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find Keyword "低温保存" 14 results
  • THE INJURY OF CALCIUM OVERLOAD TO THE HEPATOCYTES IN COLD STORAGE

    【Abstract】Objective To investigate the injury of calcium on the liver. Methods By using collagenase-containing solution to perfuse the rat livers, the rat liver suspension with different viability was prepared, preserved hypothermically, and the cytosolic calcium comcentration was detected with Fura2/AM. Results ① The concentration of cytosolic calcium after 2-hour storage: Experiment group 1(viability 5%) (1055.0±30.79) nmol/L, experiment group 2(viability 10%) (853.0±20.42) nmol/L, experiment group 3(viability 30%) (616.0±13.20) nmol/L, experiment group 4(viability 50%) (562.0 ±26.06) nmol/L, experiment group 5(viability 70%-80%) (318.0±13.01) nmol/L, experiment group 6(viability 90%) (114.6±6.11) nmol/L. ②The concentration of cytosolic calcium after 24hour storage: Experiment group A(viability 10%) (1704.0±70.11) nmol/L, experiment group B(viability 50%) (1125.0±23.22) nmol/L. The results showed that the lower was the viability, the higher was the cytosolic calcium concentration.With the same viability the cytosolic calcium concentration elevated more than two times higher than the original concentration with the time lengthened. Conclusion Calcium overload is one of the main factors which attribute to the ischemiareperfusion injury of the hepatocytes.

    Release date:2016-08-28 05:30 Export PDF Favorites Scan
  • 同种瓣的制作与临床应用

    目的报告液氮深低温下保存同种带瓣血管的制作方法、组织活性及临床应用效果。方法制作同种瓣24个、抗生素灭菌、梯度降温后置于液氮中保存,并测定冷冻保存后同种瓣的组织活性。同种瓣临床应用5例,其中法洛四联症、肺动脉闭锁2例,先天性主动脉瓣狭窄1例,法洛四联症术后发生室间隔缺损残余漏伴肺动脉瓣重度关闭不全1例,Bentall术后发生感染性心内膜炎1例。结果抗生素灭菌、液氮深低温技术保存同种瓣具有良好的组织活性,糖代谢测定24h葡萄糖消耗大于16mg/dl,组织培养见成纤维细胞生长良好。临床移植5例均成功,术后随访3~8个月,同种瓣无狭窄或关闭不全。结论液氮深低温保存同种瓣安全可靠,临床应用早期效果良好。

    Release date:2016-08-30 06:25 Export PDF Favorites Scan
  • 低温保存对山羊大动脉细胞形态的影响

    目的 观察低温保存过程对山羊动脉细胞形态的影响. 方法 将12段山羊动脉按不同的降温速率分成4组,分别用透射电子显微镜和扫描电子显微镜观察冻存前后的动脉细胞形态. 结果 与正常血管组织相比,0.5 K/min组和0.1 K/min组虽细胞间连接紧密,但内皮细胞附着松散,出现大的空泡,有的平滑肌细胞肿胀,细胞质苍白多水,线粒体收缩;还有的虽平滑肌细胞轻微肿胀,但细胞器肿胀严重破坏. 5 K/min组和2 K/min组内皮细胞与基膜大量脱离,内皮细胞和平滑肌细胞破坏严重,出现核溶解及线粒体一些不能恢复的变化. 结论 经低温保存后的山羊主动脉与新鲜组织的区别明显,细胞组织形态被破坏,低温保存的降温速率越快,破坏越严重.

    Release date:2016-08-30 06:32 Export PDF Favorites Scan
  • 同种带瓣大动脉重建右心室流出道

    摘要目的 应用自行采集和冷冻保存的同种带瓣大动脉(VHC)完成先天性心脏病右心室流出道的重建,并观察其疗效及存在的问题。方法 用VHC材料治疗先天性心脏病95例,90例手术根治,5例行VHC右心室流出道与肺动脉连接姑息手术。结果 院内死亡13例;术后随访68例,死亡2例,均为感染。66例长期生存者中25例胸部X线片示VHC有钙化,多为主动脉材料,仅5例有轻中度压力阶差(35~60mmHg)。结论 VHC可广泛用于治疗复杂先天性心脏病。程序降温、超低温保存和两步化冻是VHC使用质量的保证。VHC的长期通畅比合成管道好,随植入时间延长钙化率增加,肺动脉VHC优于主动脉。有肺动脉高压者宜尽早手术。为了预防VHC植入后感染,应重视、改进其收集和保存的方法。

