Zhixian Wu

Umbilical Cord Mesenchymal Stromal Cell With Autologous Bone Marrow Cell Transplantation in Established Type 1 Diabetes: A Pilot Randomized Controlled Open-Label Clinical Study to Assess Safety and Impact on Insulin Secretion

OBJECTIVE To determine the safety and effects on insulin secretion of umbilical cord (UC) mesenchymal stromal cells (MSCs) plus autologous bone marrow mononuclear cell (aBM-MNC) stem cell transplantation (SCT) without immunotherapy in established type 1 diabetes (T1D). RESEARCH DESIGN AND METHODS Between January 2009 and December 2010, 42 patients with T1D were randomized (n = 21/group) to either SCT (1.1 × 106/kg UC-MSC, 106.8 × 106/kg aBM-MNC through supraselective pancreatic artery cannulation) or standard care (control). Patients were followed for 1 year at 3-month intervals. The primary end point was C-peptide area under the curve (AUCC-Pep) during an oral glucose tolerance test at 1 year. Additional end points were safety and tolerability of the procedure, metabolic control, and quality of life. RESULTS The treatment was well tolerated. At 1 year, metabolic measures improved in treated patients: AUCC-Pep increased 105.7% (6.6 ± 6.1 to 13.6 ± 8.1 pmol/mL/180 min, P = 0.00012) in 20 of 21 responders, whereas it decreased 7.7% in control subjects (8.4 ± 6.8 to 7.7 ± 4.5 pmol/mL/180 min, P = 0.013 vs. SCT); insulin area under the curve increased 49.3% (1,477.8 ± 1,012.8 to 2,205.5 ± 1,194.0 mmol/mL/180 min, P = 0.01), whereas it decreased 5.7% in control subjects (1,517.7 ± 630.2 to 1,431.7 ± 441.6 mmol/mL/180 min, P = 0.027 vs. SCT). HbA1c decreased 12.6% (8.6 ± 0.81% [70.0 ± 7.1 mmol/mol] to 7.5 ± 1.0% [58.0 ± 8.6 mmol/mol], P < 0.01) in the treated group, whereas it increased 1.2% in the control group (8.7 ± 0.9% [72.0 ± 7.5 mmol/mol] to 8.8 ± 0.9% [73 ± 7.5 mmol/mol], P < 0.01 vs. SCT). Fasting glycemia decreased 24.4% (200.0 ± 51.1 to 151.2 ± 22.1 mg/dL, P < 0.002) and 4.3% in control subjects (192.4 ± 35.3 to 184.2 ± 34.3 mg/dL, P < 0.042). Daily insulin requirements decreased 29.2% in only the treated group (0.9 ± 0.2 to 0.6 ± 0.2 IU/day/kg, P = 0.001), with no change found in control subjects (0.9 ± 0.2 to 0.9 ± 0.2 IU/day/kg, P < 0.01 vs. SCT). CONCLUSIONS Transplantation of UC-MSC and aBM-MNC was safe and associated with moderate improvement of metabolic measures in patients with established T1D.

Simultaneous Islet and Kidney Transplantation in Seven Patients With Type 1 Diabetes and End-Stage Renal Disease Using a Glucocorticoid-Free Immunosuppressive Regimen With Alemtuzumab Induction

OBJECTIVE—The aim of this study was to evaluate the efficiency and safety of simultaneous islet and kidney transplantation in patients with type 1 diabetes and end-stage renal disease using a glucocorticoid-free immunosuppressive regimen with alemtuzumab induction. RESEARCH DESIGN AND METHODS—Seven patients with type 1 diabetes and end-stage renal failure were transplanted with allogenic islets and kidneys procured from brain-dead donors. To prevent organ rejection, patients received alemtuzumab for induction immunosuppression, followed by sirolimus and tacrolimus. No glucocorticoids were given at any time. RESULTS—The median duration of follow-up was 18.3 months (range 13–31). Kidney survival was 100%. Four patients became insulin independent at 1 year. The other three reduced insulin use to less than 25% of the amount required before transplantation. Serum C-peptide levels were significantly greater posttransplant in all patients, indicating continued islet function. No major procedure-related complications were observed. CONCLUSIONS—Our results demonstrate that a steroid-free immunosuppressive regimen consisting of alemtuzumab, sirolimus, and tacrolimus is feasible for simultaneous islet and kidney transplantation. The question of whether this induction regimen is superior to more standard induction deserves large studies.

