Clinical application of human egg cryopreservation

Clinical egg cryopreservation has been applied during a 4-year period with some limited success. Mostly mature and a few immature eggs were frozen slowly and thawed rapidly in 1,2-propanediol and sucrose, and subsequently inseminated by intracytoplasmic sperm injection (ICSI). Three studies were performed in which: (i) it was established that 55% of aged unfertilized mature eggs survive freezing; (ii) in 22 cycles of thawed donated eggs cryosurvival was 24% with 15 cycles reaching transfer, and five pregnancies were initiated, one of which went to term at 39 weeks with fraternal twin boys, and one remains ongoing at 37 weeks; and (iii) in five cycles, where in-vitro fertilization patients had some of their own eggs frozen/ thawed, cryosurvival of mature eggs was poor at only 2.2%, although 44% sibling germinal vesicle (GV) stage eggs survived. A normal female infant delivered at 40 weeks arose from transfer of two embryos where GV eggs underwent in- vitro maturation post-thaw and were fertilized by ICSI. Pregnancies reported here and by others indicate a burgeoning awareness of the potential benefits of egg cryopreservation, prompting cautious optimism for the future of this technology.      

海藻酸钠载体中成年兔关节软骨细胞生长情况

目的:软骨细胞单层培养条件下增殖较快,但极易失分化,在生物载体材料构成的立体环境中培养,则能较好的维持表型。实验以海藻酸钠为载体,观察兔关节软骨细表型及增殖情况。方法:实验于2005-09/2006-12在山西医科大学第二医院骨科实验室完成。①实验动物和材料:6月龄新西兰大白兔24只,雌雄各半。实验过程中对动物处置符合动物伦理学要求。海藻酸钠由青岛明月海藻公司提供。②实验方法:以培养液配制的0.4% Pronease酶、0.025%Ⅱ型胶原酶顺序消化兔软骨分离细胞,以4×109 L-1海藻酸钠的浓度制成细胞悬液。③实验评估:倒置显微镜观察细胞在海藻酸钠中的形态及增殖情况,检测细胞收获效率和存活率。苏木精-伊红染色和AB-PAS染色观察软骨细胞的生长情况及评价软骨分泌胶原的情况。免疫组织化学定性观察Ⅱ型胶原的含量变化及有无Ⅰ型胶原的产生。Alcian blue染色法测定细胞盘中蛋白多糖的含量变化。结果:①软骨经两步酶消化软骨基质逐步解离和降解,细胞被完全分离,消化分离的软骨细胞总数达到5×106,细胞成活率达95.5%。②软骨细胞与海藻酸钠复合后体外培养,细胞生长旺盛、增殖活跃,增殖成株状或岛状,株状细胞团周围有类似的软骨陷窝形成,细胞排列密集,核圆形。③Ⅱ型胶原免疫组化和AB-PAS染色均呈阳性,细胞盘中蛋白多糖含量随培养时间延长逐渐增加。结论:海藻酸钠凝胶能与软骨细胞完全嵌合,是一种良好的软骨组织工程材料。     

焦虑抑郁对慢性阻塞性肺疾病急性加重和住院的影响

本研究旨在通过为期1年的多中心、前瞻性随访评价焦虑抑郁作为独立危险因素对慢性阻塞性肺疾病急性加重和住院的影响.研究随机选取来自北京10所综合医院的491例稳定期COPD患者,所有患者进入研究前均签署知情同意书.     

Transjugular intrahepatic portosystemic shunt-placement increases arginine/asymmetric dimethylarginine ratio in cirrhotic patients