    Release date:2016-08-30 06:33 Export PDF Favorites Scan
  • EFFECT OF DIFFERENT TEMPERATURES ON SYSTEM OF IN VITRO PHYSIOLOGICAL ENVIRONMENT FOSTERING LIMBS

    Objective To investigate the effects of different temperatures on the system of in vitro physiological environment fostering limbs. Methods Twenty-four limbs were harvested from 6 adult Bama mini pigs and were randomly divided into 4 groups (n=6) according to different temperatures: limbs were placed in in vitro physiological environment foster-ing limbs at 26℃ (group A), 4℃ (group B), 10℃ (group C), and 18℃(group D). After 12 hours of perfusion, the morphology observation was done for the structure and ultrastructure changes of the skeletal muscle by light microscope and transmission electron microscope. The mRNA levels of tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) were detected by real-time fluorescent quantitative PCR (RT-qPCR). Results Histological results showed that the skeletal muscle exhibited mild edema, integrity of the sarcolemma, and occasional perivascular inflammatory cell infiltration in groups B, C, and D, meanwhile, the cells of group C had normal morphology; however, muscle fibers degenerated, muscle cells were seriously damaged, a great number of inflammatory cells infiltrated in the fractured muscle fibers in group A. Transmission electron microscope results showed as follows: the muscle fibers arranged in disorder, and many focal solubility necrosis occurred in group A; the muscle fibers arranged in order relatively and sarcolemma was still intact, with mild swelling and flocculent degenerative mitochondria in group B; a large number of muscle fibers arranged in order and regularity with clear sarcomere in group C; and the muscle fibers arranged in disorder and irregularity and partly dissolved in group D. RT-qPCR results showed that the expressions of inflammatory factor TNF-α and IL-1β mRNA in group A were significantly higher than those in groups B, C, and D (P lt; 0.05); the expressions were significantly lower in groups B and C than in group D, and in group C than in group B (P lt; 0.05). Conclusion In the system of in vitro physiological environment fostering limbs, temperature plays an important role in the preservation of amputated limbs. It is suggested that 10℃ can significantly attenuate the reperfusion-induced skeletal muscle cell injuries in this system.

    Release date:2016-08-31 04:05 Export PDF Favorites Scan
  • EXPERIMENTAL STUDY OF TISSUE ENGINEERED BONE WITH CRYOPRESERVATON ON HEALING OF BONE DEFECTS

    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.

    Release date:2016-09-01 09:29 Export PDF Favorites Scan
  • PRELIMINARY STUDY ON RESEARCH METHOD OF CELL SURVIVAL RATE AT PROCEDURE OF CRYOPRESERVATION OF TISSUE ENGINEERED TENDONS

    Objective To study the research method of cell survival rate at the procedure of cryopreservation of tissue engineered tendons.Methods In the 4thgeneration of human fibroblasts, the dead cells were stained with propidium iodine (PI), while the living cells with Hoechst 33342(Ho). The living cells and dead cells emitted fluorescence of red and blue respectively after they were stimulated by suitable ultra-violet, then flow cytometry was applied to distinguishthem. The seeding cells were collected to make them to be the cell suspension of suitable concentration(6.0×105 cell/ml) before they were divided into two parts. We cryopreserved and defrosted one part three times to kill the cells and didnot cryopreserve the other part, then we made cell suspension at different ratios of cryopreserved cell to noncryopreserved cells. The fluorescence staining and flow cytometry were used to study the correlation between cell ratios of cryopreservedcell to non-cryopreserved cell and the cell survival rates. We compared the cll survival rates between immediate flow cytometry and that 2 hours after fluorescence staining. Results The results of flow cytometry showed that correlation between the ratio of cryopreservation and the cell survival rate was significant (r=0.970,Plt;0.05), image analysis study also showed the correlation was significant (r=0.982,Plt;0.05).The cell survival rate decreased by use of flow cytometry twohours after fluorescence staining, but there was no significant difference when compared with that of immediate flow cytometry (Pgt;0.05). We could also observe the cells on the tissue engineered tendons by fluorescence image directly.Conclusion Flow cytometry and fluorescence image afterPI and Ho staining is a good way in study cell survival rate at the procedure of cryopreservationof tissue engineered tendons.

