Recipient-derived EDA fibronectin promotes cardiac allograft fibrosis

Advances in donor matching and immunosuppressive therapies have decreased the prevalence of acute rejection of cardiac grafts; however, chronic rejection remains a significant obstacle for long-term allograft survival. While initiating elements of anti-allograft immune responses have been identified, the linkage between these factors and the ultimate development of cardiac fibrosis is not well understood. Tissue fibrosis resembles an exaggerated wound healing response, in which extracellular matrix (ECM) molecules are central. One such ECM molecule is an alternatively spliced isoform of the ubiquitous glycoprotein fibronectin (FN), termed extra domain A-containing cellular fibronectin (EDA cFN). EDA cFN is instrumental in fibrogenesis; thus, we hypothesized that it might also regulate fibrotic remodelling associated with chronic rejection. We compared the development of acute and chronic cardiac allograft rejection in EDA cFN-deficient (EDA(-/-)) and wild-type (WT) mice. While EDA(-/-) mice developed acute cardiac rejection in a manner indistinguishable from WT controls, cardiac allografts in EDA(-/-) mice were protected from fibrosis associated with chronic rejection. Decreased fibrosis was not associated with differences in cardiomyocyte hypertrophy or intra-graft expression of pro-fibrotic mediators. Further, we examined expression of EDA cFN and total FN by whole splenocytes under conditions promoting various T-helper lineages. Conditions supporting regulatory T-cell (Treg) development were characterized by greatest production of total FN and EDA cFN, though EDA cFN to total FN ratios were highest in Th1 cultures. These findings indicate that recipient-derived EDA cFN is dispensable for acute allograft rejection responses but that it promotes the development of fibrosis associated with chronic rejection. Further, conditions favouring the development of regulatory T cells, widely considered graft-protective, may drive production of ECM molecules which enhance deleterious remodelling responses. Thus, EDA cFN may be a therapeutic target for ameliorating fibrosis associated with chronic cardiac allograft rejection.     

CCR5 blockade in combination with rapamycin prolongs cardiac allograft survival in mice

Both chemokine receptor 5 (CCR5) blockade and rapamycin (rapa) are effective in modulating transplant immunity and led to prolonged allograft survival, yet a great many grafts were ultimately lost to acute rejection. In this study we examined the inhibition of CCR5 in combination with the treatment with rapa in cardiac transplantation. Fully major histocompatibility complex-mismatched murine cardiac allograft models were randomized to five groups. They were administered with anti-CCR5 antibody or control antibody and rapa or phosphate-buffered saline (PBS), respectively. An additional group was treated with anti-CCR5 antibody, rapa and anti-CD25 antibody. Allograft rejection was investigated by flow cytometric analyses and enzyme-linked immunospot assay. Allografts treated with anti-CCR5 antibody plus rapa showed significantly prolonged survival (83 ± 3 days, P < 0·001) compared with control antibody plus PBS-treated allografts (6 ± 1 days). Treatment with anti-CCR5 monoclonal antibody (mAb) plus rapa inhibited significantly the progression of chronic rejction. Further analysis of donor hearts in the anti-CCR5 antibody plus rapa-treated group demonstrated increased infiltration of CD4⁺CD25⁺forkhead box P3⁺ regulatory T cells, and depletion of CD25⁺ cells resulted in acute rejection of allografts in 18 ± 1 day. CCR5 blockade in combination with rapa is effective in preventing acute and chronic rejection in a robust murine model. This effect is mediated by CD25⁺ T cell recruitment and control of T lymphocyte proliferation.     

