Heterotopic heart transplantation as a left ventricular biological assistance: a new two-stage method proposal

Abstract Since Barnard's first heterotopic heart transplant in 1974, Copeland's method has been the greatest contribution to heterotopic transplants but has the drawback of donor's right ventricular atrophy. This new method proposes a modification in the anastomosis of the superior vena cava aiming to pre-serve donor's right ventricular function by decompressing the pulmonary territory and reducing the pulmonary arterial pressure, as a biological ventricular assist device. Finally, a second intervention is proposed, where a "twist" is performed to place the donor's heart in an orthotopic position after re-moval of the native heart. A pioneering research on this method received approval from the ethics committee of the Heart Institute of São Paulo. We believe that this method has the potential to im-prove quality of life in a selected group of patients.

Review of Heterotopic Thyroid Autotransplantation

Total thyroidectomy is increasingly accepted for the management of bilateral benign thyroid disorders. Postoperatively, patients require lifelong levothyroxine replacement therapy to avoid postoperative hypothyroidism, which besides the burden of compliance, has been proven to be associated with several long-term side effects. Heterotopic thyroid autotransplantation was proposed several decades ago to avoid the need for life-long postoperative replacement therapy with maintaining the autoregulatory mechanism of thyroxin production inside the body according to its needs. Available data regarding this topic in literature is relatively poor. Before applying thyroid autotransplantation on humans, several studies have been done on animals, where the autologous transplantations were found to be successful in almost all the cases, proved by follow up postoperative 8-week measurements of thyroid hormones and histopathological examination of the removed autografts. Regarding the clinical application, few trials have been done using cryopreserved in vivo, in vitro or immediately autotransplanted thyroid autografts. Satisfactory results were obtained, however, the number of these studies and the number of patients per each study was very low. Besides the study methodologies were not so consistent.

Establishment and comparison of stoma and stoma-free heterotopic small intestine transplantation models in mice / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To establish stoma and stoma-free murine models of heterotopic small intestine transplantation in order to choose a more effective and reliable model.</p><p><b>METHODS</b>A total of 140 male 8-10 weeks age C57BL/6(B6) mice weighted 25-30 g were enrolled in the experiment. Syngeneic heterotopic small intestine transplantation was performed between C57BL/6 mice, and recipient mice were divided into either stoma or stoma-free group. Heterotopic small intestine transplantation was performed in 70 mice, with 35 mice in each group. After closing the proximal end of the graft by ligation, the distal end of graft was exteriorized as a stoma then secured to the skin of the abdominal wall in stoma group. In stoma-free group, the distal end of graft was anastomosed end-to-side to the recipient ileum. Successful rate of operation, two-week survival rate, operation time, associated complications, postoperative care time and body weight change were recorded and compared between two groups.</p><p><b>RESULTS</b>The successful rate of stoma group was 65.7%, while it was 80.0% of stoma-free group (χ(2)=1.806, P=0.179). The operation time of donor in stoma group was (48.1±6.6) minutes, while it was (47.2±5.9) minutes in stoma-free group (t=0.598, P=0.552). The operation time of recipient in stoma group was (77.9±9.1) minutes, while it was (76.4±8.3) minutes in stoma-free group (t=0.683, P=0.497). The cold ischemic time of graft in stoma group was (34.7±4.0) minutes, while it was (33.9±4.6) minutes in stoma-free group(t=0.667, P=0.507). The two-week survival rate of stoma group was 45.7%, and it was 77.1% of stoma-free group(χ(2)=7.295, P=0.007). The stoma group had more complications[54.3%(19/35) vs. 22.9%(8/35), χ(2)=7.295, P=0.007], which needed more postoperative care time(191 min vs. 35 min). The weight loss in stoma group in the third day after operation was more significant [(81.52±5.20)% vs. (85.46±4.65)%, t=2.856, P=0.006]. By 2 weeks after operation, the weight of mice in both groups retruned to 95% of the postoperative wight.</p><p><b>CONCLUSION</b>The murine heteropotic small intestine transplantation model with stoma-free appears to be more reasonable and reliable.</p>

Cell therapy in the treatment of bronchiolitis obliterans in a murine model / Terapia celular no tratamento da bronquiolite obliterante em modelo murino

OBJECTIVE: To evaluate the importance of stem cells derived from adipose tissue in reducing graft inflammation in a murine model of allogeneic heterotopic tracheal transplant. METHODS:We performed a heterotopic tracheal allografting in dorsal subcutaneous pouch and systemically injected 5x105 mesenchymal stem cells derived from adipose tissue. The animals were divided into two groups according to the time of sacrifice: T7 and T21. We also carried out histological analysis and digital morphometry. RESULTS:The T7 animals treated with cell therapy had median obstructed graft area of 0 versus 0.54 of controls (p = 0.635). The treated T21 subjects had median obstructed graft area of 0.25 versus 0 in controls (p = 0.041). CONCLUSION:The systemically injected cell therapy in experimental murine model of bronchiolitis obliterans did not reduce the severity of the allograft inflammation in a statistically significant way in seven days; Conversely, in 21 days, it increased the allograft inflammatory process.

