鞘内注射吗啡对切口疼痛大鼠脊髓背角蛋白激酶Cγ免疫反应的影响

目的 研究鞘内注射吗啡对切口疼痛大鼠脊髓背角蛋白激酶Cγ(PKCγ)免疫反应的影响。方法 SD雄性大鼠24只,随机分为4组(每组6只):假手术组(Ⅰ组)、对照组(Ⅱ组)术前30 min鞘内注射人工脑脊液20μl,术后吗啡治疗组(Ⅲ组)、术前吗啡治疗组(Ⅳ组)分别于术后和术前30min鞘内注射吗啡5μg(10 μl)。所有大鼠均按Brennan法制成切口疼痛模型,用免疫组织化学方法观察脊髓背角PKCγ的表达。结果 Ⅱ组术侧脊髓背角PKCγ-IR表达高于非术侧及Ⅰ组(P<0.01)。与Ⅱ组比较,Ⅲ组、Ⅳ组脊髓背角PKCγ-IR表达降低(P<0.05或0.01),而Ⅲ组、Ⅳ组比较差异无显著性。结论 在大鼠切口疼痛模型中,鞘内注射吗啡的镇痛作用可能与抑制脊髓背角的PKCγ-IR免疫反应有关。     

Decreased expression of DMPK: correlation with CTG repeat expansion and fibre type composition in myotonic dystrophy type 1

Abstract Myotonic dystrophy type 1 (DM1) is an autosomal dominant disease caused by a trinucleotide repeatexpansion, cytosine-thymine-guanine (CTG)_n, in the 3′ untranslated region of a gene encoding the myotonic dystrophy protein kinase (DMPK). To correlate CTG expansion and protein expression, we studied muscle specimens from 16 adult DM1 patients using three anti-DMPK antibodies for immunoblotting. We estimated the amount of the full-length DMPK (85 kDa) in muscle biopsies from normal controls and from DM1 patients carrying different (CTG)_n expansions. We found that DMPK concentration was decreased to about 50% in DM patients’ muscles; the protein decrease did not seem correlated with the CTG repeat length. However, the fibre type composition in skeletal muscle seemed somehow to affect DMPK decrease, as the lowest level of the enzyme was found in patients with the lowest content of type 1 fibre.     

Effects of chelating agents on the rheological property of cervical mucus

As an ongoing effort to elucidate the mechanisms involved in the calcium-dependent fertility regulation process, the viscoelastic properties of the mucus obtained from lamb cervix and human semen, as well as their water and total protein contents after exposure to EDTA, a chelating agent, or Nonoxynol-9 (N-9), a spermicidal agent, were examined. The viscosity was measured using a Cone Plate Digital Viscometer, while the water and total protein contents were determined by the lyophilization process and the Lowry method, respectively. The significant changes in the rheological properties of mucus, such as its viscosity and the water content, upon exposure to EDTA were demonstrated. The viscosity of cervical mucus and human semen were significantly increased by EDTA treatment (as compared to the controls): lamb cervical mucus (2.9 ± 0.3 vs. 2.2 ± 0.3 cps) and human semen (5.0 ± 0.3 vs. 4.3 ± 0.3 cps), respectively. The hydration rate was decreased by EDTA treatment as compared with the control (93.6 ± 0.7 vs. 96.8 ± 0.8%). Among tested samples, the reduction in the percentage of sperm penetration through the cervical mucus was the highest in the mucus containing EDTA, which had the lowest water content (93.6 ± 0.7%), indicating that there is a positive relationship between the hydration rate of the cervical mucus and its ability to permit the penetration of spermatozoa. This result indicates that spermicidal activity exerted by high concentrations of EDTA is in part due to its effect on the rheological properties of cervical mucus or semen.     