Cotransplantation of Bone Marrow Mononuclear Cells and Umbilical Cord Mesenchymal Stem Cells in Avascular Necrosis of the Femoral Head

OBJECTIVE We sought to investigate the therapeutic effects of cotransplantation of autologous bone marrow mononuclear cells (BMMNCs) and allogeneic umbilical cord mesenchymal stem cells (UC-MSCs) on avascular necrosis of the femoral head (ANFH). METHODS In all, 30 patients (49 hips; 24 males and 6 females) with ANFH were enrolled. According to the system of the Association Research Circulation Osseous, there were 24 hips in phase II and 25 hips in phase Ⅲ. Blood supply to the femoral head was evaluated by using digital subtraction angiography. Generally, 60 to 80 mL of autologous BMMNCs and 30 to 50 mL of UC-MSCs were infused into the femoral head artery. Harris scores including pain and joint function were used to evaluate the effects before and 3, 6, 9, and 12 months after transplantation. Computed tomography and radiographs were performed before and 12 months after the treatment. RESULTS Clinical symptoms of pain and claudication were gradually improved. After the treatment, 93.3% (28/30), 86.7% (26/30), and 86.7% (26/30) of patients showed relief of hip pain, improvement of joint function, and extended walking distances, respectively. The Harris scores were increased significantly at 3, 6, and 12 months posttransplant compared with those pretransplant. In addition, the bone lesions in 89.7% of hips (44/49) were improved as showed on computed tomography after transplantation. CONCLUSION Cotransplantation of autologous BMMNCs and allogeneic UC-MSCs showed therapeutic effect on ANFH without severe adverse effects.

The long-term outcomes of pediatric kidney transplantation: a single-centre experience in China.

Abstract:  To explore the long‐term outcomes of paediatric kidney transplantation and the effects of renal allograft on growth, education, employment, marriage and procreation. Twenty‐seven children with ESRD received the renal allograft from 1985 to 2001. The patient and kidney survival rate, renal function, growth and employment, etc., were reviewed retrospectively. The average follow‐up period was 10.3 ± 4.4 yr. The one‐, three‐, five‐ and 10‐yr graft survival rates were 96.3%, 88.9%, 81.5% and 66.7%, respectively, and the corresponding patient survival rates were 100%, 92.6%, 85.2% and 68.8%. The body weight gain was 4–10 kg in one‐yr post‐operative and the height increased 0–2 cm for girls and 2–5 cm for boys. A total of 44.4% of the recipients accomplished their education above junior high school. The employment rate was 46.2% in males, and 57.2% in females. Twelve patients were married. Non‐adherence occurred in 30% of the recipients. Forty percent of the surviving recipients developed complications. Seven patients died. More attention should be paid to non‐adherence of medications and more supports from the society are required to improve the life quality of paediatric recipients, especially in employment and education.

Protective effects of mesenchymal stromal cells on adriamycin-induced minimal change nephrotic syndrome in rats and possible mechanisms

BACKGROUND AIMS Minimal change nephrotic syndrome is the most frequent cause of nephrotic syndrome in childhood. Current treatment regimes, which include glucocorticoid hormones and immunosuppressive therapy, are effective and have fast response. However, because of the side effects, long treatment course, poor patient compliance and relapse, novel approaches for the disease are highly desired. METHODS The adriamycin-induced nephrotic rat model was established. Rats were allocated to a model group, a prednisone group or mesenchymal stromal cell (MSC) group. Clinical parameters in each treatment group were determined at 2 weeks, 4 weeks and 8 weeks. The messenger RNA (mRNA) levels of synaptopodin, p21 and monocyte chemoattractant protein-1 were determined through the use of quantitative real-time-polymerase chain reaction. Protein levels were determined by means of Western blot or enzyme-linked immunosorbent assay. Podocytes were isolated and apoptotic rate after adriamycin with or without MSC treatment was analyzed by means of flow cytometry. RESULTS MSC intervention improved renal function as assessed by urinary protein, blood creatinine and triglyceride levels. MSC intervention reduced adriamycin-induced renal tissue damage visualized by immunohistochemistry and light and electron microscopic analysis and reduced adriamycin-induced podocyte apoptosis. After MSC intervention, mRNA and protein levels of synaptopodin and p21 in renal cortex were significantly increased. MSCs also restored synaptopodin mRNA and protein expression in isolated podocytes. In addition, monocyte chemoattractant protein-1 mRNA in renal cortex and protein level in serum of the MSC treatment group were significantly decreased compared with that in the adriamycin-induced nephropathy model group. CONCLUSIONS Our data indicate that MSCs could protect rats from adriamycin-induced minimal change nephrotic syndrome, and the protective effects of MSCs are mediated through multiple actions.