AIM：To analyze the change of dimethylarginine plasma levels in cirrhotic patients receiving transjugular intrahepatic portosystemic shunt（TIPS）.METHODS：To determine arginine,asymmetric dimethylarginine（ADMA）,symmetric dimethylarginine（SDMA）,and nitric oxide（NO） plasma levels,blood samples were collected from the superior cava,hepatic,and portal vein just before,directly after,and 3 mo after TIPS-placement.RESULTS：A significant increase in the arginine/ADMA ratio after TIPS placement was shown.Moreover,TIPS placement enhanced renal function and thereby decreased systemic SDMA levels.In patients with renal dysfunction before TIPS placement,both the arginine/ADMA ratio and creatinine clearance rate increased significantly,while this was not the case in patients with normal renal function before TIPS placement.Hepatic function did not change significantly after TIPS placement and no significant decline in ADMA plasma levels was measured.CONCLUSION：The increase of the arginine/ADMA ratio after TIPS placement suggests an increase in intracellular NO bioavailability.In addition,this study suggests that TIPS placement does not alter dimethylarginine dimethylaminohydrolase（DDAH） activity and confirms the major role of the liver as an ADMA clearing organ.     

Histidine-rich glycoprotein is present in human platelets and is released following thrombin stimulation

Histidine-rich glycoprotein, and alpha 2-glycoprotein in human plasma, has been shown to interact with heparin, with the high-affinity lysine- binding site of plasminogen, with divalent cations, and is associated with the rosette formation between erythrocytes and lymphocytes. A specific enzyme-linked immunosorbent assay for histidine-rich glycoprotein has been developed and used to demonstrate that histidine- rich glycoprotein is present in human platelets. Histidine-rich glycoprotein was detected and quantified in detergent extracts of washed human platelets, with a mean level of 371 ng/10(9) platelets. Plasma histidine-rich glycoprotein, either in the platelet suspending medium or on the surface of the platelets, accounted for less than 3.4% of the detectable platelet histidine-rich glycoprotein. Histidine-rich glycoprotein was also demonstrated in human bone marrow megakaryocytes by immunofluorescence. The extent of histidine-rich glycoprotein release from platelets was dependent on the thrombin dose and correlated directly with the extent of serotonin release. The platelet and plasma histidine-rich glycoprotein were similar by immunochemical analysis. Anti-histidine-rich glycoprotein IgG did not inhibit platelet aggregation. Histidine-rich glycoprotein released by platelets following thrombin stimulation may play a significant role in modulating inflammatory events in the microenvironment of the platelet plug.     

Combined procoagulant activity and proteolytic activity of acute promyelocytic leukemic cells: reversal of the bleeding disorder by cell differentiation

In the human promyelocytic cell line HL60, we observed both a strong procoagulant activity (PCA) on the cell membrane and proteolytic activity in the lysate of these cells. Because these cell-line cells are susceptible to differentiation to either a more mature granulocytic or monocytic form, we were able to study the hypothesis that the combination of PCA and proteolytic activity is confined to the promyelocyte. This may explain the severe coagulopathy seen in patients with acute promyelocytic leukemia. Cell differentiation in a myeloid direction induced by retinoic acid or DMSO led to a diminished PCA, while not affecting the fibrinolytic activity. On the other hand, monocytic differentiation obtained by culturing the cells in the presence of 1; 25 dihydroxy vitamin D3 led to the complete disappearance of the proteolytic activity of the cell lysate, although the procoagulant activity was still present. Furthermore, we found that the elastase activity almost disappeared after monocytic differentiation. We also studied the PCA, proteolytic activity, and elastase activity of blast cells of patients with acute myeloid leukemia. Only in patients with acute promyelocytic leukemia did we observe both a strong PCA and fibrinolytic activity. This supports our hypothesis that the combination of these activities is unique to the promyelocyte and may explain the observed bleeding complications in patients with acute promyelocytic leukemia.     

治疗颅下颌关节强直的新观点

治疗颅下颌关节强直有多种方法,但任一治疗方式都不能令人完全满意。作者基于强直骨块的形成是病理性反应结果,它不是新生物,无持续生长的能力,解除关节强直不一定要去除强直骨块,而提出了治疗颅下颌关节强直的新观点。 材料和方法 采取向颞部延伸之耳前切口,沿整个升支的宽度,显露强直骨块的侧面及正下方,在强直骨块下正常骨块区行水平截骨。剥离附于下颌升支上的翼肌一嚼肌链,拉开截骨之断端形成间隙,使下颁活动自如,被动开口度不少于3cm。以颞肌