OBJETIVO: Avaliar a importância das células-tronco derivadas de tecido adiposo na redução do processo inflamatório no enxerto em modelo murino de transplante traqueal heterotópico alogênico. MÉTODOS:Foi realizado alotransplante traqueal heterotópico em bolsa dorsal subcutânea e injetado 5x105 células-tronco mesenquimais, derivadas de tecido adiposo, sistemicamente. Os animais foram distribuídos em dois grupos, conforme o tempo de sacrifício: T7 e T21. Procedida a análise em HE e morfometria digital. RESULTADOS:Os T7 tratados com terapia celular apresentaram mediana de área obstruída do enxerto de 0 contra 0,54 dos controles (p=0,635). Os T21 tratados apresentaram mediana de área obstruída da luz do enxerto de 0,25 nos tratados e 0 nos controles (p=0,041). CONCLUSÃO: A terapia celular injetada sistemicamente em modelo experimental murino de bronquiolite obliterante não reduziu a gravidade do processo inflamatório no aloenxerto de forma estatisticamente significativa em sete dias; de modo contrário, em 21 dias, aumentou o processo inflamatório no aloenxerto.

Transforming Growth Factor -beta 1 (TGF-beta 1) immunoreactivity in heterotopic grafts of adult dental apical papilla / Inmunoreactividad del Factor Transformador del Crecimiento-beta 1 (TGF-beta 1) en la papila apical dental adulta heterotópicamente trasplantada

To analyze the expression of transforming growth factor-beta 1 inheterotopic grafts of adult dental apical papilla. Methodology: The apical papilla of adult Wistar rats was grafted in the ear of the same donor rats. 1, 3, 7 and14 days after grafting, rats were perfused and the tissue containing the graft was processed for histological conventional technique and for immunohistochemical detection of transforming growth factor-beta 1. Results: Heterotopically grafted apical papilla developed osteoid dentine. In an early post-grafting stage, odontoblast-like cells organized themselves in palisade and synthesized dentine. However, newly formed dentine possessed the structural appearance of reactive osteoid dentine, which was systematically destroyed by the activity of osteoclaste-like cells. Transforming Growth Factor-beta 1 was observed in mesenchymal cells, extracellular matrix of the graft and surrounding host tissue, while odontoblast-like cells were systematically devoid of immunoreactivity. Conclusion: The different expression of transforming growth factor-beta 1 between normal tissue and grafted tissue development suggests that in heterotopic graft conditions the inflammatory mediation of the transforming growth factor-beta 1 prevails against its morphogenetic role...

Analizar la expresión del factor transformador del crecimiento-beta1 en trasplantes heterotópicos de papila dental del incisivo de la rata adulta. Metodología: La papila apical del incisivo de 12 ratas Wistar adultas fue trasplantada en la oreja de las mismas ratas donantes, y perfundidas 1, 3, 7 y 14 días postrasplante. El tejido fue procesado para histología convencionaly para la detección inmunohistoquímica del factor transformador del crecimiento-beta1. Resultados: La papila apical trasplantada desarrolló osteodentina. En fases tempranas postrasplante se observaron células parecidas a los odontoblastos que se organizaron en empalizada y segregaron dentina que se depositó sobre su superficie apical o secretora. Esta dentina evolucionó a osteodentina caracterizada por perder su estructura tubular e incluir a las células odontoblásticas en lagunas de su matriz. Finalmente, la osteodentina presentó procesos líticos mediados por células de tipo osteoclasto. Durante todo el proceso la expresión del factor transformador del crecimiento-beta1 se restringió a las células mesenquimales, a la matriz del trasplante y a las zonas circundantes del huésped, estando ausente en los odontoblastos, a diferencia de lo que sucede durante la odontogénesis normal. Conclusión: La diferente localización de la expresión del Factor Transformador de crecimiento beta1 entre el tejido hospedero y el trasplantado sugieren que en condiciones de trasplante heterotópico de papila dental la mediación inflamatoria del Factor Transformador de crecimiento beta1 prevalece sobre su papel morfogenético...

