Fresh Transplantation Research Articles

The effects of fresh embryo transfer and frozen-thawed embryo transfer on the perinatal outcomes of single fetuses from mothers with PCOS.

To investigate the effects of fresh embryo transfer and frozen-thawed embryo transfer on perinatal outcomes among patients with PCOS. Patients who underwent in vitro fertilization and embryo transfer at the reproductive medicine center of the Affiliated Hospital of Guangdong Medical University from February 2013 to March 2021 were retrospectively analyzed. Patients were divided into the fresh embryo transfer group and frozen-thawed embryo transfer group according to whether fresh embryo transfer was performed. According to their conditions, patients were further classified into the ET-PCOS group (group A, n = 104), ET-non-PCOS group (group B, n = 212), FET-PCOS group (group C, n = 102), or FET-non-PCOS group (group D, n = 148); the general data, laboratory indicators and pregnancy outcomes of the patients were statistically analyzed, and the perinatal outcomes and related factors between the groups were compared and analyzed. The level of E2 on the HCG test day in the ET group was lower than that in the FET group. The natural birth rate of group D was lower than that of group A and group B, and the cesarean section rate was higher than that of group A and group B; the clinical pregnancy rate of group A was higher than that of group B and group D, and the difference was statistically significant (P < 0.05). There was no significant difference in the total abortion rate, early abortion rate or late abortion rate between the groups (P > 0.05). There was no significant difference in gestational age, neonatal sex or neonatal weight between the groups (P > 0.05). The incidence of placenta previa in Group B was significantly lower than that in Group D, and the difference was significant (P < 0.05). The incidence of fetal distress in Group B was significantly lower than that in Groups C and D, and the incidence of neonatal jaundice in Group D was significantly higher than that in Groups A and B (P < 0.05). In the multivariate analysis, the number of high-quality embryos was independent factors affecting clinical pregnancy, and the embryo transfer method was an independent factor affecting fetal distress and neonatal jaundice. Young PCOS patients without risk of OHSS have a high clinical pregnancy rate with fresh transplant cycles. PCOS disease itself has no significant effect on the perinatal outcomes of the mother or singleton infant. Frozen-thawed embryo transfer may increase the incidence of low placenta, fetal distress and neonatal jaundice.

First successful ovarian cortex allotransplant to a Turner syndrome patient requiring immunosuppression: wide implications

ObjectiveTo determine whether we can safely and successfully transplant an ovary tissue allograft from a non-identical donor to her Turner syndrome sister. DesignTransplantation of cryopreserved ovary tissue, as well as fresh transplantation of ovarian tissue between identical twins, is now well established with numerous reported successful cases. However there have not yet been any ovary transplants between non-identical women requiring immunosuppression (ovary allotransplant). This could be a much more common indication for ovary tissue transplantation if safe and reliable immunosuppression were available. SubjectsA 20 year old amenorrheic woman with non-mosaic 45-XO Turner syndrome requested ovary tissue transplantation from her fertile 22 year old 46-XX sister. They were an HLA match but were ABO incompatible, a well known contra-indication to solid tissue or organ transplantation. The Turner syndrome sister strongly preferred to be able to become pregnant naturally without donor egg IVF, and to avoid HRT (hormone replacement therapy). In her religious group, that would also be important for finding a marital match. Despite the poor prognosis associated with ABO incompatibility, an ovary from her 22 year old non-identical fertile sister was transplanted to her employing the immunosuppression protocol now used for kidney transplant patients in our centers at Washington University and Johns Hopkins. Main Outcome MeasuresPost-operatively at 5 months she developed normal monthly menstrual ovarian function, and she became spontaneously pregnant with a normal baby girl. The relation between her post-op FSH and AMH levels continues to support the theory that tissue pressure controls primordial follicle recruitment. The fact that ABO incompatibility did not prevent success suggests that diffusion and not revascularization may be all that is required for successful long term ovarian cortex transplant survival with spontaneous pregnancy. ResultsOvary allotransplantation with safe immunosuppression allows natural conception, and also normal hormone function obviates the need for HRT. Orthotopic placement of the graft and surgical technique is critical for natural conception and a higher pregnancy rate. ConclusionAllotransplantation requiring safe immunosuppression, if successful, may be a much more commonly used indication for ovary transplantation in the future than frozen ovary grafts or grafts between identical twins.

