Influence of up-regulation of Notch ligand DLL4 on biological behaviors of human gastric cancer cells

AIM: To investigate the potential roles of Delta-like ligand 4 (DLL4) on the biological behavior of gastric cancer cells and its molecular mechanisms. METHODS: A recombinant eukaryotic expression vector containing human DLL4 gene was constructed and transfected into the human gastric cancer cell line SGC7901. Clones with up-regulated DLL4 were selected and amplified. The effect of DLL4 up-regulation on gastric cancer cell growth was assessed using cell growth assay. The migration and invasion were assessed using a transwell migration assay and matrigel invasion assay. Matrix metalloproteinases were detected using the zymogram technique. Cells were implanted subcutaneously into male BALB/c nu/nu mice. Tumor volumes were then calculated and compared. DLL4 staining in the implanted tumor was performed using immunohistochemistry technique. RESULTS: Growth curves over a six-day time course showed significantly promoted cell proliferation of SGC7901 cells with up-regulated DLL4. DLL4 up-regulation in SGC7901 cells promoted the migration (205.4 ± 15.2 vs 22.3 ± 12.1, P < 0.05) and invasion (68.8 ± 5.3 vs 18.2 ± 6.0, P < 0.05) in vitro and tumorigenicity in vivo (2640.5 ± 923.6 mm 3 vs 1115.1 ± 223.8 mm 3 , P < 0.05). Furthermore, significantly increased mRNA level and increased secretion of matrix metalloproteinase-2 (MMP-2) proenzyme were observed in SGC7901 cells with up-regulated DLL4. However, increased MMP-9 mRNA level but decreased extracellular MMP-9 proenzyme level was observed. CONCLUSION: Our observations indicated a mechanism by which activation of DLL4-mediated Notch signaling promotes the expression and secretion of MMP-2 proenzyme and influences the progress of gastric cancer.

Influence of CO_2 pneumoperitoneum on intracellular pH and signal transduction in cancer cells

Object: The authors studied the influence of CO2 pneumoperitoneum on intracellular pH and signal transduction arising from cancer cell multiplication in laparoscopic tumor operation. Method: They set up a simulation of pneumoperitoneum under different CO2 pressure, and then measured the variation of intracellular pH (pHi) at different time and the activity of protein kinase C (PKC) and protein phosphatase 2a (PP2a) at the end of the pneumoperitoneum. After 1 week, the concentration of cancer cells in the culture medium was calculated. Result: When the pressure of CO2 pneumoperitoneum was 0, 10, 20, 30 mmHg respectively, the average pHi was 7.273, 7.075, 6.783, 6.693 at the end of the pneumoperitoneum; PKC activity was 159.4, 168.5,178.0, 181.6 nmol/(g.min) and PP2a was 4158.3, 4066.9, 3984.0, 3878.5 nmol/(g.min) respectively. After 1 week, the cancer cells concentration was 2.15×105, 2.03×105, 2.20×105, 2.18×105 L-1. Conclusion: CO2 pneumoperitoneum could promote acidosis in cancer cells, inducing the activation of protein kinase C and deactivation of protein phosphatase 2a, but it could not accelerate the mitosis rate of the cancer cells.

Diagnosis and management of colonic injuries following blunt trauma

AIM: To retrospectively evaluate the preoperative diagnostic approaches and management of colonic injuries following blunt abdominal trauma. METHODS: A total of 82 patients with colonic injuries caused by blunt trauma between January 1992 and December 2005 were enrolled. Data were collected on clinical presentation, investigations, diagnostic methods, associated injuries, and operative management. Colonic injury-related mortality and abdominal complications were analyzed. RESULTS: Colonic injuries were caused mainly by motor vehicle accidents. Of the 82 patients, 58 (70.3%) had other associated injuries. Laparotomy was performed within 6 h after injury in 69 cases (84.1%), laparoscopy in 3 because of haemodynamic instability. The most commonly injured site was located in the transverse colon. The mean colon injury scale score was 2.8. The degree of faecal contamination was classified as mild in 18 (22.0%), moderate in 42 (51.2%), severe in 14 (17.1%), and unknown in 8 (9.8%) cases. Sixty-seven patients (81.7%) were treated with primary repair or resection and anastomosis. Faecal stream diversion was performed in 15 cases (18.3%). The overall mortality rate was 6.1%. The incidence of colonic injury- related abdominal complications was 20.7%. The only independent predictor of complications was the degree of peritoneal faecal contamination (P = 0.02). CONCLUSION: Colonic injuries following blunt trauma are especially important because of the severity and complexity of associated injuries. A thorough physical examination and a combination of tests can be used to evaluate the indications for laparotomy. One stage management at the time of initial exploration is mostoften used for colonic injuries.