Assessment of ovarian tissues autografted to various body sites followed by IVM in mouse

Ovarian tissue transplantation is emerging technologies for fertility preservation. In addition, in vitro maturation [IVM] of oocytes retrieved from ovarian tissues may overcome the fertility defects in certain cases. The aim was to evaluate the best site for ovarian tissue transplantation in mice. Also, feasibility of IVM of oocytes retrieved from auto grafted ovarian tissues was freshly assessed. Hemi-ovaries from 6 weeks old mice were auto grafted into kidney capsule [K] versus the back muscle [B] and leg muscle [L] in a mouse auto graft model which was stimulated with gonadotrophins. Then ovarian grafts were recovered and processed histologically for follicle assessment compared with control, also the ability of oocytes to mature with IVM was studied 14 days after transplantation. Total follicle count was significantly higher in K-graft [3.5 +/- 3.17] and the antral follicles were only observed in K-site model. The number of retrieved immature oocytes as well as successful IVM in K-grafts was significantly higher than other groups [p=0.008, p=0.016]. The kidney capsule is a promising site for ovarian tissue auto graft in mice. This resulted in better follicular survival and IVM outcomes

A gastrocnemius heterotopical transplant model with end-to-side neurorraphy

PURPOSE: To present an animal model to assess the effects of end-to-side innervation in the heterotopically transplanted model with reduced chances of neural contamination. METHODS: The medial portion of the gastrocnemius muscle in wistar male rats was isolated and its pedicle dissected and performed a flap in the abdominal portion. To prevent neural contamination in the abdominal region, the muscle was wrapped with a Goretex(r) sheet. The specimens were divided into 2 groups (G). In G1 was performed an end-to-end suture between tibial nerve of the gastrocnemius and femoral motor nerve and between the saphenous sensory nerve and the motor nerve. In G2 was performed a end-to-side suture between the tibial nerve and the motor femoral and between the tibial nerve and saphenous motor nerve. The specimens were evaluated 60 days later to check the structure of the neurorraphy. Sections were obtained proximal and distal to the coaptation site. RESULTS: The medial gastrocnemius muscle had the advantage of maintaining visible mass after 60 days. No disruption of the coaptation site was found. No major injury to the donor nerve was seen in group 2. CONCLUSION: The proposed model is simple, reproduciple and prevent the neural contamination in the flap in end-to-side suture. .

Re-epithelializaiton by epithelial inoculation with recipient phenotype in heterotopically transplanted rat allografts / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Re-epithelialization has remained a major obstacle in both tracheal and lung transplantations. This study examines the realization of re-epithelialization by epithelial inoculation in a rat heterotopic tracheal transplantation model.</p><p><b>METHODS</b>The original epithelia of tracheas from donor Wistar rats were removed and the tracheas were then inoculated with 10(6)/ml in vitro cultured epithelial cells of the Spraque-Dawley (SD) rat phenotype. These allo-tracheas were then heterotopically transplanted into SD rats. After 28 days, the allo-trachea tissues were recovered and assessed for epithelial morphology and cellular differentiation using immunohistochemical analysis. An additional experimental group was used to compare the outcomes of re-epithelialization in immunosuppressed animals.</p><p><b>RESULTS</b>Histological examination showed that allografts with epithelial inoculation maintained patent tracheal lumens, which were obliterated in controls. Recipient immunosuppression facilitated the formation of an integrated ciliated epithelial layer, further demonstrated by the presence of a dense cilia population, a well-developed plasma membrane, and readily recognizable intercellular junctions. Epithelial cellular differentiation markers such as cytokeratin 14 and 18, and cystic fibrosis transmembrane conductance regulator (CFTR) were all positive in allografts under immunosuppression.</p><p><b>CONCLUSION</b>Concurrent recipient-derived epithelial inoculation with immunosuppression can result in complete re-epithelialization with the recipient phenotype and suppress the luminal obliteration process in heterotopic transplantations.</p>

Establishment of a novel abdominal heart transplantation model of mice / 生物医学工程学杂志

This paper is aimed to establish a novel abdominal heart transplantation model in mice and to generalize the experience of the successful cases. The thoracic inferior vena cava instead of pulmonary artery was employed to reconstruct the outflow tract of the graft heart (in the new method group, 82 cases). Meanwhile, in other 47 cases as the control group, traditional anastomosis was used between pulmonary artery of the graft and vena cava of the recipient. The recipient surgery time, vena cava-vena cava anastomosis time, graft cold ischemia time and graft re-beating time were (41.5 +/- 1. 5) min, (8.4 +/- 0.6) min, (32.3 +/- 0.4) min and (1.5 +/- 0.2) min respectively. All the above data were statistically superior to those in the traditional method group (P < 0.001 or P < 0.05). The survival rate of 100 d post surgery in the new method group was 93. 9%. Meanwhile, the cardiac tissue remained almost normal examined by HE and Picro-sirus red staining. Therefore, the novel model can facilitate the anastomosis of the outflow tract in recipient operation in mouse heart transplantation model.

