The Thirty-Six Year Experience in Kidney Transplantation at Changzheng Hospital.

Between June 1978 and June 2014, 4,199 kidney transplants were performed at the Transplantation Center of PLA, Changzheng Hospital, Second Military Medical University. In our initial practice period (1978-1985), graft and patient survivals were 48.2% and 56.5%, 27.3% and 31.7%, 22.5% and 24.4%, 20.1% and 23.2%, and 16.5% and 20.8%, at 1, 5, 10, 15, and 20 years, respectively. These results improved tremendously after cyclosporine A (1986-1998) was used at our center. The rates of 1-, 5-, 10-, 15-, and 20-year graft and patient survival were 84.3% and 88.5%, 72.3% and 76.7%, 60.4% and 65.4%, 55.1% and 58.2%, and 49.0% and 51.8%, respectively. Tacrolimus (1999-2014) further increased graft survival to 95.1%, 84.4%, 77.1%, and 70.9%, and patient survival to 98.3%, 90.4%, 80.7%, and 73.4%, at 1, 5, 10, and 15 years, respectively. Multivariate Cox analysis suggested that transplant year, delayed graft function, rejection, immunosuppressive regimen, and original disease were independent predictors of graft survival and that poor HLA matching with 5-6 mismatches had an adverse effect on graft survival compared with 1-2 mismatches. The major causes of patient death included infection (38.1%), cardio-cerebral accident (30.2%), and malignancy (16.3%). As one of the pioneer transplant centers in China, our greatest contribution to organ transplantation in China is our self-developed organ preservation solution (HC-A), which has been used in more than 100,000 grafts and for more than 30 years.

Measurements of serum pituitary-gonadal hormones and investigation of sexual and reproductive functions in kidney transplant recipients.

Objective. To investigate changes in serum pituitary-gonadal hormones and restoration of sexual and reproductive functions after successful kidney transplantation. Patients and Methods. Serum pituitary-gonadal hormones before and after kidney transplantation were measured in 78 patients with end-stage renal disease (ESRD) and in 30 healthy adults. Pre- and postoperative semen specimens of 46 male recipients and 15 male controls were collected and compared. Additional 100 married kidney transplant recipients without children were followed up for 3 years to observe their sexual function and fertility. Results. Serum PRL, LH, and T or E(2) levels gradually restored to the normal ranges in all kidney transplant recipients, and sperm density, motility, viability, and morphology significantly improved in the male recipients 4 months after successful kidney transplantation (P < .05). Thirty-three male recipients (55.93%) reobtained normal erectile function, and 49 kidney transplant recipients (61.25%) had children within the 3-year follow-up period. Conclusion. Successful kidney transplantation could effectively improve pituitary-gonadal hormone disturbance and sexual and reproductive dysfunctions of ESRD patients.

Impact of tension-free vaginal tape procedure on dysfunctional voiding in women with stress urinary incontinence.

OBJECTIVES: To determine the prevalence of dysfunctional voiding (DV) in female stress urinary incontinence (SUI) and its modification after tension-free vaginal tape (TVT) procedure. METHODS: Three hundred and sixty women with SUI were enrolled and underwent urodynamics from 2002 to 2008. DV was determined when non-neurogenic detrusor-sphincter dyssynergia occurred during voluntary voiding. It was further quantitatively analyzed using the tense/loose value, a parameter derived from external anal sphincter electromyogram. The distribution of other urodynamic variables was also evaluated. One hundred and fifty patients underwent the TVT procedure and forty of them were studied with urodynamics after surgery during follow up. RESULTS: Overall, DV was diagnosed in ninety-nine patients, with a prevalence of 27.5%. The functional profile length in SUI women with DV was significantly shorter than that in SUI women without DV (3.13 +/- 0.76 vs 3.32 +/- 0.65, P = 0.017). After the TVT procedure, the recovery of SUI between cases with and without DV showed no significant difference. The rate of DV state change after the surgery, namely from with to without DV or from without to with DV, significantly differed between the female patients with and without DV (66.7% vs 3.6%, P < 0.05) during follow up. The DV improved after the surgery in SUI women with DV. CONCLUSIONS: DV might represent a coexistent finding in women with SUI. The main difference of women with SUI and DV, as compared with those without DV, is a shortened functional profile length. In such cases, TVT procedure can improve DV along with the treatment of SUI.

A modified CZ-1 preserving solution for organ transplantation: comparative study with UW preserving solution.

