KM－1d突变株小鼠的模型建立及遗传分析

在饲养经剖腹产净化后的ＫＭ种群时,我们发现了一些后肢畸形的动物、通过挑选后已成为一个稳定遗传的突变株,由于此群体产生的后代１００％均为后肢畸形动物,因此可以认定为１ｄ／１ｄ纯合子动物。命名为ＫＭ—１ｄ小鼠。将ＫＭ－１ｄ纯合子动物与近交系ＤＢＡ／２小鼠杂交得到Ｆ１代,再经Ｆ１代互交或与双亲回交。通过对后代的形态学观察及遗传方式的分析,证明为常染色体隐性单基因遗传。另外,对１３８只ＫＭ－１ｄ畸形小鼠进行解剖观察还发现有４２％的动物在骨骼畸形的同时伴有左侧泌尿系统畸形。因此可以认定ＫＭ－１ｄ小鼠是一种患有骨－肾先天性畸形综合症的动物模型。     

侵袭性牙周炎伴错牙合畸形患者牙周-正畸联合治疗前后血清SAA、leptin的变化及与牙周指标和辅助性T细胞亚群的相关性分析

摘要 目的：探讨侵袭性牙周炎伴错牙合畸形患者牙周-正畸联合治疗前后血清淀粉样蛋白A（SAA）、瘦素（leptin）的变化及与牙周指标和辅助性T细胞（Th）亚群的相关性。方法：选择2020年6月-2022年8月解放军总医院京中医疗区黄寺门诊部口腔科收治的80例侵袭性牙周炎伴错牙合畸形患者（牙周炎组）和65例于口腔门诊检查的健康志愿者（对照组）。所有患者均接受牙周-正畸联合治疗,治疗前后分别检测血清SAA、leptin水平以及外周血中Th1、Th2、Th17细胞占比,并评估牙周指标变化。Pearson相关性分析血清SAA、leptin水平与牙周指标以及外周血中Th1、Th2、Th17细胞占比的相关性。结果：牙周炎组治疗前血清SAA、leptin水平,外周血Th1、Th17细胞占比,出血指数（SBI）、菌斑指数（PLI）、附着丧失（AL）、牙周探诊深度（PD）高于对照组（P＜0.05）,外周血Th2细胞占比低于对照组（P＜0.05）。牙周炎组治疗后血清SAA、leptin水平,外周血Th1、Th17细胞占比,PLI、SBI、AL、PD较治疗前降低（P＜0.05）,外周血Th2细胞占比较治疗前增高（P＜0.05）。牙周炎组血清SAA、leptin与PLI、SBI、AL、PD,外周血Th1、Th17细胞占比呈正相关,与外周血Th2细胞占比呈负相关（P＜0.05）。结论：侵袭性牙周炎伴错牙合畸形患者血清SAA、leptin水平增高,经牙周-正畸联合治疗后下降,高水平SAA、leptin与牙周组织破坏程度以及Th亚群紊乱有关,检测血清SAA、leptin水平可评估侵袭性牙周炎牙周组织破坏程度以及细胞免疫状态。     

Stem deformity inPinus radiata plantations in south-eastern Australia

Plantations of radiata pine (P. radiata D.Don) on soils previously under legume based pastures have a high incidence of stem deformity compared with forest soils. A comparison of soil properties and tree nutrition of 5 to 7 year-old radiata pine on former pastures in the first part of the study showed that stem deformity was strongly correlated with mineralisation of soil N and in particular with nitrification. Other soil properties that have changed as a result of pasture improvement, e.g. pH, available P and Mn, were only partially correlated with stem deformity. In the second part of the study, the role of N availability and other soil properties in the expression of deformity was further investigated in a separate field experiment on soils formerly under native eucalypt forest, tobacco cropping, and improved pasture. Young radiata pine plantings were treated with lime, phosphorus, and nitrogen applied as urea and sodium nitrate. Liming increased soil pH by around 1.5 units, raised exchangeable Ca²⁺ and decreased available Mn. Soil mineral N content was only marginally affected by liming. Superphosphate increased soil available P and raised levels of P in foliage. Changes in soil pH, availability of P, Mn, and B did not affect growth or stem deformity at any of the sites. In contrast, application of N fertilisers at 200 and 600 kg N ha^-1 increased mineral N content and stimulated nitrification, particularly at the forest site. The high rate of N fertiliser increased basal area at the forest site by 45%, but also raised the level of stem deformity from 12% to 56%. At the tobacco and pasture sites, this treatment did not increase growth and did not significantly raise stem deformity above the already high basic level of deformity (63%). Implications of stem deformity in young plantations of radiata pine on potential utilisation later in the rotation are discussed.     

Effects of micronutrients on the growth form of two families of Pinus radiata (D.Don) seedlings

Micro-element deficiencies have been implicated in the development of poor stem form in Pinus radiata plantations. Cu, Mn and B have been implicated in previous studies as influencing tree form and/or the process of lignification in plants. Therefore an experiment was initiated to examine the individual and interactive effects exhibited by these trace elements on stem form and lignification.The investigation showed that Cu deficiency reduced both lignification and stem form in seedlings. Mn, competing with Cu for uptake, enhanced the symptoms of Cu deficiency. Boron addition did not alleviate the expression of deformity or increase levels of lignification. There was a clear family influence on the development of seedling stem form in response to variations in mineral nutrition.     