Teste farmacológico pré-implante na hipertensão pulmonar elevada e fixa em candidatos a transplante heterotópico / Farmacologic test pre implants in hypertension pulmonary elevated and still in candidates to heterotopic transplantation

OBJETIVOS: Avaliar a pressão da artéria pulmonar, nos momentos que precedem ao transplante, e verificar se o nitroprussiato de sódio pode possibilitar a conversão para técnica ortotópica. MÉTODOS: Entre 1992 e 2007, foram realizados 228 transplantes e esta sistemática foi empregada em sete pacientes que apresentavam na avaliação hemodinâmica pré-operatória: Pré NP (mmHg) Pós NP (mmHg) Pressão Sistólica Arterial Sistêmica (PSAS) 108 - 78 (101,7 ±10,9) 90 - 74 (79,5 ± 15,2) Pressão Sistólica Arterial Pulmonar (PSAP) 88 - 51 (69,8 ± 13,2) 70 - 40 (57,8 ± 9,9) Gradiente Transpulmonar (GTP) 16 11 (14,2 ± 1,7) 14 - 11 (12,4 ± 1,2) Resistência Vascular Pulmonar (RVP/w) 7,9 - 4,8 (6,2 ± 1,0) 5,9 - 4,1 (5,0 ± 0,8). RESULTADOS: Os achados intra-operatórios foram: Pré NP (mmHg) e Pós NP (mmHg), repectivamente, PSAS 91-78 (8,5 ± 5,2) e 65-59 (63,8 ± 4,2) (P = 0,017), queda 19,9 por cento, Queda 29,3 por cento; PSAP 71-52 (61,8 ± 6,1) e 43-32 (37,5 ± 3,3) (P = 0,018), queda 28 por cento, Queda 41 por cento Diante destes dados, os pacientes foram transplantados pela técnica ortotópica, não sendo constatada mortalidade a curto e a longo prazo em evolução de 5 meses a 6 anos. CONCLUSÃO: A aplicação desta metodologia permitiu a conversão da técnica heterotópica para ortotópica, com bons resultados imediatos e tardios.

BACKGROUND: Evaluation of pulmonary artery pressure just before transplanting with sodium nitroprusside may allow conversion to orthotopic technique. METHODS: Between 1992 and 2007, 228 transplants were performed systematically and this was used in seven patients with preoperative hemodynamic evaluation: Pre NP (mmHg) Post NP (mmHg) Systolic systemic blood pressure (PSAS) 108-78 (101.7 ± 10.9) 90-74 (79.5 ± 15.2) pulmonary arterial systolic pressure (PASP) 88-51 (69.8 ± 13.2) 70-40 (57.8 ± 9.9) Gradient transpulmonary (GTP) 16-11 (14.2 ± 1.7) 14-11 (12.4 ± 1.2) pulmonary vascular resistance (PVR/w) 7.9 to 4.8 (6.2 ± 1 0) 5.9-4.1 (5.0 ± 0.8). RESULTS: The intraoperative findings were: Pre NP (mmHg) e Post NP (mmHg), respectively, PSAS 91-78 (8.5 ± 5.2) and 65-59 (4.2 ± 63.8) (P = 0.017), decrease 19.9 percent, decrease 29.3 percent; PSAP 71-52 (61.8 ± 6.1) and 43-32 (37.5 ± 3.3) (P = 0.018), decrease 28 percent, decrease 41 percent. In light of these data, patients were transplanted by orthotopic technique not being observed mortality in the short and long-term evolution from 5 months to 6 years. CONCLUSION: This methodology allowed the conversion of the technique for heterotopic orthotopically, with good early and late otcomes.

