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丙酮酸乙酯对脓毒症保护作用研究进展

丙酮酸是糖酵解的终产物及三羧酸循环的起始物，是机体能量代谢的主要中间产物，当机体氧供应充足时丙酮酸在线粒体内氧化为乙酰辅酶A，维持三羧酸循环；当氧供应不足时在乳酸脱氢酶作用下直接转化成乳酸，迅速为机体供能。丙酮酸在细胞内可能作为内源性抗氧化物，无需酶就能降解过氧化氢，产生二氧化碳和水。丙酮酸消除过氧化氢的反应高效迅速。除了清除过氧化氢，丙酮酸也能清除羟基、一氧化氮等其他活性氧。丙酮酸乙酯（EP）是丙酮酸亲脂性的酯化物，在抗炎、抗凝及安全性方面明显优于丙酮酸[1]

Value of Tumor Type M2 Pyruvate Kinase in Differential Diagnosis of Pleural Effusion

Objective To investigate the value of tumor type M2 pyruvate kinase ( M2-PK) in the differential diagnosis of pleural effusion. Methods A total of 146 patients with pleural effusion during January 2006 to December 2008 were recruited at the department of respiratory medicine of the Shantou Affiliated Hospital and the First Affiliated Hospital of Sun Yat-sen Medical College. Pleural effusion was malignant in 72 cases ( 52 cases with lung cancer and 20 cases with metastatic lung cancer) and benign in 74 cases ( 54 cases with infective pleural effusion and 20 with transudation effusion) . The patients were divided into a malignant pleural effusion group, an infective pleural effusion group, and a transudation group.Then the infective group was further divided into subgroups of tuberculosis pleural effusion group andparapneumonic effusion group. The concentration of tumor M2-PK in pleural fluid obtained during the first thoracocentesis was measured by enzyme-linked immunosorbent assay( ELISA) . Results The concentration of tumor M2-PK was significantly higher in the malignant pleural effusion group compared with the benignpleural effusion groups ( P lt; 0. 01) . Significant differences were also found in the concentration of tumor M2-PK between malignant pleural effusion caused by lung cancer and metastatic lung cancer( P lt; 0. 05) .When the cutoff value of tumor M2-PK was set at 18. 68 U/mL, the sensitivity, specificity, and accuracy for the diagnosis of malignant pleural effusion was 87. 6% , 86. 0% , and 87. 4%, respectively. Furthermore,the detection of tumor M2-PK in combination with CEA showed better diagnostic sensitivity( 96. 0% ) ,specificity ( 85. 0% ) , and accuracy ( 91. 1% ) . Conclusions The detection of tumor M2-PK in pleural effusion is of some clinical significance in the differential diagnosis of benign and malignant pleural effusion.The detection of tumor M2-PK in combination with CEA is a good diagnostic tool with high sensitivity andspecificity.

肿瘤M2型丙酮酸激酶对肺癌诊断价值的临床研究

目的 探讨肺癌患者血浆肿瘤M2 型丙酮酸激酶( TU M2-PK) 水平变化对其诊断的临床意义。方法 用酶联免疫吸附试验( ELISA) 分别检测40 例肺癌患者( 肺癌组) 、40 例良性肺部疾病患者( 良性肺部疾病组) 和40 例正常人( 正常对照组) 血浆TU M2-PK 水平, 并与癌胚抗原( CEA)进行比较分析。结果 癌组患者血浆TU M2-PK 和血清CEA 与良性肺部疾病组和正常对照组比较差异有统计学意义( TU M2-PK:18.6 U/mL比9. 6 和8. 5 U/mL; CEA: 12.02 μg/L比2. 4 和2. 6 μg/L;P 均lt;0.05) 。随病程加重, 肺癌患者血浆TU M2-PK 阳性率逐渐升高, 肺癌Ⅲ、Ⅳ 期患者血浆TU M2-PK和血清CEA 阳性率( 78. 1% 与75. 0% ) 明显高于Ⅰ、Ⅱ 期患者( 50. 0% 与50. 0% , P 均lt;0. 05) 。单独检测血浆TUM2-PK 对肺癌的敏感性为72. 5% , 特异性为90. 2% , 联合CEA 检测敏感度为81. 3% , 特异性为93. 4% 。结论 TU M2-PK 对诊断肺癌有较高的临床价值, 联合CEA 检测可提高肺癌的敏感性。