去细胞异体神经的形态结构及移植后的组织相容性

目的:为面神经缺损寻找一种理想的异体神经移植物。方法:取Wistar大鼠胫神经,经Triton X-100和脱氧胆酸钠溶液进行化学去细胞处理。将处理后的神经行组织学染色和免疫组织化学染色;并行异体移植修复面神经缺损,观察其组织相容性。结果:去细胞神经为一中空的神经基质管,其中的细胞和髓鞘成分被有效清除,神经基底膜被保留;异体移植后无明显炎症反应,无排斥和吸收反应,能引导宿主轴突和Schwann细胞增殖。结论:去细胞异体神经移植物具有良好的仿生性和组织相容性,可能用于修复面神经缺损。     

高压蒸气灭菌同种异体骨移植的实验研究

目的：观察高压蒸气灭菌同种异体骨在骨移植中的作用，探索简便、经济的骨移植方法。方法：用家用压力锅，对同种异体鼠骨高压蒸气灭菌３０ｍｉｎ后，将其植入２５只成年大白鼠双侧胫骨（新生鼠骨植入右侧１２例，成年鼠骨植入左侧２５例）前上部一个长１０ｍｍ，深２ｍｍ的缺损中，于术后３、６、９、１２周取材，制片，在普通、荧光光镜和透射电镜下进行观察。结果：发现经高压蒸气灭菌后的同种异体骨有较好的生物相容性，可以引导骨生长。结论：高压蒸气灭菌处理同种异体骨，是一种简便、经济的骨移植方法，易于在农村和基层开展     

膝关节移植中应用同种异体骨软骨有效性与安全性的Meta分析

文题释义：同种异体移植：是指同种不同基因型个体之间的移植,是临床最常见的移植类型,也是移植免疫学研究的重点所在。同种异体移植的出现,有效解决了自体移植所面临的移植物数量不足、延长手术时间以及手术伤口面积增加等问题。单组率的Meta分析：对于某一项结局指标,所纳入的全部研究只提供一组人群的总人数和事件发生人数(患病率、检出率、手术率、病死率、感染率等)的研究(常见于病例系列研究)进行定量综合,为后续研究提供更可靠证据的科研方法。背景：同种异体骨软骨移植治疗膝关节软骨缺损是临床应用最久的方法之一。虽然该方法在临床中应用广泛且深受骨科医师信赖,但仍缺乏循证医学方面的支持。目的：分析同种异体骨软骨在膝关节移植术中的疗效,对所有已获得的数据进行系统综述和Meta分析,评价同种异体骨软骨在膝关节移植过程中作为移植替代物的有效性及安全性。方法：在PubMed/Medline、EMBASE、Cochrane协作网图书馆、中国知网(CNKI)以及万方数据库中进行文献检索。检索同种异体骨软骨在膝关节移植术中的应用,根据文献纳入标准进行选择,重点选择数据可以被提取以及能够进行Meta分析的文章。结果与结论：①25项研究符合纳入标准,均为病例系列研究,患者报道的结果是同种异体骨软骨可以作为修复膝关节缺损的移植材料;共1 081例患者(1 111膝)纳入研究,发病年龄11-75岁,平均年龄约为 34.41岁,女性比例约为40.81%,随访时间4-384个月,平均76.8个月;②在这些研究中,接受的供者年龄为10-65岁,而根据供者年龄范围与研究数目进行统计发现,15-45岁的供者为主要的获取目标;③术后总体的成功率为74%,总体二次手术率17%;单极移植成功率74.44%(501/673),双极移植成功率50.94%(27/53),单极移植成功率明显高于双极移植(χ²=13.679,P < 0.05);④67例(13.14%,67/510)出现治疗并发症,常见并发症为手术部位持续疼痛(15例)和移植物骨折或碎裂(12例);⑤提示同种异体骨软骨是膝关节移植中一种有效且安全的移植替代物,整体成功率74%,二次手术率17%,是一种成功率高、再次手术风险低的治疗方式。ORCID: 0000-0001-6786-6595(华堃池)中国组织工程研究杂志出版内容重点：人工关节;骨植入物;脊柱;骨折;内固定;数字化骨科;组织工程     

Changes of enzyme activities in ovine fetal fluids and maternal blood serum with gestational age