Experiência com transplante cardíaco heterotópico em pacientes com resistência pulmonar elevada: seguimento tardio / Experiencia con trasplante cardíaco heterotópico en pacientes con resistencia pulmonar elevada: seguimiento tardío / Experience with heterotopic heart transplantation in patients with elevated pulmonary vascular resistance: late follow-up

FUNDAMENTO: Nos últimos anos o numero de artigos sobre transplante cardíaco heterotópico tem sido escasso na literatura, inclusive internacional, e em particular do seguimento de longo prazo destes pacientes, o que levou ao presente relato. OBJETIVO: Relatar a experiência clínica inicial e evolução tardia de quatro pacientes submetidos a transplante cardíaco heterotópico, sua indicação e principais complicações. MÉTODOS: As cirurgias ocorreram entre 1992 e 2001, sendo que a indicação de transplante heterotópico, em todas, foi pela RVP, variável de 4,8UW a 6.5UW, com gradiente transpulmonar acima de 15mmHg. No 3º paciente, foi realizada uma anastomose direta entre as artérias pulmonares sem emprego de tubo protético e, no coração nativo, foi realizada uma valvoplastia mitral e aneurismectomia de ventrículo esquerdo (VE). O esquema imunossupressor imediato foi duplo com ciclosporina e azatioprina nos três primeiros pacientes e ciclosporina e micofenolato mofetil no 4º paciente. RESULTADOS: Um óbito imediato por falência do enxerto, um óbito após dois anos e meio por endocardite em trombo intraventricular no coração nativo, e um terceiro óbito seis anos após o transplante, por complicações pós-operatórias de cirurgia na valva aórtica do coração nativo. O remanescente, 15 anos após o transplante, encontra-se bem, em classe funcional II (NYHA), seis anos após a oclusão cirúrgica da valva aórtica do coração nativo. CONCLUSÃO: O transplante cardíaco heterotópico é um procedimento com resultado inferior ao transplante cardíaco ortotópico, por apresentarem maior RVP. Os trombos intraventriculares no coração nativo, que exigem anticoagulação prolongada, bem como as complicações de válvula aórtica, também no coração nativo, podem exigir tratamento cirúrgico. Entretanto, em um paciente, a sobrevida de 15 anos mostrou a eficácia de longo prazo desse tipo de alternativa, para pacientes selecionados.

BACKGROUND: Along the past few years the number of papers on heterotopic cardiac transplant has been very scarce in the medical literature, including at the international level; this is particularly true in reference to the long term follow-up of these patients and the reason which led to the presentation of our report. OBJECTIVE: To report the initial clinical experience and late evolution of 4 patients undergoing heterotopic heart transplantation, indications for this procedure and its major complications. METHODS: The surgeries were performed between 1992 and 2001, and all had as indication for heterotopic transplantation the PVR, which ranged from 4.8 WU to 6.5WU, with a transpulmonary gradient above 15mmHg. In the 3rd patient, a direct anastomosis between the pulmonary arteries was performed without the use of a prostetic tube, and a mitral valvuloplasty and a LV aneurysmectomy were performed in the native heart. The immediate immunosuppressive regimens were double, with cyclosporine and azathioprine in the first 3 patients, and cyclosporine and mycophenolate mofetil in the 4th patient. RESULTS: One immediate death occurred from graft failure, one death occurred after 2 ½ years, from endocarditis in an intraventricular thrombus in the native heart, and a third death occurred 6 years after transplantation, from post-operative complications of the aortic valve surgery in the native heart. The remaining patient is well, 15 years after the transplantation. This patient is in functional class II (NYHA), 6 years after a surgical occlusion of the native heart aortic valve. CONCLUSION: Heterotopic heart transplantation results are inferior to those of orthotopic heart transplantation because they present higher RVP. The intraventricular thrombi, in the native heart, which require prolonged anticoagulation, and aortic valve complications, also in the native heart, may require surgical treatment. However, a patient's 15-year survival has demonstrated ...

FUNDAMENTO: En los últimos años el número de artículos sobre trasplante cardíaco heterotópico y, en particular, del seguimiento a largo plazo de estos pacientes, ha sido escaso en la literatura, inclusive internacional, lo que llevó al presente relato. OBJETIVO: Relatar la experiencia clínica inicial y la evolución tardía de cuatro pacientes sometidos a trasplante cardíaco heterotópico, su indicación y principales complicaciones. MÉTODOS: Las cirugías se realizaron entre 1992 y 2001, y la indicación de trasplante heterotópico, en todas, fue mediante RVP, variable de 4,8 UW; a 6.5 UW, con gradiente transpulmonar superior a 15 mmHg. En el tercer paciente, se realizó una anastomosis directa entre las arterias pulmonares sin empleo de tubo prostético, y, en el corazón nativo, se realizó una valvuloplastia mitral y aneurismectomía de VI. El esquema inmunosupresor inmediato fue doble, con ciclosporina y azatioprina en los tres primeros pacientes y ciclosporina y micofenolato mofetil en el cuarto paciente. RESULTADOS: Un óbito inmediato por falla del injerto, un óbito luego de dos años y medio por endocarditis en trombo intraventricular en el corazón nativo, y un tercer óbito seis años después del trasplante, por complicaciones postoperatorias de una cirugía en la válvula aórtica del corazón nativo. El restante, 15 años después del trasplante, se encuentra bien, en clase funcional II (NYHA), seis años después de una oclusión quirúrgica de la válvula aórtica del corazón nativo. CONCLUSIÓN: El trasplante cardíaco heterotópico es un procedimiento con resultado inferior al trasplante cardíaco ortotópico, por presentar mayor RVP. Los trombos intraventriculares en el corazón nativo, que exige anticoagulación prolongada, así como las complicaciones de válvula aórtica, también en el corazón nativo, pueden exigir tratamiento quirúrgico. Sin embargo, en un paciente, la sobrevida de 15 años mostró la eficacia a largo plazo de este tipo de alternativa, ...

