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視網膜母細胞瘤房水烯醇化酶定量檢測的臨床研究

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.

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.

丙酮酸乙酯對膿毒癥保護作用研究進展

丙酮酸是糖酵解的終產物及三羧酸循環的起始物，是機體能量代謝的主要中間產物，當機體氧供應充足時丙酮酸在線粒體內氧化為乙酰輔酶A，維持三羧酸循環；當氧供應不足時在乳酸脫氫酶作用下直接轉化成乳酸，迅速為機體供能。丙酮酸在細胞內可能作為內源性抗氧化物，無需酶就能降解過氧化氫，產生二氧化碳和水。丙酮酸消除過氧化氫的反應高效迅速。除了清除過氧化氫，丙酮酸也能清除羥基、一氧化氮等其他活性氧。丙酮酸乙酯（EP）是丙酮酸親脂性的酯化物，在抗炎、抗凝及安全性方面明顯優于丙酮酸[1]

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）

丙酮酸脫氫酶 E1α 亞基基因突變所致 Leigh 綜合征的臨床進展

Leigh 綜合征（Leigh syndrome，LS）是一種罕見的致死性亞急性腦病，是兒童線粒體病的常見表型，核基因及線粒體基因上多種基因位點突變均可導致這一罕見病，其中丙酮酸脫氫酶 E1α 亞基（PDHA1）基因所致的 LS（PDH-LS）約占比 10%，主要臨床表現為肌無力、共濟失調、神經退行性障礙等，但卻缺乏特異性。該疾病預后較差，目前尚無特異性治療方案，早期診斷及干預有助于改善 PDH-LS 預后，除了傳統的藥物治療及改善營養狀態外，生酮飲食亦是一種有益的治療選擇，因此早期明確診斷及相關基因型改變具有重大意義。LS 臨床表現多變且缺乏一致的特異性，為早期診斷治療帶來了挑戰。文章討論了 PDH-LS 的臨床表現，并總結了文獻報道的 PDH-LS 患者的表現及有預警意義的表現，以期提高其早期診斷率和相關的治療建議，希望能夠對此類患者的診治有所幫助。

腫瘤M2型丙酮酸激酶對肺癌診斷價值的臨床研究

目的 探討肺癌患者血漿腫瘤M2 型丙酮酸激酶( TU M2-PK) 水平變化對其診斷的臨床意義。方法 用酶聯免疫吸附試驗( ELISA) 分別檢測40 例肺癌患者( 肺癌組) 、40 例良性肺部疾病患者( 良性肺部疾病組) 和40 例正常人( 正常對照組) 血漿TU M2-PK 水平, 并與癌胚抗原( CEA)進行比較分析。結果 癌組患者血漿TU M2-PK 和血清CEA 與良性肺部疾病組和正常對照組比較差異有統計學意義( 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) 。隨病程加重, 肺癌患者血漿TU M2-PK 陽性率逐漸升高, 肺癌Ⅲ、Ⅳ 期患者血漿TU M2-PK和血清CEA 陽性率( 78. 1% 與75. 0% ) 明顯高于Ⅰ、Ⅱ 期患者( 50. 0% 與50. 0% , P 均lt;0. 05) 。單獨檢測血漿TUM2-PK 對肺癌的敏感性為72. 5% , 特異性為90. 2% , 聯合CEA 檢測敏感度為81. 3% , 特異性為93. 4% 。結論 TU M2-PK 對診斷肺癌有較高的臨床價值, 聯合CEA 檢測可提高肺癌的敏感性。

PKM2 在肝細胞肝癌中的表達及其與預后的關系

目的 探討 M2 型丙酮酸激酶（PKM2）在肝細胞癌（以下簡稱“肝癌”）組織中的表達，并分析其與肝癌患者臨床病理特征及預后的關系。 方法 收集 2008 年 1 月至 2011 年 1 月期間重慶市第九人民醫院和重慶市九龍坡第二人民醫院病理診斷為肝癌的石蠟標本，采用免疫組織化學方法檢測肝癌組織中 PKM2 蛋白的表達，采用Kaplan-Meier 生存分析法、Cox 回歸法分析 PKM2 蛋白表達與肝癌患者臨床預后的關系。 結果 共收集 196 例肝癌患者的石蠟組織標本，其中 129 例（65.8%）肝癌患者的肝癌組織中 PKM2 蛋白高表達，其高表達與肝癌患者的性別（P=0.049鴐、腫瘤分化程度（P=0.000鴐、血清甲胎蛋白水平（P=0.010鴐、腫瘤數目（P=0.002）及 TNM 分期（P=0.001）有關，而與患者的年齡、有無肝硬化、腫瘤最大直徑及乙肝表面抗原無關（P>0.05鴐。PKM2 蛋白高表達患者的 1、3、5 年總體生存率和無瘤生存率均明顯低于 PKM2 低表達患者（P<0.05鴐，且多因素 Cox 回歸分析結果顯示 PKM2 蛋白高表達是肝癌患者總體生存率（HR=1.661，P=0.024）及無瘤生存率（HR=1.662，P=0.013）的獨立危險因素。 結論 PKM2 可能對肝癌患者的生物學行為有重要影響，可作為判斷肝癌預后的一個新指標。

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.