Our objectives were threefold: (1) to assess the activities of tissue enzymes, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and creatine kinase (CK) in the fetal fluids (amniotic and allantoic) collected from the gravid sheep uteri obtained from the abattoir and also in the maternal blood serum at various stages of pregnancy, (2) to compare the enzyme activities of fetal fluids relative to the maternal serum, and (3) to compare the enzyme activities in serum of pregnant ewes to nonpregnant ones. The stages of gestation, viz., stage I (30–60 days), stage II (61–90 days), and stage III (91–120 days) were identified based on the crown anus length of the fetus. As the gestational ages increased, AST significantly (p < 0.01) increased in the amniotic fluid and maternal blood serum but decreased in allantoic fluid; ALT had no changes in fetal fluids and the maternal blood serum; ALP decreased significantly (p < 0.01) in allantoic fluid but had no changes in the amniotic fluid and maternal serum; LDH increased significantly (p < 0.01) in amniotic and allantoic fluids but decreased in maternal serum; CK decreased significantly (p < 0.01) in fetal fluids and maternal serum. The enzyme activities were significantly higher in maternal sera than fetal fluids but were successively less in allantoic and amniotic fluids (p < 0.01). The activity of enzymes in maternal sera of pregnant and nonpregnant ewes were nearly the same. Except for LDH and CK, the greatest activities were found in the maternal serum in stage I and for AST in stage III of pregnancy (p < 0.01). These findings may have appreciable diagnostic significance in prenatal detection of disease status in both the dam and the fetus.     

Pregnancy duration and fetal number: Effects on maternal behavior in rats

It was shown previously that in rats postpartum mothers are much more likely than pup sensitized virgin mothers to retrieve pups from a T-maze extension of their home cage. In the present experiment, the physiological basis of this difference was explored further by inducing maternal behavior (retrieving, crouching, licking) following hysterectomy-ovariectomy on Days 21, 16, and 10 of pregnancy. In general, the longer the duration of pregnancy, the shorter the latency to become maternal and the greater the propensity to retrieve pups from the T-maze. An additional group of Day 16 pregnancy terminated rats carrying six or fewer fetuses were not impaired with respect to onset of maternal behavior compared to their large-litter counterparts, but only a small percentage of these females retrieved in the T-maze. These findings suggest that both the late pregnancy estrogen surge and progesterone withdrawal after its peak and sustained secretion enhance maternal responsiveness and that T-maze pup retrieval (but not onset latency) may be additionally affected by the level of placental hormones.     

IDO、树突状细胞与免疫耐受

吲哚胺2,3双加氧酶(IDO)是一种免疫调节酶,可催化色氨酸分子中吲哚环氧化裂解,从而沿犬尿酸途径分解代谢的限速酶。而树突状细胞(DC)是一种功能强大的抗原递呈细胞(APC),是唯一能够激活初始型T细胞的APC。IFN-γ等可刺激其表面表达IDO,并通过降解色氨酸,使局部组织中的色氨酸耗竭,代谢产物犬尿氨酸含量增加,从而抑制T细胞的增殖。因此DC表面表达IDO可能在诱导免疫耐受中起着重要的作用。     

An immunohistochemical study of the secretory immune system in human fetal membranes and decidua of the first trimester of pregnancy

PROBLEM: We analyzed the presence and distribution of components of the secretory immune system (SIS) in human fetal membranes (amnion, yolk sac, chorion) and decidua from the first trimester of pregnancy. MATERIALS AND METHODS: Specimens from 17 embryos (4-8 weeks of pregnancy) and nine fetuses (9-12 weeks) were divided into those that had not been exposed to massive foreign antigenic effects (Group I, n = 18) and those that had suffered acute chorioamnionitis (Group II, n = 8). RESULTS: Positive immunostaining for secretory component (SC), joining (J) chain, IgG, IgA, and macrophages was seen from 4 to 5 weeks of development and then during the whole first trimester of pregnancy in the syncytio- and cytotrophoblasts, the amniotic epithelium, the yolk sac endoderm and decidual cells. Macrophages with J chain, IgG and IgA were found in embryonic tissues on week 4, whereas lymphocytes, including those synthesizing IgA and IgM, appeared only at the end of the first trimester of pregnancy. In the decidua, lymphocytes and macrophages were recognized during the whole period of study. In cases with chorioamnionitis (Group II), reactivity of IgG and IgA in the mentioned cells of fetuses decreased sharply while the rate of immunoreactivity of SC and J chain as well as the number of T and B lymphocytes did not change. In the decidua, the number of immune reactive cells sharply increased with the appearance of plasma cells. CONCLUSIONS: In the fetal membranes and decidua all the SIS components are present. We suggest that two SIS, maternal and fetal, participate in the formation of the barrier between a mother and the fetus. Both these systems have different origin and cellular content as well as different immune reactions.     

