目的 探讨一个遗传性无纤维蛋白原血症家系的分子发病机制.方法 应用Clauss法测定血浆纤维蛋白原活性,应用免疫比浊法测定纤维蛋白原抗原.提取先证者及其家系成员外周血DNA,PCR扩增纤维蛋白原FGA、FGB和FGG基因所有外显子和侧翼序列,构建纤维蛋白原野生型和突变型表达载体.在先证者血浆中加入凝血酶进行纤维蛋白聚集曲线测定；应用Western blot分析血浆纤维蛋白原,用野生型或FGB突变型质粒转染COS-7细胞,用Western blot和ELISA法检测转染后的细胞裂解液及细胞培养上清中纤维蛋白原的表达.结果 先证者APTT、PT、凝血酶时间明显延长；血浆纤维蛋白原活性及纤维蛋白原抗原检测结果均为0；先证者FGB基因2号外显子核苷酸2833～2834之间插入GTTT(纯合突变),先证者父亲、母亲、胞弟和儿子为杂合突变；凝血酶诱导的血浆纤维蛋白聚集曲线显示患者血浆无纤维蛋白聚集；Western blot分析显示,非还原条件下先证者血浆缺乏完整的纤维蛋白原分子和纤维蛋白原半分子,在还原条件下未检出截短的Bβ链.在转染突变型质粒的COS-7细胞裂解液中检出异常纤维蛋白原分子(相对分子质量＞340 000),细胞培养上清中未检出异常纤维蛋白原.野生型和突变型质粒转染的COS-7细胞裂解液中纤维蛋白原含量差异无统计学意义[(2.47±0.30)μg/ml对(2.65±0.60)μg/ml,P=0.0889];转染突变型质粒的COS-7细胞培养上清中纤维蛋白原含量低于转染野生型质粒的COS-7细胞,差异有统计学意义[(0.01±0.01)tg/ml对(3.80±0.80)μg/ml,P=0.0001].结论 纤维蛋白原FGB基因插入突变是该家系遗传性无纤维蛋白原血症的分子发病机制；该突变导致纤维蛋白原分子合成异常、组装及分泌障碍.

Objective To investigate the genetic defect and its mechanism in a patient with congenital afibrinogenemia.Methods The plasma fibrinogen activity and antigen of the patient was determined using the Clauss method and immuno-nephelometric assay,respectively.Genomic DNA was isolated from peripheral blood of the proband and his related family members.All exons and exon-intron boundaries of the three fibrinogen genes (FGA,FGB,FGG) were amplified by PCR followed by direct sequencing.Thrombin fibrin aggregation curve were detected in the plasma of the patient.Wild-type and mutation type fibrinogen vectors were constructed,and then transfected into COS-7 cells.The wild-type and mutant proteins from the culture media and cell lysates were tested by Western blot and ELISA.Results APTT,PT,TT were significantly longer in the proband.Plasma fibrinogen activity and antigen of the patient could not be detected using the Clauss method and immuno-nephelometry,respectively.Gene analysis revealed that a

为了探讨海藻糖对冷冻草鱼鱼糜肌原纤维蛋白冷冻作用的影响，以冷冻鱼糜盐溶性蛋白含量、肌原纤维蛋白Ca2＋ -ATPase活性以及肌原纤维蛋白表面疏水性为指标。结果表明，加入3％，6％及9％海藻糖的草鱼鱼糜冻藏60d后，盐溶性蛋白含量比空白组分别提高6．11％，22．01％，23．43％；肌原纤维蛋白Ca2＋ -ATPase活性比空白组分别提高14．82％，35．40％，36．04％；肌原纤维蛋白表面疏水性比空白组分别下降13．63％，31．63％，31．45％。添加海藻糖能有效延缓草鱼鱼糜肌原纤维蛋白的冷冻变性，其中以添加6％海藻糖的综合效益为最佳。

The effect of trehalose on the myofibrillar protein freezing denaturation of grass carp surimi during frozen storage is studied. In this paper, salt-solubility of myofibrillar protein, Ca2+-ATPase activity and surface hydrophobicity are used as indexes. The results indicate that after 60-day frozen storage, the contents of salt-soluble protein of the 3%, 6% and 9%trehalose treatment group are enhanced respectively by 6.11%, 22.01% and 23.43% when compared with the control group, the Ca2+-ATPase activity of myofibrillar proteins is enhanced respectively by 14.82%, 35.40% and 36.04%, the surface hydrophobicity of myofibrillar protein is reduced respectively by 13.63%, 31.63% and 31.45% at the end of the storage. Therefore, the addition of trehalose can effectively prevent myofibrillar protein denaturation during frozen storage, and the additive amount of 6%trehalose can achieve the best effect.

