Histocompatibility studies in pigs from outbred and semi-inbred families

Skin graft experiments with pig siblings from semi-inbred and outbred families confirmed the existence of a pig main histocompatibility system (MHS). Grafts exchanged between serological (lymphocytotoxicity) MHS incompatible siblings from outbred matings survived 5.6 days and between compatible animals 9.8 days. In compatible pairs from semi-inbred matings they survived 22.4 days and in incompatible individuals 6.1 days. Skin grafts exchanged between individuals with E blood group compatibility led to the formation of haemagglutinating and lym-phocytotoxic antibodies reacting with some E antigens. The results indicate that the survival of skin grafts in semi-inbred pigs is apparently influenced also by differences in the E system.     

Growth and carbon partitioning in compatible and incompatible peach/plum grafts

The shoot growth of compatible ( Prunus persica L. Batsch cv. Springtime grafted on Prunus cerasifera L. Ehrh cv. Myrobolan P2032) and incompatible ( Prunus persica L. Batsch cv. Springtime grafted on Prunus cerasifera L. Ehrh cv. Myrobolan P18) peach/plum grafts was observed over a period of 100 days after grafting under controlled conditions. Leaf and root activities were determined by studying carbon assimilation and partitioning, leaf mineral contents and water relations. Shoot length and leaf number were not significantly affected in the incompatible combination during the first 55 days after grafting, but then, shoot growth rate was significantly reduced. Final total dry weights of the shoot were similar in both graft combinations. The incompatible combination did not show any water stress. Soluble sugar and starch contents increased in the leaves of the incompatible combination, accounting for about 36% of the increase of leaf dry weight per unit area. Photosynthesis was affected by the compatibility of the grafts. Leaf nitrogen content (% dry weight) fell in the incompatible graft combination 65 days after grafting. However, nitrogen content on an area basis was not affected. The possibility of nitrogen stress is discussed.     

Time course of angiogenesis in solid pineal autografts

Angiogenesis and reperfusion of blood vessels were analysed qualitatively, at the light- and electron-microscopical levels, in solid pineal autografts placed intracerebrally in adult rats (post-transplantation survival times: 1, 3, 7, 10, 14 and 28 days). Reperfusion of blood vessels was studied in sections from immersion-fixed brains incubated to demonstrate the endogenous peroxidase activity of erythrocytes within the lumen of blood vessels. The possible presence of the blood-brain barrier (BBB) within the grafts was also investigated by injecting native horseradish peroxidase (HRP) intravenously into the rats. Angiogenesis, the morphological and functional properties of blood vessels vascularizing the grafts and the survival of pineal tissue were analysed ultrastructurally following transplantation. Revascularization of pineal autografts placed into the adult host central nervous system occurred very slowly, requiring 7–10 days to establish anastomoses between graft and host blood vessels. During this process, signs of angiogenesis in pineal and cerebral capillaries were evident, suggesting that both contributed to graft revascularization. Morphological and functional studies with HRP revealed that, following transplantation, blood vessels at the graft-host interface or within pineal autografts maintained their morphological and functional properties: they were fenestrated and did not present a BBB to blood-borne peroxidase. Thus, after grafting, the presence or absence of the BBB is graft-determined. Revascularized pineal tissue showed good survival and pinealocytes revealed structural features of active secretory cells.     

Effect of six weekly transfusions on canine marrow grafts: tests for sensitization and abrogation of sensitization by procarbazine and antithymocyte serum.