Establishment of a predictive nomogram for clinical pregnancy rate in patients with endometriosis undergoing fresh embryo transfer

To establish a nomogram model for predicting clinical pregnancy rate in patients with endometriosis undergoing fresh embryo transfer. We retrospectively collected the data of 464 endometriosis patients undergoing fresh embryo transfer, who were randomly divided into a training dataset (60%) and a testing dataset (40%). Using univariate analysis, multiple logistic regression analysis, and LASSO regression analysis, we identified the factors associated with the fresh transplantation pregnancy rate in these patients and developed a nomogram model for predicting the clinical pregnancy rate following fresh embryo transfer. We employed an integrated learning approach that combined GBM, XGBOOST, and MLP algorithms for optimization of the model performance through parameter adjustments. The clinical pregnancy rate following fresh embryo transfer was significantly influenced by female age, Gn initiation dose, number of assisted reproduction cycles, and number of embryos transferred. The variables included in the LASSO model selection included female age, FSH levels, duration and initial dose of Gn usage, number of assisted reproduction cycles, retrieved oocytes, embryos transferred, endometrial thickness on HCG day, and progesterone level on HCG day. The nomogram demonstrated an accuracy of 0.642 (95% CI: 0.605-0.679) in the training dataset and 0.652 (95% CI: 0.600-0.704) in the validation dataset. The predictive ability of the model was further improved using ensemble learning methods and achieved predicative accuracies of 0.725 (95% CI: 0.680-0.770) in the training dataset and 0.718 (95% CI: 0.675-0.761) in the validation dataset. The established prediction model in this study can help in prediction of clinical pregnancy rates following fresh embryo transfer in patients with endometriosis.

What dead seaweeds can tell us about metal uptake and their application to control marine pollution

The mechanisms of trace element uptake by seaweeds are still unknown, despite being key to understand the impact of pollution in coastal environments. This knowledge gap, in addition to the lack of standardization, have also hindered the use of seaweeds to monitor seawater pollution. To address these shortcomings, we tested the use of devitalization as a pre-exposure treatment for brown seaweed transplants, and we compared devitalized and fresh transplants to gain some insights into the mechanisms of element uptake. We exposed four types of Fucus vesiculosus transplants in 6 sites for 4, 8 and 20 days: fresh and devitalized (dried or boiled) algal segments held in mesh bags, and whole algal thalli imitating natural conditions. We then determined he concentrations of 11 trace elements in the algal tissues. The element concentrations were highest in the devitalized transplants, but the material lost consistency and weight throughout the exposure period, limiting their use to short periods. We proposed several factors that may contribute to the different accumulation patterns between treatments, and examined the implications for the uptake mechanisms, revealing that two of the most important are surface adsorption of sediment particles and chemical bounds to extracellular components.

A French single-center experience on allogeneic stem cell transplant cryopreservation during severe acute respiratory syndrome coronavirus 2 pandemic