成熟的弹性纤维主要由弹性蛋白、原纤维蛋白微纤维及与弹性纤维有关的蛋白组成.内源性老化的皮肤弹性和柔韧度降低,弹性纤维网断裂和衰退.光老化皮肤不仅是富含原纤维蛋白的微纤维在表皮真皮交界处丢失,弹性蛋白变性,更重要的是在真皮深层混乱的弹性纤维蛋白物质的沉积,弹性蛋白的功能也受到影响.弹性纤维的修复可归纳为促进组成蛋白表达、改善组装条件、减少破坏因素3方面.

Mature elastic fibers are mainly composed of elastin,fibrillin microfibrils and some proteins.Endogenously aged skin is manifested as a decrease in skin elasticity and flexibility as well as rupture and degeneration of elastic fibers,while photoaged skin not only involves disappearance of fibrillin-rich microfibrils in the epidermal-dermal junction,denaturation of elastic protein,but also deposition of disarranged elastic fibrous protein materials in the deep dermis with impaired function of elastic protein.The repair of elastic fibers may be realized by promoting component protein expression,improving assembly condition and reducing damage-related factors.

目的 首次报告3例长期应用蛇毒血凝酶致低纤维蛋白原血症患者.方法 对3例长期应用蛇毒血凝酶致低纤维蛋白原血症患者的临床资料进行分析并复习相关文献.结果 例1,女,2岁,闭合性肝脏损伤,予大量输血及输注纤维蛋白原、基因重组凝血因子Ⅶa、蛇毒血凝酶(2 U/d)等治疗,患儿肝脏创面出血很快停止,伤后18d血浆纤维蛋白原0.12 g/L;例2,男,3岁,闭合性肝脏损伤,行肝脏创口修补术,术后给予蛇毒血凝酶2 U/d,术后8d腹腔引流管和静脉穿刺处大量渗血,血浆纤维蛋白原0.24 g/L;例3,男,45岁,因下颌恶性肿瘤行全下颌切除术,术后给予蛇毒血凝酶4U/d,术后12d手术切口出现持续渗血,血浆纤维蛋白原0.25 g/L.在停用蛇毒血凝酶并补充纤维蛋白原后,3例患者血浆纤维蛋白原与凝血检查恢复正常,例2、例3出血停止.结论 蛇毒血凝酶长期应用可导致低纤维蛋白原血症与严重出血.

Objective A first report of 3 patients who developed hypofibrinogenemia due to longterm administration of hemocoagulase.Methods The clinical data of three patients with hypofibrinogenemia due to long-term administration of hemocoagulase were analyzed,and the related literature was reviewed.Results Case 1,a two-year old girl,had liver traumatic rupture and then treated with massive transfusion and fibrinogen infusion in addition to intravenous recombinant factor W a (two times) and hemocoagulase (2 U/d).The liver wound bleeding was soon stopped.However,her plasma fibrinogen level decreased to 0.12 g/L after continuous administration of hemocoagulase for 18 days.Case 2,a three-year old boy,had liver traumatic rupture and was treated with surgical repair,and then received hemocoagulase (2 U/d).On the 8th day,a large amount of blood was found to exude from abdominal cavity drainage tube and indwelling venous catheter,and his fibrinogen dropped to 0.24 g/L.Case 3 was a 45 year-old man who u

目的研究乙型肝炎患者血浆纤维蛋白原及血浆D-二聚体水平的变化。方法以Clauss法测定血浆纤维蛋白原,免疫比浊法测定D-二聚体定量水平。分别检测125例乙型肝炎患者及68例正常人血浆纤维蛋白原和D-二聚体的含量。结果与正常对照组比较,乙型肝炎患者纤维蛋白原和D-二聚体定量水平有极显著性差异(P0.001)。结论乙型肝炎患者体内呈现不同程度的继发性纤溶,提示联合血浆纤维蛋白原和D-二聚体检测能反映乙型肝炎患者肝功能受损的凝血变化,对预后的判断有良好的临床应用价值。

Objective To study the changes of plasma fibrinogen and D-dimer in patients with hepatitis B. Method plasma fibrinogen were measured by Clauss,D-dimer were quantitated by immunoassay turbidimetry ,the two experiment were determined in 125 patients with hepatitis B and 68 normal controls. Result There were significant differences between the patients group and normal controls group ,such as the level of plasma fibrinogen and D-dimer(P<0.001). Conclusion Different degree of hyperfibrinolysis exists in hepatitis B ,indicating plasma fibrinogen and D-dimmer is clinically useful for judgement of the outcome prediction of the patients with hepatitis B.