缺血预处理对大鼠小体积供肝一氧化氮合酶的影响及意义

目的:探讨缺血预处理（IPC）对大鼠小体积肝移植术后肝脏组织eNOS和 iNOS mRNA表达的影响及意义。 方法:60只SD大鼠随机分为3组（每组10对）:无热缺血组（NWI）、单纯缺血再灌注组（WI）和缺血预处理组（IPC）。用双袖套法建立大鼠小体积肝移植模型。肝组织eNOS mRNA和iNOS mRNA的表达检测用荧光定量PCR法。 结果:IPC后肝组织eNOS mRNA表达较IPC前高(P<0.05)。供肝再灌注后0.5、1、2及 3 h，各组肝组织eNOS mRNA表达均高于术前(P<0.05)，NWI组和WI组eNOS mRNA表达无显著差异（P>0.05），IPC组eNOS mRNA表达高于其它两组(P<0.05或P<0.01)。各组肝组织iNOS mRNA均在供肝再灌注后1h开始表达，术后 2 h 和 3 h IPC组iNOS mRNA表达低于WI组(P<0.05或P<0.01)，NWI组iNOS mRNA表达又低于IPC组(P<0.05或P<0.01)。 结论:IPC可能通过促进供肝再灌注后早期eNOS mRNA表达和抑制再灌注后晚期iNOS mRNA表达而保护小体积供肝。     

原发性肝癌患者血清AFP及外周血AFP mRNA结果分析

目的:探讨血清AFP及外周血AFP mRNA联检对原发性肝癌(PLC)患者的早期诊断和微小转移的临床意义。方法:采用电化学发光法和实时荧光定量PCR分别检测40例PLC患者血清AFP和外周血AFPmRNA。结果:PLC患者血清AFP和外周血AFP mRNA阳性率分别为72.50%(29/40)和57.50%(23/40),显著高于肝硬化患者(8.33%和4.17%,P<0.05)。其中血清AFP阳性和阴性的PLC患者分别有21例(72.41%)和2例(18.18%)检出AFP mRNA。结论:外周血AFP mRNA可作为血循环中存在肝癌细胞的一个早期标志物,与血清AFP联检有助于提高PLC的确诊率,更好地预测肿瘤转移和复发。同时为开展PLC靶向治疗提供依据。     

甲胎蛋白异质体L3对低浓度甲胎蛋白肝癌诊断的临床意义

目的探讨在低浓度甲胎蛋白（AFP）肝病患者中,甲胎蛋白异质体L3（AFP-L3）的百分含量（AFP—L3％）对肝癌的早期诊断和疗效评估的临床意义。方法收集245例血清低AFP含量（5～40ng／m1）的肝病患者样本（其中肝硬化患者100例、肝癌患者145例）,检测AFP—L3％,并对其中100例肝硬化患者和20例肝癌术后患者分别进行3个月和12个月的随访。结果以AFP—L3％≥10％作为阳性判断标准,100例肝硬化患者中阳性为23例,经3个月随访后其中8例诊断为肝癌;145份肝癌患者血清AFP-L3％阳性率为46．2％（67／145）。低浓度AFP肝癌组的AFP—L3％水平显著高于低AFP肝硬化组（t=7．318,P=0．001〈0．01）;20例肝癌患者术后有5例AFP．L3％仍为阳性,其在12个月内的生存率为0,而术后AFP—L3％阴性患者生存概率为15／15。结论AFP—L3％在低浓度AFP肝病患者中对肝癌的早期诊断和术后疗效评估都具有一定的临床意义。     