BACKGROUND: The University of Wisconsin colloid based preserving solution (UW solution) is the most efficient preserving solution for multiorgan transplantation. Unfortunately, unavailability of delayed organ preserving solutions hindered further progression of cardinal organ transplantation in China. In this study, we validated an organ preserving Changzheng Organ Preserving Solution (CZ-1 solution) and compared it with UW solution. METHODS: A series of studies were conducted on how and how long CZ-1 solution could preserve the kidneys, livers, hearts, lungs and pancreas of New Zealand rabbits and SD rats. Morphology of transplanted organs was studied by visible microscopy and electron microscopy; biochemical and physiological functions and the survival rate of the organs during prolonged cold storage were studied. RESULTS: There was no significant difference between CZ-1 and UW solutions in preserving the kidneys, livers, hearts or lungs of rabbits; kidneys, livers, intestinal mucosa or pancreases of SD rats or five deceased donors' testicles. In some aspects, such as preserving rabbits' hearts, rats' intestinal mucosa and pancreases, the effect of CZ-1 solution was superior to UW solution. CZ-1 could safely preserve kidneys for 72 hours, livers for 24 hours, hearts for 18 hours and lungs for 8 hours for SD rats. Twelve kidneys preserved in cold CZ-1 solution for 22 - 31 hours were transplanted successfully and the mean renal function recovery time was (3.83 +/- 1.68) days. CONCLUSIONS: CZ-1 solution is as effective as UW solution for organ preservation. The development of CZ-1 solution not only reduces costs and improves preservation of organs, but also promotes future development of organ transplantation in China.

Dysfunctional voiding confirmed by transdermal perineal electromyography, and its effective treatment with baclofen in women with lower urinary tract symptoms: a randomized double-blind placebo-controlled crossover trial.

OBJECTIVES: To assess the activity of baclofen, a potent gamma-aminobutyric acid-ergic agonist, against dysfunctional voiding (DV) and lower urinary tract symptoms (LUTS) in a clinical trial, as DV is a leading cause of LUTS in women, but there is no effective treatment. PATIENTS AND METHODS: We conducted a randomized double-blind placebo-controlled crossover trial in 60 women with DV and LUTS between January 2003 and January 2006; patients were randomly assigned either baclofen 10 mg three times daily, then matching placebo for 4 weeks, or matching placebo then baclofen 10 mg three times daily for 4 weeks, separated by a 2-week washout period. Voiding diaries and multichannel urodynamics (at baseline, 4 and 10 weeks) were used to record the changes of voids/24 h and urodynamic variables. The primary efficacy endpoint was the change in voids/24 h, and the TL value (log[T/L]), a new electromyographic variables showing the extent of DV. Efficacy outcomes at 4 and 10 weeks were compared with the baseline data, using a crossover-designed analysis of variance model. RESULTS: The efficacy analysis of the treatment showed that baclofen was associated with significantly fewer voids/24 h than placebo (mean difference from baseline 5.53 vs 2.70; P = 0.001) and a significant increase in TL (mean difference from baseline -1.78 vs 0.01, P = 0.001). No significant adverse events were reported. CONCLUSIONS: A 4-week course of baclofen significantly reduced the number of voids/24 h and increased the TL value in women with DV confirmed by transdermal perineal electromyography. These encouraging results suggest that baclofen could be used for treating DV in women.

Urological malignancy as a complication of renal transplantation: a report of twelve clinical cases.