Surgical management of gastric stump cancer:a report of 37 cases

Objective:To observe the clinicopathological characteristics of gastric stump cancer(GSC)and evaluate the benefitsof radical surgery of GSC.Methods:The clinicopathological characteristics and postoperative survival time of 37 GSC patientswho underwent surgery were investigated retrospectively.The survival time was compared according to the type of surgicaloperation(radical resection vs palliative operation).Twenty-one cases that received radical resection were analyzed based on thepTMN stage.Survival curves were traced by using Kaplan-Meier methods.Results:Most GSC(32/37)was detected in patientswho had received Billroth Ⅱ reconstruction after partial gastrectomy for benign gastric disease.The lesser curvature side and thesuture line of anastomosis were the most frequent sites where GSC occurred(27/37).Differentiated adenocarcinoma was thedominant histopathological type(24/37).The postoperative 5-year survival rate of early stage GSC patients(n=9)was significantlyhigher than advanced stage GSC(n=12)(55.6% vs 16.5%,x_L^2=11.48,P<0.01).Five-year survival rate of 21 GSC patients withradical resection were 75%(3/4)for stage Ⅰ,60%(3/5)for stage Ⅱ,14.2%(1/7)for stage Ⅲ,and 0%(0/5)for stage Ⅳ respectively.The median survival time of 21 GSC patients who underwent radical resection was longer than those undergoing palliative op-eration(43.0 m vs 13.0 m,X_L^2=36.31,P<0.01),the median survival time of stage Ⅳ patients with radical resection was 23.8months.Conclusions:Without remote metastasis,radical resection for GSC is possible,and is an effective way to improve theprognosis of GSC.Even in stage Ⅳ GSC,radical resection can still prolong the survival time.It is necessary for the patients withbenign gastric diseases who received partial gastrectomy to carry out the endoscopy follow-up,especially in patients with BillrothⅡ reconstruction procedure at 15-20 years.

Detection of pancreatic cancer with normal carbohydrate antigen 19-9 using protein chip technology

AIM: To develop a method to differentiate pancreatic cancer patients from healthy or benign individuals when carbohydrate antigen (CA) 19-9 is normal.

Conservative resection for benign tumors of the proximal pancreas

AIM:To evaluate the safety and long-term prognosis of conservative resection (CR) for benign or borderline tumor of the proximal pancreas.METHODS: We retrospectively analyzed 20 patients who underwent CR at the Second Affi liated Hospital of Zhejiang University School of Medicine between April 2000 and October 2008. For pancreaticojejunostomy, a modified invagination method, continuous circular invaginated pancreaticojejunostomy (CCI-PJ) was used. Modified continuous closed lavage (MCCL) was performed for patients with pancreatic fistula.RESULTS: The indications were: serous cystadenomas in eight patients, insulinomas in six, non-functional islet cell tumors in three and solid pseudopapillary tumors in three. Perioperative mortality was zero and morbidity was 25%. Overall, pancreatic fistula was present in 25% of patients. At a mean follow up of 42.7 mo, all patients were alive with no recurrence and no new-onset diabetes mellitus or exocrine dysfunction.CONCLUSION: CR is a safe and effective procedure for patients with benign tumors in the proximal pancreas, with careful CCI-PJ and postoperative MCCL.