臂丛损伤再生中GAP-43 mRNA及其蛋白的表达

目的：分析臂丛损伤后脊髓前角运动神经元表达GAP-43 mRNA及其蛋白的变化规律，探讨神经损伤再生的机制。方法：建立3种臂丛损伤模型：右C₇前根撕脱（A组）；右C₇前根撕脱＋同侧C5-T₁后根断离（B组）；右C₇前根撕脱+右C₅C₆间脊髓半横断（C组）。用荧光定量RT-PCR方法检测术后14 d时 C₇前角GAP-43 mRNA的表达量。用免疫组化方法检测术后1、 3、 7、14 d脊髓前角GAP-43免疫阳性运动神经元的表达。结果：对照组C₇前角GAP-43 mRNA呈低表达，损伤组GAP-43 mRNA表达显著上调。损伤组术后1 d、3 d时均未见C₇前角 GAP-43免疫阳性神经元，术后7 d各损伤组GAP-43免疫阳性神经元开始出现，14 d时免疫阳性神经元数目达到高峰。3组间比较，C组表达量最高，B组最低，A组居中。结论：臂丛损伤诱导运动神经元GAP-43 mRNA及其蛋白表达上调，GAP-43合成增加是神经元蛋白重组所致，与轴索再生和神经功能重建有关。     

miR-144和miR-502—3p在原发性肝细胞癌术后复发患者中的表达及临床意义

目的探讨miR-144和miR-502-3p在原发性肝细胞癌术后复发患者中的表达及临床意义。方法从我院肝癌数据标本库中挑选82例原发性肝癌深低温冻存组织标本。其中50例早期复发组（术后1年内肝内出现复发病灶）；32例非早期复发组（术后超过2年未出现肝内复发病灶）。运用实时荧光定量PCR验证miR-144、miR-502—3p在两组肝癌组织中的表达水平。结果相对于非早期复发组，miR-144在肝癌早期复发组中表达上调[（43．893±107．890）VS（6．321±6．845），P=0．018]；其在肝癌组织中的表达仅与肝癌术后早期复发有关（P〈0．05）。而miR-502—3p在肝癌早期复发组中表达下调[（6．702±9．775）VS（26．467±39．613），P=0．009]；其在肝癌组织中的表达与肿瘤直径、肝癌术后早期复发、脉管癌栓、肝硬化、Edmonson病理分级有关（P〈0．05）。结论miR-144、miR-502-3p与肝癌的早期复发转移密切相关，并且可能是肝癌早期复发的分子标记物以及未来肝癌靶向治疗的有效靶点。     

血清CRP和AFP联合检测在良恶性肝病鉴别诊断中的意义

目的探讨血清C反应蛋白（CRP）和甲胎蛋白（AFP）联合检测在良恶性肝病鉴别诊断中的临床意义。方法对158例肝病患者及30例健康人用免疫比浊法定量测定血清CRP含（x）量及用化学发光法定量测定血清AFP含量。结果肝病患者各组血清CRP水平和AFP含单独检测血清CRP和AFP对原发性肝癌诊断的敏感性分别是85．7％和78．6％特异性分别是74．0％和89．7％：以AFP和CRP至少一项阳性作为判断标准,敏感性上升为96．9％,二者均阳性对原发性肝癌诊断的敏感性为67．4％,特异性为97．3％。结论血清AFP检测是诊断原发性肝癌的理想指标,但在原发性肝癌患者中有20~30％的病例AFP为阴性,部分良性肝病患者AFP也呈不同程度阳性,血清CRP和AFP联合检测对肝癌诊断有互补作用．对良恶性肝病鉴别诊断有重要意义。     

血清S-CD105与VEGF联合监测复发转移性乳腺癌的意义

目的：探讨联合检测血清S-CD105和VEGF表达对乳腺癌术后复发、转移的临床意义。方法：用酶联免疫吸附实验（ELISA）分别检测50例复发转移性乳腺癌、20例原发性乳腺癌、20例乳腺良性疾病及15例健康女性血清S-CD105和VEGF的浓度;用微粒子化学发光法检测复发转移患者同期血清CEA、CA15-3的浓度。结果：复发转移性乳腺癌组血清S-CD105、VEGF浓度明显高于原发性乳腺癌组、乳腺良性疾病组及健康对照组（P〈0．05）。血清S—CD105与VEGF联合表达阳性率高于CEA与CA15-3联合表达率,差异有统计学意义（P〈0．05）。结论：复发转移性乳腺癌患者血清S-CD105与VEGF联合检测优于CEA与CA15-3联合检测,可能对乳腺癌患者术后复发转移的早期诊断有重要的临床价值。     