Background aimsAllogeneic hematopoietic stem cell transplantation (allo-SCT) is a curative treatment for chemo-resistant hematological malignancies. Because of transport restriction imposed by the coronavirus disease 2019 pandemic, regulatory bodies and societies recommended graft cryopreservation before recipient conditioning. However, the freezing and thawing processes, including washing steps, might impair CD34+ cell recovery and viability, thereby impacting the recipient engraftment. Over 1 year (between March 2020 and May 2021), we aimed to analyze the results of frozen/thawed peripheral blood stem cell allografts in terms of stem cell quality and clinical outcomes. MethodsTransplant quality was evaluated by comparing total nucleated cells (TNCs), CD34+ cells and colony-forming unit–granulocyte/macrophage (CFU-GM)/kg numbers as well as TNC and CD34+ cell viabilities before and after thawing. Intrinsic biological parameters such as granulocyte, platelet and CD34+ cell concentrations were analyzed, as they might be responsible for a quality loss. The impact of the CD34+ cell richness of the graft on TNC and CD34 yields was evaluated by designing three groups of transplants based on their CD34 /kg value at collection: >8 × 10 6/kg, between 6 and 8 × 106/kg and <6 × 106/kg. The consequences of cryopreservation were compared in the fresh and thawed group by evaluating the main transplant outcomes. ResultsOver 1 year, 76 recipients were included in the study; 57 patients received a thawed and 19 patients a fresh allo-SCT. None received allo-SCT from a severe acute respiratory syndrome coronavirus 2–positive donor. The freezing of 57 transplants led to the storage of 309 bags, for a mean storage time (between freezing and thawing) of 14 days. For the fresh transplant group, only 41 bags were stored for potential future donor lymphocyte infusions. Regarding the graft characteristics at collection, median number of cryopreserved TNC and CD34+ cells/kg were greater than those for fresh infusions. After thawing, median yields were 74.0%, 69.0% and 48.0% for TNC, CD34+ cells and CFU-GM, respectively. The median TNC dose/kg obtained after thawing was 5.8 × 108, with a median viability of 76%. The median CD34+ cells/kg was 5 × 106, with a median viability of 87%. In the fresh transplant group, the median TNC/kg was 5.9 × 108/kg, and the median CD34+ cells/kg and CFU-GM/kg were 6 × 106/kg and 276.5 × 104/kg, respectively. Sixty-one percent of the thawed transplants were out of specifications regarding the CD34+ cells/ kg requested cell dose (6 × 106/kg) and 85% of them would have had this dose if their hematopoietic stem cell transplant had been infused fresh. Regarding fresh grafts, 15.8% contained less than 6 × 106 CD34+ cells /kg and came from peripheral blood stem cells that did not reach 6 × 106 CD34+ cells /kg at collection. Regarding the factor that impaired CD34 and TNC yield after thawing, no significant impact of the granulocyte count, the platelet count or the CD34+ cells concentration/µL was observed. However, grafts containing more than 8 × 10 6/kg at collection showed a significantly lower TNC and CD34 yield. ConclusionsTransplant outcomes (engraftment, graft-versus-host disease, infections, relapse or death) were not significantly different between the two groups.

Development of a Cryopreservation Technique for Xenogeneic Kidney Grafts: Evaluation Using a Mouse Model.

To align the xeno-metanephros and renal progenitor cell timing for transplantation treatments, cryopreservation techniques and an efficient transportation of regenerated renal products such as xeno-metanephroi and renal progenitor cells should be established. Therefore, we propose a novel method of xenogeneic regenerative medicine for patients with chronic kidney disease by grafting porcine fetal kidneys injected with human renal progenitor cells. To develop a useful cryopreserve system of porcine fetal kidney and human renal progenitor cells, we examined the cryopreservation of a fetal kidney implanted with renal progenitor cells in a mouse model. First, we developed a new method for direct cell injection under the capsule of the metanephros using gelatin as a support for unzipped fetal kidneys. Then, we confirmed in vitro that the nephrons derived from the transplanted cells were regenerated even after cryopreservation before and after cell transplantation. Furthermore, the cryopreserved chimeric metanephroi grew, and regenerated nephrons were observed in NOD. We confirmed that even in cryopreserved chimeric metanephroi, transplanted cell-derived nephrons as well as fresh transplants grew.

Influence of the sperm DNA fragmentation index on the outcome of rescue ICSI and the clinical value of rescue ICSI.

