Objective To prepare humanized monoclonal antibodies(Mabs)targeting Acinetobacter baumannii(Ab)based on single memory B cell sequencing technology,construct the immune repertoire of the core protein of Ab,hemolysin-coregulated protein(Hcp),and express its Mabs with binding activity.Methods E.coli BL21 harboring the recombinant plasmid pGEX-6p-1-Hcp was constructed.Hcp protein was obtained using protein expression and affinity chromatography.Female SPF BALB/c mice(6～8 weeks old,weighing 18～20 g)were immunized intramuscularly with antigen Hcp to generate specific memory B cells.Single antigen-specific memory B cells were sorted using flow cytometry.The immune repertoire of Hcp was constructed using single-cell sequencing technology,and bioinformatics analysis was performed on the sequencing results.Mabs were obtained using antibody humanization techniques.The in vitro binding activity of the antibodies was detected by ELISA.Results The target protein Hcp with a purity>95%was obtained after expression and purification.The immune repertoire of Hcp was successfully constructed,and the results of BCR clonotype identification and analysis,CDR3 region characteristic analysis,and V-J gene pairing characteristic analysis were achieved.Antibody humanization got 7 Mabs,that is,IgG1-1,IgG1-2,IgG2-1,IgG2-2,IgG3-1,IgG4-1 and IgG4-2.ELISA results showed IgG1-1,IgG3-1,IgG4-1,and IgG4-2 had an antibody binding titer of 1∶1 280,IgG2-2 of 1∶10 240,IgG2-1 of 1∶5 120,and IgG1-2 of 1∶160.Conclusion Single-cell sequencing technology enables rapid,accurate,and efficient construction of an Hcp protein immune repertoire containing extensive antibody information.Utilizing this immune repertoire allows for the expression of Mabs with binding activity.

Cancer-associated thromboembolism(CAT)is one of the most common complications and the second direct cause of mortality in patients with malignant tumors,which seriously affect patients'life quality and prognosis.In recent years,with the deepening of mechanism researches of cancer and thrombosis,treatment strategies of CAT were also improved.The current status and progresses of clinical treatment of CAT were reviewed in this article.

Objective To identify the miR-217 targeted gene ANLN by experiment.Methods Bioinformatic algorithms were used to predict the potential targets of miR-217.Then,ANLN binding with miR217 and mutant ANLN (mutANLN) sequence were designed and synthesized,and their amplified fragments were inserted into plasmid psiCHECK-2,and recombinant plasmid psiCHECK-2-ANLN and psiCHECK-2-mutANLN were reconstructed.The two recombinant plasmids were co-transfected into pancreatic cancer cell line PANC1 with miR-217,miR-217 inhibitor,NC,NC inhibitor by liposome,respectively.Dual luciferase reporter system was used to determine the luciferase activity,and Western blot was used to measure the expression of ANLN protein.Results The luciferase activities of psiCHECK-2-ANLN,psiCHECK-2-ANLN +miR-217,psiCHECK-2ANLN + miR-217 inhibitor,psiCHECK-2ANLN + NC,psiCHECK-2-ANLN + NC inhibitor were 2.221 ± 0.188,0.769 ± 0.061,3.764 ± 0.371,2.265 ± 0.201,2.242 ± 0.018,and the difference among these groups was statistically significant (F =77.405,P ＜0.001),but the difference among psiCHECK-2ANLN group,psiCHECK-2-ANLN + NC group and psiCHECK-2-ANLN + NC inhibitor group was not statistically significant.However,luciferase activities of psiCHECK-2-ANLN + miR-217 group were significantly decreased when compared with other 3 groups,and luciferase activity of psiCHECK-2-ANLN +miR-217 inhibitor group were significantly increased when compared with other 4 groups (all P ＜0.001).Luciferase activities of groups transfected with psiCHECK-2-mutANLN was not significantly different (P =0.053).The expression of ANLN protein in PANC1 with psiCHECK-2-ANLN + miR-217 co-transfection was significantly down-regulated when compared with that with psiCHECK-2-ANLN transfection alone.Conclusions ANLN is one of the direct target genes of miR-217 in PANC1 cells.

OBJECTIVETo study the application of three-dimensional visualization technology for laparoscopic resection of cystic carcinoma in the pancreatic body and tail.

METHODSSix cases of cystic carcinoma in the pancreatic body and tail treated between Nov, 2009 and Mar, 2011 were retrospectively analyzed. The original image data of 64-slice spiral CT were collected and using adaptive region growing algorithm, the serial CT images were segmented and automatically extracted to obtain the 3-dimensional reconstruction images with customized image manipulation software. The specific surgical approach (the trocar position) and surgical procedure were planned based on the reconstructed mode.