Role of CD97^(stalk) and CD55 as molecular markers for prognosis and therapy of gastric carcinoma patients

Objectives： To explore the mechanism of development and aggressiveness in gastric carcinomas by investigating the expression and role of CD97 and its cellular ligand CD55 in gastric carcinomas. Methods： Tumor and corresponding normal mucosal tissue, collected from 39 gastric carcinoma patients, were examined by immunohistochemistry and RT-PCR for the expression of CD97 and CD55. Results： CD97^stalk was strongly stained on scattered tumor cells or small tumor cell clusters at the invasion front of gastric carcinomas. The expression of CD97^stalk was frequently observed in tumors of stage Ⅰ and T1 gastric carcinoma patients. The expression of CD97^stalk between Stage Ⅰ and Stage Ⅱ, Ⅲ, Ⅳ specimens showed significant difference （P〈0.05）, between T1 and T2, T3, T4 specimens also showed significant difference （P〈0.05）. Specimens with tumor invasion depth limited in mucosa of T1 specimens showed higher positive CD55 expression than specimens with the same tumor invasion depth in T2, T3, T4 specimens, the expression of CD55 between T1 and T2, T3, T4 specimens was significantly different （P〈0.05）. There was strong correlation between the distribution patterns of CD97^stalk and CD55 on tumor tissues （r=0.73, P〈0.05）. Signet ring cell carcinomas frequently contained strong CD97^stalk and CD55-staining. Conclusions： Our results suggest that CD97^stalk is probably involved in the growth, invasion and aggressiveness of gastric carcinomas by binding its cellular ligand CD55. CD97^stalk and CD55 could be useful as molecular markers for prognosis and therapy of gastric carcinoma patients.

Patients' quality of life after laparoscopic or open cholecystectomy

Objective: This study was aimed at evaluating and comparing the quality of life in patients who underwent laparo-scopic and open cholecystectomy for chronic cholecystolithiasis. Methods: The study included 25 patients with laparoscopic cholecystectomy (LC group) and 26 with open cholecystectomy (OC group). The quality of life was measured with the Gastrointestinal Quality of Life Index (GLQI) preoperatively, thereafter regularly at 2, 5, 10 and 16 weeks after the operation. Results: The mean preoperative overall GLQI scores were 112.5 and 110.3 in LC and OC group respectively (P>0.05). In the LC group, the mean overall GLQI score reduced slightly to 110.0 two weeks after the operation (P>0.05). The LC group showed significant improvement in overall score and in the aspects of symptomatology, emotional and physiological status from 5 to 16 weeks postoperatively. In the OC group, the GLQI score reduced to 102.0 two weeks after surgery (P<0.05). Significant reductions were shown in the aspects of symptomatology, physiological and social status. The GLQI scores returned to the preoperative level of 115.6 ten weeks after the operation (P>0.05). The patients experienced significant improvements of GLQI sixteen weeks after OC operation (P<0.01-0.05). Within the 10 postoperative weeks, the LC group had significantly higher GLQI scores than the OC group (P<0.05). Conclusions: LC can improve the quality of life postoperatively better and more rapidly than OC. The assessment of quality of life assessment is a valid method for measuring the effects of surgical treatment.

Resection of non-cystic adenocarcinoma in pancreatic body and tail

AIM: To report the outcome of Chinese patents with non-cystic adenocarcinoma in pancreatic body and tail (NCAPBT) after resection and to discuss its surgical strategy. METHODS: Resection of NCAPBT was performed in eight Chinese patients with complete clinical-pathological data in our hospital from January 2000 to May 2004. The surgical strategy was explored by analyzing the results of these patients. RESULTS: The resection rate of NCAPBT in patients without back pain was higher than that in patients with back pain (66.67% vs 20%, 2/3 vs 1/5). The prognosis in the group receiving palliative resection was poorer than that in the group receiving curative resection. The median survival time was 12 mo in the curative resection group and 6 mo in the palliative resection group, respectively. CONCLUSION: The overall survival time of the Chinese patients with NCAPBT is dismal. The Chinese patients after curative resection of NCAPBT have a longer survival time. The Chinese NCAPBT patients with back pain trend to have a lower curative resection rate, but back pain should not be considered a contraindication for curative resection.

