Clinicopathological and prognostic implications of endoglin (CD105) expression in hepatocellular carcinoma and its adjacent non-tumorous liver

AIM: The expression pattern of endoglin (CD105) in hepatocellular carcinoma (HCC) has not been reported so far. We hypothesized that CD105 could differentially highlight a subset of microvessels in HCC, and intratumoral microvessel density (IMVD) by CD105 immunostaining (IMVD-CD105) could provide better prognostic information than IMVD by CD34 immunostaining (IMVD-CD34).METHODS: Paraffin blocks of tumor and adjacent nontumorous liver tissues from 86 patients who underwent curative resection of HCC were used for this study. Serial sections were stained for CD105 and CD34, respectively,to highlight the microvessels. IMVD was counted according to a standard protocol.RESULTS: In the HCC tissues, CD105 was either negatively or positively stained only in a subset of microvessels. In contrast, CD34 showed positive and more extensive microvessel staining in all cases examined. However, in the adjacent non-tumorous liver sections, CD105 showed a diffuse pattern of microvessel staining in 20 of 86 cases,while CD34 showed negative or only focal staining of the sinusoids around portal area. Correlation with clinicopathological data demonstrated that lower scores of IMVD-CD105 were found in larger sized tumors [mean 41.4/0.74 mm2 (＞5 cm tumor) vs 65.9/0.74 mm2(≤ 5 cm tumor), P = 0.043] and more aggressive tumors,as indicated by venous infiltration [36.8/0.74 mm2 (present)vs 64.2/0.74 mm2 (absent), P = 0.020], microsatellite nodules [35.1/0.74 mm2 (present) vs 65.9/0.74 mm2(absent), P = 0.012], and advanced TNM tumor stage [38.8/0.74 mm2 (stage 3 or 4) vs 68.3/0.74 mm2 (stage 1or 2), P = 0.014]. No prognostic significance was observed when median values were used as cut-off points using either IMVD-CD105 or IMVD-CD34. However, the presence of the diffuse pattern of CD105 expression in the adjacent non-tumorous liver tissues predicted a poorer disease-free survival (median 8.6 vs 21.5 mo, P = 0.026).CONCLUSION: Our data demonstrate that a lower IMVDCD105 is associated with larger and more aggressive tumors. In this study, IMVD-CD105 did not provide significant prognostic information. However, active angiogenesis as highlighted by diffuse CD105 staining of the microvessels in the adjacent non-tumorous liver tissues is predictive of early recurrence.

Downregulation of Hes1 expression in experimental biliary atresia and its effects on bile duct structure

AIM To analyze the expression and function of the notchsignaling target gene Hes1 in a rhesus rotavirusinduced mouse biliary atresia model. METHODS The morphologies of biliary epithelial cells in biliary atresia patients and in a mouse model were examined by immunohistochemical staining. Then, the differential expression of Notch signaling pathway-related molecules was investigated. Further, the effects of the si RNAmediated inhibition of Hes1 expression were examined using a biliary epithelial cell 3 D culture system.RESULTS Both immature(Ep CAM+) and mature(CK19+) biliary epithelial cells were detected in the livers of biliary atresia patients without a ductile structure and in the mouse model with a distorted bile duct structure. The hepatic expression of transcripts for most Notch signaling molecules were significantly reduced on day 7 but recovered to normal levels by day 14, except for the target molecule Hes1, which still exhibited lower m RNA and protein levels. Expression of the Hes1 transcriptional co-regulator, RBP-Jκ was also reduced. A 3 D gel culture system promoted the maturation of immature biliary epithelial cells, with increased expression of CK19+ cells and the formation of a duct-like structure. The administration of Hes1 si RNA blocked this process. As a result, the cells remained in an immature state, and no duct-like structure was observed.CONCLUSION Our data indicated that Hes1 might contribute to the maturation and the cellular structure organization of biliary epithelial cells, which provides new insight into understanding the pathology of biliary atresia.