As a remedy for the failure of in vitro fertilization (IVF), rescue intracytoplasmic sperm injection (R-ICSI) has been widely carried out, but it has failed to significantly improve the fertilization rate and clinical pregnancy rate. Sperm DNA fragmentation index (DFI) was highly correlated with pregnancy outcome of artificial assisted reproduction. This study aims to investigate the effect of the sperm DFI on the outcome of R-ICSI and the clinical value of R-ICSI. This retrospective analysis was conducted among 140 infertile couples receiving R-ICSI in from January 2014 to December 2019. The subjects were assigned into a total fertilization failure (TFF)+low DFI group (R-ICSI after TFF and DFI<30%) (n=63), a TFF+high DFI group (R-ICSI after TFF and DFI≥30%) (n=16), a partial fertilization failure (PFF)+low DFI group (R-ICSI after PFF and DFI<30%) (n=52), a PFF+high DFI group (R-ICSI after PFF and DFI≥30%) (n=9). All transferred embryos were come from R-ICSI. The general clinical data [infertility duration, male age, female age, basal serum level of follicle stimulating hormone (FSH), basal serum level of luteinizing hormone (LH), antral follicle count, endometrial thickness of human chorionic gonadotropin (HCG) day, and eggs] and R-ICSI cycle outcomes (fertilization rate, normal fertilization rate, cleavage rate, good embryo rate, implantation rate, clinical pregnancy rate and live birth rate) were analyzed. In addition, the effect of R-ICSI on the fertilization outcome of conventional IVF total fertilization failure and partial fertilization failure was explored. There was no significant difference in the general clinical data and R-ICSI cycle outcome between the TFF+low DFI group and the TFF+high DFI group (all P>0.05). There was no significant difference in the general clinical data between the PFF+low DFI group and the PFF+high DFI group (all P>0.05). The fertilization rate and normal fertilization rate in the PFF+low DFI group were significantly higher than those in the PFF+high DFI group (85.40% vs 72.41%, 71.90% vs 58.62%, respectively; both P<0.05). However, there was no significant difference in cleavage rate, good embryo rate, implantation rate, clinical pregnancy rate, and live birth rate between the 2 groups (all P>0.05). The R-ICSI cycle of TFF: A total of 79 fresh cycles, 57 fresh transplant cycles, a total of 761 unfertilized oocytes, and 584 M II oocytes were treated with R-ICSI, the fertilization rate was 83.22%, the normal fertilization rate was 75.51%, the cleavage rate was 98.15%, the good embryo rate was 40.74%, the implantation rate was 30.56%, and the clinical pregnancy rate was 43.86%; 29 live births were obtained. The R-ICSI cycle of PFF: A total of 61 fresh cycles, 31 fresh transplant cycles, a total of 721 unfertilized oocytes, and 546 M II oocytes were treated with R-ICSI; the fertilization rate was 83.33%, the normal fertilization rate was 69.78%, the cleavage rate was 97.36%, the good embryo rate was 44.39%, the implantation rate was 25.42%, and the clinical pregnancy rate was 45.16%; 12 live births were obtained. In the case of partial fertilization failure of IVF, the sperm DFI affects the fertilization rate and normal fertilization rate of R-ICSI; whether it is a TFF of IVF or PFF of IVF, ICSI can be used as an effective remedy way.

Subcutaneous transplantation of human thyroid tissue into a pre-vascularized Cell Pouch™ device in a Mus musculus model: Evidence of viability and function for thyroid transplantation.

This study aimed to investigate the survival and efficacy indicators of human thyroid tissue transplantation into a retrievable, prevascularized implanted Sernova Corp Cell Pouch™ (CP) device. Thyroid tissue from human donors was transplanted subcutaneously into the pre-implanted CP device or into the subcutaneous (SC) space alone as a control in a nude Mus musculus model. Transplanted M. musculus were monitored for human serum thyroglobulin (TG) levels for 3 months until the transplants were removed for histological assessment. Human thyroid tissue survived and continued to produce TG in transplanted nude M. musculus in the CP, with no adverse events. CP transplants exhibited more persistent and robust production of human TG than tissue placed in the SC space alone from 3 to 13 weeks post transplantation. Fresh thyroid transplants had better survival and function compared to cryopreserved transplants. Thyroid transplant viability correlated with TG levels at 3 months post-transplant (p = 0.03). Immunofluorescence staining of transplants for TG and TPO localized in thyroid follicles. Human thyroid tissue transplanted into the subcutaneously implanted pre-vascularized CP in nude M. musculus survived and continued to produce robust and persistent human TG and warrants further investigation as a treatment for postoperative hypothyroidism.

Endocrine Function of Ovarian Tissue Grafts After Normo- and Hypothermic Storage

Morphological characteristics and function of ovarian tissue after 2–120 hrs of storage at 4 and 37°C in incubation media with different composition (mannitol-containing solution (MСS) and phosphate buffered saline (PBS)) were investigated. The function of ovarian tissue after normo- and hypothermic storage was studied by heterotopic transplantation to recipient animals simultaneously with bilateral ovariectomy. After 2-hour storage at 37ºC and 24-hour one at 4ºC in all the studied media, the estradiol and progesterone levels in plasma of recipient animals were experimentally proved to be 18.2–27.5 and 13.5–18.5 ng/ml, respectively, (after fresh tissue transplantation these values for estradiol and progesterone made (28.1 ± 3.6) pg/ml and (19.1 ± 2.8) ng/ml, respectively). Prolongation of tissue storage time at 37ºC increased the number of degenerative follicles, augmented TBARS concentration and reduced sex hormone levels after transplantation using both MCS and PBS. A protective effect of MCS only, manifested in endocrine function restoration in recipient animals after transplantation ((13.5 ± 2.5) pg/ml of estradiol) was found after 48 hrs of ovarian tissue hypothermic storage. If PBS was used, the estradiol level was within the indices for ovariectomized animals ((7.1 ± 1.5 (pg/ml)).