We have previously shown that blood transfusions can immunize a dog and lead to rejection of a subsequent marrow graft despite lethal total body irradiation (TBI). Sensitization to histocompatibility antigens induced by two prior transfusions of whole blood could be overcome by a regimen of procarbazine and anti-thymocyte serum (ATS) preceding TBI. The current study investigated a) whether this regimen could abrogate sensitization induced by six weekly transfusions given from days --50 to --15 preceding a marrow graft, and b) whether platelet survival studies and two in vitro tests of immunity could predict marrow graft rejection. All donor-recipient pairs were histoincompatible, unrelated, and of different breed. Twenty-two recipients received platelet concentrate transfusions and eight received whole blood transfusions. Recipients were given 1200 R TBI and a graft of marrow and peripheral blood leukocytes from the transfusion donor on day 0. Three of 15 recipients (20%) given procarbazine, 12.5 mg/kg i.v. on days --8, --6, and --4, and ATS, 0.6 ml/kg subcutaneously on days --7, --5 and --3, Rejected their grafts, whereas 11 of 15 dogs (73%) not given procarbazine and ATS rejected their grafts (p less than 0.01). Serum lymphocytotoxic antibodies, peripheral leukocyte migration inhibition, and in vivo donor platelet recovery and survival were studied in those recipients receiving six weekly transfusions and in 18 other recipients receiving a single donor transfusion 3 months before marrow grafting. No significant correlation was found among these in vitro and in vivo tests of sensitization. Sensitization to marrow grafts was not reliably detected by the presence of cytotoxic antibodies or leukocyte migration inhibition. Platelet survival, however, was positively correlated with the results of marrow grafting in 12 of 15 (80%) evaluable recipients (p approximately 0.15).     

The alleles of theH-13 locus

Seven congenic strains differing from C57BL/10Sn at theH-13 locus have been produced which define fourH-13 alleles. Isografting, exchanging of grafts between sublines, F(1) testing, and linkage testing demonstrate the presence of additionalH genes in four of these strains. The medial survival times (MSTs) of skin grafts fromH-13(a) to unimmunizedH-13(b) recipients ranged from 69 to 83 days. Rejection across all other barriers was extremely weak with most MSTs being > 100 days. Preinjection of donor strain thymocytes caused accelerated rejection of skin grafts fromH-13(a) toH-13(b) mice, but had only minimal effect on skin grafts across other barriers. Rejection ofH-13 incompatible grafts was significantly stronger when the donor and host areH-3(a) than when they wereH-3(b).     

负压封闭引流技术在躯干部皮肤恶性肿瘤切除术中的应用

目的:探讨负压封闭引流技术应用于躯干部皮肤恶性肿瘤切除术的临床效果,并分析其临床应用价值。方法:回顾性分析2012年5月~2015年3月我院收治的11例躯干部皮肤恶性肿瘤患者的临床资料,均予行肿物切除术并同时行自体皮游离移植术,并外用持续负压封闭引流术于植皮术区。结果:所有患者均接受肿物切除+植皮+封闭负压引流技术治疗。9例患者于4~9天后拆除负压材料,皮片全部成活。1例患者治疗后24小时内吸引管堵塞,经术后于24小时内行管路冲洗疏通后再次行负压吸引,于术后5天首次拆除负压材料,见皮片部分成活,部分皮片仍有浮动,血运未明显建立,予以扩大皮片引流孔后再次行负压封闭引流术,术后9天再次拆除负压材料,见皮片成活较好。1例患者治疗4天后出现新鲜渗血经引流管引出,予拆除负压材料,所植皮片下有积血块,有新鲜渗血,予以清创后再次行负压封闭引流术,术后9天再次拆除负压材料,见皮片成活较好。随访半年到3年,所植皮片无破溃,肿瘤无复发。结论:躯干部皮肤恶性肿瘤切除术中,植皮联合封闭式负压引流技术可使所植皮片固定确实,充分引流渗液积血,利于皮片成活,对无法有效包扎固定肢体特殊部位(肩部、臀部、躯干部)等术区植皮提供了有效方法,具有一定的推广应用价值。     

