脑梗死患者血浆蛋白C、蛋白S动态观察

目的　动态观察脑梗死患者血浆蛋白C抗原 (PC :Ag)和蛋白S(PS)水平。方法　测定 94例脑梗死患者血浆PC :Ag、总蛋白S(TPS)和游离蛋白S(FPS)并与 5 2名健康者相对照 ,动态观察 41例脑梗死患者PC :Ag和PS浓度。结果　脑梗死组PC :Ag、TPS、FPS降低发生率明显高于对照组 (P <0 0 5 )。PC :Ag浓度动态观察提示脑梗死急性期PC :Ag浓度呈非线性改变 ,在发病第二周出现一“反跳”性增高。结论　部分脑梗死患者存在PC和PS缺乏现象 ,多数为继发性缺陷 ,发现这些患者并区别其为原发性或继发性对明确脑梗死的病因、治疗和预防有重要意义。     

急性脑梗死患者蛋白S基因多态性和血浆蛋白S水平与其家族史关系的研究

目的探讨急性脑梗死(ACI)患者蛋白S基因(PROS1)多态性和血浆总蛋白S(tPS)和游离蛋白S(fPS)水平与其家族史的关系。方法对93例ACI患者,其中散发(NFCI)组83例,有家族史(FCI)组10例,采用聚合酶链式反应-限制性片段长度多态性分析法(PCR-RLFP)检测PROS1A2148G多态性;采用酶联免疫吸附双抗体夹心法(ELISA)测定血浆tPS和fPS水平;并与正常对照组进行比较。结果ACI组PROS1A2148G的AA、AG和GG型频率分别为18.3%、48.4%和33.3%,与正常对照组(22.0%、48.0%和30.0%)差异无统计学意义(均P>0.05)。FCI组A2148GGG基因型频率(50%)高于正常对照组(30%),但差异无统计学意义(P>0.05)。ACI组GG基因型患者血浆tPS水平和fPS水平[(13.67±1.49)、(6.07±0.61)μg/ml]显著低于AA型和AG型[(15.73±1.24)、(6.72±0.55)和(15.15±1.48)、(6.63±0.66)μg/ml](均P<0.01),且FCI组血浆fPS水平[(6.12±0.69)μg/ml]显著低于NFCI组[(6.48±0.75)μg/ml](P<0.05)。结论PROS1A2148GGG基因型导致FCI患者血浆fPS水平降低,其基因型可能与ACI家族史有关。     

脑梗死后抑郁症患者血清S100B蛋白浓度的变化及意义

目的探讨脑梗死后抑郁症患者血清S100B蛋白浓度的变化及意义。方法选取2010-01—2012-01在我院接受治疗的300例脑梗死患者,采用汉密尔顿抑郁量表HAMD进行抑郁症筛查,脑梗死后抑郁症患者125例为实验组（A组）,余下单纯脑梗死患者175例中随机选取125例为对照组（B组）。观察2组治疗前后血清S100B蛋白浓度变化和病情改变情况。结果 A组患者治疗前HAMD评分（21.80±6.01）分,治疗后为（14.45±3.82）分,差异有统计学意义（P〈0.05）;治疗前血清S100B蛋白浓度（0.32±0.083）μg/L,治疗后为（0.21±0.056）μg/L,差异有统计学意义（P〈0.05）。治疗前后A组血清S100B蛋白浓度均高于同期B组,差异有统计学意义（P〈0.05）;B组治疗前后血清S100B蛋白浓度变化不明显（P〉0.05）。结论脑梗死后抑郁症患者抗抑郁治疗后血清S100B蛋白浓度显著降低,抑郁症病情改善,血清S100B蛋白浓度可作为脑梗死病情、预后、抑郁症发作和抗抑郁治疗效果的预测指标。     

脑梗死后抑郁症患者血清 S100B 蛋白浓度的变化及意义

目的：探讨脑梗死后抑郁症患者血清S100B蛋白浓度的变化及意义。方法选取2010-01-2012-01在我院接受治疗的300例脑梗死患者,采用汉密尔顿抑郁量表HAMD进行抑郁症筛查,脑梗死后抑郁症患者125例为实验组（A组）,余下单纯脑梗死患者175例中随机选取125例为对照组（B组）。观察2组治疗前后血清S100B蛋白浓度变化和病情改变情况。结果A组患者治疗前HAMD评分（21.80±6.01）分,治疗后为（14.45±3.82）分,差异有统计学意义（P＜0.05）;治疗前血清S100B蛋白浓度（0.32±0.083）μg/L,治疗后为（0.21±0.056）μg/L,差异有统计学意义（P＜0.05）。治疗前后A组血清S100B蛋白浓度均高于同期B组,差异有统计学意义（P＜0.05）;B组治疗前后血清S100B蛋白浓度变化不明显（P＞0.05）。结论脑梗死后抑郁症患者抗抑郁治疗后血清S100B蛋白浓度显著降低,抑郁症病情改善,血清S100B蛋白浓度可作为脑梗死病情、预后、抑郁症发作和抗抑郁治疗效果的预测指标。     

