信号转导与转录活化因子3蛋白在小鼠实验性三硝基苯磺酸结肠炎发病中的作用

背景：克罗恩病（CD）的病因尚不清楚，有研究认为肠黏膜免疫功能异常导致了CD的发病。目的：研究信号转导与转录活化因子3（STAT3）蛋白在小鼠实验性三硝基苯磺酸（TNBS）结肠炎发病中的作用。方法：建立小鼠实验性TNBS结肠炎模型，实验第3天予STAT3反义寡核苷酸（ASON）灌肠，第7天处死。分离肠黏膜同有层单个核细胞（LPMC），流式细胞仪检测LPMC细胞凋亡，蛋白质印迹分析检测LPMCSTAT3、磷酸化STAT3（pSTAT3）以及凋亡相关蛋白Bcl-2、Bax的表达，酶联免疫吸附测定（EUSA）检测肠组织匀浆中肿瘤坏死因子（TNF）-αγ、干扰素（INF）-γ等炎性细胞因子的表达。结果：与对照组相比，TNBS结肠炎组炎症肠黏膜LPMC pSTAT3的表达增高，Bcl-2表达增高而Bax表达降低，炎症肠组织TNF—α、INF-γ高表达。以STAT3ASON治疗后，肠黏膜LPMCSTAT3、pSTAT3表达降低，Bcl-2表达降低而Bax表达增高，LPMC凋亡增加，炎症肠组织中TNF—α、INF-γ的表达也明显降低。结论：STAT3通过调节炎性细胞因子的分泌和调控LPMC凋亡相关蛋白的表达，参与了小鼠实验性TNBS结肠炎的发病；抑制STAT3的激活能明显缓解小鼠结肠炎的病情。     

急性胰腺炎后胰腺功能和形态变化的研究现状

急性胰腺炎作为临床上常见的疾病，关于其治愈或好转后胰腺功能和形态改变的研究较少。目前认为急性胰腺炎发病后只要去除诱发病因以及相关并发症，胰腺功能和形态就可恢复，但这一观点仍存争议。本文就急性胰腺炎后胰腺功能和形态的变化作一综述。     

5-脂氧合酶在急性胰腺炎发病中的作用

5-脂氧合酶（LOX）是将花生四烯酸、亚油酸和其他多不饱和脂肪酸转变为具有生物活性的代谢产物如白三烯类的限速酶，从而影响细胞信号转导、结构和代谢。研究发现5-LOX在急性胰腺炎的发生、发展中起重要作用，本文就此作一综述：     

SOCS3在实验性急性胰腺炎急性肺损伤大鼠肺组织中的表达和作用

背景：细胞因子信号转导抑制分子3（SOCS3）在多种疾病的各种器官损伤中起重要的调节作用,可通过对JAK/STAT信号通路的负反馈作用调节炎症因子的释放,从而起一定的抑炎作用。目前关于SOCS3在重症急性胰腺炎（SAP）急性肺损伤中的表达和作用尚未见报道。目的：探讨SOCS3在实验性急性胰腺炎（AP）合并急性肺损伤大鼠肺组织中的表达变化及其可能的作用。方法：32只Sprague-Dawley大鼠随机分为对照组和AP 6 h、12 h、18 h组。以4%牛磺胆酸钠胰胆管逆行注射诱导AP模型。动态测定各组血清淀粉酶（AMY）水平、肺湿/干重比;光学显微镜下观察肺组织学表现;ELISA法检测血清白细胞介素（IL）-6、IL-18含量;免疫组化法和蛋白质印迹法检测肺组织中SOCS3的定位和表达。结果：与对照组相比,各AP模型组血清AMY水平、肺湿/干重比均明显升高（P〈0.05）;肺组织损伤随病情进展而逐渐加重;血清IL-6、IL-18水平显著上调（P〈0.05）;肺组织SOCS3表达逐渐增强（P〈0.05）,于18 h时达高峰。结论：SAP急性肺损伤导致的炎症反应可诱导SOCS3在肺组织中表达,并随着肺组织损伤和炎症反应严重程度的增加而逐渐增高,提示可能与其负反馈调节JAK/STAT信号通路介导的炎症反应的作用存在一定的联系。     