臂丛损伤脊髓运动神经元与神经根GAP-43 mRNA表达

目的：探讨臂丛根性撕脱伤后脊髓腹角运动神经元胞体及其神经根GAP-43 mRNA的表达变化及其影响因素,为臂丛损伤的修复治疗提供理论依据.方法：本实验创立三种臂丛根性撕脱伤模型：C7前根撕脱（Ⅰ组）;C7前根撕脱＋切断同侧C5～T1后根（Ⅱ组）;C7前根撕脱＋C5和C6之间作同侧脊髓半横断（Ⅲ组）.术后2周按CBS评分标准检查动物神经缺失症状,用SYBR Green荧光定量RT-PCR方法检测脊髓腹角运动神经元胞体及其神经根GAP-43 mRNA的表达改变.结果：根据CBS评分标准,对照组计为0分,Ⅰ组计分较低、Ⅲ组计分最高.对照组C7神经元胞体和C7神经根中GAP-43 mRNA表达量相近,但三种损伤组术后2周神经元胞体内GAP-43 mRNA表达均上调,而神经根内表达却下调.结论：（1）臂丛根性撕脱伤后脊髓腹角运动神经元胞体GAP-43 mRNA表达受突触前机制的调控;（2）臂丛损伤2周时神经元胞体内GAP-43 mRNA表达呈现高峰期,此时进行神经移位术将显著提高神经修复的效果.     

Tiam-1 mRNA及其蛋白的表达与鼻咽癌浸润转移的关系

目的探讨Tiam-1 mRNA及其蛋白表达与鼻咽癌浸润转移的关系及意义。方法41例不同临床分期人鼻咽癌组织以及对照组20例鼻咽慢性炎症黏膜组织标本，用RT-PCR方法检测Tiam-1 mRNA表达水平，用免疫组化检测Tiam-1蛋白表达。结果Tiam-1 mRNA在鼻咽癌组织的平均表达水平为I．83±0．73，明显高于鼻咽黏膜慢性炎组织（0．87±0．45）（P〈0．01）；而且鼻咽癌组Tiam-1 mRNA高表达率为46．34％，明显高于对照组（P〈0．01）。Tiam-1蛋白在鼻咽癌组阳性率为65．85％（27／41），明显高于对照组（15％，3／20）（P〈0．01）；其高表达率为43．90％（18／41），明显高于对照组（P〈0．01）。Tiam-1 mRNA及其蛋白高表达率在临床TNM各分期间的差异均有明显统计学意义（P〈0．01）；在各T分期之间的差异亦有明显统计学意义（P〈0．01）。鼻咽癌中有淋巴结转移组Tiam-1 mRNA及其蛋白高表达率分别为59．26％（16／27）及55．56％（15／27），均明显高于无淋巴结转移组的21．43％（3／14）及14．28（2／14），（P〈0．05）。结论Tiam-1 mRNA及其蛋白高表达与鼻咽癌浸润转移有密切关系，提示Tiam-1基因可能是鼻咽癌的重要促浸润转移因子，有望成为治疗的靶标及有价值的预后判断指标。     

组织因子在肝细胞癌的发生及侵袭转移中的研究

目的：检测组织因子（TF）在肝细胞癌患者中的表达并探讨其意义。方法:采用酶联免疫法检测肝细胞癌患者50例及对照组30例的血浆TF水平，采用RT-PCR法检测其中27例肝癌、癌旁、正常肝组织TF mRNA表达。结果:①肝细胞癌患者血浆TF显著高于对照组（P<0.05）。血浆TF在分化高低、肿瘤大小及是否合并肝硬化组间表达有显著差异（P<0.05），在有淋巴转移、肝外脏器转移及门脉癌栓组高于无转移及癌栓组(P<0.05)，而在不同癌灶数目及包膜情况组间无显著差异（P＞0.05）。②TF mRNA在肝癌组织中阳性率及相对表达强度分别为62.96%（17/27）、0.567±0.268，均显著高于癌旁组织及正常组织。阳性患者相对表达强度在肿瘤大小及部分侵袭转移指标中表达有显著差异（P<0.05） 。结论：TF在肝癌患者血浆及组织中升高且与部分侵袭转移指标相关，提示其TF可能促进了肝癌的发生与侵袭转移。     