蛋白C、活化蛋白C抵抗与脑梗死相关性探讨

目的探讨蛋白C(PC)、活化蛋白C抵抗(APCR)与脑梗死(CI)的相关性。方法对92例急性期CI患者及99例非脑血管病患者血浆PC和APCR进行检测。结果病例组血浆PC水平[(108.10±37.30)%]低于对照组[(124.32±51.75)%,P0.05],复发组CI患者血浆PC水平[(90.39±41.69)%]明显低于初发组[(114.75±33.49)%,P0.05],病例组的≥60岁亚组血浆PC水平[(103.77±37.58)%]明显低于60岁亚组[(127.24±30.06)%]以及对照组的≥60岁亚组[(125.78±56.63)%,均P0.05],病例组的60岁亚组与对照组的60岁亚组血浆PC水平比较差异无统计学意义。各组间APCR阳性率比较差异无统计学意义。结论 PC缺乏与CI的发生相关;PC活性降低是CI复发的重要因素,尤其与老年人CI的发生密切相关;APCR并未增加CI的发生率。     

蛋白C、蛋白S、活化蛋白C抵抗与缺血性脑血管病的相关性研究

目的研究蛋白C(PC)、蛋白S(PS)、活化蛋白C抵抗(APCR)与急性缺血性脑血管病间的关系.方法对804例急性脑血管患者及168例非脑血管病患者行PC、PS、APCR检测.结果在缺血性卒中组PC、PS活性分别为(115.8±12.5)%和(112.6±13.8)%,明显低于出血性卒中组的(119.2±11.3)%和(112.1±16.5)%及对照组的(120.2±12.8)%和(122.4±15.8)%(P＜0.01).进一步比较发现,＜45岁的缺血性卒中组的PC、PS分别为(54.67±8.9)%和(40.49±9.1)%,明显低于其他各组,PS在45岁以上的缺血性卒中组为(119.2±15.6)%,明显低于对照组的(122.4±15.8)%,(P＜0.01),其余各组间无显著差异.APCR总发生率6.6%(64/972),其中缺血性卒中组为7.14%(46/644),出血性卒中组为6.25%(10/160),两组比较无明显差异(P＞0.05),45岁以下缺血性卒中组APCR的发生率为46.3%(25/54),明显高于其他各组.结论PC、PS活性降低,APCR是缺血性脑血管病发生的重要因素,特别与45岁以下的缺血性卒中的发生密切相关.     

不同类型缺血性脑血管病与血同型半胱氨酸、超敏C反应蛋白及静息心率的关系

目的观察急性期不同血管病变类型缺血性脑血管病与血同型半胱氨酸、超敏C反应蛋白及静息心率的关系。方法对183例急性期缺血性脑血管病患者进行血浆Hcy、hs-CRP水平和静息心率的测定,将患者分为腔隙性脑梗死(LI)组和动脉粥样硬化血栓形成性脑梗死(ACI)组,比较2组患者的血浆Hcy、hs-CRP水平和静息心率的变化。结果 ACI组血浆Hcy水平(26.60±14.43)μmol/L显著高于LI组的(17.44±4.04)μmol/L(P<0.05),ACI组静息心率(82.72±8.26)次/min显著高于LI组的(73.26±12.59)次/min(P<0.05),ACI组hs-CRP水平(4.12±11.94μg/mL)高于LI组的(1.33±1.91)μg/mL(P<0.05)。2组年龄、性别、吸烟史、血脂、体质量指数以及高血压、糖尿病患病率差异无统计学意义。结论血浆Hcy和hs-CRP水平及静息心率的升高同血管变病类型密切相关,而且是AS的危险因素。     

单纯性糖耐量异常脑梗死患者血同型半胱氨酸及超敏C反应蛋白水平的改变

目的 研究单纯性糖耐量异常(IGT)脑梗死患者血同型半胱氨酸(Hcy)及超敏C反应蛋白(hs-CRP)水平的改变.方法 根据血糖水平及糖耐量试验将756例脑梗死患者分为糖耐量正常组、IGT组及糖尿病组.采用免疫透射比浊法测定血清hs-CRP水平,荧光偏振免疫分析法测定血浆Hcy水平.结果 血糖正常组331例,IGT组142例,糖尿病组283例.血Hcy及hs-CRP水平IGT组[(19.17±9.35)μmol/L,(8.0±11.9) μg/ml]及糖尿病组[(20.46±10.56) μmol/L,(7.7±20.7) μg/ml]明显高于血糖正常组[(16.17 ±7.35) μmol/L,(3.5 ±9.2) μg/ml](均P＜0.001),IGT组与糖尿病组间的差异无统计学意义.结论 IGT脑梗死患者的血 Hcy和hs-CRP水平明显升高.     