胎儿内镜手术对孕妇的近远期并发症

母胎医学对于提高围产儿的生存质量和提高出生人口素质具有重要意义,胎儿内镜手术是母胎医学的重要组成部分。虽然是微创手术,但无论对于胎儿还是孕妇来说,手术的风险始终存在。本文阐述了在胎儿内镜手术中,孕妇近远期并发症的发生率、病因、病理生理机制以及简要的处理,旨在手术矫正胎儿畸形的同时,最大限度地降低受术健康孕妇的风险。     

孕妇外周血中胎儿有核红细胞用于产前诊断的研究进展

本文概述孕妇外周血中胎儿有核红细胞的分离与检测用于产前诊断的研究状况,描述母体外周血中的胎儿细胞的特点,胎儿有核红细胞的富集和分离方法,并对各方法的优点、缺点、病理性妊娠的意义、临床应用、存在的问题及研究进展作一综述。     

Two-year follow-up of bone mineral density changes in the knee after meniscal allograft transplantation: Results of an explorative study

Background

The potential chondroprotective effect of meniscal allograft transplantation (MAT) is unclear. Subchondral bone mineral density (BMD) and subchondral bone remodeling play important roles in osteoarthritis development. Evaluation of subchondral BMD after MAT might give more insight into the potential chondroprotective effect. The purpose of this study was to determine early BMD changes in the knee after MAT.

Embryonic resistance to tumour necrosis factor-{alpha} mediated cytotoxicity: novel mechanism underlying maternal immunological tolerance to the fetal allograft

The cytokine tumour necrosis factor- (TNF) has been postulatedto play an essential role in the cytotoxic activity of cell-mediatedimmunity against allogenic or tumour cells invading the host.Several tumour cell lines, however, are resistant to TNF mediatedcytotoxicity and respond paradoxically by cellular proliferationand by autocrine secretion of TNF. In view of the metastaticcharacter of the mammalian embryo, the aim of this study wasto assess the potential of murine embryos to secrete TNF invitro, to express TNF receptors and to resist TNF mediated cytotoxicityduring their in-vitro development to the blastocyst stage. Thepotential of human embryos to secrete TNF in vitro until theblastocyst stage was also investigated. From a total of 11 humanembryos, which were allowed to proceed to blastocyst formation,seven secreted TNF in the range of 2–117 pg/ml/24 h. Atotal of 123 C57BL/6J mouse embryos were studied of which 55%secreted TNF in the range of 1.25–3.95 mg/ml/24 h. Thepresence of high levels of exogenous TNF (10–300 IU) wasnot detrimental to the in-vitro development of murine embryos.Using immunohistochemical techniques, we were not able to detectthe presence of type I or II TNF receptors on the surface ofmurine embryos. Our findings suggest that human and C57BL/6Jmurine embryos have the potential to secrete TNF in vitro duringthe developmental stages leading to blastocyst formation. Inboth species, the presence of TNF in the culture medium didnot cause subsequent necrosis of the conceptus, suggesting thatmammalian embryos may be TNF resistant cell lines. The observedembryonic resistance to TNF may be explained by the absenceof TNF receptors by which the cytotoxic effect is usually mediated.It is suggested that embryonic resistance to physiological concentrationsof TNF released by effectors of the host's immune system, couldbe via a mechanism underlying the maternal immunological toleranceto the fetal allograft.     