O-178 Reproductive and endocrine outcomes after fresh and frozen-thawed ovarian tissue transplantation based on age and anti-cancer therapy: A systematic review and individual patient data meta-analysis

Abstract Study question Do reproductive and endocrine outcomes from fresh and frozen-thawed ovarian transplants differ based on age and anti-cancer therapy before cryopreservation? Summary answer There was a significant difference in reproductive outcomes of women who have their tissue cryopreserved before or at the age of 35 years. What is known already Ovarian tissue cryopreservation (OTC) and transplantation is emerging as a new fertility preservation method. Despite being available for two decades, there is a marked variation in the delivery of this procedure worldwide. Most of the data are based on case reports from specialised centres with expertise in providing this procedure, but there are many unreported cases. Through this review, we aim to collate reproductive and endocrine outcomes from ovarian tissue transplantation. In particular the outcomes in women based on age at cryopreservation and whether they had anti-cancer therapy before cryopreservation were explored. Study design, size, duration This study was a systematic review and individual participant level meta-analysis to synthesize the existing evidence on the use of fresh and cryopreserved ovarian tissue transplantation. The review protocol was registered with PROSPERO (CRD42018115233) in November 2018 and the review was concluded in December 2020, including 87 studies (768 women). Participants/materials, setting, methods Literature search was performed using MEDLINE, EMBASE, CINAHL and Cochrane Central Register of Controlled Trials from inception to October 2020. After screening 20,566 abstracts, 87 studies (768 women) were included in the review. Patient-level data was extracted for 388 women and study-level data for 380 women. Authors were also contacted for data if relevant outcomes were not reported in published manuscripts. Meta-analysis was performed using inverse-variance weighting to calculate summary estimates using a fixed-effects model. Main results and the role of chance Age at cryopreservation was provided for 319 out of 388 (82%) women at participant level data. Of these, 283 (88.7%) had ovarian tissue retrieved at ≤ 35 years of age. A subgroup of four studies that reported data on participants age at cryopreservation and transplantation were included in meta-analysis. Pregnancy rates were higher in participants at ≤ 35 years of age at cryopreservation, with results being statistically significant (OR, 0.35; 95% CI: 0.13 to 0.92; z = 2.13; P 0.03, I2= 0%). Return of hormonal function shown as a decrease in FSH (IU/L) was also lower in this group (MD, 4.38; 95% CI: -4.29 to 13.05; z = 0.99; P 0.32, I2= 0%. Whether a participant had received chemotherapy before cryopreservation was explicitly reported in 122 out of 388 (31%) participants and 56 of them (46%) had received anti-cancer treatment before OTC. Thirty-five pregnancies and twenty-four live births were reported in these women. A further meta-analysis from 5 studies showed that although the results were not statistically significant for return of endocrine function, a decrease in FSH, an increase in oestrogen and increased pregnancy rates were noted in participants who did not receive anti-cancer therapy before cryopreservation. Limitations, reasons for caution Although we gathered 768 cases of ovarian transplants published in the literature, most were case reports and therefore not included in the meta-analysis. Of the studies included in the meta-analysis, information such as age and anti-cancer therapy were not always provided for individual participants but as an aggregate. Wider implications of the findings There was no difference in reproductive and endocrine outcomes for anti-cancer therapy before OTC. Previous chemotherapy alone should therefore not be a deterrent in offering young girls and women OTC. Furthermore, the ideal age to achieve higher pregnancy and live birth rates from OTC is less than 35 years. Trial registration number PROSPERO (CRD42018115233)

Cost-effectiveness analysis of GnRH-agonist long-protocol and GnRH-antagonist protocol for in vitro fertilization