Participation of CD45,NKR-P1A and ANK61 antigen in rat hepatic NK cell(pit cell)-mediated target cell cytotoxicity

AIM Several triggering receptors have beendescribed to be involved in natural killer（NK）cell-mediated target cytotoxicity.In these studies,NKcells derived from blood or spleen were used.Pitcells are liver-specific NK cells that possess ahigher level of natural cytotoxicity and a differentmorphology when compared to blood NK cells.The aim of this study was to characterize the roleof the NK-triggering molecules NKR-P1A,ANK61antigen,and CD45 in pit cell-mediated killing oftarget cells.METHODS 51Cr-release and DNA fragmentationwere used to quantify target cell lysis andapoptosis,respectively.RESULTS Flow cytometric analysis showed thatpit cells expressed CD45,NKR-P1A,and ANK61antigen.Treatment of pit cells with monoclonalantibody（mAb）to CD45（ANK74）not onlyinhibited CC531s or YAC-1 target lysis but alsoapoptosis induced by pit cells.The mAbs to NKR-P1A（3.2.3）and ANK61 antigen（ANK61）had no effect on pit cell-mediated CC531s or YAC-1 targetcytolysis or apoptosis,while they did increase theFcγ receptor positive（FcγR+）P815 cytolysis andapoptosis.This enhanced cytotoxicity could beinhibited by 3,4-dichloroisocoumarin,an inhibitorof granzymes.CONCLUSION These results indicate that CD45participates in pit cell-mediated CC531s and YAC-1target cytolysis and apoptosis.NKR-P1A andANK61 antigen on pit cells function as activationstructures against FcγR+ P815 cells,which wasmediated by the perforin/granzyme pathway.

Role of CD56-expressing immature biliary epithelial cells in biliary atresia

AIM: To analyze the clinical and pathological parameters and expression of the neural cell adhesion molecule(CD56) in patients with biliary atresia(BA).METHODS: Established clinical laboratory markers of hepatic function, including enzyme activity, protein synthesis, and bilirubin metabolism, were evaluated in patients with BA and compared with those in patients with choledochal cysts and neonatal hepatitis. Pathological changes in tissue morphology and fibrosis were examined by histological and tissue collagen staining. Immunohistochemical staining for the biliary epithelial cell markers CD56 and CK19 together with the Notch signaling related molecules Notch1 and Notch2 was performed in the context of alterations in the structure of intrahepatic biliary ducts.RESULTS: Differences in some clinical laboratoryparameters among the three diseases examined were observed, but they did not correlate with the pathological classification of fibrosis in BA. Immunohistochemical staining showed the presence of CD56-positive immature bile ducts in most patients(74.5%) with BA but not in patients with choledochal cysts or neonatal hepatitis. The number of CD56-expressing cells correlated with disease severity, with more positive cells present in the later stages of liver damage(81.8% vs 18.2%). Furthermore, bile plugs were mainly found in CD56-positive immature biliary ducts. Notch signaling was a key regulatory pathway in biliary duct formation and played a role in tissue fibrosis. Notch1 was co-expressed in CD56-positive cells, whereas Notch2 was found exclusively in blood vessels in the portal area of patients with BA. CONCLUSION: The maturation of biliary epithelial cells and the expression of Notch may play a role in the pathogenesis of BA.

Biochemically curative surgery for gastrinoma in multiple endocrine neoplasia type 1 patients

AIM: To search for the optimal surgery for gastrinoma and duodenopancreatic neuroendocrine tumors in patients with multiple endocrine neoplasia type 1. METHODS: Sixteen patients with genetically confirmed multiple endocrine neoplasia type 1 (MEN 1) and Zollinger-Ellison syndrome (ZES) underwent resection of both gastrinomas and duodenopancreatic neuroendocrine tumors (NETs) between 1991 and 2009. For localization of gastrinoma, selective arterial secretagogue injection test (SASI test) with secretin or calcium solution was performed as well as somatostatin receptor scintigraphy (SRS) and other imaging methods such as computed tomography (CT) or magnetic resonance imaging (MRI). The modus of surgery for gastrinoma has been changed over time, searching for the optimal surgery: pancreaticoduodenectomy (PD) was first performed guided by localization with the SAST test, then local resection of duodenal gastrinomas with dissection of regional lymph nodes (LR), and recently pancreas-preserving total duodenectomy (PPTD) has been performed for multiple duodenal gastrinomas. RESULTS: Among various types of preoperative localizing methods for gastrinoma, the SASI test was the most useful method. Imaging methods such as SRS or CT made it essentially impossible to differentiate functioning gastrinoma among various kinds of NETs. However, recent imaging methods including SRS or CT were useful for detecting both distant metastases and ectopic NETs; therefore they are indispensable for staging of NETs. Biochemical cure of gastrinoma was achieved in 14 of 16 patients (87.5%); that is, 100% in 3 patients who underwent PD, 100% in 6 patients who underwent LR (although in 2 patients (33.3%) second LR was performed for recurrence of duodenal gastrinoma), and 71.4% in 7 patients who underwent PPTD. Pancreatic NETs more than 1 cm in diameter were resected either by distal pancreatectomy or enucleations, and no hepatic metastases have developed postoperatively. Pathological study of the resected specimens revealed co-existence of pancreatic gastrinoma with duodenal gastrinoma in 2 of 16 patients (13%), and G cell hyperplasia and/or microgastrinoma in the duodenal Brunner's gland was revealed in all of 7 duodenal specimens after PPTD. CONCLUSION: Aggressive resection surgery based on accurate localization with the SASI test was useful for biochemical cure of gastrinoma in patients with MEN 1.