蛋白C、蛋白S与脑血管病关系的研究

目的:探讨蛋白C(PC)、蛋白S(PS)与脑血管病(CVD)的关系。方法:检测94例缺血性脑血管病(ICVD)患者和26例脑出血(CH)患者的血浆蛋白C:抗原(PC:Ag)、总蛋白S(TPS)和游离蛋白S(FPS)。结果:ICVD患者PC: Ag(17％)、TPS(26．6％)、FPS(23．4％)异常降低者多于对照组(2％)(P＜0．01)。CH患者PC:Ag(19．2％)异常降低者多于对照组(P＜0．05)。结论:17％～26．6％ICVD患者存在PC、PS缺陷;19．2％CH患者存在PC缺陷,多数为继发性缺陷,发现这些患者并区别其为原发性或继发性对明确CVD的病因、治疗和预防有重要意义。     

抑郁症患者血清S100B蛋白及其分泌型糖基化终产物受体的研究

目的 探讨首发和复发抑郁症患者血清S100B蛋白及其分泌型糖基化终产物受体(esRAGE)浓度的改变.方法 采用酶联免疫方法检测34例抑郁症患者(首发15例,复发19例) 和34名正常对照的血清S100B、esRAGE蛋白浓度,比较患者组和对照组间的差异;采用17项汉密尔顿抑郁量表(Hamilton Depression Rating Scale-17,HAMD-17)评定患者的症状,分析上述两个生化指标与症状之间的关系.结果 总体患者组血清S100B浓度高于对照组[(0.57±0.04) ng/mL vs(0.38±0.12)ng/mL,P<0.01],esRAGE浓度低于对照组[(0.52±0.11) ng/mL vs(0.66±0.10)ng/mL,P<0.01];首发和复发患者组之间血清S100B浓度与esRAGE的差异均有统计学意义[(0.46±0.01) ng/mL vs(0.57±0.02)ng/mL,P<0.01;(0.44±0.02)ng/mL vs(0.53±0.10)ng/mL,P<0.01];与对照组相比,首发及复发患者组的血清S100B浓度均升高而esRAGE浓度均下降,差异均有统计学意义(P<0.01).未见患者组的血清S100B、esRAGE浓度相关或二者与HAMD评分相关(P>0.05).结论 血浆S100B、esRAGE可能在抑郁症的发病中起了一定的作用,复发患者神经胶质细胞分泌S100B能力可能更强.     

Regulation of bovine activated protein C by protein S: the role of the cofactor protein in species specificity

The anticoagulant activity of bovine activated Protein C was observed to exhibit species specificity when tested in the plasmas from cow, dog, rabbit, and human. The relative effectiveness in descending order was cow, dog, human, and rabbit. When bovine Protein S was added to each of these plasmas, the species specificity of bovine activated Protein C was lost. No activated Protein C cofactor activity could be detected in either rabbit or human plasma. Bovine activated Protein C enhanced the rate of inactivation of both rabbit and human Factor Va in serum. The rate of activated Protein C catalyzed inactivation of rabbit and human Factor Va could be enhanced by the addition of bovine Protein S. These results indicated that the species specificity of the anticoagulant activity of bovine activated Protein C is mediated by a cofactorenzyme interaction rather than an enzyme-substrate interaction. These results further demonstrated that the anticoagulant activity of activated Protein C is due, in part, to the inactivation of Factor Va.     