The gonadotropin releasing hormone agonist (GnRH-a) long-protocols and the GnRH-antagonist protocols are two commonly used protocols for in vitro fertilization (IVF), but their cost-effectiveness has not been studied, especially in China. A retrospective study involving 1638 individuals in GnRH-a long-protocol and 621 in GnRH-antagonist protocol were conducted and a decision tree model analysis was used to analyze the cost-effectiveness. Both direct and indirect costs were calculated. As a result, during the fresh embryo transplantation cycles, there was no significant difference in the rate of ongoing pregnancy between the two protocols, the average cost of per ongoing pregnancy in the GnRH-antagonist protocol was $ 16970.85, and that in the GnRH-agonist long-protocol was $19902.24. The probability of cumulative ongoing pregnancy per start cycle was estimated at 60.65% for the GnRH-antagonist protocol and 71.6% for the GnRH-agonist long-protocol (P < 0.01). Considering the cumulative ongoing pregnancy rate, the mean costs per ongoing pregnancy were estimated at $8176.76 and at $7595.28 with GnRH-antagonist protocol and GnRH-agonist long protocol, respectively. In conclusion, in fresh embryo transplantation cycle, the GnRH-antagonist protocol has economic advantage. However, the GnRH-agonist long protocol is more cost effective considering the cumulative ongoing pregnancy rate in the fresh embryo and frozen embryo transplantation cycles.

Функция трансплантатов овариальной ткани после гипотермического хранения: значение состава среды инкубации

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P387 Efficacy of combination therapy of fresh faecal microbiota transplantation and triple-antibiotic therapy for ulcerative colitis

P387 Efficacy of combination therapy of fresh faecal microbiota transplantation and triple-antibiotic therapy for ulcerative colitis

Ectopic Organogenesis after Allotransplantation of Freshly Removed or Cryopreserved Neonatal Testicle under the Renal Capsule in Rats.

In experiments on white outbred male rats, a freshly removed (20 experiments) or cryopreserved (10 experiments) testicle from newborn rats (1-2 days after birth) was transplanted under the renal capsule after bilateral orchiectomy. In all experiments with transplantation of freshly removed testicle, it was engrafted. In 3 months, histological examination revealed the formation of mature seminiferous tubules, but spermatogenesis was blocked at the stage of spermatogonia; groups of proliferating Leydig cells in the loose connective tissue between the tubules were also seen. In 6 and 12 months, the status of the seminiferous tubules remained unchanged, but structures typical of the epididymis and developing vas deferens were revealed. The number of proliferating Leydig cells increased. The initially low testosterone concentration in the blood of castrated males increased significantly as soon as in 1 month after transplantation and grew up to 3 months, remaining at a level ~50% of normal. Engraftment of cryopreserved neonatal testicular tissue was observed in 60% cases, however, engrafted tissue, similar to the fresh one, retained the ability for organogenesis with the formation of mature seminiferous tubules, epididymis, and groups of proliferating Leydig cells. The dynamics of blood testosterone concentration in rats with cryopreserved and fresh transplantation was similar. Subcapsular transplantation did not adversely affect the kidneys, which was seen from normal histological structure of the kidneys and creatinine and urea concentrations in the blood.

Xenotransplantation of goat ovary as an alternative to analyse follicles after vitrification.

The aim of this study was to evaluate the caprine preantral follicles enclosed on vitrified/warmed ovarian cortex grafted to nude BALB/mice during 1month. The ovarian cortex from goats was fragmented (3×3×0.5mm) and divided into four groups: fresh control, vitrified control, fresh transplant and vitrified transplant. Follicular morphology, development and density, fibrosis as well as apoptosis, and tissue revascularization were evaluated. It was also observed a significant decrease in morphologically normal preantral (primordial, transition, primary and secondary) follicles in both vitrified control and vitrified transplant treatments when compared with both fresh control and fresh transplant. However, fresh control and fresh transplant exhibited a similar percentage of developing follicles. Additionally, Vitrified control showed a significant increase in developing follicles in comparison with both fresh control and fresh transplant. Follicular density significantly decreased in all treatments in comparison with fresh control. We observed high fibrosis in both fresh transplant and vitrified transplant. The mRNA expression of caspase 3 was lower in both fresh transplant and vitrified transplant in comparison with vitrified control. In conclusion, xenotransplantation is an excellent strategy to maintain normal preantral follicle morphology after vitrification/warming of goat ovarian tissue. Yet, in order to ensure the survival and development of these follicles, it is essential to improve the revascularization of the graft.

Does embryo vitrification affect the mice offsprings’ growth, development and the nervous system?

