检测结直肠癌患者血清巨噬细胞集落刺激因子(M-CSF)的含量并探讨其临床意义。方法:采用酶联免疫吸附分析法(ELISA)对62例经病理证实的术前结直肠癌患者、40例结直肠良性病患者和40例健康体检者血清M-CSF水平进行检测。结果:结直肠癌患者血清M-CSF水平明显高于结直肠良性病患者和健康体检者(Plt;0.01);结直肠癌患者血清M-CSF水平与肿瘤分期、淋巴结转移及远处转移有关(Plt;0.05),与性别、年龄、分化程度不相关(Pgt;0.05)。结论:M-CS与结直肠癌的肿瘤分期、淋巴结转移及远处转移有关,可能是一个判断结直肠癌预后的生物学指标。
OBJECTIVE: To sum up the clinical results of allogeneic humeral transplantation with vascular anastomosis, and evaluate the clinical significance. METHODS: From September to November 1979, 1 case with humeral shaft defect of 10 cm in length and 2 cases with tibia shaft defect of 12 cm in length were repaired by allogeneic humeral transplantation with vascular anastomosis. Azathiopurine and prednisone were applied for 3 months postoperatively. All cases were followed up for 20 years. RESULTS: Case 1 recovered well with good bone union and reconstruction after operation, and could work normally. In case 2, five chronic rejections were occurred during 3 years after operation, and recovered after treatment, the allograft bone was fractured after 2 years of operation, and unioned by autogeneous iliac bone transplantation. In case 3, the distal part of allograft bone was fractured after 46 months, and unioned by autogeneous iliac bone transplantation. The middle part of allograft bone was non-unioned after 20 years follow-up in case 3, but the patient could still work normally. CONCLUSION: The clinical results of allogeneic long bone transplantation can be improved by rational tissue matching test, application of effective immunosuppressive drugs in a certain period according to the principles of modern transplantation immunology.
Objective To study the differentiation of the human osteoblasts during the construction of the tissue engineered periosteum with the human acellular amniotic membrane(HAAM).Methods To construct the tissue engineered periosteum (n=60) with HAAM, the human fetal osteoblasts were used. The fetal osteoblasts were cultured for 2, 4, 6, 8, and10 days, and then their total RNA was extracted, which were reversely transcripted to cDNA. The realtime PCR analysis was used to reveal Cbfal and Osterix, and the cycle threshold (Ct) was also measured. The simplycultured osteoblasts were used as the control group (n=20).Results The expression of Cbfa1 was higher in the experimental group on the 2nd day when compared with that on the 4th, 6th, and 8th day(P＜0.05). The same result existed on the 10th day when compared with that on the 4th and 8th day. The expression of Osterix increased and was highest on the 8th day when compared with the other results(P＜0.05). Both of the 2 gene expressions were decreased in the control group when compared with those in the experimental group, but with no significant difference(P＞0.05). Conclusion Cbfa1 and Osterix can be normally expressed by the osteoblasts after their integration with HAAM. As a scaffold, HAAM can be used to keep the osteoblast phenotype and differentiation with an osteoconductive ability. Such a cell-scaffold complex may provide a basis for the osteogenesis.
OBJECTIVE: To explore the SV40-mediated immortalization, the related factors and their roles in cell immortalization. METHODS: The original articles about cell immortalization and replicative senescence in recent decade were reviewed. RESULTS: Cell immortalization was a multifaceted phenomenon, it was involved in viral DNA integration, activation of telomerase, inactivation of growth suppressors, and so on, and their roles were closely related. CONCLUSION: The research on cell immortalization may be expected to provide important insights into a broad range of cellular biological phenomenon, and the immortalized cells can play important roles in the research of cell engineering and tissue engineering as standard cells.
Objective To investigate the effect of WO-1 on the proliferation and differentiation of human embryonic osteoblasts (HEO) and to provide research methods of bone tissue engineering. Methods HEO were isolated from periosteum and calvaria and then cultrued in vitro. The doseeffect relationship between WO-1 concentration and biological effect of HEO was evaluated by growth curve and 3 H-TdR count. The effect of WO-1 on cell activity and proliferation was investigated by cloning efficiency,cell cycle analysis was determined by flow cytometer and morphological was examined through transmission electron microscope. Moreover, the effect of WO-1 on osteoblastic function was evaluated at protein and mRNA levels by ALP activity, 3 H-proline incorporation, osteocalcin secretion (RIA) and mRNA expression of type I collagen and osteocalcin (RT-PCR). Results The proliferation of HEO was inhibited in high concentration of WO-1,while it was promoted in low concentration of WO-1. The optimal dose was 8 μg/ml, and there was dose-effect relationship in the certain range of WO-1 concentration (0.25 μg/ml to 8 μg/ml). In 8 μg/ml of WO-1, the cloning efficiency and cloning volume of HEO were inereased, population doubling time was decreased.All indexes of ostoblastic function including ALP activity, type I collagen synthesis and osteocalcin secertion were inereased, the more sufficed cell organs were observed under transmission electron microscope than control group(P＜0.05). Conclusion WO-1 can promote the cell activity and proliferation of HEO cultured in vitro inlow concentration, enhance the synthesis of extracellular mamix, such as type Icollagen and osteocalcin, and accelerate the mineralization of osteoid. WO-1 can be used as a stimulant of proliferation and differentiation of HEO in the research of bone tissue engineering, which provide the theoretical basis in clinical application.