蛋白激酶C在软脂酸诱导HepG2细胞胰岛素抵抗中的作用

目的从蛋白激酶C(PKC)信号通路角度,探讨游离脂肪酸(FFA)引起肝脏胰岛素抵抗(IR)的可能机制。方法培养HepG2细胞,同时设立对照组、软脂酸(PA)组、高胰岛素组。软脂酸组、高胰岛素组分别用250μmol/L PA、5×10-7mol/L胰岛素处理24h。然后对照组、软脂酸组再根据胰岛素刺激前加( )与不加(-)PKC抑制剂白屈菜红碱盐酸盐(chelerythrine chloride,CC)5μmol/L预处理1h,随机分为两亚组:对照组(-)、对照组( )、PA组(-)、PA组( )。葡萄糖氧化酶法测定胰岛素刺激后12h葡萄糖消耗量,蒽酮法测定胰岛素刺激后3h点细胞内糖原含量,Western blotting技术检测15min点细胞内P-Ser473PKB、P-Ser21/9GSK-3α/β水平。结果PA组(-)与高胰岛素组葡萄糖消耗量无统计学差异(P=0.523)。葡萄糖消耗量、细胞内糖原含量、P-Ser473PKB、P-Ser21GSK-3α、P-Ser9GSK-3β水平均显示,PA组(-)与对照组(-)比较显著降低(P值依次为0.000,0.000,0.004,0.004,0.028),对照组( )与对照组(-)比较略有升高但无显著性差异;PA组( )与PA组(-)比较显著升高(P值依次为0.000,0.014,0.043,0.041,0.035)。结论PA(250μmol/L)体外成功诱导了HepG2细胞产生IR,PKC信号通路在FFA引起肝脏IR中起着重要作用。     

血浆蛋白C、蛋白S与脑梗死的相关性研究

目的研究血浆蛋白C、蛋白S与脑梗死发病的关系。方法对64例脑梗死患者及15例正常对照组进行血浆蛋白C(PC)、蛋白S(PS)检测,并分析其与脑梗死严重程度、年龄及其它脑梗死危险因素之间的关系。结果实验组血浆PC浓度(4.97±1.82μg/ml)与对照组血浆PC浓度(4.74±1.95μg/ml)之间差别无统计学意义;首发组血浆PC浓度(5.28±1.85μg/ml)与复发组血浆PC浓度(4.03±1.40μg/ml)之间差别有统计学意义(P=0.016);实验组血浆PS浓度(7.47±3.87μg/ml)与对照组血浆PS浓度(12.06±3.99μg/ml)之间差别有统计学意义,中老年组血浆PS浓度(6.58±3.33μg/ml)与青年组血浆PS浓度(9.28±4.33μg/ml)之间差别有统计学意义。结论PC降低是复发性脑梗死的危险因素之一;游离PS降低是脑梗死的危险因素;PS水平的降低导致PC功能的降低,进而凝血功能障碍,导致脑梗死的发生。     

Amadori糖化白蛋白对牛视网膜血管二酯酰甘油--蛋白激酶C级联的影响

目的 :探讨Amadori糖化血清白蛋白对牛视网膜血管二酯酰甘油 (DAG)———蛋白激酶C(PKC)信号级联的影响及d-α-生育酚的干预作用。方法 :体外制备的Amadori糖化的牛血清白蛋白 (AGSA)分别在生理浓度葡萄糖 (5 .5mmol/L)和高浓度葡萄糖 (2 0mmol/L)培养液中作用于新鲜牛视网膜血管。采用薄层层析和放射自显影法测定DAG含量和PKC活性 ;并检测d -α-生育酚预处理后DAG、PKC改变。结果 :在生理葡萄糖浓度下 ,牛视网膜血管暴露于AGSA2 4h或 72h后细胞内DAG含量、PKC活性均较对照组显著增加 ,当AGSA和高葡萄糖同时作用于牛视网膜血管时 ,DAG -PKC级联显著激活 ,分别为正常葡萄糖组的 3.4 7倍和 4 .5 4倍 ;在正常血清培养液中 ,与 5 .5mmol/L的葡萄糖相比 ,2 0mmol/L高糖在 2 4h时并不刺激DAG含量增加 (P >0 .0 5 ) ,而 72h时则较对照组增加 4 5 % ,PKC活性较对照增加 5 6 % ,而d -α -生育酚可逆转上述生化改变。结论 :Amadori糖化血清白蛋白可在生理葡萄糖浓度下刺激DAG -PKC途径活化 ,从而影响血管一系列生理功能 ,而d -α-生育酚对血管生化改变有保护作用。     