This study focused on the effect of embryo vitrification on the growth, development and the nervous system of the offsprings. Comparative and controlled study. Mice were used as model animals, we designed three groups including natural pregnancy-born mice (control group), conventional IVF-ET pregnancy-born mice (IVF group), and embryonic vitrification-born mice (frozen group). The growth and development indicators and neurodevelopment indexes of offsprings were compared among the above groups; two-dimensional protein electrophoresis, mass spectrometry, and Western Blotting were used to determine the changes in brain tissues of the mice. Brain tissue ultrastructures of the offsprings were observed by immunohistochemical and electron microscopy. Physiological indicators such as physical strength were compared; the neural development indicators such as the intelligence, learning ability and memory were also compared through the Morris water maze test. 1. There was no significant difference in the pregnancy rate between the fresh transplantation group and the vitrification group (48.89% Vs 51.22%, P>0.05). 2. The weights of the offsprings in the vitrification group were higher than that in the conventional IVF group and the natural conception group (1.98±0.21 Vs 1.95±0.22, P<0.05). There was no significant difference in offspring weights between the conventional IVF group and the natural conception group (1.95±0.22 Vs 1.96±1.89, P> 0.05).3. Two-dimensional differentially expressed proteins in brain tissue were searched by two-dimensional protein electrophoresis, protein profiling, and Western blotting. Bioinformatics analysis revealed that most protein expression differences did not cause functional changes.4. Through immunohistochemistry and electron microscopy, it was found that vitrified frozen embryos had no obvious damage to the brain tissue ultrastructures. 5. Vitrification has not affected the structure of brain tissue in the study of mouse embryos through immunohistochemical and electron microscopy.6. Time to reach the platform,latency,the total distance and the frequency of crossing the former platform were similar among the three groups.(see the Table). The weights of the offsprings born through embryo vitrification were increased, while fertility, learning and memory ability were not changed. The technique of embryo vitrification has no significant effect on brain protein expression and tissue ultrastructures.The vitrification technique commonly used in clinic has little effect on the offsprings.Tabled 1The result of the Morris water maze testGroupsTime to reach the platform（s）latency（s）the total distance（cm）the frequency of crossing the former platform（times）frozen group13.04±1.8723.45±1.56212.74±19.381.88±0.71IVF group15.34±1.4820.67±1.79209.34±14.742.28±0.68control group13.46±1.7521.86±1.21218.53±12.471.97±0.46P value<0.05<0.05<0.05<0.05 Open table in a new tab

In vivo and in vitro strategies to support caprine preantral follicle development after ovarian tissue vitrification

The aim of the present study was to compare fresh and vitrified goat ovarian tissue after autotransplantation and in vitro culture. Adult goats were completely ovariectomised and each ovarian pair was sliced and distributed among six different treatment groups: fresh control, fresh transplant, fresh culture, vitrified control, vitrified transplant and vitrified culture. Follicular morphology, development, growth, density, revascularisation and hormone production were evaluated in all groups. Three antral follicles (two in the fresh transplant and one in the vitrified transplant groups) were observed on the surface of the graft 90 days after transplantation. The percentage of morphologically normal follicles was similar in the fresh control, fresh transplant and vitrified transplant groups. The percentage of developing (transition, primary and secondary) follicles was higher after in vitro culture of fresh or vitrified tissue. Transplantation resulted in a lower follicle density. Serum oestradiol concentrations remained constant during the entire transplantation period. In contrast, progesterone production decreased significantly. Expression of CD31 mRNA was lower in fresh culture. In conclusion, restoration of goat ovarian function can be successfully achieved following transplantation of both fresh and vitrified goat ovarian tissue. However, transplantation induced higher follicle loss than in vitro culture.

Faecal microbiota transplantation versus placebo for moderate-to-severe irritable bowel syndrome: a double-blind, randomised, placebo-controlled, parallel-group, single-centre trial