RESULTSThe reconstructed 3-dimensional model clearly displayed cystic carcinoma in the pancreatic body and tail and the adjacent organs, showing distinct relationship between the cystoma and the splenic artery and vein. All the patients successfully underwent laparoscopic resection of the pancreatic body and tail without perioperative death. The spleen was preserved in 5 cases and removed in 1 case due to mucinous cystadenocarcinoma. The overall rate of pancreatic fistulae was 33.3% without incidences of postoperative hemorrhage. The average hospital stay of the patients was 12 days.

CONCLUSIONThree-dimensional reconstruction based on pancreatic CT data provides valuable assistance for laparoscopic resection of cystic carcinoma in the pancreatic body and tail.

Adult ; Aged ; Computer Simulation ; Cystadenocarcinoma, Mucinous ; diagnostic imaging ; surgery ; Cystadenoma, Mucinous ; diagnostic imaging ; surgery ; Cystadenoma, Serous ; diagnostic imaging ; surgery ; Female ; Humans ; Imaging, Three-Dimensional ; Laparoscopy ; adverse effects ; methods ; Length of Stay ; Male ; Middle Aged ; Pancreas ; diagnostic imaging ; surgery ; Pancreatectomy ; adverse effects ; methods ; Pancreatic Fistula ; etiology ; Pancreatic Neoplasms ; diagnostic imaging ; surgery ; Retrospective Studies ; Spleen ; surgery ; Tomography, Spiral Computed

Objective To study the application of three-dimensional visualization technology for laparoscopic resection of cystic carcinoma in the pancreatic body and tail. Methods Six cases of cystic carcinoma in the pancreatic body and tail treated between Nov, 2009 and Mar, 2011 were retrospectively analyzed. The original image data of 64-slice spiral CT were collected and using adaptive region growing algorithm, the serial CT images were segmented and automatically extracted to obtain the 3-dimensional reconstruction images with customized image manipulation software. The specific surgical approach (the trocar position) and surgical procedure were planned based on the reconstructed mode. Results The reconstructed 3-dimensional model clearly displayed cystic carcinoma in the pancreatic body and tail and the adjacent organs, showing distinct relationship between the cystoma and the splenic artery and vein. All the patients successfully underwent laparoscopic resection of the pancreatic body and tail without perioperative death. The spleen was preserved in 5 cases and removed in 1 case due to mucinous cystadenocarcinoma. The overall rate of pancreatic fistulae was 33.3% without incidences of postoperative hemorrhage. The average hospital stay of the patients was 12 days. Conclusion Three-dimensional reconstruction based on pancreatic CT data provides valuable assistance for laparoscopic resection of cystic carcinoma in the pancreatic body and tail.

Objective To study the application of three-dimensional visualization technology for laparoscopic resection of cystic carcinoma in the pancreatic body and tail. Methods Six cases of cystic carcinoma in the pancreatic body and tail treated between Nov, 2009 and Mar, 2011 were retrospectively analyzed. The original image data of 64-slice spiral CT were collected and using adaptive region growing algorithm, the serial CT images were segmented and automatically extracted to obtain the 3-dimensional reconstruction images with customized image manipulation software. The specific surgical approach (the trocar position) and surgical procedure were planned based on the reconstructed mode. Results The reconstructed 3-dimensional model clearly displayed cystic carcinoma in the pancreatic body and tail and the adjacent organs, showing distinct relationship between the cystoma and the splenic artery and vein. All the patients successfully underwent laparoscopic resection of the pancreatic body and tail without perioperative death. The spleen was preserved in 5 cases and removed in 1 case due to mucinous cystadenocarcinoma. The overall rate of pancreatic fistulae was 33.3% without incidences of postoperative hemorrhage. The average hospital stay of the patients was 12 days. Conclusion Three-dimensional reconstruction based on pancreatic CT data provides valuable assistance for laparoscopic resection of cystic carcinoma in the pancreatic body and tail.

OBJECTIVETo explore the clinical significance of miRNA-216a expression in pancreatic cancer.

METHODSFourteen patients with pancreatic cancer undergoing pancreaticoduodenectomy and 6 patients with benign pancreas lesions were examined for miR-216a expressions in the tumor or lesion tissues using Agilent Human miRNA Microarray (V12.0). The relationship between miR-216a expressions and the clinicopathological features of the patients was analyzed.