Dried fruit pomace inclusion in poultry diet:growth performance, intestinal morphology and physiology

Background: Fruit pomaces are by-products rich in polyphenol compounds and dietary fiber. They seem to play an important role in regulating the gut microbiota, morphology and physiology. The aim of this study was to assess whether apple(A), blackurrant(B) or strawberry(S) pomaces could be suitable ingredients in broiler diets and their effect on gut health. A total of 480 male broilers were randomly allotted to 8 dietary treatments with lower(3%-L) or higher(6%-H) dietary fiber content: two control groups(CL/CH), two A diets(AL/AH), two B diets(BL/BH), two S diets(SL/SH). Diet and fruit pomaces were chemically analyzed to assess polyphenol concentration and fibre fraction content. After the evaluation of growth performance, 6 birds/group were slaughtered at 35 days of age. Morphometric and histopathological investigations were performed on duodenum, jejunum and ileum.Excreta were collected to perform microbiota evaluation by 16 S DNA sequencing. Weight, viscosity, enzymatic activity, short chain fatty acid(SCFAs) and ammonia concentration were determined in ileum and/or ceca content.Results: A pomace and A diets showed the lowest polyphenol content and the highest content of soluble fibre fraction. No significant differences were observed for growth performance, gut morphometry and histopathology(P > 0.05). Dietary fruit pomace inclusion increased the weight of ileum and ceca and the ileum digesta viscosity(P < 0.05). In the ileum, A and S groups showed lower bacterial α-glucosidase activity than C groups. Moreover,small intestine SCFAs concentration was higher in fruit pomaces diets(P < 0.05). In ceca, B and S groups showed lower ammonia concentration and higher SCFAs than C. Dietary treatments also influenced the activity of α-glucosidase, α-galactosidase, β-galactosidase β-glucuronidase and xylase. Regarding microbiota, at phylum level,Firmicutes were differentially abundant across treatment(maximum for C and minimum in S, FDR > 0.05). At genus level, an increase of Weissella in AH and Erwinia in S/B diets, as well as a decrease of Lactobacillus in all fruit pomace groups were recorded(P < 0.05).Conclusions: Fruit pomaces could be suitable ingredients in poultry nutrition even if further studies are needed to better understand which doses is more recommended to avoid negative effects on gut microbiota.

Rehabilitation training improves exercise tolerance after percutaneous coronary intervention

The aim of this present study was to investigate the effects of training on exercise tolerance of patients with coronary heart disease after percutaneous coronary intervention.Fifty-seven cases of coronary heart disease after percutaneous coronary intervention were divided randomly into the rehabilitation training group（26 cases） and control group（31 cases）.Patients in the rehabilitation training group received rehabilitation training at different stages and exercise intensities 3 d after percutaneous coronary intervention for 3 months.The heart rate,blood pressure,ECG changes in treadmill exercise test,and the frequency of anginal episodes were observed.The results showed that NST and ΣST of ECG and the frequency of anginal episodes were significantly reduced in the rehabilitation training group.In addition,exercise tolerance was improved and the total exercise time was lengthened in these patients.Moreover,ST segment depression time and emergence time of angina with exercise were also lengthened compared with controls（P 〈 0.05,or 0.01）.However,the heart rate and blood pressure before and after exercise of the two groups were similar.The study indicated that rehabilitation training could significantly relieve angina,amend ischemic features of ECG,and improve exercise tolerance of coronary heart disease patients after percutaneous coronary intervention.