CD97 promotes gastric cancer cell proliferation and invasion through exosome-mediated MAPK signaling pathway

AIM: To investigate the mechanism underlying the promoting role of CD97 in gastric cancer cell proliferation and invasion. METHODS: Two types of exosomes released by gastric cancer cells with high(SGC/wt) or low(SGC/kd) CD97 expression were isolated by ultracentrifugation and identified by electron microscopy and western blot analysis. The influences of the two exosomes on gastric cancer cell proliferation and invasion were investigated by proliferation and Matrigel invasion assays. Exosomal mi RNAs were subsequently isolated from the two samples and their mi RNA profiles were compared via microarray assay analysis. Reverse transcriptionquantitative real-time polymerase chain reaction was used to validate the microarray assay. Target genes of the differently expressed micro RNAs were predicted based on five independent algorithms and were then subjected to gene oncology enrichment and Kyoto encyclopedia of genes and genomes(KEGG) pathway analysis. After identifying the pathway that was the most likely altered, tumor cells were treated with the two exosomes at different concentrations, and the pathway activation was identified through western blot analysis.RESULTS: Exosomes isolated from SGC/wt cells significantly promoted tumor cell proliferation in a dose-dependent manner in vitro. SGC/wt exosomesalso significantly elevated the invasiveness of both SGC/wt(129.67 ± 8.327 vs 76.00 ± 5.292, P < 0.001) and SGC/kd(114.52 ± 9.814 vs 45.73 ± 4.835, P < 0.001) cells as compared to the exosomes released by SGC/kd cells. Microarray assay of the two exosomes revealed that 62 mi RNAs were differently regulated with a signal intensity of > 500 and a false discovery rate < 0.05. The following KEGG analysis defined the MAPK signaling pathway as the most likely candidate pathway that regulated tumor cell proliferation and invasion. Through western blot analysis, significant up-regulations of phosphorylated MAPKs, including extracellular signal-regulated kinase, Jun NH2-terminal kinase, and p38 mitogen-activated protein kinase, were detected in a dose-dependent manner in the SGC/wt exosomes treated groups, confirming activation of the MAPK signaling pathway stimulated by SGC/wt exosomes.CONCLUSION: CD97 promotes gastric cancer cell proliferation and invasion in vitro through exosomemediated MAPK signaling pathway, and exosomal mi RNAs are probably involved in activation of the CD97-associated pathway.

Risk factors for operative morbidity and mortality in gastric cancer patients undergoing total gastrectomy

AIM： To study the risk factors for morbidity and mortality following total gastrectomy. METHODS： We retrospectively reviewed the records of 125 consecutive patients who underwent total gastrectomy for gastric cancer at the Second Affiliated Hospital of Zhejiang University School of Medicine between January 2003 and March 2008. RESULTS： The overall morbidity rate was 20.8% （27 patients） and the mortality rate was 3.2% （4 patients）. Morbidity rates were higher in patients aged over 60 [odds ratio （OR） 4.23 （95% confidence interval （CI） 1.09 to 12.05）], with preoperative comorbidity [with vs without, OR 1.25 （95% CI 1.13 to 8.12）], when the combined resection was performed [combined resection vs total gastrectomy only, OR 2.67 （95% CI 1.58 to 5.06）]. CONCLUSION： Age, preoperative comorbidity and combined resection were with the rate of morbidity gastric cancer. independently associated after total gastrectomy for

Gastric foregut cystic developmental malformation: Case series and literature review

