黄芪注射液对病毒性心肌炎患者血清胶原前肽的影响

目的研究黄芪注射液对心肌炎患者心脏胶原代谢的影响。方法83例临床心肌炎患者与20例健康志愿者作为研究对象,心肌炎患者予黄芪注射液治疗2周,在治疗前、后以ELISA法分别检查其血清中I型胶原羧基末端肘(ICTP)和Ⅲ型前胶原氨基末端肽(PⅢNP)浓度。健康志愿者不予用药作为空白对照。结果心肌炎患者血清胶原前肽水平较健康志愿者明显增高。经连续2周黄芪注射液治疗后,心肌炎患者ICTP和PIIINP血清浓度均下降,其中ICTP接近正常水平。结论黄芪注射液治疗心肌炎疗效可靠,黄芪注射液可能通过干预胶原代谢,抑制胶原过度增生而改善患者预后,减少并发症的发生。     

The Effect of Carboxyl-Terminal Propeptide of Type I Collagen (c-Propeptide) on Collagen Synthesis of Preosteoblasts and Osteoblasts

Recently we found that the carboxyl-terminal propeptide of type I collagen (c-propeptide) is a major secretory protein of osteoblasts. Mature osteoblasts secreted 64 nM c-propeptide, and it was reported that 40 nM c-propeptide inhibited collagen synthesis at 80% of the control level. In this study, we investigated the effect of c-propeptide on collagen synthesis of preosteoblasts and osteoblasts, and found that preosteoblasts downregulated collagen synthesis by 40 nM c-propeptide, but osteoblasts were not affected by the same condition. When the binding activities of c-propeptide for preosteoblasts and osteoblasts were compared, osteoblasts showed weak affinity to c-propeptide compared with preosteoblasts, and the number of receptors for c-propeptide decreased in osteoblasts. These results imply that a decrease of receptors in osteoblasts might reduce the sensitivity of osteoblasts to c-propeptide. Received: 6 August 1999 / Accepted: 9 May 2000 / Online publication: 22 September 2000     

Characterization of Demineralized Bone Matrix-Induced Osteogenesis in Rat Calvarial Bone Defects: III. Gene and Protein Expression

Our previous studies of rat cranial defect repairs after the implantation of demineralized bone matrix (DBM) have demonstrated that healing occurs initially and principally by the direct induction and proliferation of osteoblasts derived principally from resident mesenchymal stem cells of the dura, and to a lesser extent by resident mesenchymal stem cells of the connective tissues beneath the skin flap. A small amount of cartilage is also synthesized after the direct process of ossification occurs. To further confirm the molecular phenotypes of the repair cells in rat cranial defects, the present study evaluated mRNA expression and synthesis of collagens I, II, and X and osteocalcin in the DBM-induced repair tissue by Northern blot analyses, autoradiography after in vivo ³H-proline labeling of collagen, and immunohistochemistry. The results demonstrated that osteocalcin mRNA appeared in small amounts by day 4 and continued to increase over the experimental period. Much lesser quantities of collagen types II and X mRNAs appeared by day 6 and day 8, respectively. Collagen type I mRNA was present at all times examined but its expression significantly increased by day 5. Autoradiographic and immunohistochemical studies showed that type II collagen was not detected whereas type I collagen was synthesized on days 3–5. The data provide definitive molecular evidence confirming that the initial and by far the major pathway of cranial defects repair induced by implantation of DBM is by the direct induction of resident mesenchymal stem cells to osteoblasts and the direct formation of bone, which is spatially and temporarily distinct from the later formation of cartilage. Received: 30 November 1999 / Accepted: 21 March 2000     

Mineralization and alkaline phosphatase activity in collagen lattices populated by human osteoblasts

Adult human osteoblastic cells were grown in a native type I collagen gel. Proliferation and viability analyses showed that cells rapidly stopped dividing and became blocked in the G0G1 phase (91% on day 13). Carboxyfluorescein diacetate cell staining and flow cytometry showed that osteoblasts were viable for the first 16 days and then viability decreased (58% viable cells on day 22). Osteoblasts were able to retract the matrix. Betaglycerophosphate (βGP) stimulated the deposition of mineral particles in the collagen network, and electron probe microanalysis showed that they were principally calcium and phosphorus, with a Ca/P ratio of about 1.7. Various times of βGP supply were tested. We compared 10 mM βGP added only once at day 0, or continuously from day 0, day 8, or day 21. Mineralization was observed in conditions where βGP was added at day 0. Furthermore, 10 mM βGP added once during gel preparation was sufficient to induce mineralization with mineral accumulation up to day 15 whereas the speed of the gel contraction decreased. In every condition, cultures expressed high alkaline phosphatase (ALP) levels as early as day 3, which decreased afterwards. These kinetics might explain why the other conditions did not prove favorable to the mineralization process. The model was used to study the influence of blocking gel retraction. Blocking retraction delayed the ALP activity decrease, but had no effect on mineralization. In conclusion, human adult osteoblasts cultured in native collagen gel stopped proliferation and underwent mineralization very early. This model should be used to investigate the influence of effectors on the early stages of culture. Received: 15 October 1997 / Accepted: 1 July 1999     

