Objective To determine the most appropriate method in the treatment of choledocholithiasis. Methods The clinical data of 135 patients who received either laparoscopic common bile duct exploration (LCBDE) or laparoscopic transcystic common bile duct exploration (TC-BDE) from May 2005 to July 2009 were retrospectively studied. Results All patients were cured of choledocholithiasis. In 13 patients three trocars were used, in 89 patients four trocars were used, and in 33 patients conversion to open operation was done. Of the 102 patients in whom minimal invasive surgery was successfully carried out, TC-BDE was done in 6 patients and LCBDE in 96 patients. Primary closure of common bile duct (CBD) was carried out in 33 patients while T-tube drainage was done in 63 patients. Six patients had a transient bile leak which healed spontaneously in 3 to 4 days. Retained stone was detected in 2 patients which was successfully retrieved through choledochoscopy. Conclusion An individualized treatment approach is the most reasonable way to manage CBD stones. The choice of the procedure depends on the number and size of stones, the surgeons experience and the patient's choice.

Objective:To evaluate the antitumor activity of recombinant SEA for therapy of B16 melanoma established in C57BL/6 mice. Methods:C57BL/6 mice with melanoma were treated with the purified rSEA. The tumors were isolated and weighted. Results:Tumor growth was apparently inhibited by rSEA at high, middle, and low doses intraperitone-ally, whose inhibition ratio were 79.3% , 75.6 % and 73. 8% respectively. rSEA treatment in situ could inhibit tumor growth more effectively(90.6% ). Further study showed that numerous CD8+ and CD4+ T cell were infiltrated in tumor tissues, which were consistent with tumor growth inhibition induced by rSEA. Conclusions: rSEA could inhibit tumor growth effectively, especially the treatment in situ. This study paves the way for tumor immunotherapy with targeted SEA.

Objective To analyze the property of natural polysaccharides(P) from Porphyra haitanensis T. J. Chang et B. F. Zheng and its degraded polysaccharides(P3),and to investigate their effects on proliferation of lymphocytes. Methods P3 and P were identified by IR spectra. Their effects on immunological function were studied by cell culture technique. Results P3 and P displayed significant effects on promoting the proliferation of T-lymphocytes,and had inhibitory effects on the proliferation of B-lymphocytes in a dose-dependent manner. The effects of P3 were more powerful than P. Conclusion The degraded and natural polysaccharides from Porphyra haitanensis(P3 and P) can regulate the proliferation of lymphocytes,which may be related to their molecular weights.

Objective To detect the impact of reactive oxygen species ( ROS) on mitochondrial morphology and distri-bution during mitosis.Methods A viral vector in which the fluorescence gene was specifically under the control of mito-chondrial promoter was constructed and confirmed through DNA sequencing and Western blotting.After transfecting HeLa s3 cell with packaged virus, the HeLa s3-COX4tp-EGFP cell line stably expressing the mitochondrial fluorescence signal was obtained.With immunofluorescent staining, the impact of ROS on the morphology and distribution of mitochondria dur-ing mitosis was inspected.Result The cell line constantly expressing mitochondrial fluorescence signals was successfully constructed.Meanwhile,it was found that H2 O2 treatment could significantly change the morphology and distribution of mi-tochondria during mitosis by confocal microscopy.Conclusion Our study demonstrates that ROS can affect the morphology and distribution of mitochondria during mitosis.This research help study the relationship between the mitochondrial function and the regulation of mitosis in the future.

Objective To study the impact of regional lymph node metastasis on the prognosis of patients with hepatocellular carcinoma (HCC) after hepatectomy.Methods Three hundred and twenty-two HCC patients who underwent elective hepatectomy in the Heze Municiple Hospital were retrospectively studied from January 1999 to December 2013.The clinical data and follow-up results were analyzed.The diagnosis of lymph node metastasis was based on preoperative imaging data and intraperative findings.Results There were 16 patients (5.0％) with lymph node metastasis (the metastasis group) and 306 patients (95.0％) without lymph node metastasis (the no-metastasis group).On Kaplan-Merier analysis,the overall survival (OS) rates at 1,2,5 years were 43.8％,31.3％,7.5％ in the metastasis group and 83.7％,70.9％,46.1％ in the no-metastasis group,respectively (Log-rank test:P ＜ 0.05).The corresponding disease-free survival (DFS) rates were 30.0％,22.5％,6.3％ in the metastasis group and 69.8％,52.8％,34.5％ in the no-metastasis group,respectively,(P ＜0.05).On follow-up,the mid-and shortterm HCC recurrence rates (≤6 months) were 71.4％ in the metastasis group (10 patients) and 31.6％ in the no-metastasis group (142 patients,x2 =9.754,P ＜ 0.05),respectively.Absence of cirrhosis,hepatectomy extent of ＞ 1 Section,max-diameter of tumor ＞ 5 cm,poor differentiation and absence of tumor encapsulation were significant factors of DFS in the metastasis group (P ＜ 0.05).On COX regression analysis,absence of tumor encapsulation was the only independent risk factor of DFS.Conclusions Regional lymph node metastasis was one of the significant factors for OS and DFS in HCC patients who underwent hepatectomy.Resection of metastatic lymph nodes and post-operation adjuvant treatment should be helpful to improve patients' prognosis.