Origins and Effects of Extracellular α-synuclein: Implications in Parkinson’s Disease

Misfolding and abnormal aggregation of the neuronal protein α-synuclein has been implicated in the pathogenesis of Parkinson’s disease and related neurological disorders, such as dementia with Lewy bodies. α-synuclein is a conventional cytosolic protein and is thought to exert its pathogenic function exclusively in the neuronal cytoplasm in a cell-autonomous manner. However, the current model is being challenged by a series of new observations that demonstrate the presence of α-synuclein and its aggregated forms in the extracellular fluid both in vivo and in vitro. Extracellular α-synuclein appears to be delivered by unconventional exocytosis of intravesicular α-synuclein, although the exact mechanism has not been characterized. Compared to the cytosolic α-synuclein, intravesicular α-synuclein is prone to aggregation and the potential source of extracellular aggregates. A number of tissue culture studies suggest that exposure to extracellular α-synuclein aggregates induces microglial activation, release of pro-inflammatory cytokines from astrocytes, and neurotoxicity. Thus, exocytosis of α-synuclein may be an important mechanism for amplifying and spreading degenerative changes from a small group of cells to its surrounding tissues, and it potentially provides therapeutic targets for halting the progression of the disease.     

Calcium-activated, phospholipid-dependent protein kinase and protein substrates in primary cultures of astrocytes

Phosphoinositide-linked transmembrane signaling in the brain involves calcium-activated, phospholipid-dependent protein kinase (protein kinase C), but little is known about the glial contribution to this system. We observed that phosphorylation of several proteins in a cytosal fraction of rat astrocytes in primary culture was increased by the addition of calcium and phosphatidylserine. These agents also stimulated phosphate incorporation into lysine-rich histone, a substrate for protein kinase C. Addition of diacylglycerol, an activator of protein kinase C, further increased histone phosphorylation, whereas polymyxin B, an inhibitor of protein kinase C, blocked the stimulatory effect of calcium and phosphatidylserine. Based on enzyme units per mg protein, the activity of protein kinase C in astrocytes appears similar to that in whole brain cytosol. These results indicate that astrocytes display protein kinase C activity and suggest that the glial enzyme may be an important component of the receptor-linked phosphoinositide response system in the brain.     

Cyclic AMP-dependent protein phosphorylation in the rat anterior pituitary

The activation of cyclic adenosine 3'5'-monophosphate (cAMP)-dependent protein kinases has been implicated as an integral mechanism in stimulus-secretion coupling in the anterior pituitary. Therefore, we have investigated phosphorylation of endogenous protein substrates both in the presence and absence of cAMP in cell-free extracts of the rodent anterior pituitary. Specific phosphoprotein substrates in the rat anterior pituitary, which are phosphorylated by a cAMP-dependent protein kinase in vitro, were identified. Cyclic AMP potentiated the phosphorylation of proteins with apparent molecular weights of 85,000, 77,000, 63,000, 53,000, 39,000, and 33,000 as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins with apparent molecular weights of 124,000, 93,000, 48,000, and 43,000 were phosphorylated only in the presence of cAMP and not in the basal condition. The results highlight endogenous protein substrates that may potentially be involved in cAMP-dependent stimulus-secretion coupling in the anterior pituitary.     

胶质瘤PTEN和细胞周期蛋白D1表达及其意义

目的　探讨胶质瘤组织中PTEN和细胞周期蛋白D1(CyclinD1)表达的意义。 方法　应用免疫组化结合计算机图像分析方法检测PTEN和CyclinD1蛋白在 80例胶质瘤的表达及其相关关系。结果　PTEN阳性率为 6 3.8% (5 1/ 80 ) ,低分级胶质瘤 (I～II)阳性率显著高于III级和IV级 (P<0 .0 1) ;胶质瘤不同病理分级间CyclinD1蛋白表达存在显著性差异 (P <0 .0 1)。胶质瘤细胞CyclinD1与PTEN蛋白表达呈显著性负相关关系 (r=- 0 .5 4 3,P <0 .0 1)。结论　PTEN蛋白表达可能在胶质瘤细胞周期调节和细胞增殖中发挥重要作用     

Decline of proteins C and S and factors II, VII, IX and X during the initiation of warfarin therapy

Oral anticoagulants achieve an antithrombotic effect only several days after initiation of treatment. A rapid decline of the vitamin-K dependent natural anticoagulants (proteins C and S) during this period might result in a prothrombic phase. We addressed this question by measuring the rates of decline of these proteins, as well as the vitamin K dependent procoagulants, in two groups of patients: A “high dose group” (n=7), who received a single 40 mg dose of warfarin, and a “low dose group” (n=20), who received daily individually adjusted doses. In the high dose group an early and marked decline of factor VII:C and protein C antigen was observed, while levels of the other vitamin K dependent factors were still relatively high. In the low dose group, all these proteins declined more gradually. Mean ± SD of protein C antigen level at 46 hr was 56±12% in the low dose group, and only 44±6% (p<0.05) in the high dose group. We conclude that during the initiation of warfarin therapy there is a transient prothrombotic phase, which is less marked in patients given daily adjusted doses.     