糖尿病视网膜病变与血清Ⅳ胶原、层粘连蛋白的关系

目的 探讨糖尿病视网膜病变(DR)与血清IV胶原(IVC)、层粘连蛋白(LN)之间的关系。 方法 143例 2型糖尿病患者作为病例组,并依据视网膜病变情况分为三组:无视网膜病变组(NDR)58例,背景期视网膜病变组(BDR)47例,增殖期视网膜病变组(PDR)38例。同时设健康体检者55人为对照组。采用放射免疫法测其血清IVC、LN的含量。所有检测者均测定空腹血糖、糖化血红蛋白、血脂、尿白蛋白排泄率、体重指数,分析病例组血清IVC、LN水平与视网膜病变的关系。结果 (1)血清IV-C水平:三组糖尿病患者明显高于对照组(P<0.01)。病例组间比较,PDR组高于BDR组,BDR组高于NDR组,差异均有非常显著意义(P<0.01)。(2)血清LN水平:BDR组、PDR组高于对照组,差异均有非常显著意义(P<0.01),NDR组与对照组比较,增高不明显,无统计学意义(P>0.05)。病例组间比较,PDR组高于BDR组,BDR组高于NDR组,差异均有非常显著意义(P<0.01)。(3)Logistic多元回归分析显示,血清IVC、LN、病程是DR发病的独立风险因素。结论 血清IVC、LN与DR发生发展密切相关,联合检测血清IVC、LN水平,有助于估计DR病变程度及预后。     

Tissue engineered artificial skin composed of dermis and epidermis

Abstract: We made an artificial skin comprised of a stratified layer of keratinocytes and a dermal matrix with a type I collagen containing fibroblasts. In this work, we showed keratinocyte behavior under primary culture, gel contractions varying with concentration of collagen solution, and cell growth plots in the collagen gel. The optimum behavior of dermal equivalent could be obtained using 3.0 mg/ml collagen solution and attached gel culture. The attached gel culture had a jumping effect of growth factor on cell growth at the lag phase. To develop the artificial skin, 1× 10⁵ cells/cm² of keratinocytes were cultured on the dermal equivalent at air-liquid interface. Finally, to overcome the problem that artificial skin of collagen gel was torn easily during suturing of grafting, we prepared histocompatible collagen mesh and attached the mesh to the bottom of the gel. Cultured artificial skins were successfully grafted onto rats.     

Ultrastructural organization of proteoglycans and fibrillar matrix of the tectorial membrane

The molecular and supramolecular structure of the tectorial membrane (TM) was studied by transmission electron microscopy (TEM). Collagen (type A) fibrils in the TM were found associated with proteoglycans (PGs) and type B fibrils. Most PGs were orthogonally oriented and attached D-periodically to collagen fibrils. Computer averaged projections of PG particles and linear aggregates of PGs in crystalline arrays, stained with Cuprolinic blue, showed an elongated, electron-dense structure 50–65 nm in length and 10 nm in width. Image analysis of type B fibrils showed that they are constructed of globular domains arranged with a periodicity of 12–14 nm. Each globular domain contains two thin ‘arms', extended in opposite directions, which contact the ‘arms' of adjacent fibrils. Numerous type B fibrils were found between collagen fibrils. They are attached to adjacent collagen fibrils by the ‘arms' of their globular domains. An association of type B fibrils and PGs with collagen seems to result in the local ordered arrangement of the TM matrix. A hypothetical model of the TM matrix supramolecular structure is presented.     

吡啶-2,6(1H,3H)二酮生物碱对ADP、AA、Collagen诱导的兔血小板聚集的影响

目的观察吡啶－２，６（１Ｈ，３Ｈ）二酮生物碱（ＳＨ１）对ＡＤＰ、ＡＡ、Ｃｏｌｌａｇｅｎ诱导的兔血小板聚集的影响。方法用比浊法测定了ＳＨ１体外对兔血小板聚集的影响。结果ＳＨ１对３种诱导剂的最大抑制率分别为６２．１６％、４５．２５％、５３．６７％。大剂量组明显加快ＡＤＰ诱导的兔血小板聚集后的解聚速度。ＳＨ１显著延长Ｃｏｌａｇｅｎ的诱导起聚时间。结论ＳＨ１０．８～４．０ｍｍｏｌＬ－１范围内明显抑制ＡＡ、ＡＤＰ、Ｃｏｌａ－ｇｅｎ诱导的兔血小板聚集。其抑制作用有明显的量效关系。其机制待进一步研究。     

藻鳖软肝汤对实验性肝纤维化大鼠作用的研究

目的：研究藻鳖软肝汤抗纤维化的作用。方法：用CCl4复合因素制备大鼠肝纤维化模型，同时用藻鳖软肝汤治疗，用生理盐水作对照，观察其对肝功能及纤维化指标的影响。结果：藻鳖软肝汤对肝纤维化大鼠，有显著降低血清透明质酸（HA）、Ⅲ型前胶原（PCⅢ）、Ⅳ型胶原（Ⅳ-C）；明显改善肝功能的作用。结论：藻鳖软肝汤具有确切的抗纤维化作用。     

牛磺酸抑制新生大鼠心肌重塑的体外研究

目的 通过牛磺酸对新生大鼠心肌成纤维细胞增殖和胶原合成及对I／Ⅲ型胶原比值的影响，从细胞水平探讨其在体外抑制心肌重塑的作用。方法 在培养的心肌成纤维细胞中加入不同浓度的牛磺酸，用羟脯氨酸法测定胶原量，MTT比色法检测细胞增殖情况，SDS-PAGE电泳测胶原I／Ⅲ型比值。结果 100mmol／L、200mmol／L牛磺酸用药24h和48h及50mmol／L用药72h后能明显抑制心肌成纤维细胞的增殖及胶原的合成，I／Ⅲ型胶原比值明显下降。结论 牛磺酸能通过抑制心肌成纤维细胞增殖和胶原合成，降低I／Ⅲ型胶原比值起到抗心肌重塑的作用。