The Role of Gut Microbiota and Genetic Susceptibility in the Pathogenesis of Pancreatitis

Pancreatitis is one of the most common inflammatory diseases of the pancreas caused by autodigestion induced by excessive premature protease activation. However, recognition of novel pathophysiological mechanisms remains a still challenge. Both genetic and environmental factors contribute to the pathogenesis of pancreatitis, and the gut microbiota is a potential source of an environmental effect. In recent years, several new frontiers in gut microbiota and genetic risk assessment research have emerged and improved the understanding of the disease. These investigations showed that the disease progression of pancreatitis could be regulated by the gut microbiome, either through a translocation influence or in a host immune response manner. Meanwhile, the onset of the disease is also associated with the heritage of a pathogenic mutation, and the disease progression could be modified by genetic risk factors. In this review, we focused on the recent advances in the role of gut microbiota in the pathogenesis of pancreatitis, and the genetic susceptibility in pancreatitis.     

抑制JAK／STAT信号通路对实验性急性胰腺炎大鼠肝损伤的保护作用

背景：JAK／STAT细胞内信号通路广泛参与细胞的增殖、分化、凋亡以及炎症、肿瘤的发生等多种生理、病理生理过程，然而关于其在重症急性胰腺炎（SAP）急性肝损伤中作用的研究尚少。目的：观察抑制JAK〈STAT通路对实验性急性胰腺炎（AP）大鼠肝损伤的保护作用。方法：56只Sprague-Dawley大鼠随机分为正常对照组、3组AP模型组和3组JAK特异性抑制剂AG490干预组。以4％牛磺胆酸钠胰胆管逆行注射诱导AP模型。分批处死各组大鼠，动态测定血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）水平，观察肝脏大体和组织学表现，以免疫组化染色和蛋白质印迹法检测肝组织中JAK2的定位和表达。结果：与正常对照组相比，AP模型组各时间点血清ALT、AST水平均显著升高；肝组织大体和组织学损伤随病情进展而逐渐加重；肝组织JAK2表达逐渐增强，于18h时达高峰。经AG490预处理的大鼠，上述各项指标均较同时间点AP模型组显著改善。结论：JAK2参与了大鼠实验性AP肝损伤的病理过程，抑制肝组织JAK／STAT通路活化有助于SAP急性肝损伤的防治。     

胰腺癌和慢性胰腺炎中弹性蛋白酶3B(ELA3B)基因甲基化状态检测

背景：DNA甲基化指DNA中的核苷酸与甲基基团共价结合，这是一种表观遗传修饰，在许多生物学过程中发挥重要作用。目的：探讨弹性蛋白酶3B（ELA3B）基因在胰腺癌中异常表达的潜在机制。方法：以逆转录聚合酶链反应（RT-PCR）检测30例胰腺癌、20例慢性胰腺炎和10例正常胰腺组织中ELA3B mRNA的表达，以甲基化特异性PCR（MSP）检测该基因启动子区甲基化水平。结果：胰腺癌、慢性胰腺炎和正常胰腺组织ELA3B mRNA的表达率依次为46．7％、85．0％和100％。胰腺癌组织ELA3B基因启动子区甲基化指数（MI）显著高于慢性胰腺炎和正常胰腺组织（7．02±6．31对2．33±0．97和3．31±0．75．P〈0．05），其中ELA3B mRNA表达阴性胰腺癌组织的MI显著高于ELA3B mRNA表达阳性组织（11．83±7．82对3．82±1．18，P〈0．05）。结论：本实验首次检测了ELA3B基因在胰腺癌和慢性胰腺炎组织中的甲基化状态，首次揭示ELA3B基因启动子区高甲基化是导致该基因在胰腺癌组织中低表达的原因之一。     