Demonstration of the dorsal pancreatic artery by CTA to facilitate superselective arterial infusion of stem cells into the pancreas

PURPOSE To investigate the diagnostic performance of 64-section CTA in the detection of dorsal pancreatic artery before interventional therapy for patients with diabetes. MATERIALS AND METHODS The study was approved by the institutional ethics committee; written informed consent was obtained. Forty-two consecutive patients with diabetes received an experimental treatment of autologous bone marrow-derived stem cell transplantation by means of infusion into the dorsal pancreatic artery. All cases underwent abdominal CTA before angiography of pancreatic arteries in order to locate the origin and course of dorsal pancreatic artery. Angiography of coeliac artery, splenic artery, common hepatic artery and superior mesenteric artery were performed both in CTA and DSA. Superselective catheterization of dorsal pancreatic artery was carried out for the infusion of stem cell. Sensitivity, specificity and accuracy for the detection of dorsal pancreatic artery with CTA were calculated using DSA images as the reference standard. RESULTS Thirty-five and thirty-six dorsal pancreatic arteries were detected by CTA and DSA respectively. Dorsal pancreatic artery was not visualized in either CTA or DSA in 5 patients. The sensitivity, specificity and accuracy for CTA were 94.4%, 83.3% and 92.9%. CONCLUSION 64-section CTA is accurate for the detection of dorsal pancreatic artery. It may be useful for the facilitation of superselective arterial infusion of stem cells to pancreas.

Digital Subtraction Angiography and Computed Tomography Angiography of Predominant Artery Feeding Pancreatic Body and Tail

BACKGROUND Recently a considerable number of promising clinical trials have been designed to perform infusion of stem cells by pancreatic arterial intervention to improve the endocrine function of the pancreas for better diabetes control. It is necessary to investigate the pancreatic body and tail (PBT) arterial system for human islets located mostly in the PBT and identify the predominant artery or arteries. However, the arterial system in the PBT is complicated and variable. In this study we comprehensively investigated the anatomical characteristics of arteries feeding the PBT. RESEARCH DESIGN AND METHODS One hundred two patients with diabetes underwent 64-slice computed tomography angiography (CTA) and digital subtraction angiography (DSA). The target artery was catheterized, and DSA was performed to show the PBT. All images were documented for later analysis. RESULTS DSA demonstrated that the feeding arteries for the PBT included the dorsal pancreatic artery (DPA) alone (n = 51 [50%]), combined DPA and great pancreatic artery (GPA) (n = 22 [21.6%]), GPA alone (n = 16 [15.7%]), and transverse pancreatic artery (TPA) (n = 11 [10.8%]). DPA was observed to originate from the initial segment of the splenic artery (n = 34 [46.6%]), common hepatic artery (n = 17 [23.3%]), or superior mesenteric artery (n = 14 [19.2%]). The GPA was mostly from the middle (n = 36 [94.7%]), and only two were found to originate from the initial segment of the splenic artery. The TPA (n = 11) was from either the pancreatoduodenal artery (n = 5 [54.5%]) or the gastroduodenal artery (n = 4 [36.4%]). In most case, the predominant artery of the PBT (95.1%, 97 of 102) could be revealed by 64-slice CTA. CONCLUSIONS The origins and identities of the predominant artery in the PBT are variable. DSA is superior to CTA for preoperative imaging in arterial intervention therapy.