Irritable bowel syndrome (IBS) is a common condition characterised by abdominal pain, bloating, and poor quality of life. IBS might be caused by a gut dysbiosis. We aimed to compare faecal microbiota transplantation (FMT) with placebo in patients with IBS. In this double-blind, randomised, placebo-controlled, parallel-group, single-centre study, we enrolled patients with IBS with diarrhoea or with diarrhoea and constipation (excluding dominating constipation) defined by the ROME III criteria, scored as moderate to severe according to the IBS severity scoring system (IBS-SSS; a score of ≥175). Eligible participants were aged 18-75 years and were recruited locally by general practitioners in northern Norway. We randomly assigned participants (2:1) in blocks of six to active or placebo FMT. Personnel not involved in the clinical performance of the trial generated the randomisation sequence using a randomisation website. Non-study personnel performed the final allocation and standardised the active and placebo transplants to make them identical in appearance and temperature. The faeces were freshly processed, and were used the same day (fresh transplant) or were stored in a freezer for later use (frozen transplant); participants' own faeces served as placebo. A dose of 8 mg loperamide was administered orally 2 h before endoscopy to retain the transplant. The transplant (50-80 g of faeces mixed with 200 mL of isotonic saline and 50 mL of 85% glycerol) was administered by a colonoscope to the caecum. The primary endpoint was symptom relief of more than 75 points assessed by IBS-SSS, 3 months after FMT. The primary analysis was done in the modified intention-to-treat population, excluding participants who did not undergo treatment or who were diagnosed with any other disease by pinch biopsies obtained during the treatment procedure. For the safety analysis, only participants who did not undergo treatment were excluded. The study is registered with ClinicalTrials.gov, number NCT02154867. The trial has been extended with an open-labelled study treating the placebo group with frozen FMT for further exploratory studies. Between Jan 1, and Oct 30, 2015, we recruited 90 participants and randomly assigned them to active treatment (n=60) or placebo (n=30). Three participants did not undergo FMT and four were excluded after diagnosis of microscopic colitis, leaving 83 for final modified intention-to-treat analysis (55 in the active treatment group and 28 in the placebo group). 36 (65%) of 55 participants receiving active treatment versus 12 (43%) of 28 receiving the placebo showed response at 3 months (p=0·049). One participant had transient nausea and vertigo (active group) and was observed at the hospital for a few hours after the procedure. Two participants had soiling of transplant on their way home from treatment (one in each group) and three experienced self-limiting intermittent abdominal pain (one in the active group and two in the placebo group). No serious adverse events could be attributed to FMT. FMT induced significant symptom relief in patients with IBS. However, larger multicentre studies are needed to confirm the results. HelseNord and the Norwegian Centre of Rural Medicine, University of Tromsø.

Prospective Evaluation of Structural Allograft Transplantation for Osteochondral Lesions of the Talar Shoulder

Category: Ankle. Introduction/Purpose: The management of large osteochondral lesions of the talar shoulder remains a clinical challenge. Their size, three-dimensional geometry, and subchondral cystic degeneration often preclude treatment with traditional measures such as microfracture and osteochondral autograft transplantation. Structural or bulk osteochondral allograft transplantation has demonstrated efficacy in several retrospective reviews. The purpose of this study was to prospectively evaluate patients who received fresh structural allograft transplantation to the talus. Methods: A prospective evaluation of consecutive patients who underwent fresh structural allograft transplantation for an OLT form 2010 to 2013 was performed under Institutional Review Board approval. All patients failed a minimum of 6 months of conservative management. Preoperative MRI and/or CT, as well as plain radiographs were obtained on all patients. The following patient reported outcomes questionnaires were administered preoperatively and yearly after surgery: 100 mm VAS pain scale, AOFAS Ankle-Hindfoot Scale, SF-36, and the Short Musculoskeletal Functional Assessment (SMFA). Pre- and postoperative radiographs were assessed for allograft incorporation and the development of arthritis. Results: Fourteen patients underwent fresh osteochondral allograft transplantation. The mean follow-up was 47 months (range 24-73). The average size of the OLT was 2,269 mm3 (range 813-8,366) based on CT imaging and 5,797 mm3 (range 1,136-12,489) based on MRI imaging. There was significant improvement in the VAS pain score, AOFAS Ankle-Hindfoot Scale score, the SF-36 total score, and the SMFA functional and bother indices. Five (36%) of the patients required subsequent arthroscopy and removal of the screw(s) used to secure the graft. Three of these second-look patients had stable grafts without chondral damage. Two grafts demonstrated cartilage delamination. One of these patients had continued pain and progression of arthritis without additional surgery and one was converted to an ankle replacement. Therefore, the failure rate was 14%. Conclusion: Significant improvement in pain and function can be achieved with structural allograft transplantation for large OLTs. However, it is important to council patients that painful hardware and stiffness can occur in approximately one-third of patients. An unstable graft and cartilage delamination are indicators of subsequent failure. The use of a structural allograft does not preclude subsequent ankle arthrodesis or arthroplasty.

P463. Combination therapy of fresh faecal microbial transplantation and antibiotics for ulcerative colitis

P463. Combination therapy of fresh faecal microbial transplantation and antibiotics for ulcerative colitis