负压创面治疗技术对皮肤移植成活的临床观察及实验研究

目的：采用负压固定移植皮片方法,观察负压创面治疗技术（negative-pressure wound therapy,NPWT）对游离皮片成活的影响,初步探讨微血管形成与皮肤成活之间的关系。方法：采用回顾性研究的方法,对65例皮肤缺损的患者,根据皮肤移植术后皮片固定方法的不同,分为两组,其中Ⅰ组为NPWT治疗组,有35例患者,刃厚游离皮片移植术后行创面负压吸引治疗;Ⅱ组为常规治疗组。有30例患者,刃厚游离皮片移植术后用打包或加压包扎的方式固定。Balb／c小鼠20只,按皮片移植后不同固定加压方式,分为实验组：负压创面治疗技术使用组（10只）,对照组：打包加压组（10只）,于皮片移植术后第5天,大体观察移植皮片颜色、有无水疱、有无皮下积液及质地,计算并比较皮片成活率,以免疫组化染色标记毛细血管内皮,检测皮片中微血管情况。结果：临床观察表明：Ⅰ组术后皮片成活时间平均较Ⅱ组缩短,有统计学差异（P〈0．01）,Ⅰ组术后住院治疗时间平均较Ⅱ组缩短5天,有统计学差异（P〈0．01）,Ⅰ组术后抗生素费用、换药次数及换药费用较Ⅱ组减少,有统计学差异（P〈0．01）。动物实验结果表明：术后第5天,实验组小鼠移植皮片中微血管增生较对照组明显增多（P〈0．05）。结论：与常规打包或加压包扎固定皮片的方式相比,负压创面治疗技术的应用可以缩短皮片成活时间,缩短患者住院治疗时间,减少抗生素的使用及换药次数,促进移植皮片中毛细血管增生,提高皮片成活率。     

负压创面治疗技术对皮肤移植成活的临床观察及实验研究

目的:采用负压固定移植皮片方法,观察负压创面治疗技术(negative-pressure wound therapy,NPWT)对游离皮片成活的影响,初步探讨微血管形成与皮肤成活之间的关系。方法:采用回顾性研究的方法,对65例皮肤缺损的患者,根据皮肤移植术后皮片固定方法的不同,分为两组,其中I组为NPWT治疗组,有35例患者,刃厚游离皮片移植术后行创面负压吸引治疗;II组为常规治疗组,有30例患者,刃厚游离皮片移植术后用打包或加压包扎的方式固定。Balb/c小鼠20只,按皮片移植后不同固定加压方式,分为实验组:负压创面治疗技术使用组(10只),对照组:打包加压组(10只),于皮片移植术后第5天,大体观察移植皮片颜色、有无水疱、有无皮下积液及质地,计算并比较皮片成活率,以免疫组化染色标记毛细血管内皮,检测皮片中微血管情况。结果:临床观察表明:I组术后皮片成活时间平均较II组缩短,有统计学差异(P<0.01),I组术后住院治疗时间平均较II组缩短5天,有统计学差异(P<0.01),I组术后抗生素费用、换药次数及换药费用较II组减少,有统计学差异(P<0.01)。动物实验结果表明:术后第5天,实验组小鼠移植皮片中微血管增生较对照组明显增多(P<0.05)。结论:与常规打包或加压包扎固定皮片的方式相比,负压创面治疗技术的应用可以缩短皮片成活时间,缩短患者住院治疗时间,减少抗生素的使用及换药次数,促进移植皮片中毛细血管增生,提高皮片成活率。     

The effect of heat-killed streptococci on the survival of heart grafts in inbred strains of mice.

Studies with inbred strains of mice revealed that exposure to type 12, Group A, beta-hemolytic streptococci affects the host response to heart grafts implanted in the ear. Intraperitoneal injection of streptococci 10 days before grafting led to curtailed survival of syngrafts without altering the normal rejection time of allografts. Similar sensitization, combined with local injection of streptococci into the graft site at the time of grafting, was followed by rapid rejection of both syngrafts and allografts. The time interval between exposure and grafting was critical. Injection with streptococci 5 days before grafting led to a prolonged survival of allografts and no demonstrable effect on syngrafts. In contrast, injection of streptococci 15 days before grafting did not alter survival of either type of graft. The data indicate heart grafts implanted in the ear may serve as a useful model for the study of the host responses to streptococcal antigens.     