OBJECTIVE: To analyze the epidemiographic features of urological malignancy in renal allograft recipients (RAR) in a single center METHODS: A retrospective analysis was made on 3150 patients who received renal allografts from June 1978 until the autumn of 2006 and anti-rejection treatment for at least 3 months. RESULTS: Of the 3150 recipients, 33 (1.05%) developed malignancies, including 12 patients (0.38%; eight males and four females) with urological tumors and 21 patients with skin carcinoma, right liver lobular cystic adenocarcinoma, hepatocellular carcinoma, gastric cancer, colorectal carcinoma, ileocecal adenoma, lip cancer, nasopharyngeal carcinoma, Kaposi's Sarcoma, pulmonary lymphoma, and breast cancer. The 12 cases of urological malignancies included one case of renal cell carcinoma, one case of transplanted kidney carcinoma, two cases of bilateral pelvic transitional cell carcinoma (TCC), three cases of unilateral pelvic TCC, one case of bilateral ureter TCC, one case of unilateral ureter TCC, and three cases of bladder TCC. The age at which the diagnosis was made ranged from 48 to 66 years with a mean of 58.3 +/- 4.6 years, and the mean course of immunosuppressive therapy ranged from 26 to 120 months with a mean of 62 +/- 18 months. Of the 12 patients who developed urological malignancies, six had been on a Oyclosporine A + Azaithioprine + Prednisone (OsA + Aza + Pred) protocol; five on a Oyclosporine A + Mycophenolate Mofetil + Prednisone (OsA + MMF + Pred) protocol; and one on a Tacrolimus + Mycophenolate Mofetil + Prednisone (FK506 + MMF + Pred) protocol. One of the 12 patients died soon after the diagnosis was made, and the remaining 11 patients received surgical resection. Of them, 10 patients survived well, and the other one died from cerebral hemorrhage soon after operation. CONCLUSION: Urological malignancies, especially TCC, are an important complication in renal transplanatation found in this center. The incidence of urological malignancy in renal allograft recipients (RAR) is about 10 times that found in the general population of Shanghai versus two times for other malignancies. The occurrence of the malignancies in PAR seems to be closely related to the use of immunosuppressive agents. lmmunosuppression results in the weakening of immnuologic surveillance function, leading to mutation, aberration, and carcinogenesis. The immunological status of patients after renal transplantation should be assessed regularly. Painless macroscopic hematuria should be considered a significant sign in assessing the potential occurrence of urological malignancy in RAR. Treatment includes early diagnosis, timely surgical resection, and reduction of immunosuppressive agents.

[The influence of mycophenolate mofetil upon the maturation and allostimulatory activity of cultured dendritic cell progenitors and the effects of tolerance induction in allograft recipients].

OBJECTIVE: To investigate the influence of mycophenolate mofetil (MMF) on the maturation and allostimulatory activity of cultured dendritic cell progenitors (DCPs) and to evaluate the efficacy of pretreatment of donor dendritic cells (DCs) with MMF in tolerance induction in allograft recipients and its possible mechanism. METHODS: DCPs of Balb/c mice were cultured in culture fluid containing recombinant mouse granulocyte-macrophage colony stimulating factor (GM-CSF), and then divided into 4 groups: control group, MMF group, lipopolysaccharide (LPS) group, and MMF + LPS group. Seven days later, flow cytometry was used to analyze the phenotypes of the DCs. ELISA was used to examine the level of IL-12 in the supernatant. T cells from the spleens of C57/BL6 mice were cultured together with inactivated DCs from Balb/c mice, and added with [(3)H]-TdR. Mixed lymphocyte reaction (MLR) was analyzed. The DCs and MMF-DCs cultured for 5 days were co-cultured with T hybridoma cells of the line MF2.2D9 in culture fluid containing ovalbumin (OVA). Twenty-four hours later, the supernatant was collected and ELISA was used to measure the level of interleukin (IL)-12 so as to reflect the antigen-presenting ability of the DCs. OVA immunized C57/BL6 mice for 4 times. 21 days after T cells were collected from the spleens and co-incubated with DCs and MMF-DCs, [(3)H]-TdR was added and the values of counts per minute (cpm) were calculated so as to analyze the antigen-specific proliferation. Twenty-four Balb/c mice were randomly divided into 3 groups: group A (without treatment), group B (treatment with intravenous injection of untreated DCs of Balb/c mice), and group C (treatment with intravenous injection of DCs of Balb/c mice treated with MMF), and then transplanted with the hearts of C57/BL6 mice. The functions of the transplanted hearts were evaluated by touching the arterial pulse and histological examination. ELISA was used to detect the levels of Th1 cytokines, such as IL-12, IL-4, IL-10, IL-2, and IFN-gamma, in the cultured DCs and in the sera of the recipients 7 and 14 days after culture or transplantation. RESULTS: The immunophenotypic analysis showed that in comparison with those in the control group and the LPS group the expressions of the costimulatory molecules, Ia(d), CD80, and CD86, of the DCPs were significantly weaker in the MMF-group and MMP + LPS group. The IL-12 levels in the supernatant of the MMF and MMF + LPS groups of DCPs were significantly lower than those in the other groups (P < 0.01). The IL-12 level of the MF2.2D9 cells treated with MMF-treated DCs was significantly lower than control group (P < 0.01). The ability to stimulate proliferation of T cells of the same genotype in the MMF-DC group was significantly inhibited (P < 0.01). The survival time of the transplanted heart was 30.50 +/- 3.25 days in the C57/BL6 mice injected with untreated DCPs of Balb/c mice (21.25 +/- 2.12, P < 0.01) and that in the control C57/BL6 mice (8.63 +/- 1.06 days, P < 0.01) and with a significant difference between the latter 2 groups too (P < 0.01). The levels of, such as IL-2 and IFN-gamma, 7 and 14 days after heart transplantation of the group B were all significantly lower than those of the group A (both P < 0.05). The IL-2 and IFN-gamma levels 7 days after the heart transplantation were similar to those in the group B (both P > 0.05) and even lower than those of the group C (both P < 0.05). The IL-10 level in the groups B and C were all higher than those in the group A (all P < 0.05) with a significant difference between the group B and group C. The level of IL-4 was not significantly different among the 3 groups. CONCLUSION: MMF has a significant suppressive effect on the maturation and function of DCs, which leads to a donor-specific tolerance in transplant recipients.