Acinar cell injury induced by inadequate unfolded protein response in acute pancreatitis

Acute pancreatitis (AP) is an inflammatory disorder of pancreatic tissue initiated in injured acinar cells. Severe AP remains a significant challenge due to the lack of effective treatment. The widely-accepted autodigestion theory of AP is now facing challenges, since inhibiting protease activation has negligible effectiveness for AP treatment despite numerous efforts. Furthermore, accumulating evidence supports a new concept that malfunction of a self-protective mechanism, the unfolded protein response(UPR), is the driving force behind the pathogenesis of AP. The UPR is induced by endoplasmic reticulum(ER) stress, a disturbance frequently found in acinar cells, to prevent the aggravation of ER stress that can otherwise lead to cell injury. In addition, the UPR's signaling pathways control NFκB activation and autophagy flux, and these dysregulations cause acinar cell inflammatory injury in AP, but with poorly understood mechanisms. We therefore summarize the protective role of the UPR in AP, propose mechanistic models of how inadequate UPR could promote NFκB's pro-inflammatory activity and impair autophagy's protective function in acinar cells, and discuss its relevance to current AP treatment. We hope that insight provided in this review will help facilitate the research and management of AP.

Long-term prognosis in patients with severe late radiation enteropathy:A prospective cohort study

AIM: To assess persistent symptoms and mortality in a cohort of patients with severe (grade 3-4) radiation enteropathy,59 patients were followed up after 15-18 years. METHODS: Fifty-nine patients were prospectively enrolled by twelve surgical departments. Primary malignant disease,radiation therapy and surgical management were recorded at inclusion. The cause of death or persistence of symptoms was examined in public death records or by interview of survivors. RESULTS: Thirty-nine patients had received radiation therapy for gynaecological cancers,twelve for urological cancers,four for gastrointestinal cancers and four for other malignancies. Forty-five patients (76%) required surgical intervention. Complications occurred in 11 (25%) operated patients. Forty-seven patients had died at the time of follow-up,seven (12%) died as a direct result of radiation enteropathy,while radiation enteropathy contributed to death in an additional seven patients. Four of the twelve surviving patients suffered from chronic debilitating symptoms of radiation enteropathy,while three had moderate symptoms. CONCLUSION: Patients with severe delayed radiation enteropathy have a high risk of persistence of symptoms after surgery. At least one in ten patients dies from radiation-induced bowel injury.

Epithelial-mesenchymal transition in prostate cancer： providing new targets for therapy

The ability of epithelial cells to undergo phenotypic transitions during embryo-genesis, wound healing and malignant pro- gression is now widely accepted as a core biological process termed epithelialmesenchymal transition （EMT）. During cancer progression, the process of EMT has been associated with the acquisition of stemness properties, treatment resistance and metastatic progression, hallmarks of malig- nancy.

Postsurgical radiation therapy for gastric carcinosarcoma with c-kit expression: A case report

Gastric carcinosarcomas are rare morphologically biphasic tumors, consisting of carcinoma and sarcoma components, with a poor clinical course.Here we report the case of a 70-year-old man with advanced Borrmann type Ⅲ carcinosarcoma arising from the upper body of the stomach with extensive lymph node metastasis who underwent a total, but palliative, gastrectomy.Histology showed the tumor consisted of a biphasic structure of tubular adenocarcinoma and spindle cellsarcoma.Immunohistochemistry revealed sarcoma cells expressing c-kit(CD117) and CD34, which are criteria for gastrointestinal stromal tumors.Nine months after the surgical operation, tumor metastases had extended to the hepatohilar, retroperitoneal and mediastinal lymph nodes.Radiation therapy of 50 Gy markedly decreased the size of each of these nodes and reduced the risk of respiratory complications and jaundice.However, the patient died of respiratory failure due to bronchopneumonia with multiple lung metastases 22 mo after resection.Autopsy revealed severe necrosis in most of the lymph nodes with tumor metastases.Radiation therapy combined with gastrectomy should be considered to improve survival in patients with gastric carcinosarcomas that express c-kit.