The preservation of the ability of cultured quail wing bud mesoderm to elicit a position-related differentiative response

A previous study showed that grafting wedges of fresh anterior quail wing mesoderm into posterior slits of chick wing buds resulted in the formation of rods and nodules of cartilage in a high percentage of cases (B. Carlson, 1983, Dev. Biol. 101, 97-105). The purpose of the present study was to determine if a similar response could be elicited by grafting pieces of mesoderm that had been cultured in vitro. When pieces of 1-day cultured anterior mesoderm from stage 17-24 donors were grafted into standard posterior slits of chick wing buds, the percentages of supernumerary structures differed little from those which formed after the grafting of pieces of fresh mesoderm. In a time series, grafts of stage 22-23 anterior mesoderm which had been cultured for 1-4 days retained the ability to form cartilage after being grafted into posterior locations. A time series showed that the duration of this retention was longer in cultured mesoderm than it was in mesoderm that remains in the donor wing bud.     

GRAFT UNION FORMATION IN DOUGLAS-FIR

Greenhouse-grown Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco) graft unions were examined between 2 and 84 days after grafting. Room temperature was maintained at 60-70 F throughout the growing season. In most respects grafts of Douglas-fir followed development patterns previously reported for spruce and pine grafts, but specific differences were noted in contributing cell types, time of formation, and mode of healing. The time interval from first occurrence to occurrence in 80% of the grafts is as follows: contact layers, 2 days; callus bridges, 10-14 days; periderm, 10-17 days; cambia, 17-23 days. Callus bridges were generally of secondary phloem or cortex origin. Callus lignification began along cut edges of the union at 14 days and was completed across the entire length of the union by 17 days. Lignified tracheids were continuous across union zones at 35 days. When proper grafting techniques were used, all tissue systems necessary for a successful union were present 35 days after grafting. Poor grafting techniques at times retarded cambium formation for 3 months or more.     

Application of Electric Coupling on Basic Research of Grafting

Electrical resistance changes in concert with the histological changes during the development of graft union. Four phases of change in electrical resistance of the compatible graft were demonstrated. Phase Ⅰ: There was a slight decrease of the electrical resistance, lasting about several hours. Phase Ⅱ The resistance values increase rapidly, lasting for two days or so. Phase Ⅲ: In the next four days, the resistance values decreased progressively. Phase Ⅳ: The resistance values levelled off and approximated equal to the level before grafting. In contrast to the compatible grafts, three types of resistance change were observed in the incompatible graft: (1) The resistance values increased rapidly on the first day after grafting but only slightly afterwards decreased. (2) The electrical resistance rose rapidly to its peak on the first day after grafting and decreased rapidly in 2--3 days followed by steady increase again. (3) The change of resistance follows the track as in (2) for 2--3 days and then increased rapidly afterwards. The results suggested that measurement of electrical resistance could be useful tool for the detection of graft compatibility/incompatibility in commercial practice and basic research of plant grafting.     

The effects of macrophage-mediated inflammatory response to the donor site on long-term retention of a fat graft in the recipient site in a mice model