    Release date:2016-09-01 09:33 Export PDF Favorites Scan
  • REPAIR OF CARTILAGE DEFECT IN JOINT WITH TRANSPLANTATION OF CRYOPRESERVED HOMOLOGOUS EMBRYONIC PERIOSFEUM OF RABBITS

    In order to repair cartilage defect in joint with transplantation of cryopreserved homologous embryonic periosteum, 30 rabbits were used and divided into two groups. A 4 mm x 7 mm whole thickness cartilage defect was made in the patellar groove of femur of each rabbit. The homologous embryonic rabbit skull periosteum (ERSP), preserved in two-step freezing schedule, was transplanted onto the cartilage defect of joints of one group and autogenous periosteal graft was done in the joint defect of the other group. The knees were not immobilized, following operation and 16 weeks later, the newly formed tissue in the defects were assessed by gross observation, histochemical examination and biochemical analysis. The results showed that new hyaline-like cartilage was formed in the cryopreserved ERSP grafted knee, and had no significant difference from that of the knee receiving autogenous periosteal graft, but had significant difference from that of the fresh ERSP grafted knee and the non-grafted knee. Furthermore, the new hyaline-like cartilage had the biochemical characteristics of a fibrous cartilage. The conclusion was that this method might be feasible to repair articular cartilage defects.

    Release date:2016-09-01 11:07 Export PDF Favorites Scan
  • Short-, mid- and long-term preservation of human fetal retina

    Objective To observe the configuration and viability of full thickness human fetal retina after short-, mid- and long-term preservation. Methods Twenty-two full thickness human fetal retinae of gestational age of 12-24 weeks were coated by glutin and cut into 88 pieces, and then preserved in Ames' solution, DX solution, -80℃ refrigerator or under cryopreservation condition. The cell viability of retinal neuroepithelial layer was determined by trypan blue staining, retinal configuration was determined by light microscope and electromicroscope. Results The viability of neuroepithelial layer was (94.79plusmn;2.85) % in fresh fetal retina, gt;80% in Ames' solution within 4 hours, and gt;77% in DX solution within 2 days. There was no significant difference between those solution-preservations and the fresh fetal. In -80℃ refrigerator, the viability was (65.83plusmn;5.06)% after 7 days, and then dropped to (57.54plusmn;16.18)% at the end of the first month. Under the cryopreservation condition, the viability was (69.46plusmn;9.31)% at the end of first month. Light and transmission electron microscopy had not deteced any abnormals in the full thickness human fetal retina preserved in Ames' solution within 2 hours, but showed clear retinal layers with bigger intercellular space after preserved in DX solution for 2 days, in -80℃ refrigerator for 7 days and under cryopreservation condition for 1 month. Conclusion Ames' solution and DX solution can preserve good viability and configuration of full thickness human fetal retina in a certain time period.

    Release date:2016-09-02 05:46 Export PDF Favorites Scan
  • An observation on enzymic histochemistry and ultrastructure of cryopreserved human retinal epithelium

    Objective To observe the enzymic histochemical and ultrastructral changes of cryopreserved human retina. Methods To compare the activity of lactate dehydrogenase (LDH), succinate dehydrogenase (SDH) and ATPase in cryopreserved retina with those in fresh retina and to observe the histological and ultrastructural changes of cryopreserved retina. Results There was no statistical difference between the activity of LDH,SDH and ATPase in fresh and in cryopreserved retina. Histologically, in the cryopreserved retina, fluid in neural fiber and outer plexiform layers, as well as in cone and rod layer, was sligthly more than normal. The ultrastructure is normal except that the mitochondria was swollen in different degree. Conclusion Cryopreservation may be an effective method for keeping the retinal cells alive for a long period and might free the transplantation from dependance on aviability of fresh dornor tissue. (Chin J Ocul Fundus Dis,2000,16:139-212)

    Release date:2016-09-02 06:05 Export PDF Favorites Scan
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