Malignant phyllode tumor metastatic to the duodenum

叶状叶柄肿瘤(磅) 是胸的极其稀罕的肿瘤。远转移与恶意的叶状叶柄肿瘤发生在病人的 10%-20% 。转移的最普通的地点是肺和骨头。尽管理论上任何机关可以有转移，孤立的十二指肠转移没在英语语言的文学迄今为止被记录。我们此处在 31 year-old-woman 从胸的磅与孤立的十二指肠的转移报导一个案例因为周期性的恶意的磅，以前恰好经历了乳房切除术 4 年。她介绍了给我们的医院与巨大上面的胃肠的流血。临床的评估表明一个巨大的团从十二指肠发源。迫切剖腹术和 pancreaticoduodenectomy 被执行以便移开流血十二指肠的质量。resected 标本的病理学的考试显示出与变形的恶意的磅一致的一个恶意的锭子房间肿瘤。我们从胸的恶意的叶状叶柄肿瘤的由于造血的传播的由于孤立的十二指肠的转移的胃肠的流血的案例在英语文学是唯一的， pancreaticoduodenectomy 是为有孤立的十二指肠的参与的病人的祛病疗法。

Mechanistic insights into cancer immunity and immunotherapy

While immunotherapy has had an important impact on cancer treatment,only approximately a quarter of patients respond to these treatments.Why does it work in some patients but not in other patients?How can we improve the therapeutic efficacy of current immunotherapy?Are there potential new strategies to tackle this problem?The answers to these questions may rely on our in-depth understanding of the basic mechanisms of the cancer immune response,immunotherapeutic efficacy,and resistance.

BRAF mutation in multiple primary cancer with colorectal cancer and stomach cancer

Aims:Recently,BRAF mutation testing has been introduced as a marker in differentiating Lynch syndrome from sporadic colorectal cancers or in predicting colorectal cancers with worse prognosis.Individuals with hereditary predisposition to cancer development are at an increased risk of developing multiple primary cancers.The purpose of this study is to identify mutation in the BRAF gene in multiple primary cancers with colorectal cancer and stomach cancer.Methods:BRAF mutation was analysed in 45 patients with colorectal cancer and stomach cancer,synchronously or metachronously.Results:Mean age was 64.07 years(range:47–83 years).For the colorectal cancer,tumors were located at the sigmoid colon in eight patients(17.8%)and at the rectum in 22 patients(48.9%).Twenty-three patients(51.1%)had synchronous cancer.Four patients(8.9%)had family members with cancer.BRAF mutation was identified in three patients(6.7%).All three of these patients had metachronous cancers.The colorectal cancers were located in the sigmoid colon(1 patient)and the rectum(2 patients).Conclusions:BRAF mutation rate was low in the multiple primary cancer with colorectal cancer and stomach cancer.With only BRAF gene study,it was not possible to identify any correlation with family history of colorectal cancer.Further study means considering other genes–MSI,MSH2,MLH1,MSH6.

啮齿类动物出血性休克模型中血浆对内皮细胞多糖包被的恢复作用

背景在出血性休克的创伤患者应用血浆为主的复苏与死亡率下降有关。尽管有学者提出通过替代凝血蛋白可产生有益的效果，但是输入血浆的保护作用可能机制尚不明了。我们以前在细胞培养模型中的研究显示，与晶体液相比，血浆可降低内皮细胞的通透性。多糖包被由粘附于多配体聚糖骨架上的蛋白多糖和糖蛋白组成，共同保护下层的内皮细胞。我们假设，血浆对内皮细胞的保护作用，部分是由于出血性休克后内皮细胞多糖包被的恢复和多配体聚糖-1的维持所致。方法在大鼠建立出血性休克模型使其平均动脉压在30mmHg维持90分钟，然后以乳酸林格（1actatedRinger＇s，LR）液或新鲜血浆复苏，使其平均动脉压达到80mmHg，相当于假手术组或单纯休克组动物。2小时后，提取肺组织多配体聚糖的mRNA，使用抗多配体聚糖-l免疫染色或使用苏木精伊红染色。为了特异地检测出血浆对内皮细胞的影响，我们通过小肠系膜灌注镧溶液，然后应用电子显微镜鉴别出小静脉，观察其多糖包被。所有数据以均数±标准差来表示。运用单尾方差分析合并Tukey事后检验来分析结果。结果电子显微镜显示出血性休克后多糖包被发生降解，输注血浆可部分恢复多糖包被水平而LR无该效应。血浆复苏组动物肺组织多配体聚糖．1的mRNA表达（2．76±0．03）明显高于单纯休克组（1．39±0．22）或ut组（0．82±0．03），并且与细胞表面多配体聚糖．1的免疫染色相关。休克也可导致组织病理学评分方面肺损伤显著（1．63±0．26），应用血浆复苏可减轻该损伤（0．67±0．17），而LR无该效应（2．0±0．25）。结论血浆的保护作用可能部分是由于出血性休克后血浆能恢复内皮细胞多糖包被水平，并维持多配体聚糖-1水平。