Determination of functional and antigenic protein C inhibitor and its complexes with activated protein C in plasma by ELISA''s

Enzyme-linked immunosorbent assays (ELISA's) were developed for the measurement of protein C inhibitor (PCI) antigen and activity and for its complexes with activated protein C (APC) in plasma. For PCI activity and antigen, APC or anti-PCI, respectively, was immobilized to microtiter plates and PCI bound was detected with labelled anti-PCI antibodíes. For APC:PCI complexes, two different antibodies directed against protein C and PCI were used. The assays for PCI were calibrated with pooled normal human plasma (NHP) and with purified PCI, and for APC:PCI complexes with known concentrations of purified pre-formed complexes added to buffer or to plasma. The lower limit of sensitivity of the PCI activity and antigen assays was 10 ng/ml and 0.5 ng/ml, respectively and for plasma APC:PCI complexes 12 ng/ml. Mean coefficients of variation of 1.5 % to 5.8 % (intro-assay) and 2.1 % to 9.8 % (inter-assay) were found for the assays. For PCI antigen, a range of 56 % to 162 % of the NHP value was obtained in samples from 70 healthy donors (mean ±SD = 98.6 % ±23.1%). For PCI activity, the range was 59 % to 148 % (94.3 % ± 20.2). A good correlation (0.92) was obtained when both assays were compared. Plasma levels of APC:PCI complexes in 30 normals were under the detection limit (< 12 ng/ml). In plasma samples from 10 patients with disseminated intravascular coagulation (DIC) PCI antigen concentrations were decreased (55.6% ±20%) and 8 of the patients had APC-PCI complex levels between 32 and 240 ng/ml (median, 35 ng/ml). After addition of 20 ωg/ml APC to NHP or to protein C depleted plasma, 6.1 μg/ml complexes were recovered after 90 min incubation. Incubation of 10 μg/ml APC with NHP in the presence of 10 U/ml heparin yielded 11 μg/ml complexes after 90 min, which represent more than 90 % of the maximum possible value. Thus, the method should be adequate to study complexes of APC in vivo in clinical conditions in which activation of protein C pathway may occur.     

Protein C inhibitor: Purification and proteinase reactivity

Protein C inhibitor was purified from human plasma by a modification of a published procedure (Suzuki, K., Nishioka, J., and Hashimoto, S. J. Biol. Chem. 258, 163–168, 1983). Approximately 1 mg of pure protein was obtained from 1 L plasma, a yield of about 17%. The protein C inhibitor preparation did not lose activity over 4 weeks at 4°C. Second order rate constants were measured for the inhibition of activated protein C, thrombin, and urokinase, and bimolecular complexes of protein C inhibitor with activated protein C and thrombin were visualized by denaturing polyacrylamide gel electrophoresis. Heparin accelerated the inhibition of the three proteinases in a manner consistent with a template mechanism. Plasma or pure protein C inhibitor (at the same concentration) showed the same effect of heparin on activated protein C inhibition, indicating that protein C inhibitor accounts for all the heparin-dependent inhibition of activated protein C in vivo.     

Orientation of the myelin proteolipid protein C-terminus in oligodendroglial membranes.

The topology of the integral membrane proteolipid protein (PLP) has important structural and functional implications for central nervous system myelin. To determine the orientation of the carboxyl-terminal portion of PLP, cultured mouse oligodendrocytes were probed with polyclonal antibodies raised against a synthetic terminal peptide corresponding to PLP residues 264-276 and with ten separate monoclonal antibodies that react with this region. Cells were examined by double-label indirect immunofluorescence for the presence of the PLP C-terminus and either oligodendrocyte-specific surface or intracellular antigens. To detect surface antigens, both living and paraformaldehyde-fixed cells were incubated with primary antibodies and then stained with fluorochrome-conjugated second antibodies. Antigens located within the cytoplasmic space were identified after fixation and permeabilization of cells. Live-labeled oligodendrocytes were stained brightly for myelin-oligodendrocyte glycoprotein, galactocerebroside, and other surface markers but did not stain for the PLP C-terminus or the intracellular proteins myelin basic protein and beta-tubulin. Fixation alone was sufficient for partial permeabilization of oligodendrocytes to antibodies and resulted in limited staining of the PLP C-terminus and intracellular proteins. The permeabilized oligodendrocytes stained intensely for the PLP C-terminus, myelin basic protein, and beta-tubulin. Finally, trypsinization of living oligodendrocytes eliminated surface myelin-oligodendrocyte glycoprotein staining but did not change the immunostaining properties of the PLP C-terminus. These results provide evidence that the carboxyl-terminus of PLP is located at the cytoplasmic face of oligodendroglial membranes.