Midkine mRNA Is Overexpressed in Pancreatic Cancer

Purpose Midkine (MK) has been reported to be a possible molecular marker for the diagnosis of pancreatic cancer. We investigated the feasibility of quantitative analysis of MK mRNA by quantitative real-time RT-PCR (qRT-PCR) as a promising tool for the diagnosis of pancreatic cancer. Results We found that pancreatic cancer tissues expressed significantly higher levels of MK mRNA than intraductal pancreatic mucinous neoplasm (IPMN) and non-neoplastic pancreatic tissues (P < 0.05); in contrast, we did not find any differences in MK mRNA expression between IPMN and non-neoplastic pancreatic tissues. Additionally, we observed that poorly differentiated carcinoma samples expressed higher levels of MK mRNA than well-differentiated carcinoma samples, although a significant difference was not observed. Conclusions The present data suggests that quantitative analysis of MK mRNA provides an objective and sensitive evaluation and may be a promising modality for the diagnosis of pancreatic cancer and the prediction of its prognosis.     

Pancreatitis and pancreatic cancer: A case of the chicken or the egg

Acute pancreatitis (AP), chronic pancreatitis (CP) and pancreatic cancer are three distinct pancreatic diseases with different prognoses and treatment options. However, it may be difficult to differentiate between benign and malignant disease. AP may be a first symptom of pancreatic cancer, particularly in patients between the ages of 56 and 75 with presumed idiopathic AP who had a concomitant diagnosis of new-onset diabetes mellitus or patients who present with CP at diagnosis of AP. In these patients, additional imaging is warranted, preferably by endoscopic ultrasonography. CP may lead to pancreatic cancer through oncogenic mutations, mostly in patients with hereditary CP, and in patients in whom risk factors for pancreatic cancer (e.g., nicotine and alcohol abuse) are also present. Patients with PRSS1-mediated CP and patients with a history of autosomal dominant hereditary CP without known genetic mutations may be considered for surveillance for pancreatic cancer. Pancreatic inflammation may mimic pancreatic cancer by appearing as a focal mass-forming lesion on imaging. Differentiation between the above mentioned benign and malignant disease may be facilitated by specific features like the duct-penetrating sign and the duct-to-parenchyma ratio. Research efforts are aimed towards developing a superior discriminant between pancreatitis and pancreatic cancer in the form of imaging modalities or biomarkers. This may aid clinicians in timely diagnosing pancreatic cancer in a potentially curable stage.     

雷帕霉素抑制JAK/STAT通路对急性肝损伤大鼠肝组织HMGB1表达的影响

目的：探讨雷帕霉素抑制JAK/STAT通路对急性肝损伤大鼠肝组织HMGB1表达的影响．方法：采用D-Galn/LPS复制急性肝损伤(ALI)模型．大鼠随机分为正常对照组(n=30)、ALI组(n=30)、STAT抑制剂雷帕霉素(RPM)处理组(n=30)．用Western blot方法测定肝组织HMGB41蛋白,全自动生化分析仪测定肝功能指标．结果：与正常对照组HMGB1表达水平(1．00±0．02)相比,ALI组24．72 h HMGB1表达显著升高(3．12±0．06,3．9±0．08,2．83±0．04,t值分别为16．01,3．86,10．46,均P＜0．01),血清丙氨酸转氨酶(ALT)6和24 h有两个峰值,且24 h峰值高于6 h．与ALI组相比,RPM预处理组24-72h HMGB1蛋白表达均显著抑制(1．67±0．05 vs 3．12±0．06：1．93±0．06 vs 3．9±0．08：1．47±0．04 vs 2．83±0．04：t值分别为20．11,41．90,26．02,均P＜0．01),ALT在24,48,72 h也均有不同程度下降(P＜0．01)．ALT与HMGB1呈正相关(r=0．741,P＜0．01)．结论：抑制JAK/STAT可明显下调肝组织中HMGB1蛋白表达,并有助于减轻D-Galn/LPS所致的急性肝损伤．     

k-ras基因寡核苷酸芯片的研制及在胰腺癌组织中检测应用研究

目的建立ras基因寡核苷酸芯片对胰腺癌基因突变的检测系统。评估16例胰腺癌组织k-ras基因12、13、61位密码子变突检测。方法采用双重或单独不对称PCR扩增标本中的目的DNA。扩增产物加杂交液后与芯片进行杂交、清洗、扫描。结果16例胰腺癌组织中k-ras为75%,突变都发生在12密码子,k-ras以12密码子第2位核苷酸突变发生率高(8/12)。结论k-ras基因芯片系统具有高度的灵敏性和准确性、快速简便、自动化程度高等优点,可同时检测胰腺癌k-ras多个突变位点基因,有利于临床应用。     