Epidermal growth factor receptor (EGFR) and its ligands (EGF and TGFalpha) are over-expressed in a variety of tumors. Immunization EGF-carrier protein inhibits tumor growth through abrogating binding of EGF to EGFR. Here, a chimeric protein of EGF and TGFalpha (E5T) was genetically fused to Staphylococcal enterotoxin A (SEA), a bacterial superantigenic protein which promotes humoral B cell response through enhancement of Ag-specific CD4 T cells activity. The resulted fusion proteins were expressed in Escherichia coli and purified though metal chelating affinity chromatography. Immunization of E5T-mSEA fusion protein in mice induced production of high titers antibodies, which recognize both EGF and TGFalpha. Anti- E5T-mSEA serum at dilution of 1:10 significantly inhibited growth of A431 cell lines but had little effect on 293T cell lines.
Amino Acid Sequence ; Animals ; Cancer Vaccines ; biosynthesis ; immunology ; Cell Line, Tumor ; Enterotoxins ; biosynthesis ; genetics ; Epidermal Growth Factor ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Humans ; Immunization ; Mice ; Mice, Inbred C57BL ; Molecular Sequence Data ; Random Allocation ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Transforming Growth Factor alpha ; biosynthesis ; genetics

Objective:To study the prognostic factors in patients with occult ruptured hepatocellular carcinoma (HCC) after hepatectomy.Methods:The clinical data of 31 patients with occult ruptured HCC who underwent hepatectomy at the Heze Multiple Hospital from January 2001 to December 2014 were retrospectively analyzed. There were 27 males and 4 females, with ages ranging from 29 to 73 years. Kaplan-Meier survival analysis was carried out for survival analysis, and compared by the log-rank test. Multivariate analysis was conducted using the Cox proportional hazards regression model.Results:Of 31 patients in this study, 27(87.1%) had died, and 25(80.6%) had developed tumor recurrence and metastasis. The median overall survival was 8 months and the median tumor-free survival was 4 months. On Cox regression analysis, non-R 0 resection was an independent risk factor of overall survival ( RR=2.816, 95% CI: 1.006-7.887) and disease-free survival ( RR=3.295, 95% CI: 1.215-8.940). Poor tumor differentiation was an independent risk factor of overall survival ( RR=3.120, 95% CI: 1.193-8.160). Conclusions:The prognosis of patients with occult rupture HCC who underwent no R 0 resection was poor. However, the prognosis of patients who underwent non-R 0 resection was even poorer.

Arabidopsis thaliana is an important and long-established model species for plant molec-ular biology,genetics,epigenetics,and genomics.However,the latest version of reference genome still contains a significant number of missing segments.Here,we reported a high-quality and almost complete Col-0 genome assembly with two gaps(named Col-XJTU)by combining the Oxford Nanopore Technologies ultra-long reads,Pacific Biosciences high-fidelity long reads,and Hi-C data.The total genome assembly size is 133,725,193 bp,introducing 14.6 Mb of novel sequences compared to the TAIR1 0.1 reference genome.All five chromosomes of the Col-XJTU assembly are highly accurate with consensus quality(QV)scores＞60(ranging from 62 to 68),which are higher than those of the TAIR10.1 reference(ranging from 45 to 52).We completely resolved chro-mosome(Chr)3 and Chr5 in a telomere-to-telomere manner.Chr4 was completely resolved except the nucleolar organizing regions,which comprise long repetitive DNA fragments.The Chr1 cen-tromere(CEN1),reportedly around 9 Mb in length,is particularly challenging to assemble due to the presence of tens of thousands of CEN180 satellite repeats.Using the cutting-edge sequencing data and novel computational approaches,we assembled a 3.8-Mb-long CEN1 and a 3.5-Mb-long CEN2.We also investigated the structure and epigenetics of centromeres.Four clusters of CEN180 monomers were detected,and the centromere-specific histone H3-like protein(CENH3)exhibited a strong preference for CEN 180 Cluster 3.Moreover,we observed hypomethylation patterns in CENH3-enriched regions.We believe that this high-quality genome assembly,Col-XJTU,would serve as a valuable reference to better understand the global pattern of centromeric polymorphisms,as well as the genetic and epigenetic features in plants.