The clinicopathological and prognostic relevance of cytokeratin 7 and 19 expression in hepatocellular carcinoma. A possible progenitor cell origin

AIMS: Cytokeratin (CK) 7 and CK19 expression, present in hepatic progenitor cells (HPCs) and in cholangiocytes but not in normal hepatocytes, has been reported in some hepatocellular carcinomas (HCCs); however, the incidence and relevance of this expression in HCC in Caucasians is not known. Therefore, our aim was to study the occurrence and clinicopathological characteristics of HCC expressing CK7 and/or CK19 in 109 Caucasian patients. METHODS AND RESULTS: The expression of hepatocellular differentiation markers (Hepar, canalicular polyclonal carcinoembryonic antigen), biliary/progenitor cell markers (CK7, CK19), alpha-fetoprotein (AFP), p53 and beta-catenin in HCC was semiquantitatively assessed by immunohistochemistry. Of 109 HCCs, 78 were CK7-/CK19- (72%), 13 CK7+/CK19- (12%), seven CK7-/CK19+ (6%), 11 CK7+/CK19+ (10%). CK19 expression was significantly associated with elevated serum AFP (400 ng/ml) (P = 0.023), tumour AFP expression (P < 0.0001), presence in serum of anti-hepatitis B core (P = 0.016), less fibrosis in non-neoplastic parenchyma (P = 0.009) and less nuclear beta-catenin expression (P = 0.021). CK7 expression was significantly associated with elevated serum bilirubin (> 2 mg/dl) (P = 0.0005) and less nuclear beta-catenin expression (P = 0.003). HCC expressing CK19 had a higher rate of recurrence (P = 0.009, hazard ratio 12.5, n = 31) after liver transplantation compared with CK19- tumours. CONCLUSIONS: In our series, 28% of HCCs contained cells expressing CK7 and/or CK19. They potentially derive from HPCs. The higher recurrence rate of CK19+ HCC after transplantation suggests a worse prognosis for these HCCs compared with CK19- HCC.     

Peri-Transplant Change in AFP Level: a Useful Predictor of Hepatocellular Carcinoma Recurrence Following Liver Transplantation

Pretransplant alpha-fetoprotein (AFP) is a useful tumor marker predicting recurrence of hepatocellular carcinoma (HCC). Little is known, however, about the relationship between changes in AFP concentration and prognosis. This study investigated the clinical significance of change in peri-transplant AFP level as a predictor of HCC recurrence. Data from 125 HCC patients with elevated pretransplant AFP level who underwent liver transplantation (LT) between February 2000 and December 2010 were retrospectively reviewed. Patients with AFP normalization within 1 month after LT were classified into the rapid normalization group (n = 97), with all other patients classified into the non-rapid normalization group (n = 28). Tumor recurrence was observed in 17 of the 97 patients (17.5%) with rapid normalization; of these, 11 patients had high AFP levels and six had normal levels at recurrence. In contrast, tumor recurrence was observed in 24 of the 28 patients (85.7%) without rapid normalization, with all 24 having high AFP levels at recurrence. Multivariate analysis showed that non-rapid normalization (harzard ratio [HR], 4.41, P < 0.001), sex (HR, 3.26, P = 0.03), tumor size (HR, 1.15, P = 0.001), and microvascular invasion (HR, 2.65, P = 0.005) were independent risk factors for recurrence. In conclusion, rapid normalization of post-LT AFP level at 1 month is a useful clinical marker for HCC recurrence. Therefore, an adjuvant strategy and/or intensive screening are needed for patients who do not show rapid normalization.     