Effect of dietary protein on Masheri-induced drug metabolising enzymes

The modulation of drug metabolising enzymes by Masheri extract (ME) and Benzo(a)Pyrene [B(a)P] was studied in male Sprague Dawley rats fed different dietary protein levels. Two groups of 21 days old male Sprague Dawley rats were put on a high protein diet (SHP) with 20% Casein, and a low protein diet (SLP) with 3% Casein semisynthetic based diets for 12 weeks. The SLP fed animals showed lower basal levels of the Phase I activating enzymes viz. Cytochrome P450, Benzo(a)Pyrene hydroxylase, Benzphetamine demethylase and Phase II glutathione detoxification system viz. Glutathione (GSH) and Glutathione-S-transferase. ME and B(a)P treatment significantly depleted the glutathione detoxification system in the SLP group whereas an opposite effect was observed in the SHP group. Interstingly, ME and B(a)P treated rats in the SLP group showed a higher percent increase in the hepatic and pulmonary Phase I enzyme activities than those observed in the treated ME/B(a)P treated SHP rats. Furthermore, both ME and B(a)P significantly decreased the hepatic pool of vitamin A while a concomittant increase in that of vitamin C was observed.     

CD117阳性小肠间质瘤中Ki67和p53蛋白的表达及在预后中的意义

目的 :研究小肠间质瘤 (smallbowelstromaltumors,SBST)Ki6 7和p5 3免疫组织化学的表达 ,探讨两者在肿瘤良恶性划分和预后中的作用。方法 :选用CD117阳性的SBST 33例进行光镜观察 ,用EnVisionTM+二步法免疫组化方法检测Ki6 7和 p5 3蛋白在肿瘤中的表达情况并与形态学进行比较。 结果 :33例SBST患者 ,男 2 0例 ,女13例 ,年龄 2 1～ 71岁 ,发生于十二指肠 5例 ,空肠 2 2例 ,回肠 6例。肿瘤最大径的范围在 1.5～ 2 0cm之间。在光镜下出现肿瘤内坏死的有 11例。肿瘤核分裂象计数每 5 0个高倍镜视野 0个有 6例 ,1～ 2个 5例 ,≥ 3个的有 2 2例。 33例中良性 4例 ,交界性 4例 ,恶性 2 5例。随访时间为 3～ 180个月 (平均 73.3个月 ) ,结果为良性组 4例均无瘤生存 ;交界性组 2例无瘤生存 ,1例失访 ,1例在无瘤生存 18个月后失访 ;恶性组无瘤生存有 7例 ,带瘤生存者及死亡有 16例 ,2例失访。Ki6 7染色阴性 2例 ,均为良性 ;其余 31例阳性中 ,增殖指数 (阳性细胞数 )大于 5 %的有恶性 16例 (94 .1% ) ,交界性 1例。在免疫组化p5 3染色中 ,恶性组 2 5例均呈阳性表达 (平均阳性细胞数 4 2 .9% ) ,其中 12例阳性细胞数大于 5 0 %。当Ki6 7和 p5 3均为阴性或两者阳性细胞数分别是 <1%和 <10 %时 ,肿瘤呈良性经过 ,预后     

胃肠道肿瘤病人围手术期细胞因子及蛋白质代谢的变化

目的 :探讨胃肠道肿瘤病人手术后肿瘤坏死因子 α (TNF α)、白介素 6 (IL 6 )、白介素 10 (IL 10 )的变化以及与蛋白质代谢的关系。　方法 :用酶联免疫法 (ELISA)检测 19例胃肠道肿瘤根治术病人术前和术后 1、3、5天血清TNF α、IL 6和IL 10水平 ,同时检测蛋白质分解代谢相关指标、2 4h尿中尿素和肌酐排泄量。　结果 :胃肠道肿瘤根治术后病人TNF α呈下降趋势 ,术后第 1天较术前明显下降 (P <0 .0 5 )。IL 6和IL 10术后升高 (P <0 .0 1) ,尤以术后第 1天升高明显。术后 2 4h尿中尿素和肌酐排泄量明显升高。　结论 :胃肠道肿瘤根治术后病人IL 6和IL 10的升高支持手术应激导致的细胞因子释放变化 ,但TNF α呈下降趋势 ,可能与胃肠道肿瘤病人术前TNF α的高表达有关。术后蛋白质分解代谢增加 ,可能与细胞因子IL 6和IL 10的升高有关。