Inflammatory responses mediated by macrophages play a role in tissue repair. However, it is unclear whether the repair in the donor site after liposuction would have any effects on fat graft retention in the recipient site. This study is designed to evaluate the effects of a macrophage-mediated inflammatory response in donor sites on long-term retention of fat grafting. In this study, mice were randomly divided into two groups. One underwent simulated liposuction, called the fat procurement plus grafting (Pro-Grafting) group, and the other underwent sham surgery, called the fat grafting only (Grafting Only) group. The prepared fat (0.3 ml each) was engrafted and cellular events over a 90-day period were assessed. We found macrophages were infiltrated into adipose tissue at the recipient site in the Grafting Only group within 7 days and the repair essentially completed within 30 days. By contrast, few macrophages infiltrated the recipient site in the Pro-Grafting group within 7 days and the entire remodeling process took 30 days longer in the Pro-Grafting than the Grafting Only group. Moreover, C-reactive protein levels were immediately upregulated after surgery, and the inflammatory factors' expression was higher at the donor rather than the recipient site. However, the repair processes and the long-term retention rate became normal when the adipose tissue was grafted after the donor site did not require macrophages for repair. Therefore, we suggest higher inflammatory factors promote macrophage infiltration and the adipose tissue regeneration process at the donor site. This process is delayed at the recipient site, which may affect long-term retention of fat grafts.     

Description of and treatment to inhibit the rejection of human split-thickness skin grafts by congenitally athymic (nude) rats

Gradual rejection of topically engrafted human split-thickness skin grafts (HSTSG) occurred in greater than 90% of congenitally athymic (nude) rats between 21 and 42 days of grafting. Engraftment and rejection of HSTSG is accompanied by a partial restoration of some cell-mediated immune components, the mixed lymphocyte response and lysis of human target cells. Histologic features of the rejection process were those seen in a host-versus-graft reaction. Immunofluorescent analysis of skin undergoing rejection demonstrated IgG at the basement membrane zone in most grafts. Nude rats rejecting HSTSG had circulating IgG which bound to the basement membrane zone and blood vessels of human skin. Nude rats treated with cyclosporine injections for 21 days had an enhanced survival of HSTSG, 120 or more days.     

Prolongation of skin xenograft survival with modified cultured fibroblasts

Cyclosporine A, one of the most potent immunosuppressive drugs, mediates some of its immunosuppressive and nephrotoxic effects by enhancing transforming growth factor-beta secretion and receptor expression. In this experimental study, the effect of cyclosporine pretreatment of cultured dermal fibroblasts on xenogeneic tissue rejection after microimplantation beneath skin grafts was investigated. The effects of the site-specific immunosuppressive strategy on skin xenograft survival were tested. Because the skin is an immunological indicator of the rejection of composite tissue allografts, it was considered that this strategy could be used as a supportive therapy for composite tissue allotransplantation in the future. In the first stage of the study, fibroblast cultures obtained from skin biopsy samples from five rats were treated with different single doses of cyclosporine (100 to 3000 ng/ml), and transforming growth factor-beta levels were measured in culture supernatants after 72 hours. In the second stage, 60 Sprague-Dawley rats were divided into six groups, as follows. For group I (sham), only the standard grafting procedure was performed. For group II, after the standard grafting procedure, rats were treated with intraperitoneal injections of 30 mg/kg (n = 5) or 10 mg/kg (n = 5) cyclosporine for 10 days. For group III, cultured fibroblasts obtained from skin biopsy samples from rats were treated with 100 or 500 ng/ml cyclosporine, and the cells were collected by light trypsinization and centrifugation after 72 hours. After the standard skin grafting procedure, modified fibroblasts were implanted between the graft and the recipient bed with a Pasteur pipet. For group IV, the same procedures as for group III were performed and then rats were treated with 10 mg/kg cyclosporine, administered intraperitoneally, for 10 days. For group V, in addition to standard grafting, unmodified fibroblasts (not treated with cyclosporine) were implanted between the graft and the recipient bed. For group VI, the same procedures as for group V were performed and then rats were treated with 10 mg/kg cyclosporine, administered intraperitoneally, for 10 days. The rejection process was observed macroscopically, and statistical significance was determined with the Mann-Whitney test (p < 0.01). Graft survival times were significantly prolonged in groups III and IV, compared with groups I, II, V, and VI (p < 0.001). No difference between groups III and IV was observed. The novel finding of this investigation is that xenogeneic skin graft survival times could be prolonged with microimplantation of cyclosporine-treated cultured fibro-blasts.     

