IL-6R (rats in CP 12 h group were administrated with anti-rat soluble IL-6 receptor antibody), and control group. At 1, 3, 7, 14 d postoperative, IL-6 concentration in liver homogenate and cholangiocyte proliferation were detected by enzyme linked immunosorbent assay and histochemistry respectively. Expressions of IL-6 mRNA, phosphorylated-STAT3 and cyclin D1 protein in cholangiocytes were determined by real-time PCR or Western blot analysis. Serum concentrations of ALP and GGT and histology analysis were also performed.</p><p><b>RESULTS</b>Minimal expressions of IL-6, p-STAT3 and cyclin D1 were detected in CP 1 h group, with a slight cholangiocytes proliferation. Cholangiocytes responded to extended cold preservation with severe bile duct injures and marked increase in IL-6 secretion, p-STAT3 and cyclin D1 protein expression, followed by compensatory cholangiocytes regeneration. Parallel to this observation, biochemical index and morphology indicated that bile duct injury was recovery at 14 d postoperative. However, anti-sIL-6R inhibited cholangiocytes proliferation and reduced the expressions of IL-6, STAT3 and cyclin D, with the cellular injury and increase of serum ALP or GGT.</p><p><b>CONCLUSIONS</b>IL-6/STAT3 pathway might participate to initiate cholangiocytes regeneration after cold ischemia and preservation injury, which might benefit biliary recovery after liver transplantation.</p>

The safety analysis of living-related kidney donors in short term after transplantation / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the safety of living related donors in short term after transplantation.</p><p><b>METHODS</b>Two hundred and fifty-one cases of living related donor kidney transplantation from May 2000 to July 2007 were analysed retrospectively. There were 117 male and 134 female aged from 22 to 72 years old, with a mean of 46.6 years old. The indexes were compared including serum creatinine (SCr), creatinine clearance (CCr), glomerular filtration rate (GFR) and quality of life before and after donation. Surgical complications were followed-up.</p><p><b>RESULTS</b>Donors' SCr was (75.9 +/- 17.2) micromol/L before donation, (107.4 +/- 21.2) micromol/L on 7 d after donation, (130.4 +/- 58.2) micromol/L at the 1(st) month and (116.1 +/- 24.1) micromol/L at the 3(rd) month. There were significant difference between any 2 time points (P < 0.01). CCr was (94.4 +/- 17.5) ml/min before donation and (63.5 +/- 17.8) ml/min on 10 d after donation (P < 0.01). In 62 donors, total GFR was (82.4 +/- 21.8) ml/min before donation. On 10 d after donation, GFR of remaining kidney was (57.4 +/- 14.1) ml/min which was 34.7% higher than GFR of this kidney before donation (42.6 +/- 11.8) ml/min. There was no significant difference in quality of life before living related donors and non-donor populations (P = 0.116). Surgical complications included splenic rupture in 1 case, descending colon rupture in 1 case and wound infection in 5 cases.</p><p><b>CONCLUSION</b>Living donor kidney transplantation is safe for donors, although part of indexes would vary within normal range during the early time after donation.</p>

Endothelial cell chimerism by fluorescence in situ hybridization in gender mismatched renal allograft biopsies / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The blood vessels of a transplanted organ are the interface between donor and recipient. The endothelium in the blood vessels is thought to be the major target for graft rejection. Endothelial cells of a transplanted organ can be of recipient origin after transplantation. In this study, we tested whether endothelial chimerism correlated with the graft rejection and cold ischemia.</p><p><b>METHODS</b>We studied the biopsy samples from 34 renal transplants of female recipients who received the kidney from a male donor for the presence of endothelial cells of recipient origin. We examined the tissue sections of renal biopsy samples by fluorescence in situ hybridization (FISH) for the presence of endothelial cells containing two X chromosomes using a biotinylated Y chromosome probe and digoxigenin labelled X chromosome probe, and then analyzed the relationship between the endothelial cell chimerism and the rejection and cold ischemia.</p><p><b>RESULTS</b>Endothelial chimerism was common and irrespective of rejections (P > 0.05). The cold ischemic time of chimerism group was longer than no chimerism group ((14.83 +/- 4.03) hours vs (11.27 +/- 3.87) hours, P < 0.05).</p><p><b>CONCLUSIONS</b>There is no correlation between the percentage of recipient endothelial cells in vascular endothelial cells and the type of graft rejection. The endothelium damaged by ischemic injury might be repaired by the endothelial cells from the recipient.</p>

Analysis of complicated malignant tumor following renal transplantation / 中国组织工程研究

A total of 1 100 patients underwent renal transplantation in the Organ Transplantation Center,Second Affiliated Department of General Hospital of Chinese PLA between 1988 and 2008 were collected,and retrospective analysis was performed in five female patients with malignant tumor,which appeared at 68 months (20-132 months) following renal transplantation,including 2 with renipelvic and uretal cancer and 3 with bladder cancer. Two of the 3 patients with bladder cancer presented homolateral renipelvic and uretal metastasis. Three cases and 1 case of upper uretal cacer were observed at the homolateral or heterolateral of kidney grafts respectively. The main characteristic of sign was iterative and painless gross hematuria. The 5 patients underwent renal transplantation with intravesical instillation therapy and nephrectomy. All patients were survived without rejection in the 1-62 months follow-up. The incidence of malignancy in renal allograft recipients is much higher than that in normal ones,which is related to the long term use of immunosuppressants. Urinary epithelial cancer is the main complicating carcinoma and the first choice of treatment is surgical operation. Based on the normal renal grafts function,the dose of immunosuppressants should be as low as possible,Moreover,radiotherapy or chemiotherapy should be adopted according to the types and stages of tumor complicating renal transplantation.