PARP-1 inhibitor-AG14361 suppresses acute allograft rejection via stabilizing CD4+FoxP3+ regulatory T cells

Acute allograft rejection is the most common complication in organ transplantation leading to organ loss. Treg cells play an important role in preventing acute rejection, but they are unstable and easily lose function. Poly(ADP-ribose) polymerase 1(PARP-1) is involved in the differentiation stabilization of Treg cells, it has been suggested that PARP-1 inhibition could prevent acute rejection and prolong allograft survival. This study investigated AG14361 effects on acute allograft rejection. We used a fully MHC-mismatched murine heart transplantation model to compare the effect of PARP-1 inhibitor-AG14361 on alloimmunity to the control. Mice treated with PARP-1 inhibitors showed a longer median survival time of allografts (MS14 compared with the control group, MST was 8 days, and AG14361 was 6 days, P = 0.019). The combination of sirolimus and AG14361 significantly delayed allograft MST (AG14361 + sirolimus for 30 days, sirolimus for 16 days, P = 0.002). AG14361 markedly augmented the number of the CD25+FoxP3+ Treg cells in the graft and periphery. In addition, it could enhance the suppressive function of Treg cells by upregulating the level of CTLA-4, PD-1 and ICOS. In vivo, the Treg/Th17 ratio increased significantly in the AG14351 group compared to the control. In the combination with sirolimus treatment, AG14361 promoted the long-term allograft survival. Our results highlight novel effects of a PARP-1 inhibitor. PARP-1 inhibitor AG14361 may be a promising agent to attenuate acute allograft rejection as it can maintain the number and function of Treg cells in allografts.     

Percutaneous needle biopsies of renal allografts: the relationship between morphological changes present in biopsies and subsequent allograft function

In the Cambridge renal transplant unit percutaneous needle biopsies of renal transplants have been extensively used to help identify the cause of impaired allograft function. During the period 1966--1973, 154 of the 269 renal allografts transplanted were biopsied at least once during the first 90 days after transplantation. In this survey the relationship between morphological changes in these biopsy specimens and allograft function 1, 3 and 5 years after transplantation is assessed. A highly significant direct relationship exists between early graft failure and the presence of medial necrosis of arteries, acute glomerular lesions and interstitial haemorrhage. Less than 10% of grafts with one or more of these changes and none in which all three types of lesion were present were capable of supporting life at 1 year. There is a significant association between poor subsequent graft function and mononuclear cell infiltration of the intima of arteries. No clear relationship exists, however, between the function of grafts at 1 and 3 years and the degree of mononuclear cell infiltration of the interstitial tissue. Tubular necrosis was frequently observed and future graft performance is related to the extent and cause of the tubular damage.     

Exosomes in the Immune Response and Tolerance

Exosomes are small vesicles with endocytic origin secretedby many live cells. They are flattened or round sphere se-questered by a lipid bi-layer visualized by under electronmicroscopy (EM), with sizes from 30 run to 100 nm. Ex-osomes were first described…     

The dexamethasone suppression test, the Hamilton Depression Rating Scale and the DSM-III depression categories

The Hamilton Depression Rating Scale (HDRS) score and plasma cortisol values were measured in 100 depressed patients at 8 a.m., 4 p.m. and 11 p.m. after oral administration of 1 mg dexamethasone the previous night. The patients were categorized according to DSM-III as suffering from either minor depression (including dysthymic disorder, 300.40; adjustment disorder with depressed mood, 309.00; atypical depression, 296.82) or major depression (without melancholia, 296.X2; with melancholia, 296.X3; with psychotic features, 296.X4). Plasma cortisol levels of 3.5 μ/dl at 8 a.m. were found to be the most sensitive (56.9%) and specific (94.3%) discriminator between minor and major depression. Plasma cortisol levels at 4 p.m. and 11 p.m. or the combination of several cortisol values also differentiated between minor and major depression; however, the results were not so conclusive. According to the rating on the Hamilton Depression Scale the patients with major depression were more severely depressed (P < 0.001) than patients suffering from minor depression. Cortisol values at 8 a.m., 4 p.m., 11 p.m. and the highest levels were significantly (P < 0.001) correlated with the HRDS score. A maximum of 20.2% of the score variance could be explained by the correlation with the highest cortisol value observed. Severity of illness does not exclusively account for the biological differences between minor and major depression.     