Some factors affecting the spread of mycoplasma in plants

Studying the spread of mycoplasma, the causal agent of potato witches' broom disease, in tomato plants after grafting with infectious grafts ofNicotiana glauca Grah., we found that after 9 days of graft symbiosis a hundred per cent infection occurred, whereas with infectious grafts ofSolanum lycopersicum this took place after 16 days. The first symptoms of the disease were manifested on tomato plants 21 days after grafting with infectiousNicotiana glauca grafts and 28 days after grafting with infectious tomato grafts. The results obtained present evidence for the possible preference of tomato plants for mycoplasma.     

Long-term survival of corneal allografts is dependent on intact CD1d-reactive NKT cells

BALB/c mice that tolerate the allogeneic grafts develop allogeneic-specific anterior chamber-associated immune deviation. Because CD1d-reactive NKT cells are essential for anterior chamber-associated immune deviation, we postulated that the survival of C57BL/6 (B6) cornea graft in BALB/c mice was also dependent on CD1d-reactive NKT cells. The B6 corneal graft rejection rate in BALB/c vs Jalpha281 knockout (KO) mice, which lack NKT cells, was measured. While there were no difference in the early phase of rejection, the survival rates at 12 wk after grafting for BALB/c and Jalpha281 KO mice were 50 and 0%, respectively. Because anti-CD1d mAb abrogated the corneal graft survival in the wild-type mice we concluded that CD1d-reactive NKT cells were essential for graft survival. Moreover, allospecific T regulatory (Tr) cells correlated with acceptance of B6 grafts in BALB/c mice, and the adoptive transfer of these allospecific Tr cells to Jalpha281 KO mice allowed a 50% survival rate of B6 cornea grafts. In conclusion, CD1d-reactive NKT cells are required for induction of allospecific Tr cells and are essential for survival of corneal allografts. Mechanisms that contribute to cornea graft acceptance may lead to new therapies for improvement in graft survival in high-risk corneas and other transplanted tissues and grafts.     

Graft establishment between compatible and incompatible Prunus spp

Graft compatibility has been studied in apricot (prunus armeniacaL.) grafted on Prunus cerasifera Ehrh. Two apricot cultivars,one compatible and one incompatible on this rootstock, wereselected for this study. In these species incompatibility isonly manifested by tree breakdown at a late phase of the tree'slife. The process of graft union formation was observed forthe first month following grafting. No differences were foundeither in the process of healing or in its kinetics. Thus, callusproliferation, callus differentiation and vascular connectionsare established in the same way and at the same time in bothcompatible and incompatible grafts. However, clear differencesexist in the level of differentiation of the callus produced.While in compatible grafts, callus quickly differentiates intocambium and vascular tissue, in incompatible grafts this differentiationis not complete and a portion of the tissue evolves into a parenchymatoustissue that coexists with the differentiated vascular tissue. Key words: Graft, Prunus, compatibility     

嫁接植株形成过程中接合部组织学和生长素含量的变化

本文应用植物激素间接酶联免疫技术（ＥＬＩＳＡ）,第一次定量检测了嫁接植株形成过程中生长素（ＩＡＡ）的动态变化。结果表明：嫁接植株发育的前期,亲和性与非亲和性组合其ＩＡＡ含量的变化相似。在后期,不亲和组合ＩＡＡ含量急骤减少,而亲和性的组合在第八天即维管束桥分化形成的这一天,可见到ＩＡＡ高峰值的出现。     

嫁接植株形成过程中接合部组织学和生长素含量的变化

本文应用植物激素间接酶联免疫技术(ELISA),第一次定量检测了嫁接植株形成过程中生长素(IAA)的动态变化。结果表明：嫁接植株发育的前期,亲和性与非亲和性组合其IAA含量的变化相似。在后期,不亲和组合IAA含量急骤减少,而亲和性的组合在第八天即维管束桥分化形成的这一天,可见到IAA高峰值的出现。