RESULTSThe expression of miRNA-216a was significantly lower in pancreatic cancer than in benign pancreas lesions (P=0.000). The expression of miRNA-216a was significantly correlated with the T stage of the tumor (P=0.002), but not with the patients' age, gender, smoking status, tumor stage, lymph node metastases, distant metastasis, tumor differentiation, nerve invasion, vessel invasion or serum CA19-9 level (P>0.05).

CONCLUSIONSThe down-regulated expression of miR-216a in pancreatic cancer suggests the involvement of miR-216a in the tumorigenesis and development of pancreatic cancer. miR-216a may potentially serve as a novel tumor marker and also a prognostic factor for pancreatic cancer.

Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; MicroRNAs ; genetics ; metabolism ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; Pancreatic Neoplasms ; metabolism ; pathology

Objective To evaluate the value of 18F-FDG PET in predicting the prognosis of pancreatic cancer. Methods Fifty-four cases of pancreatic cancer were divided into two groups. Taking standard uptake value(SUV) at 4 as the cut off point, patients (22 cases) with that not more than four were classified into group A, and those (32 cases) with SUV greater than four were into group B. The prognosis of patients by SUV was analyzed statistically. Results There was a statistically significant difference in survival between the two groups ( P =0. 01 ). The 1-,3-year survival rate was 68. 18% 、34. 91% in group A in those the SUV≤4, and 33.61% 、11.95% in group B( SUV ＞4), respectively. Cox proportional hazard model showed that tumor staging and SUV were the significantly independent prognostic factors in patients with pancreatic cancer. Conclusions 18F-FDG PET is of value in predicting the prognosis of patients with pancreatic cancer.

Objective To explore the diagnosis and treatment of cystic pancreatic tumors. Methods The clinical data of 21 cases with cystic pancreatic tumors were retrospectively analyzed. Results This group enrolled 21 cases including serous cystic neoplasm of the pancreas (SCN, 11 cases), mucinous cystic neoplasm of the pancreas (MCN, 6 cases), intraductal papillary mucinous neoplasms (IPMN,2 cases), solid posudopapillary tumor of the pancreas (SPTP,2 cases). Seven cases had certain symptoms or typical signs, while others were asymptomatic and tumors were found by regular physical examinations. All of the cases were diagnosed by CT scans, and the value of serum tumor markers was within normal range. Twenty patients underwent pancreatic tumor resections and there was no perioperative death. Pancreaticoduodenectomy was performed in 8 cases, and distal pancreatectomy was performed in 6 cases including 1 patient undergoing laparoscopic distal pancreatectomy. Middle segment resection was performed in 4 cases, and tumor enucleation was performed in 2 cases. These 20 patients were followed up for 11 to 96 months, and there was no tumor recurrence or metastasis. One patient with mucinous cystic carcinoma underwent palliative operation and survived 4 months after surgery. Conclusions Preoperative imaging was not able to confirm the definite tumor pathological category. Laparotomy should be performed in a patient with cystic pancreatic tumor. The selection of surgical approach should be individualized, and the laparoscopic operation is an alternative.

Objective To explore the clinical characteristics of huge pancreatic pseudocysts and to evaluate the effect of different surgical treatments. Method We retrospectively analyzed the clinical data of 27 patients with huge pancreatic pseudocyst managed from Feb 1991 to Feb 2008. Result Among a total of 129 patients with pancreatic pseudocyst treated during this period of time,27 (20.9% ) patients were diagnosed as with huge pancreatic pseudocyst (diameter > 10cm). As to the etiology, 51.9% of the psudoeyst was caused by acute pancreatitis, 33.3% by pancreatic trauma and previous surgery, 11.1% by chronic pancreatitis. Pseudoeysts in the majority of cases(21/27)had a history less than 6 weeks. Upper GI obstruction complicated 30% cases (8/27). Imaging showed that all huge pancreatic pseudocysts were single. ERCP showed communication with the main pancreatic duct in 9 out of 11 cases. Nine cases underwent catheter drainage, 10 cases underwent cystogastrostomy,2 cases underwent endoscopic drainage of pancreatic pseudocyst via ERCP, 17 cases underwent Roux-en-Y cystojejunostomy including 11 cases in which other previous procedures failed. All 27 cases were finally cured. Conclusions Huge pancreatic pseudocyst might have unique clinical characteristics. Anatomical changes of main pancreatic duct were found in most cases under ERCP. The proper time and indication for surgical intervention might be different from minor pancreatic pseudocyst.