Effective induction of immune tolerance by portal venous infusion with IL-10 gene-modified immature dendritic cells leading to prolongation of allograft survival.

Dendritic cells (DC) not only initiate T cell responses, but are also involved in the induction of tolerance. The functional properties of DC are strictly dependent on their state of maturation. It has been shown that immature DC can induce immune tolerance and prolong allograft survival. Interleukin-10 (IL-10) is an important immunosuppressive cytokine which inhibits maturation and function of DC. In order to improve the tolerogenicity of DC, we and others showed that adenovirus vectors can effectively mediate IL-10 genetic modification of DC, and IL-10 genetic modification can inhibit MHC II, B7.2, and CD40 expression, IL-12 secretion and the T cell stimulatory capacity of DC. The primary aim of this study is to examine the in vivo effects of this approach on allograft survival in a murine cardiac allograft transplantation model. To our surprise,we observed that infusion of immature DC genetically modified to express IL-10 (DC-IL-10) via the tail vein could not prolong allograft survival in the recipients, but shortened their survival. More interestingly, portal venous infusion of DC-IL-10 markedly prolonged allograft survival. The diverse effects of DC-IL-10 infusion through different routes may be due to the different immune responses to alloantigens in recipients that received DC-IL-10 via either the portal or the tail vein. Decreased cytotoxicity, polarization of Th2 response, poor T cell stimulating activity of liver DC and enhanced incidence of donor DC in the recipients may contribute to the more efficient prolongation of allograft survival observed after portal venous infusion of DC-IL-10. These results suggest that portal venous infusion may be an effective approach for immature DC to induce immune tolerance or hyporesponsiveness against donor antigens, and prolong allograft survival.

[Study the mechanisms and inducing transplantation immune tolerance of FTY720].

OBJECTIVE: To explore the operational mechanisms and potential approach to inducing transplantation immune tolerance of FTY720. METHODS: Mouse splenocytes were incubated with FTY720, then the DNA was extracted and analyzed using gel electrophoresis. Hearts of inbred BALB/c (H-2(d)) mice were transplanted heterotopically in C57BL/6 (H-2b) mice. Recipients were randomly divided into six groups. Group-1 (n = 6) was the nil-treated control. Groups-2, 3 and 4 were given FTY720 at the dose of 3 mg.kg(-1) by oral gavage once a day with different time courses. Group-2 (n = 14) were administrated from 3 days before transplantation (day-3) to the 11th day after the transplantation (day 11); Group-3 (n = 6) from day 0 to day 14; Group-4 (n = 6) from day-3 to day 0. Group-5 (n = 5) and 6 (n = 5) were treated with Cyclosporine A (10 mg.kg(-1)) and 40-O-(2-hydroxyethyl)-rapamycin (RAD) (3 mg.kg(-1)) respectively by daily gavage from day 3 to day 11. The long survivors (> 100 d) in Group-2 were detected with their IL-4 and IFN-gamma levels and their tolerant state was challenged with second graft: the donor type skin. RESULTS: Apoptosis changes of the mouse splenocytes incubated with FTY720 was showed by typical DNA ladders. The median survival time (MST) of Group-1 was 8 d. MST of Group-2 was 55 d and grafts in six mice survived more than 100 d. MST of Group-3 was 16.5 d. Group-4 has a MST of 14 d with one case exceeded 100 d. MST of Group-5 and 6 were 10 d and 13 d respectively. Long survivors of Group-2 can accept donor-type skin graft and the level of IL-4 in their serum is up-regulated while IFN-gamma remained stable. CONCLUSIONS: Pretreatment of FTY720 bring about effect on the early events of transplantation immune responses. This effect might be mediated by apoptosis induction in lymphocytes using this drug. We originally designed the regime of FTY720 monotherapy, which started pre-operationally and maintained for a short period of time, and induced stable tolerance the allo-graft in mouse.

[Analysis of DNA content of spermatogenic cells in the adult human testis and epididymis by flow cytometry].