FTY720-induced lymphocyte apoptosis inhibits acute graft versus host disease in rat small bowel transplantation / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the effect and mechanism of FTY720 on acute graft versus host disease (GVHD) in rat small bowel transplantation (SBTx).</p><p><b>METHODS</b>Heterotopic SBTx was performed using a parent (WF)-into-F1 (WFxACI) rat combination. Recipient rats were divided into experimental group (n=6) and control group (n=6). Rats in the experimental group were administered with FTY720 at 0.5 mg/kg for 14 days. Lymphocyte apoptosis in the liver and the mucosa of intestine and graft was detected by TUNEL and flow cytometry 15 days after transplantation. Recipient survival and lymphocyte apoptosis were compared between the two groups.</p><p><b>RESULTS</b>Recipients in the control group died of GVHD after a mean survival time of (16+/-2.1) days. FTY720-treated recipients had a significantly longer survival (>100 days). After administration of FTY720, the percentage of apoptotic lymphocytes was significantly increased in the graft as compared to that in the control group by flow cytometry. The ratio of apoptotic lymphocyte in the liver and graft was also significantly higher in the experimental group by TUNEL.</p><p><b>CONCLUSION</b>FTY720 effectively induces the lymphocyte apoptosis, inhibits the lesion of target tissues by GVHD, and prolongs the recipient survival.</p>

Comparison of Four Pancreatic Islet Implantation Sites

Although the liver is the most common site for pancreatic islet transplantation, it is not optimal. We compared kidney, liver, muscle, and omentum as transplantation sites with regard to operative feasibility, and the efficiency of implantation and glycemic control. Islets from C57BL/6 mice were transplanted into diabetic syngeneic recipients. The mean operative time and mortality were measured to assess feasibility. To assess implantation efficiency, the marginal mass required to cure diabetes and the mean time taken to achieve normoglycemia were measured. A glucose tolerance test was performed to assess glycemic control efficiency. The data are listed in the order of the kidney, liver, muscle, and omentum, respectively. The mean mortality rate was 6.7, 20.0, 7.1, and 12.5%; the mean operative time was 10.2, 27.4, 11.2, and 19.8 min; the marginal islet mass was 100, 600, 600, and 200 islet equivalence units and the mean time to reach euglycemia was 3.0, 15.1, 26.6, and 13.9 days. The glucose kinetics of omental pouch islets was the most similar to controls. Thus, a strategic approach is required for deciding on the best transplantation recipient sites after considering donor sources and islet volume. Alternatives can be chosen based on safety or efficacy.

Extracellular matrix accumulation and expression of gelatinases and their tissue inhibitors in a mechanically unloaded heart model / 中国应用生理学杂志

<p><b>AIM</b>To investigate the relationship between the expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 and ECM accumulation in rat left ventricle in a mechanical unloaded heart model.</p><p><b>METHODS</b>12-week-old male Lewis rats were subjected to abdominal heterotopic heart transplantation to achieve pressure and volume unloading(mechanical unloading). Age and sex matched in situ heart of Lewis rats were used as control. Collagen volume fraction(CVF) was analyzed by picrosiris-red staining plus polarized microscopy. MMP-2 and -9 gelatinolytic activity were measured by gelatin-zymography. mRNA level of MMP-2, MMP-9, TIMP-1 and TIMP-2 were measured by real-time quantitative PCR. TIMP-1 and TIMP-2 protein level were measured by immunoblotting.</p><p><b>RESULTS</b>Myocardial cross-sectional area of transplanted heart was significantly reduced, and accompanied by excessive ECM deposition (CVF 5.22% +/- 1.6% vs. 2.21% +/- 0.9%, P < 0.05) compared to in situ heart. MMP-2 and MMP-9 activity were significantly increased, as well as mRNA level of MMP-2, MMP-9, TIMP-1 and TIMP-2 compared to in situ heart. TIMP-1 and TIMP-2 protein level in mechanically unloaded heart were significantly upregulated compared to in situ heart, especially for TIMP-1.</p><p><b>CONCLUSION</b>Mechanical unloading of left ventricle may lead to excessive ECM deposition, accompanied by imbalance between MMPs and TIMPs system, especially the upregulation of TIMPs.</p>