Foregut cystic developmental malformation(FCDM) is a very rare lesion of the alimentary tract, especially in the stomach. We discuss the concepts of gastric duplication cyst, bronchogenic cysts, and FCDM. Nomenclature has been inconsistent and confusing, but, by some definitions, gastric duplication cysts involve gastric mucosa and submucosal glands, bronchogenic cysts involve respiratory mucosa with underlying cartilage and glands, and FCDM lacks gastric mucosa or underlying glands or cartilage but has pseudostratified ciliated columnar epithelium(PCCE). We searched our departmental case files from the past 15 years and identified 12 cases of FCDM in the alimentary tract. We summarize the features of these 12 cases including a report in detail on a 52-year-old man with a submucosal cyst lined with simple PCCE and irregular and stratified circular muscle layers that merged with gastric smooth muscle bundles near the lesser curvature of the gastric cardia. A literature review of cases with this histology yielded 25 cases. We propose the term gastric-FCDM for such cases. Our own series of 12 cases confirms that preoperative recognition of the entity is infrequent and problematic. The rarity of this developmental disorder, as well as a lack of understanding of its embryologic origins, may contribute to missing the diagnosis. Not appreciating the diagnosis preoperatively can lead to an inappropriate surgical approach. In contrast, presurgical recognition of the entity will contribute to a good outcome and reduced risk of complications.

Role of dissection of secondary branches of splenic pedicle in portal hypertension cases undergoing splenectomy

Background It is well known that conventional splenectomy, which requires careful handling and ligation of tissue of the splenic hilum, can easily cause complications such as splenic fever and pancreatic fistula. Here, we use the technique of dissection of the secondary branches of the splenic pedicle to handle the hilum in the portal hypertension patients who are subjected to splenectomy. Methods We retrospectively compared and analyzed the complications, postoperative hospital stay, operative time, and occurrence of hemorrhage in 121 patients with portal hypertension undergoing splenectomy and devascularization of the gastric cardia from January 1999 to December 2007. The selected cases consisted of 51 patients undergoing conventional splenectomy and 70 patients undergoing dissection of secondary branches of the splenic pedicle. In addition, we analyzed the relationship between size of the spleen and occurrence of complications. Results The incidence of pancreatic fistula and splenic fever （0/70 and 9/70） was lower in patients undergoing dissection of secondary branches of the splenic pedicle as compared with that of the conventional group （5/51 and 18/51 respectively）. In addition, there was no significant difference in operative time and volume of blood loss between two groups. The spleen thickness of those patients who had pancreatic fistula and splenic fever was significantly greater than those without complications. Conclusions These results indicate that dissection of secondary branches of the splenic pedicle in portal hypertension patients undergoing splenectomy can decrease the incidence of splenic fever and pancreatic fistula, and shorten the postoperative hospital stay, especially in the patients with a large spleen. So dissection of secondary branches of the splenic pedicle is a valuable technique for splenectomy.

胆囊息肉的微创外科治疗(英文)

目的 :观察没有合并结石、胆囊功能良好的良性胆囊息肉患者 ,行经皮内镜胆囊息肉切除术的疗效。方法 :85例胆囊息肉 ,硬膜外麻醉 ,胆囊底部皮肤小切口 ,插入胆镜到胆囊内 ,用自制微波电极加热凝固息肉蒂部或基底部 ,切除息肉并作病理检查。术后定期随访。结果 :全部手术均顺利 ,平均手术时间 1～ 1 5h。 6 7例平均随访 5 5年 (2～ 9年 )结果无任何症状 ,B超检查 6 4例胆囊功能完好 ,无结石或息肉复发。结论 :该方法创伤小、效果好、并发症少 。

Diagnosis and surgical treatment of pancreatic endocrine tumors in 36 patients:a single-center report