肝癌肝移植后亚砷酸化疗的应用

背景：移植后肿瘤复发是影响肝癌肝移植疗效的主要因素,如何防止肝癌肝移植后肿瘤复发是目前肝移植研究的热点问题之一。亚砷酸全身化疗被认为对中晚期肝癌具有一定作用,但在肝移植后的应用还未见报道。 目的：观察超出米兰标准的肝癌患者肝移植后应用亚砷酸全身化疗的对肿瘤复发的干预效果。 方法：对23例超出米兰标准的肝癌患者肝移植后采用亚砷酸行预防性化疗：静脉滴注10 mg/d,连续使用7 d后间隔7 d,重复4次为1个疗程,患者接受1~4个疗程。观察以上使用亚砷酸化疗患者的生存、肿瘤复发情况,以及化疗不良反应,并与同期16例未使用化疗的肝癌肝移植患者相比较。 结果与结论：经过3~32个月随访,共30例患者出现肝癌复发,化疗组16例,非化疗组14例,复发部位最常见于肺部、移植肝及骨骼。化疗组与非化疗组肿瘤复发率差异无显著性意义,但化疗组复发时间明显延迟(P=0.026);两组6个月、1年生存率差异无显著性意义,化疗组2年生存率显著高于非化疗组(P=0.037);两组6个月无瘤生存率差异无显著性意义,1年、2年无瘤生存率化疗组显著高于明显非化疗组(P=0.030,0.023)。亚砷酸使用过程中未发现严重不良反应。提示肝癌肝移植患者静脉使用亚砷酸化疗可以延迟肿瘤复发,提高生存率。     

Serum alpha-fetoprotein and its lectin reactivity in liver diseases: a review.

Increased serum concentration of alpha-fetoprotein (AFP) can be found in benign and malignant liver diseases, in yolk sac tumors, and in several nonhepatic neoplasms at advanced stage. The frequency and level of elevated serum AFP are highest in hepatocellular carcinoma (HCC) and yolk sac tumors. Most levels of serum AFP in HCC are greater than 500 ng per mL, whereas the serum AFP in most of the benign liver diseases is only moderately elevated and is transient in nature. Determination of lectin reactivity of serum AFP is helpful for the differentiation of HCC from other diseases associated with elevated serum AFP. Determination of Len culinaris agglutinin (LCA) reactivity of serum AFP is useful for the differentiation of HCC from benign liver diseases, and for early detection of hepatoma. Determination of concanavalin A (Con A) nonreactive AFP variant is useful for the differentiation of HCC from yolk sac tumors and may also allow for the differentiation of HCC from nonhepatic neoplasms. However, reaction with several lectins may be required if differentiation among various nonhepatic neoplasms is needed.     

经导管肝动脉化疗栓塞对肝癌患者外周血黑色素瘤抗原1 mRNA的影响

目的 探讨经导管肝动脉化疗栓塞(TACE)对肝癌患者外周血黑色素瘤抗原1(MAGE-1)mRNA表达水平的影响.方法 采用巢式RT-PCR方法 测定18例肝癌患者TACE治疗前后外周血MAGE-1 mRNA的表达.分析其与肝癌临床病理特征间的关系.结果 18例肝癌患者治疗前外周血MAGE-1 mRNA表达10例阳性、8例阴性;TACE治疗后11例阳性、7例阴性.治疗前后外周血MAGE-1mRNA阳性率差异无统计学意义(55.6%比61.1%.P>0.05).治疗前后肝癌患者外周血MAGE-1mRNA阳性率与肿瘤TNM分期((0%比76.9%,20%比76.9%,P<0.05)、远处转移有关(85.7%比36.4%,100%比36.4%,P<0.05),而与肿瘤大小、血清AFP无关(P>0.05).结论 TACE治疗肝癌不会促进肿瘤的血源性转移,MAGE-1 mRNA可作为评估肝癌疗效的一个重要指标.     

Combined Analysis of Serum Alpha-Fetoprotein and MAGE-A3-Specific Cytotoxic T Lymphocytes in Peripheral Blood for Diagnosis of Hepatocellular Carcinoma

We investigated the feasibility of the combined detection of HLA-A2/MAGE-A3 epitope-specific cytotoxic T lymphocytes (CTLs) and serum alpha-fetoprotein (AFP) for specific diagnosis of hepatocellular carcinoma (HCC). We detected the frequency of MAGE-A3 epitopes (p112–120, KVAELVHFL) in spontaneous CTLs in the peripheral blood of HCC patients, liver cirrhosis patients, and healthy subjects with HLA-A2/polypeptide complex (pentamer) detection technology. Eighty-five HCC cases, 38 liver cirrhosis cases, and 50 healthy cases who were HLA-A2-positive were selected from 175 HCC patients, 80 patients with liver cirrhosis, and 105 healthy volunteers, respectively. The frequency of HLA-A2-specific MAGE-A3⁺ CTLs in the HCC group was significantly higher than that in the other groups. Combined detection of MAGE-A3⁺ CTL frequency and serum AFP value had a higher specificity than either of the two indicators alone. The pentamer technique is helpful in distinguishing benign lesions and malignant lesions in the liver. Combined with serum AFP, it can improve the diagnosis performance for HCC, especially for AFP-negative cancer.     