Trasplante heterotópico de corazón en ratas: desarrollo de un modelo de microcirugía / Heterotopic heart transplantation in rats

Introducción. El trasplante de corazón heterotópico en ratas es usado en investigación experimental, particularmente en el estudio de la inmunosupresión. El objetivo del trabajo fue desarrollar un modelo de trasplante heterotópico de corazón en ratas. Material y Método. Se utilizaron ratas macho Lewis (Receptor) y Brown Norway (Donante) de 200 a 350 gr. Para el procuramiento, se realizó una incisión en mariposa del tórax y se instaló un clip metálico a la vena cava superior e inferior para infundir solución fisiológica con heparina, logrando detener los latidos cardíacos, y se seccionó la arteria pulmonar y la aorta. Las venas cavas y las pulmonares son ligadas en conjunto y seccionadas. En la rata receptora se identificó la aorta y vena cava abdominal y se instaló un clamp vascular atraumático tipo bulldog. Se realizó una anastomosis término lateral entre la aorta ascendente del donante y la aorta abdominal del receptor, y entre la arteria pulmonar del donante y la cava del receptor, con microsutura 10-0 continua. Se consideró un trasplante exitoso cuando el injerto estaba funcional por más de 24 horas, por palpación de latidos en el abdomen. Resultados. Se trasplantaron 80 ratas en total. La principal causa de pérdida del injerto fue el prolongado tiempo operatorio y la hemorragia postoperatoria. Se realizaron modificaciones en la técnica: Ligadura única de las venas cavas y de las venas pulmonares, venotomía elíptica y lateral, ligadura de un vaso paralelo a la aorta que evita una hemorragia letal y reposición de volumen postoperatorio. Discusión. Es un modelo reproducible. Modificaciones de la técnica permiten disminuir el tiempo de isquemia, permitiendo un aumento en la sobrevida del injerto.

Introduction: Heterotopic heart transplantation in rats is an experimental research model, specially used to study immunosuppression. Aim: To develop a model of heterotopic heart transplantation in rats. Material and methods: Lewis rats (as receptors) and Brown Norway rats (as donors), weighing 200 to 350 g, were used. For procurement, a butterfly chest incisión was done, a metallic clip was placed in inferior and superior vena cava to infuse a physiologic solution with heparin, stopping cardiac beats and sectioning pulmonary and aortic arteries. Pulmonary and cava veins were ligated jointly and sectioned. In the receptor rats, aorta and abdominal vena cava were identified and an atraumatic bulldog vascular clamp was placed. Termino lateral anastomoses between donor ascending aorta and receptor abdominal aorta, and between donor pulmonary artery and receptor vena cava were performed with continuous microsuture. Transplantation was considered successful when the graft was functional for more than 24 hours, determined palpating beats in the abdomen. Results: Eighty rats were transplanted. We main causes of graft loss were a prolonged operative time and postoperative hemorrhage. The technique was modified, using a unique ligation of cavas and pulmonary veins, elliptic and lateral venotomy, ligation of a vessel that is parallel to aorta that avoids lethal hemorrhages and postoperative fluid replacement. Discussion: This is a reproducible model. The technical modifications introduced, reduce the lapse of ischemia and increase graft survival.

Effect of cariporide on the expression of bcl-2 and bax genes after neck heart transplantation from non heart-beating rats caused by warm ischemia / 中南大学学报(医学版)