视网膜母细胞瘤房水烯醇化酶定量检测的临床研究

NALYSIS OF RATE-LIMITING ENZYMES RELATING TO GLYCOLYTIC PATHWAY IN RETINAL TISSUE OF DIABETIC RATS

Eighteen SD rats with streptozotocin-induced diabetes were observed for the influence of magnesium in glycolytic pathway in their retinal tissue.The diabetic rats were divided into 3 groups:6 of them drank 0.5% Mgso4 solution every day,6 received intramuscular Mgso4 (0.05/kg)in half month interval,and the another 6 drank tape water every day.Six normal rats were employed as employed as nondiabetic control.The activity of the three crucial rate-limiting enzymes ralating to glycolytic pathway-hexokinase,phosphofructokinase and pyruvate kinase in retinal tissue of the rats was investigated after a period of 30days.The results revealed that the levels of the enzymes were significantly depressed in diabetic rats not taking magnesium,while the enzyme levels maintained nearly the same in diabetic rats taking magnesium,while the enzyme levels maintained nearly the same in diabetic rats taking magnesium as in the control group.This suggested that the glycolytic pathway in retinal tissue was disturbed in early stage of diabtes,and magnesium might play an important role in maintaining the normal metabolism of glucose. （Chin J Ocul Fundus Dis,1993,9:81-83）

Research of TAP Induction The Pancreatic Acinar Cells to Release HMGB1 in Rat

Objective　To explore the secretion law of high mobility group box 1（HMGB1）in rat pancreatic acinar cells induced by trypsin activation peptide（TAP）and release of HMGB1 affected by ethyl pyruvate（EP）. Methods　The experiment was performed in 12 SD rats. The pancreatic acinar cells of rats were taken out and then separated into three groups：control group， TAP group， and EP group. TAP was added into TAP group and EP group（keep TAP at a final concentration of 3 nmol/L）， respectively， but EP was added into EP group only （keep EP at a final concentration of 28 mmol/L）. The expressions of HMGB1 mRNA and protein were detected by using real-time quantitative reverse transcription polymerase chain reaction（RT-PCR）or Western blot at 3 h， 6 h， 12 h， and 24 h time point， respectively. The relationship between HMGB1 and TAP action time was explored by rank correlation. Results　Compared with control group， the expressions of HMGB1 mRNA and protein were increased with prolongation of the TAP action in TAP group and EP group（P＜0.05）. Compared with TAP group， the expressions of HMGB1 mRNA and protein were decreased in EP group（P＜0.05）. The expressions of HMGB1 mRNA and protein were increased with prolongation of the TAP action（P＜0.05）， and were highest at 12 h time point（P＜0.01）in TAP group. There were positive correlation between the expressions of HMGB1 mRNA and protein and TAP action time（rs=0.971， P＜0.01；rs=0.966， P＜0.01）.Conclusions　TAP can induce the release of HMGB1 in pancreatic acinar cells. There is positive relationship between TAP in early stage and HMGB1 in later period of acute pancreatitis. EP can inhibit the release of HMGB1.

PKM2 在肝细胞肝癌中的表达及其与预后的关系

目的 探讨 M2 型丙酮酸激酶（PKM2）在肝细胞癌（以下简称“肝癌”）组织中的表达，并分析其与肝癌患者临床病理特征及预后的关系。 方法 收集 2008 年 1 月至 2011 年 1 月期间重庆市第九人民医院和重庆市九龙坡第二人民医院病理诊断为肝癌的石蜡标本，采用免疫组织化学方法检测肝癌组织中 PKM2 蛋白的表达，采用Kaplan-Meier 生存分析法、Cox 回归法分析 PKM2 蛋白表达与肝癌患者临床预后的关系。 结果 共收集 196 例肝癌患者的石蜡组织标本，其中 129 例（65.8%）肝癌患者的肝癌组织中 PKM2 蛋白高表达，其高表达与肝癌患者的性别（P=0.049鴐、肿瘤分化程度（P=0.000鴐、血清甲胎蛋白水平（P=0.010鴐、肿瘤数目（P=0.002）及 TNM 分期（P=0.001）有关，而与患者的年龄、有无肝硬化、肿瘤最大直径及乙肝表面抗原无关（P>0.05鴐。PKM2 蛋白高表达患者的 1、3、5 年总体生存率和无瘤生存率均明显低于 PKM2 低表达患者（P<0.05鴐，且多因素 Cox 回归分析结果显示 PKM2 蛋白高表达是肝癌患者总体生存率（HR=1.661，P=0.024）及无瘤生存率（HR=1.662，P=0.013）的独立危险因素。 结论 PKM2 可能对肝癌患者的生物学行为有重要影响，可作为判断肝癌预后的一个新指标。