Objective To develop a new tissue engineering bone material which has an antiinfective function. Methods Collagen loaded bio-derived bone material was made by using type I collagen and allograft bone. WO-1was absorbed to collagen loaded bio-derived bone, then the morphological feature of the new bone material was observed by scanning electronic microscopy.3 H tetracycline was diluted by WO-1 solution, and was absorbed to collagen loaded bio-derived bone，then the releasing kinetics of WO-1 was detected by 3 Htetracycline in vitro. WO-1 bioderived bone material was grafted into a culturemedium with staphylococcus aureus, escherichia coli, and pseudomonas aeruginosato observe its bacteriostasis ability. WO-1 bio-derived bone material was grafted into radius of defected rabbits, the concentration of WO-1 was detected onthe 9th, 16th, 23th, and 30th day byHLPC in blood, in bone and in muscle. The bacteriostasis ability of WO-1 loaded bio-derived bone was tested in vitro and in vivo. Results WO-1 loaded bioderived bone maintained natural network pore system and the surface of network pore system was coated with collagen membrane. The release of WO-1 from WO-1 loaded bioderived bone showed bursting release on the 1st day, then showed stable release. WO-1 loaded bioderived bone showed lasting and stable bacteriostasis to common pathogens of orthopaedic infections. The high concentration of WO-1 was observed in bone tissue and in muscle tissue at differenttime points and the difference among groups had no significance（P＞0.05）, while the concentration of WO-1 in blood was very low（P＜0.05). Conclusion WO-1 loaded bioderived bone has good capability of drug controlled-release and bacteriostasis.
Objective To review the latest development of amniotic membrane andits application. Methods Related literatures on the development of amniotic membrane and its application were extensively reviewed and summarized. Results There were amniotic epithelial cells and many growth factors in the outer layer of amniotic membrane and there were many kinds of collagen in the basement. The special structure promoted the growth of many kinds of cells. It was widely used in ophthalmology. Conclusion As it is easily available, compatible, cheap in price, low in antigenicity, and able to promote the growth of many kinds of cells, with few ethical problems involved, amniotic membrane will be more and more widely applied.
Objective To review and summarize the latest development of the therapy for the Duchenne muscular dystrophy (DMD). Methods Therecentlypublished articles related to the therapies for DMD were extensively reviewed and briefly summarized. Results The therapeutic approaches for DMD included the gene therapy, the cell therapy, and the pharmacological therapy. The gene therapy and the cell therapy were focused on the treatment for the cause of DMD by the delivery of the missing gene, the modification of the mutated gene, and the transfer of the normal cells including the stem cells, while the pharmacological therapy dealt with the downstream events caused by the dystrophin gene defect, slowed down the pathologic progress of DMD, and improved the DMD patient’s life quality and life span, by medication and other factor treatments. Conclusion There is still no cure for DMD because of various difficulties in replacing or repairing thedefected gene and of the multifaceted nature of the severe symptoms. Therefore,it is imperative for us to find out a more effective treatment that can solve these problems.
目的:探讨开胸心脏瓣膜替换术后胸壁瘘及慢性化脓性肋软骨炎的处理方法。方法:对单根的肋软骨炎并胸壁瘘者,在压痛最明显处直接切除受累的肋软骨及窦道组织;对伴瘘的胸部多根肋软骨炎,可在经胸壁相对正常处切开,建立以远离感染部位为蒂的开放胸部皮瓣,经瘘口加压注入美蓝使受累的肋软骨及坏死筋膜染色,沿染色部完整切除受累的肋软骨及筋膜、瘘管周围组织;在手术创面皮瓣下置放盆式多孔引流管,术后持续低负压吸引,选用敏感抗生素。结果:本组3例,术后6天拨管,10天后伤口愈合,效果良好。结论:经正常皮肤切口入路,建立开放胸壁皮瓣,彻底清除感染坏死的肋软骨及瘘管周围组织是治疗开胸心脏换瓣术后胸壁瘘及慢性化脓性肋软骨炎的可靠方法。
Objective To investigate the effect of tissue engineered bone with cryopreservation on healing of bone defects and to explore feasibility of cryopreservation for tissue engineered bone. Methods Tissue engineeredbones were constructed with osteoblasts being seeded onto bio-derived materials made from freshhuman bones,and they were preserved at 4℃ and －196℃ for 3 months and 6 monthsrespectively.They were applied to repair segmental bone defects of rabbit’s radius while the tissue engineered bone without cryopreservation and bio-derived materials were brought into control groups.The experiment was divided into groups A3,A6,B3,B6,C and D(group A3:tissue engineered bones were preserved at 4℃ for 3 months; group A6:tissue engineered bones were preserved at 4℃ for 6 months;group B3:tissue engineered bones were preserved at －196℃ for 3 months; group B6:tissue engineered bones were preserved at －196℃ for 6 months; group C: tissueengineered bones without cryopreservation; group D: bio-derived materials). Macroscopical and histologial examination were done at the 2nd,4th,6th,12th weeks, X-ray examination was done at the 6th,12th weeks and biomechanics were determined at 12th weeks after operation respectively. Results Macroscopical observation showed no significant differences among group A3, A6, B3, B6 and C, but less new bone formation and more obvious boundary in group D were observed. Histological observation showed more collagen and new bone around the edge of implant of group A3, A6, B3, B6 and C than group D, and histological evaluation showed significant differences between group D and other groups(P＜0.05). Radiographic observation showed no absorbability of the implant cortex and less new bone formation in group D, but the unity between implant and host bone, medullary cavity reopened, disappearance of fracture line and fine bone modelling were observed in other groups at 12 weeks after operation. Biomechanics between group D and other groups showed significant differences(P＜0.05). Conclusion Cryopreservation (4℃ and -196℃) were capable of preserving tissue engineered bone for long time, and tissue engineered bone withcryopreservation has significant effect on healing of bone defects. The methods f it clinical application.