Background Pancreatic endocrine tumors （PETs） are rare and their surgical treatment is often debated. The purpose of this retrospective study was to analyze the diagnosis and surgical strategy of functioning and non-functioning PETs. Methods From May 1980 to March 2006, 36 patients with pancreatic endocrine tumors at the Second Affiliated Hospital of Zhejiang University were retrospectively studied. Results Among the 36 patients, 29 （81%） had functioning tumors, and 7 （19%） had nonfunctioning tumors. Ninety-two percent of insulinomas were benign, whereas 4 （57%） of nonfunctioning PETs were malignant. The size of functioning tumors was （2.3i-0.3） cm, that of nonfunctioning tumors was less than （5.1^-0.5） cm. The combination CT and transabdominal ultrasonography resulted in a diagnostic sensitivity of 84%. Thirty-three primary lesions were precisely located in 32 patients （89%）. Atypical tumor resection was performed for 73% of functioning tumors, while typical pancreatectomy was performed for 6 （85%） of nonfunctioning tumors. Moreover, 5 liver resections and 1 lymph node dissection were performed. During the follow-up, fifteen complications occurred in 12 （36%） patients after operation. The 5-year survival rate for patients with benign tumors was 92% compared to 50% for those with malignant tumors. Surgical cure was achieved in 95% of patients with benign insulinomas. Conclusions Surgical strategy for PETs depends on the size and location of the tumor and the risk of malignancy. The optimal surgical procedure is key to prevent postoperative complication. Radical resection including initial and metastatic lesion may benefit patients with maliqnant PETs.

Splenic hamartoma: case report and review of literature

Splenic hamartoma is a benign malformation composed of an anomalous mixture of normal splenic elements.1 Approximately half of the patients with the tumor are asymptomatic and diagnosed accidentally by autopsy or splenectomy. The tumor-related renal hematological and dermatological abnormalities can be treated by removal of the tumor. Splenic hamartoma which is related to renal diseases including extramembranous glomerulonephritis, membranoproliferative glomerulonephritis and renal adenocarcinoma has been reported previously.

Changes of splenic macrophage during the process of liver cancer induced by diethylnitrosamine in rats

Background It is generally accepted that spleen plays a complex role in the tumor immunity, which would change in the different periods of cancer. In this study, we investigated the changes in the function of splenic macrophage （Mφ） in different stages of liver cancer induced by diethylnitrosamine （DEN） in rats. The aim was to support the characteristics of ＂two-way＂ and ＂phase＂ of spleen in tumor immunity. Methods The model of pulmonary metastasis of liver cancer was established in forty male SD rats by DEN. In the 8th, 13th and 16th week, 10 rats were randomly chosen and sacrificed, and divided into cirrhosis, liver cancer and pulmonary metastasis groups depending on the pathological result, respectively. The other 10 rats were taken as control group. The Mφ was isolated by anchoring cultivation. The changes in ultrastructure, phagocytosis, cytokine secretion, antigen processing and presenting, and viability of splenic Mφ） were detected by transmission electron microscopy, VybrantTM Phagocytosis Assay, DQ^TM Ovalbumin, and rat TNF-α ELISpot kits. Results Under the electron microscope, the Mφ） in the control group had some pseudopodium-like prominences, and mitochondria, ribosome, rough endoplasmic reticulum, lysosome can be found in the cytoplasm, and phagocytized RBC. In the liver cirrhosis and liver cancer group, Mφ had more prominences, meanwhile much more mitochondria, ribosome, rough endoplasmic reticulum, lysosome can be found in the cytoplasm, especially in the liver cancer group. In the pulmonary metastasis group, the Mφ was swelling, with few organelle. As compared to the control group, the function of splenic Mφ） increased in cirrhosis and cancer groups, but decreased in metastasis group （phagocytosis rate： （84.7±1.9）% （89.5±3.1）%, and （36.0±2.6）% vs （75.6±1.7）%, P〈0.05, P〈0.01; viability： （1.53±0.15）%, （1.56±0.14）%, and （1.12±0.29）% vs （1.48±0.17）%, P 〈0.05, P 〈0.01; TNF-α secretion： （741.0±52.9）%, （1126.2±174.5）%, and （313.8±50.8）% vs （626.6±24.6）%, P 〈0.05, P 〈0.01; positive cell rate of antigen processing and presenting： （24.03±1.87）%, （27.95±2.63）%, and （10.46±2.16）% vs （16.45±1.86）%, P 〈0.01）. Conclusions In the stage of cirrhosis and early cancer, the immune functions of splenic Mφ were reinforced. It may promote the non-specificity tumor immunity. On opposite, in the stage of pulmonary metastasis, the immune functions of splenic Mφ were impaired. It may lead to the decrease of tumor immunity.