The Expression and Correlation of PKM2 and YAP in Hepatocellular Carcinoma Tissue

ObjectiveTo investigate the significance of expression and correlation of pyruvate kinase M2 (PKM2) and yes association protein (YAP) in hepatocellular carcinoma (HCC) tissue, and then explore the relationship between the 2 kinds of protein. MethodsA total of 120 patients' HCC tissues and adjacent tissues were collected retrospectively, who treated in our hospital from Apr. 2010 to Oct. 2013, the expressions of PKM2 and YAP protein in these HCC tissues and adjacent tissues were detected by SP immunohistochemical method, and then analyzed the relationship between the expressions of PKM2 and YAP ptotein with the clinicopathological features of HCC. Of the 120 patients, the expressions of YAP and PKM2 protein and its mRNA in 50 cases of HCC tissues and adjacent tissues were also examined by Western blot and real-time PCR methods respectively. Results① The immunohistochemical results showed that, the positive rate of PKM2 protein and YAP protein in HCC tissues were 67.50% (81/120) and 71.67% (86/120) respectively, which were both higher than those of adjacent tissues[PKM2 protein:20.83% (25/120); YAP protein:29.17% (35/120)], P < 0.050. ② The expression of PKM2 protein was significantly positively correlated with the expression of YAP protein in HCC tissues (r=0.519, P < 0.001). ③ In HCC tissues, the expression of PKM2 protein was significantly correlated with the diameter of tumor, TNM staging, differentiation of HCC, and lever of alpha fetal protein (P < 0.050), and the expression of YAP protein was significantly correlated with the differentiation of HCC and lever of alpha fetal protein (P < 0.050). ④ Western blot results showed that, the expression levels of PKM2 protein and YAP protein in HCC tissues were 1.25± 0.11 and 1.08±0.10 respectively, which were significantly higher than those of adjacent tissues (PKM2 protein:0.38±0.01, YAP protein:0.41±0.02), P < 0.050. ⑤ Real-time PCR assays results showed that, basing the expressions of PKM2 mRNA and YAP mRNA in adjacent tissues (both as 1), expressions of PKM2 mRNA and YAP mRNA in HCC tissues were 11.38±0.35 and 19.96±0.48 respectively, which were both higher than those of adjacent tissues (P < 0.050). ConclusionPKM2 and YAP protein were related to the initiation of HCC, and they were also closely correlated with the differentiation and prognosis of HCC.

丙酮酸脱氢酶 E1α 亚基基因突变所致 Leigh 综合征的临床进展

Leigh 综合征（Leigh syndrome，LS）是一种罕见的致死性亚急性脑病，是儿童线粒体病的常见表型，核基因及线粒体基因上多种基因位点突变均可导致这一罕见病，其中丙酮酸脱氢酶 E1α 亚基（PDHA1）基因所致的 LS（PDH-LS）约占比 10%，主要临床表现为肌无力、共济失调、神经退行性障碍等，但却缺乏特异性。该疾病预后较差，目前尚无特异性治疗方案，早期诊断及干预有助于改善 PDH-LS 预后，除了传统的药物治疗及改善营养状态外，生酮饮食亦是一种有益的治疗选择，因此早期明确诊断及相关基因型改变具有重大意义。LS 临床表现多变且缺乏一致的特异性，为早期诊断治疗带来了挑战。文章讨论了 PDH-LS 的临床表现，并总结了文献报道的 PDH-LS 患者的表现及有预警意义的表现，以期提高其早期诊断率和相关的治疗建议，希望能够对此类患者的诊治有所帮助。