OBJECTIVES: To detect the changes of DNA ploidy of spermatogenic cells in testis and epididymis. METHODS: Right epididymides and testes from 15 fertile youth donors who died of accident were collected. Samples of spermatogenic cells in different regions of epididymis (caput, corpus and cauda) and tests were collected. DNA of spermatogenic cells were detected by flow cyctometry (FCM). RESULTS: The haploid(1n), diploid(2n) and tetraploid(4n) spermatogenic cells were existed in different regions of epididymis and testis. The 1n cells increased from (24.87 +/- 7.28)% in testis to (96.33 +/- 1.58)% in epididymis cauda, there were significant differences among regions of testis and epididymis caput, corpus(P < 0.01), and the difference among regions of epididymis corpus and epididymis cauda were also significant(P < 0.05). While the percentages of 2n and 4n cells decreased from (63.07 +/- 8.96)% and (9.43 +/- 3.83)% in tesits to (2.47 +/- 0.93)% and (1.17 +/- 0.95)% in epididymis respectively. There was significant difference of 2n cells between testis and epididymis caput, corpus(P < 0.01), and was also remarkable difference between epididymis corpus and cauda (P < 0.05). There was no difference of 4n cells between testis and epididymis caput(P > 0.05). There were significant difference among regions of epididymis caput, corpus and cauda(P < 0.05). CONCLUSIONS: The percentage of immature spermatogenic cells decreased along with passing through the epididymis.

[Cyclosporin and portal venous inoculation of donor splenocytes prolongs cardiac allograft survival in mice].

OBJECTIVE: To investigate the effect of portal venous inoculation of donor splenocytes combined with cyclosporin A (CsA) administration on cardiac allograft survival in mice. METHODS: Heterotopic cardiac transplantation between fully allogenic NIH/q and BALB/C strain mice was performed. A modified procedure of neonatal heart-in-ear transplantation, as originally described by Fulmer et al, was adopted. We prepared donor splenocytes from NIH/q or third-party C57BL/6 spleens for BALB/C recipients, which were injected preoperatively via the recipient portal vein or the systemic vein 1 week before the heart-in-ear transplantation. The recipients were subsequently treated with a short course of the immunosuppressive agent, CsA (4 mg/kg starting from 7 d before the operation till 5 d after it). RESULTS: Portal venous inoculation of donor splenocytes combined with CsA significantly prolonged cardiac graft survival (n=6, P<0.05) that reached 31.00+/-3.23 d, and 2 of the 6 allografts survived for more than 35 d. Donor splenocytes injected via the systemic vein or third-party C57BL/6 mice splenocytes injected via the portal vein did not prolong graft survival (P>0.05). CsA alone or portal venous inoculation of donor-specific splenocytes alone also prolonged graft survival (P<0.05), with mean graft survival time of 18.50+/-2.59 d and 16.11+/-1.97 d respectively. CONCLUSION: Combination of portal venous inoculation of donor-specific splenocytes and CsA can prolong murine cardiac allograft survival, which is donor antigen-specific.

[Cadaver renal transplantation and multivariate analysis for graft survival: a clinical review of 2 016 cases].

OBJECTIVE: To review kidney transplantation in the center and analyze the risk factors affecting long-term allograft survival. METHODS: Thirty-two relative variables were analyzed with SAS statistical software. Using Log-rank method, we investigated influence of these variables on short-and long-term survival of grafts. Kaplan-Meier analysis was used to estimate the 1-, 3-, 5-, 10-years graft survival rates and half-life. Proportional hazards regression analysis (Cox model) was used to assess and rank the relative risk of potential variables. RESULTS: The 1-, 3-, 5-, 10-years graft survival rates were 83%, 75%, 66% and 48%. After excluding the patients died with functioning grafts, the 1-, 3-, 5-, 10 years grafts survival rate increased to 89%, 82%, 75% and 69%, respectively. The mean half-life was 8.78 +/- 0.14 and 14.09 +/- 0.20 years, respectively. By Log-rank analysis, factors affecting short- and long-term graft survival were identified as: renal function, duration of graft function became normal, cold-ischemia time, presence of acute rejection, delayed graft function, immunosuppressive regimen, complication, infection, anti-rejection therapy. Cox model multivariate analysis showed that there were 18 factors affecting graft survival. CONCLUSIONS: New immunosuppressive agents not only significantly increase short-term graft survival, but also have the better long-term outcome tendency. Making assurance to get high quality donor organ and minimizing the death with graft function may be the most feasible way to prolong graft survival at present.