Role of long-term lamivudine treatment of hepatitis B virus recurrence after liver transplantation

In this study, the long-term (>3 years) efficacy of combination therapy for hepatitis B virus (HBV) recurrence and the associated factors were investigated. One hundred and sixty-five consecutive HBsAg-positive patients (92 with liver cirrhosis, 73 with hepatocellular carcinoma; HCC) who underwent liver transplantation were assessed with a median follow-up time of 40 months. One hundred and twenty-one patients (121/165, 73.3%) were treated with lamivudine before transplantation for a mean of 8.4 months (range 0.1-72 months). The post-transplantation treatment protocol consisted of a high dose intravenous hepatitis B immunoglobulin (HBIg) followed by a low dose intramuscular HBIg and lamivudine combination therapy. Seven (4.2%, 7/165) recipients experienced HBV recurrence at a median time of 19 months (range 5-36 months) following transplantation. Six of seven cases of HBV recurrence were treated with lamivudine before transplantation for a median period of 15 months (range 0.6-30 months). Eighteen (24.6%, 18/73) patients had HCC recurrences after transplantation. Of the four patients with both HCC and HBV recurrence, three experienced HBV recurrence after recurrence of HCC. The clinical factor associated with HBV recurrence in the total cohort (n = 165) was the duration of antiviral treatment (over 6 months) before transplantation (P = 0.004). In the HCC group, HCC recurrence after transplantation (P = 0.002), tumor burden before transplantation (P = 0.005), and postoperative adjuvant chemotherapy (P = 0.002), were additional factors for HBV recurrence. Combination therapy of HBIg and antiviral drugs was effective over 3 years regardless of the pretransplantation viral load. However, the possible recurrence of HBV needs to be monitored cautiously in patients treated with long-term (over 6 months) lamivudine.     

Serological biomarkers of hepatocellular carcinoma in Egyptian patients

Hepatocellular carcinoma (HCC) is one of the most aggressive cancers worldwide. In Egypt, the disease is usually detected in an advanced stage at which no treatment may be effective including surgery. Early detection of the disease is thus an important goal allowing the patient to be treated before the enlargement of the tumor or its metastasis to distant organs. Tumor markers are serological agents which serum level may be useful in predicting the presence of the tumor at early stages. Alpha fetoprotein (AFP) which is the golden marker for HCC is of low sensitivity, therefore, additional markers such as alpha-L-fucosidase (AFU), transforming growth factors alpha and beta (TGF-α and TGF-β) and interleukin-8 (IL-8) are suggested to be simultaneously evaluated in order to enhance the detection of HCC. A total of 96 patients with different liver diseases such as HCC, hepatitis C virus (HCV), hepatitis B virus (HBV) and cirrhotic patients are included in this study. Sixteen healthy volunteers are used as a control group. In patients with HCC each of AFP, AFU, TGF-α and TGF-β recorded significantly higher levels than the other patient groups and controls. HCC patients recorded significantly lower level of IL-8 compared to the other patient groups but significantly higher than the control. For AFP, AFU, TGF-α, TGF-β and IL-8, at the optimal cut-off values (obtained from the receiver operating characteristic (ROC) curves), the calculated sensitivities are 46%, 72.97%, 67.56%, 54.05% and 83.8%, respectively. The simultaneous evaluation using all of the suggested markers resulted in increasing the sensitivity up to 100%. It thus recommended that, if patients with cirrhosis, as high risk patients, are subjected to regular examination using these markers in addition to AFP, HCC may be detected by 100% sensitivity in an early stage and as a consequence an effective treatment can be achieved.