OBJECTIVE@#To detect the expression of bcl-2 and bax genes after heterotopic heart transplantation in rats that died of warm ischemia, and to explore the effect of cariporide on the protection of the ratos non heart-beating donors.@*METHODS@#One hundred and twelve clearing Sprague-Dawley male rats were divided into 7 groups at random (each group contained 16 rats): the control group (Group C), the groups of transplanted hearts after 10, 30, and 45 min of asystolia (Group S10,S30,and S45), and the groups of transplanted hearts after 10,30, and 45 min of asystolia and infused with cariporide(Group SH10,SH30, and SH45).The experimental groups were sacrificed totally by warm ischemia, and heterotopic heart transplantation was processed by the Cuff method. The heart samples of S10,SH10,S30, and SH30 groups were taken at 48 hours after the transplantation, and the heart samples of S45, and SH45 groups were taken just after transplantation. The expression of bcl-2 and bax genes were detected by RT-PCR.@*RESULTS@#The death of rats was affirmed when cardiac electric waves vanished after 9~11 minutes of transsection of abdominal aorta. On the RT-PCR test, the expression of bcl-2 gene was the highest and ROD value was maximum in the control group. The expression of bax gene was the lowest and ROD value was minimum in the control group. The ROD value of bcl-2 genes in S10 and S30 groups was less than that in SH10 and SH30 group. The ROD value was just the opposite, and there was stastistical difference (P0.05).@*CONCLUSION@#The model of heteroto-pic neck heart transplantation is a convenient animal model for the cardiac muscle protection. Cariporide can suppress the apoptosis of cardiac muscle cells in rats (within 30 min) after death caused by warm ischemia.

Reestablecimiento de la función endocrina de implantes ováricos heterotópicos no pediculizados / Reestablishment of endocrine function in heterotopic ovarian transplants

Introducción: el presente trabajo muestra los resultados de implantes ováricos heterotópicos no pediculizados como una alternativa de preservación de la función ovárica para pacientes que serán sometidas a radioterapia abdómino-pélvica. Se muestra la evolución de tres pacientes, dos portadoras de una cáncer de cuello uterino y una con un neuroependimoma sacro, en las cuales, previendo la gonadotoxicidad de la radioterapia pélvica, se les realizó ooforectomía bilateral y autoimplantes subcutáneos heterotópicos de tejido ovárico. En dos de ellas, además, se criopreservó tejido ovárico para el futuro. Material y método: se aplicó el protocolo del "Programa de preservación de la función ovárica" que lleva adelante la Facultad de Medicina de la Universidad de la República Oriental del Uruguay (UDELAR) y el Ministerio de Salud Pública de Uruguay. Resultados: en las tres pacientes se evidenció la revascularización del tejido trasplantado, la recuperación de la función hormonal y el crecimiento de folículos ováricos. Estas tres observaciones se suman a las muy escasas publicadas a nivel mundial. Conclusiones: se evidenció la recuperación de la función endocrina ovárica, una mejoría de la calidad de vida (retroceso del síndrome climatérico) y el desarrollo folicular en el tejido trasplantado. Así pues el desarrollo de esta técnica, dentro de un protocolo, puede ofrecer una alternativa para un grupo seleccionado de pacientes en las que se pretenda mantener la función ovárica luego de la radioterapia y evitar las consecuencias de fallo ovárico prematuro.

Improved Surgical Technique for Heterotopic Aortic Transplantation in Mice

Transplant arteriosclerosis is the main limitation for long-term survival of solid organ transplant recipients. Animal models would provide invaluable tools to investigate the cellular and molecular mechanisms underlying the pathogenesis of transplant arteriosclerosis, as well as for studies with novel drugs and other reagents for the prevention of the disease. We have therefore developed a modified technique for aortic transplantation in mice. The central suture ligation of the recipient abdominal aorta allowed a simpler end-to-side anastomosis of a segment of the donor thoracic aorta into the infrarenal portion of the recipient abdominal aorta. Using this technique, the overall survival rate was 94%. We also observed typical aspects of chronic rejection of the aortic allografts not observed with isografts. Our new technique is relatively easy to perform and has a low incidence of thrombosis, thus being useful for studying various aspects of transplant arteriosclerosis.

Research progress in ectopic grafting of testicular tissues / 中华男科学杂志

Since Nature published the first report in 2002 on using immunodeficient mice as recipients and allogeneous or heterogeneous testes as donor tissues to study the ectopic development of spermatogenic cells, the technique has been widely applied in various species (including human). In comparison with other in vitro maturation methods for male germ cells, testicular allografting or xenografting technique has such advantages as similar environment for the development of germ cells in physiological conditions, and better reproducibility. Up to now, sperm has been successfully produced by this technique from the testicular tisues of the immature mouse, hamster, cat, rabbit, pig, goat, bovine and rhesus monkey, and their offspring have even been generated by ICSI technique using the mouse and rabbit sperm derived from testis grafts. This article comprehensively reviews the development of the technique by discussing the influencing factors on the germ cell development in grafts including the variety and age of donors, the sex, integrity and immunity of recipients, the graft location and grafting time. And the applications of the technique and the existing problems are discussed as well.