Janus激酶2/信号转导和转录激活子3信号通路在心肌细胞缺氧损伤中的作用

目的探讨Janus激酶2/信号转导和转录激活子3(JAK2/STAT3)信号通路在心肌细胞缺氧损伤中的作用。方法体外培养的新生大鼠心肌细胞,构建缺氧模型,按随机数字表法分为正常对照组、缺氧组、JAK2抑制剂AG490处理组和STAT3抑制剂Statti c处理组。采用细胞计数试剂盒CCK-8检测心肌细胞活力,采用比色法检测细胞上清液中的乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)含量,并采用缺口末端标记法(TUNEL)检测各组细胞的凋亡率。采用蛋白质印迹(Western blot)检测JAK2及STAT3蛋白表达及磷酸化情况。结果与正常对照组相比,缺氧组心肌细胞成活率明显降低,为对照组的30.14%±6.23%(P<0.01),细胞上清液中LDH、MDA含量升高明显,分别为(50.11±2.58)U/L和(19.55±1.81)mol/L(均为P<0.01),SOD活力则显著降低,为(10.21±0.57)U/ml(P<0.01),缺氧组凋亡率明显升高,为24.24%±4.37%(P<0.01),JAK2、STAT3磷酸化水平上调。AG490及Stattic预处理后,JAK2及STAT3磷酸化水平降低,心肌细胞成活率明显升高(P<0.01),LDH、MDA含量显著低于缺氧组(均为P<0.01),SOD活力则高于缺氧组(P<0.01)。结论 JAK2/STAT3信号通路参与了缺氧所致心肌细胞损伤,抑制JAK/STAT通路有助于减轻缺氧所致心肌损伤。     

Role of Laparoscopy in the Diagnosis of Pancreatic Cancer

Abstract: The role of laparoscopy in the diagnosis of pancreatic cancer was evaluated. Twenty two patients who were suspected of having pancreatic cancer, following physical examination, laboratory tests, and imaging techniques, received a laparoscopy at our clinic. The laparoscopic and histological findings were analyzed retrospectively. Eighteen patients in whom the pancreas was observed were laparoscopically diagnosed as having pancreatic cancer, and one other patient from a laparoscopic ultrasonography. Among 14 patients in whom a direct forceps biopsy of the pancreas was done, pancreatic tissue was obtained from 11 patients (78.6%) and cancer was histologically proven in only 3. Liver metastasis were observed in 11 patients, among whom an aimed biopsy to the hepatic lesion was done in 10, and adenocarcinoma was histologically proven in all of these patients. Peritonitis carcinomatosa was confirmed in 4 patients and adenocarcinorna was proven in all of the patients by histology after a punch biopsy to the lesion. Thus, histological evidence of cancer was obtained in 13 out of the 22 patients studied (59.1%). By doing laparoscopy, histological evidence of pancreatic cancer could be obtained by a less invasive procedure than lapayotomy. Conclusively, laparoscopy seems to be useful in the diagnosis of pancreatic cancer.     

汉黄芩素调节JAK/STAT信号通路并诱导肺癌A549细胞凋亡及周期阻滞的作用及机制研究

目的 研究汉黄芩素对肺癌A549细胞凋亡、周期及对JAK/STAT信号通路的调节作用.方法取对数生长期的肺癌A549细胞,采用低(20μmol/L)、中(40μmol/L)、高(80μmol/L)剂量的汉黄芩素分别干预24 h、48、72 h,MTT法测定不同浓度汉黄芩素对肺癌A549细胞的增殖抑制率,实时定量PCR检...