Triploid Atlantic salmon Salmo salar have a higher dietary phosphorus requirement for bone mineralization during early development

The effect of a dietary phosphorus regime in freshwater on vertebra bone mineralization was assessed in diploid and triploid Atlantic salmon, Salmo salar. Fish were fed either a low phosphorus (LP) diet containing 10.5 g kg⁻¹ total phosphorus or a normal phosphorus (NP) diet containing 17.4 g kg⁻¹ total phosphorus from ∼3 to ∼65 g (day 126) in body weight. Two further groups were fed the NP diet from ∼3 g in body weight, but were then switched to the LP diet after 38 (∼10 g in body weight) or 77 (∼30 g in body weight) days. Growth, vertebral ash content (% ash) and radiologically detectable vertebra pathologies were assessed. Triploids were initially smaller than diploids, and again on day 77, but there was no ploidy effect on days 38 or 126. Vertebral ash content increased with increasing body size and those fish fed the NP diet had higher vertebral ash content than those groups fed the LP diet during the intervening time period, but this diet effect became less apparent as fish grew, with all groups having relatively equal vertebral ash content at termination. In general, triploids had lower vertebral ash content than diploids on day 38 and this was most evident in the group fed the LP diet. On day 77, those triploids fed the LP diet during the intervening time period had lower vertebral ash content than diploids. At termination on day 126, the triploids had the same vertebral ash content as diploids, irrespective of diet. There was a ploidy × diet interaction on vertebral deformities, with triploids having higher prevalences of fish with ≥1 deformed vertebra in all dietary groups except continuous NP. In conclusion, between days 0 and 77 (3–30 g body size), triploids required more dietary phosphorus than diploids in order to maintain similar vertebral ash content. A possible link between phosphorus feeding history and phosphorus demand is also discussed.     

Expression of TGFβ3 RNA during chick embryogenesis: a possible important role in cardiovascular development

Formin was originally isolated as the gene affected by the murine limb deformity (ld) mutations, which disrupt the epithelial-mesenchymal interactions regulating patterning of the vertebrate limb autopod. More recently, a rapidly growing number of genes with similarity to formin have been isolated from many different species including fungi and plants. Genetic and biochemical analysis shows that formin family members function in cellular processes regulating either cytokinesis and/or cell polarisation. Another common feature among formin family members is their requirement in morphogenetic processes such as budding and conjugation of yeast, establishment of Drosophila oocyte polarity and vertebrate limb pattern formation. Vertebrate formins are predominantly nuclear proteins which control polarising activity in limb buds through establishment of the SHH/FGF-4 feedback loop. Formin acts in the limb bud mesenchyme to induce apical ectodermal ridge (AER) differentiation and FGF-4 expression in the posterior AER compartment. Finally, disruption of the epithelial-mesenchymal interactions controlling induction of metanephric kidneys in ld mutant embryos indicates that formin might function more generally in transduction of morphogenetic signals during embryonic pattern formation. Received: 24 September 1998 / Accepted: 30 September 1998     

Expression of osteoblast marker genes in rat calvarial osteoblast-like cells, and effects of the endocrine disrupters diphenylolpropane, benzophenone-3, resveratrol and silymarin

Compelling evidence indicates that some endocrine disrupters (EDs), acting as selective estrogen-receptor modulators, interfere with osteoblast differentiation and function. Hence, we investigated whether four EDs [bisphenol-A (BSP), benzophenone-3 (BP3), resveratrol and silymarin] affect differentiation and growth of rat calvarial osteoblast-like (ROB) cells in primary in vitro culture. ROB cells were cultured for up 30 days in a medium supplemented with fetal calf serum (FCS), and conventional RT-PCR detected the expression of collagen-1alpha and osteonectin mRNAs through the entire culture period. Real time-PCR demonstrated that at days 2 and 7 of culture the expressions of collagen-1alpha and osteonectin were very low, and underwent a 192- and a 334-fold increase, respectively, at day 21 of culture. In contrast, osteocalcin expression remained unchanged from days 2 to 21 of culture. EIA showed that ROB cells secreted sizeable amounts of osteocalcin and osteopontin between days 13 and 15 of culture. EDs were added at day 13 of culture at concentrations ranging from 10(-10) to 10(-6) M, being the culture medium deprived of FCS, and their effects were tested 48 h later. None of EDs was found to affect osteocalcin and osteopontin secretion from ROB cells, suggesting that their effects were tested at a relatively earlier stage of culture, when ROB cell differentiation into osteoblats is not fully accomplished, and/or the presence of estrogens contained in FCS is needed for EDs to exert their osteoblast-differentiation modulating action. BSP and BP3, but not resveratrol and silymarin, decreased proliferative activity of cultured ROB cells, a cytotoxic effect conceivably independent of their estrogen-receptor modulating activity.     

Use of human perivascular stem cells for bone regeneration

Human perivascular stem cells (PSCs) can be isolated in sufficient numbers from multiple tissues for purposes of skeletal tissue engineering. PSCs are a FACS-sorted population of 'pericytes' (CD146+CD34-CD45-) and 'adventitial cells' (CD146-CD34+CD45-), each of which we have previously reported to have properties of mesenchymal stem cells. PSCs, like MSCs, are able to undergo osteogenic differentiation, as well as secrete pro-osteogenic cytokines. In the present protocol, we demonstrate the osteogenicity of PSCs in several animal models including a muscle pouch implantation in SCID (severe combined immunodeficient) mice, a SCID mouse calvarial defect and a femoral segmental defect (FSD) in athymic rats. The thigh muscle pouch model is used to assess ectopic bone formation. Calvarial defects are centered on the parietal bone and are standardly 4 mm in diameter (critically sized). FSDs are bicortical and are stabilized with a polyethylene bar and K-wires. The FSD described is also a critical size defect, which does not significantly heal on its own. In contrast, if stem cells or growth factors are added to the defect site, significant bone regeneration can be appreciated. The overall goal of PSC xenografting is to demonstrate the osteogenic capability of this cell type in both ectopic and orthotopic bone regeneration models.