爱情满意度与包容及情商的关系

目的 探讨爱情满意度与包容力以及情商的关系,找出提高爱情满意度的方法.方法 使用包容力测试、EQ测试与爱情满意度问卷对870位在校大学生进行调查研究.结果 不同性别之间在爱情满意度上没有显著差异,但是在情商和包容力上却差异显著(t=2.543,P＜0.05;t=5.358,P＜0.01).是否是独生子女、不同的年级和不同的年龄之间,爱情满意度、情商和包容力没有显著的差异.爱情满意度与包容力和情商之间的相关关系十分微弱(r=-0.142,0.161).结论 更多包容和更高情商并不会带来更满意的爱情,而是需要双方积极和良好的沟通才能把存在的问题和矛盾化解.     

TNF-alpha and TGF-beta1 gene polymorphisms and renal allograft rejection in Koreans

This study was performed in order to evaluate the association of tumour necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) gene polymorphisms with renal allograft rejection in Koreans. Five TNF-alpha (-1031T/C, -863C/A, -857C/T, -308G/A and -238G/A) and two TGF-beta1 (codon 10 T/C and codon 25 G/C) single-nucleotide polymorphism (SNP) sites were studied by using polymerase chain reaction (PCR) single-strand conformation polymorphism and PCR restriction fragment length polymorphism methods in 100 controls and 164 patients. The patients underwent renal transplantation, having one or more Human leukocyte antigen (HLA)-A, HLA-B and HLA-DR antigens mismatched with their donors. For the TGF-beta1 gene, we also studied the polymorphism of donors. The allele frequencies of each SNP site in controls were not different from those of patients. The frequency of TNF-alpha high-producer genotype, -308GA, and TGF-beta1 lower (intermediate)-producer genotype, codon 10 CC and codon 25 GG, were significantly higher in patients with recurrent acute rejection episodes (REs), compared to those in patients with no or one RE. The highest risk group for developing recurrent REs showed the combination of TNF-alpha high- and TGF-beta1 lower-producer genotypes. Analysis of chronic renal allograft dysfunction (CRAD) revealed that TGF-beta1 high-producer genotype of donors, codon 10 TT/TC and codon 25 GG, is associated with CRAD especially in patients with recurrent REs. The highest risk group for developing CRAD showed the combination of recipient's TNF-alpha high- and donor's TGF-beta1 high-producer genotypes. These results would be useful for predicting high-risk group for acute rejection or CRAD in renal transplantation.     

Cryopreservation,survival and function of intrastriatal fetal mesencephalic grafts in a rat model of Parkinson's disease

Summary In the present study we quantitatively assessed to what extent freeze-storage at liquid nitrogen temperature influences the survival and function of fetal mesencephalic grafts in the dopamine-depleted rat striatum. Ventral mesencephalic (VM) tissue was dissected from rat fetuses and stored overnight in a preservative medium at 4 °C (hibernation). It was grafted intrastriatally either as a fresh cell suspension or was frozen as tissue fragments or as a cell suspension after stepwise incubation in ascending concentrations of dimethyl-sulphoxide. Following a cryopreservation interval of 80 days in liquid nitrogen, the frozen samples were rapidly thawed, rinsed, and grafted. Cellular viabilities of graft cell suspensions, as assessed by ethidium bromide/acridine orange staining, were decreased from 90% in fresh tissue to 38-35% in frozen and thawed tissue. Amphetamine-induced turning behavior at 6 weeks post-grafting was significantly attenuated in hosts that had received fresh grafts or grafts that were frozen as tissue fragments. Tyrosine hydroxylase-(TH-) immunocytochemistry of recipient brains revealed significant decreases in TH-positive graft cell numbers in rats grafted with cryopreserved tissue (38–42% of fresh tissue). Moreover, the dye exclusion viability of thawed VM tissue was found to accurately predict the subsequent graft survival. There was no difference with respect to graft cell numbers between the two freezing methods employed, though block storage seems to be more simple from a practical point of view. The present study indicates that freezing in liquid nitrogen may be a feasible method for long-term storage of fetal neural tissue for grafting, although a marked decrease in graft survival and function of cryopreserved tissue must be taken into account.Present address: Dept. of Medical Physiology