Is radiofrequency ablation justified for liver metastatic VIPoma patient undergoing Whipple procedure？

Vasoactive intestinal polypeptide secreting tumors （VIPoma） are one kind of rare endocrine tumor which is mostly located in the pancreas.The annual incidence was estimated to be about 1 per 10 000 000 individuals in general population.1 The VIPoma is usually associated with Verner-Morrison syndrome following tremendous over-secretion of vasoactive intestinal polypeptide （VIP）, which consisted of water diarrhea,hypokalemia and achlorhydria.This syndrome has been termed pancreatic cholera, watery diarrhea syndrome.

Whole-exome mutational landscape of neuroendocrine carcinomas of the gallbladder

Neuroendocrine carcinoma(NEC)of the gallbladder(GB-NEC)is a rare but extremely malignant subtype of gallbladder cancer(GBC).The genetic and molecular signatures of GB-NEC are poorly understood;thus,molecular targeting is currently unavailable.Inthe present study,we applied whole-exome sequencing(WES)technology to detect gene mutations and predicted somatic singlenucleotide variants(SNVs)in 15 cases of GB-NEC and 22 cases of general GBC.in 15 GB-NECs,the C>T mutation was predominantamong the 6 types of SNVs.TP53 showed the highest mutation frequency(73%,11/15).Compared with neuroendocrine carcinomasof other organs,signifcantly mutated genes(SMGs)in GB-NECs were more similar to those in pulmonary large-cell euroendocrinecarcinomas(LCNECs),with drver roles for TP53 and RB1.Iin the COSMIC database of cancer-related genes,211 genes were mutated.Strikingly,RB1(4/15,27%)and NAB2(3/15,20%)mutations were found specifically in GB-NECs;in contrast,mutations in 29 genes,including ERB82 and ERBB3,were identified exclusively in GBC.Mutations in RB1 and NAB2 were significanty related to downregulation of the RB1 and NAB2 proteins,respectively,according to immunohistochemical(IHC)data(p values=0.0453 and0.0303).Clinically actionable genes indicated 23 mutated genes,including ALK,BRCA1,and BRCA2.Iin addition,potential somaticSNVs predicted by ISowN and SomVarlUS constituted 6 primary coSMIC mutation signatures(1,3,30,6,7,and 13)in GB-NEC.Genes carrying somatic SNVs were enriched mainly in oncogenic signaling pathways involving the Notch,WNT,Hippo,and RTK-RASpathways.In summary,we have systematically identified the mutation landscape of GB-NEC,and these findings may providemechanistic insights into the specifc pathogenesis of this deadly disease.

Assessment of 188Re marked anti MHC class II antibody by peripheral blood mononuclear cells stimulated by donor alloantigen

Background Previous studies showed that anti MHC-II monoclone antibody （MAb） only had partial inhibiting effect of alloreactive mixed lymphocyte reaction （MLR） in vitro and it was unsteady and non-persistent. The aim of this research was to determine whether radioactive isotope 188Re marked MHC-II antibody could benefit the allograft acceptance in transplantation as compared to normal MHC-II antibody. Methods 188Re was incorporated to 2E9/13F（ab＇）2 which is against swine MHC class II antigen （MAb-~88Re）. Porcine peripheral blood mononuclear （PBMC） cells were examined for proliferation and cytokine mRNA expression after stimulation with MHC-II MAb or MAb-188Re. Results The proliferative response of recipient PBMCs in mixed lymphocyte reaction （MLR） to donor alloantigen showed that the stimulation index of MAb-188Re group was significantly lower than the MHC-II MAb group and control （P 〈0.05）. mRNA expression of interleukin 2, interferon y and tumor necrosis factor a （type 1 cytokines） was lower in MAb-188Re group than the MHC-II MAb group, while interleukin 10 （type 2 cytokines） was higher in MAb-~aSRe group in the first 24 hours. Conclusion MAb-188Re could help the graft acceptance by inhibiting T cell proliferation, lowering the expression of type 1 cytokines and elevating the type 2 cytokines produced by PBMC.