Chinese Pediatric Organ Donation With Scheduled Cardiac Arrest After Brain Death: A Novel China Classification Beyond Maastricht

OBJECTIVE Organ donation with scheduled cardiac arrest after brain death (s-DBCD) is a special category in China. This study was to evaluate the procedure of pediatric s-DBCD, graft quality, and clinical outcomes of single kidney transplantation. METHODS We retrospectively analyzed the data of 8 Chinese pediatric donors. RESULTS The death causes of the donors (age 4-12 years) were cerebral hypoxia after cardiopulmonary resuscitation (n = 1), intracranial vascular malformation (n = 1), severe traumatic brain injury (n = 3), and brain malignancy (n = 3). The functional warm ischemia time of the grafts was 18 (13-26) minutes. Sixteen kidneys were recovered using liver-kidney en bloc procurement after in situ perfusion. All kidneys had a length >7 cm and were transplanted to 3 adolescent and 13 adult recipients. Two cases of delayed graft function occurred. The patients had a median serum creatinine level of 97 (55-123) μmol/L by the last visit. The median estimated glomerular filtration rate level was 85.4 (58-136) mL/min. Five episodes of biopsy-proven acute rejection occurred in 4 patients, which were reversed by methylprednisolone pulse therapy. Renal arterial stenosis was observed in 1 patient, which was cured by interventional balloon dilatation and stent implantation. CONCLUSION Pediatric s-DBCD is feasible with acceptable graft quality. Single kidney transplantation with pediatric graft size >7 cm has good clinical outcomes.

Expression levels of Notch1 and Delta-like 4 in peripheral lymphocytes and their relationship with T helper 17 (Th17) cells in renal transplant recipients.

OBJECTIVE This study aimed to investigate the role of the Notch1/Delta-like 4 signaling pathway and its relationship with T helper 17 (Th17) cells in the peripheral transplantation immune of renal transplant recipients. METHODS Fifty-two kidney transplant recipients in our hospital were selected and divided into the acute rejection group (AR), renal tubular necrosis (ATN) group, and stable renal function group, according to their postoperative recovery. Flow cytometry was used to detect the expression of Notch1 and Delta-like 4 in peripheral lymphocytes and the presence of Th17 cells in the kidney of transplant recipients. RESULTS The expression levels of Notch1 and Delta-like 4 and level of Th17 cells among the three groups before surgery and at postoperative day 1 showed no significant differences (P>0.05). At 3, 7, and 14d after surgery, these three factors in the AR group were significantly higher than in the stable renal function group (P<0.01) and ATN group (P<0.01), where the levels in the latter two groups were similar. Upon the occurrence of acute rejection, the Notch1 and Delta-like 4 expression and Th17 cell ratio were significantly increased (P<0.01) but gradually decreased after anti-rejection therapy. Notch1 and Delta-like 4 were significantly positively correlated with Th17 cells (r=0.893, P<0.01 and r=0.893, P<0.01, respectively). CONCLUSION The detection of Notch1 and Delta-like 4 expression in peripheral blood lymphocytes of renal transplant recipients can serve as a positive indicator for evaluating the diagnosis and treatment efficacy of the AR reaction.

Insulin independence after islet transplantation through an indwelling catheter in the right gastric vein

Background Current islet transplantation approaches have various defects. This study was to investigate the feasibility and safety of an indwelling catheter in the right gastric vein for intra-hepatic islet transplantation. Methods Twenty patients had islet transplants through the indwelling catheter in the right gastric vein. The catheters were placed into the portal vein trunk using open surgery. While monitored with Doppler ultrasound, the islet suspensions were infused after the catheter location was confirmed in the trunk of the portal vein. The catheter was kept indwelling and secured to the skin for optional subsequent infusions and flushed with heparinized saline once per day to avoid peri-catheter thrombosis. After one month, the catheter was removed. Adverse effects and transplant efficacy parameters were observed. Results Insulin independence was finally achieved in 17 patients; 11 patients received a second infusion. The mean surgical duration was 55 ∓ 7 minutes and the hemorrhage volume was approximately 40 ∓ 11 mL. No significant change in portal pressure was observed (before infusion 2.9 ∓ 1.5 cm H2O, after infusion 2.6 ∓ 1.7 cm H2O, p>0.05). However, peak systolic velocity (PRV) of the hepatic artery after infusion was markedly higher than that before infusion (35.1 ∓ 10.7 cm/s vs. 68.5 ∓ 46.2 cm/s, p<0.01). Neither infection nor severe hemorrhage was found after surgery. Conclusions It is feasible, convenient, and safe to use an indwelling catheter in the right gastric vein for islet transplantation.