Prognostic factors analysis of Siewert type Ⅱ and Ⅲ adenocarcinoma of esophagogastric junction after radical resection with different surgical approaches / 中华消化外科杂志

Objective:To investigate the prognostic factors of Siewert type Ⅱ and Ⅲ adenocarcinoma of esophagogastric junction (AEG) after radical resection with different surgical approaches.Methods:The retrospective case-control study was conducted. The clinicopathological data of 442 patients who were admitted to Tianjin Medical University Cancer Institute and Hospital from February 2003 to July 2011 were collected. There were 362 males and 80 females, aged from 21 to 85 years, with a median age of 64 years. Patients underwent radical resection of AEG. Observation indicators: (1) surgical situations; (2) follow-up; (3) progrostic factors analysis of AEG after radical resection; (4) survival of patients after radical resection of AEG via abdominal approach; (5) survival of patients after radical resection of AEG via thoracoabdominal approach; (6) survival of patients after radical resection of Siewert type Ⅱ type AEG; (7) survival of patients after radical resection of Siewert type Ⅲ AEG. Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival of patients up to June 2018. Measurement data with skewed distribution were described as M (range). Count data were expressed as absolute numbers or percentages. Kaplan-Meier method was used to calculate survival rates and draw survival curves, and Log-rank test was used for survival analysis. Univariate analysis was conducted using the Kaplan-Meier method. Multivariate analysis was conducted using the COX proportional hazard model. Results:(1) Surgical situations: 442 patients underwent radical resection of AEG, including 204 via abdominal approach and 238 via thoracoabdominal approach. There were 391 patients with D 2 lymphadenectomy and 51 with D 2+ lymphadenectomy. (2) Follow-up: 442 patients were followed up for 8-162 months, with a median follow-up time of 37 months. All the 442 patients survived for 2-156 months, with a median survival time of 31 months. The 1-, 3-, 5-year overall survival rates were 79.2%, 42.0%, 30.0%, respectively. (3) Prognostic factors analysis of AEG after radical resection: results of univariate analysis showed that tumor diameter, Lauren type, pathological T staging, pathological N staging, pathological TNM staging, lymphatic vessel invasion, and soft tissue infiltration were related factors for prognosis of patients after radical resection of Siewert type Ⅱ and Ⅲ AEG ( χ2=4.028, 4.885, 19.435, 17.014, 34.449, 9.707, 11.866, P<0.05). Results of multivariate analysis showed that pathological TNM staging, lymphatic vessel invasion, and soft tissue infiltration were independent influencing fators for prognosis of patients after radical resection of Siewert type Ⅱ and Ⅲ AEG ( hazard ratio=1.255, 0.486, 1.454, 95% confidence interval: 1.024-1.539, 0.325-0.728, 1.096-1.928, P<0.05). (4) Survival of patients after radical resection of AEG via abdominal approach: of the 204 patients undergoing radical resection of AEG via abdominal approach, the 1-, 3-, 5-year survival rates were 83.6%, 50.4%, 37.8% for 121 patients with Siewert type Ⅱ AEG, respectively, versus 72.0%, 39.3%, 31.8% for 83 patients with Siewert type Ⅲ AEG, showing no significant difference in the survival between the two groups ( χ2=1.854, P>0.05). (5) Survival of patients after radical resection of AEG via thoracoabdominal approach: of the 238 patients undergoing radical resection of AEG via thoracoabdominal approach, the 1-, 3-, 5-year survival rates were 79.6%, 38.8%, 23.8% for 183 patients with Siewert type Ⅱ AEG, respectively, versus 79.1%, 37.6%, 29.3% for 55 patients with Siewert type Ⅲ AEG, showing no significant difference in the survival between the two groups ( χ2=0.215, P>0.05). (6) Survival of patients after radical resection of Siewert type Ⅱ AEG: of the 304 patients with Siewert typeⅡAEG, the postoperative 1-, 3-, 5-year survival rates were 83.6%, 50.4%, 37.8% for 121 patients undergoing radical resection of AEG via abdominal approach, respectively, versus 79.6%, 38.8%, 23.8% for 183 patients undergoing radical resection of AEG via thoracoabdominal approach, showing no significant difference in the survival between the two groups ( χ2=2.406, P>0.05). (7) Survival of patients after radical resection of Siewert type Ⅲ AEG: of the 138 patients with Siewert type Ⅲ AEG, the postoperative 1-, 3-, 5-year survival rates were 72.0%, 39.3%, 31.8% for 83 patients undergoing radical resection of AEG via abdominal approach, respectively, versus 79.1%, 37.6%, 29.3% for 55 patients undergoing radical resection of AEG via thoracoabdominal approach, showing no significant difference in the survival between the two groups ( χ2=0.640, P>0.05). Conclusions:Pathological TNM staging, lymphatic vessel invasion, and soft tissue infiltration are independent fators for prognosis of patients after radical resection of Siewert type Ⅱ and Ⅲ AEG. Siewert types and surgical approach are not related factors for prognosis of patients after radical resection of AEG. There is no significant difference in the survival between patients with different Siewert types of AEG undergoing radical resection via different surgical approaches.

Comparing the characteristics of tissue culturein vitro and different cell types of cancer microenvironment in tissues at different distances from colorectal cancer lesions / 中国癌症杂志

Background and purpose:Cancer microenvironment has become a hot topic of cancer research. It is important in the initiation of colorectal cancer. This study aimed to discuss the correlation between the characteristics of tissue culturein vitro and different cell types in cancer microenvironment.Methods:Samples were collected at different distances from the colorectal cancer lesions, which were named as positions 1, 2 and 3 from distal to proximal. Tissues were cut into 1-2 mm3 forin vitro culturing. HE staining was used to observe the structure of crypts. Immunohistochemistry was used to detect the expressions of Cyclin D1 (CD1), CD133, cytokeratin18 (CK18), vimentin and α-smooth muscle actin (α-SMA).Results:Position 3 grew faster than position 2 and position 1. As getting closer to the colorectal cancer lesions, expressions of CD1, CD133, vimentin and α-SMA were increased while expression of CK18 was decreased.Conclusion:The tissue structure and the expression of different cell types in cancer microenvironment changed more seriously as get-ting closer to the colorectal cancer lesions. This indicated that the change of cancer microenvironment may contribute to the initiation of colorectal cancer.

Study on the association of gastric conduit width and postoperative early delayed gastric emptying in middle-lower esophageal cancer patients undergoing Ivor-Lewis procedure / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To examine the relationship between gastric conduit width and postoperative early delayed gastric emptying (DGE) in patients with middle-lower esophageal carcinoma who underwent Ivor-Lewis operation.</p><p><b>METHODS</b>Clinical data of 282 consecutive patients with middle-lower esophageal cancer who underwent the Ivor-Lewis operation by same surgical team in our department from January 2013 to June 2015 were retrospectively analyzed. Patients were divided into three groups according to the width of gastric conduit: width > 5.0 cm as broad group (n=93); width 3.0-5.0 cm as moderate group (n=70); width < 3.0 cm as narrow group (n=119). The gastric conduits of patients in narrow group were completely positioned the esophageal bed and fixed to the pericardium posterior wall. None of patients received pyloroplasty or pylorotomy. Perioperative data, operation-associated complications, and postoperative upper gastrointestinal radiographic results(1 week and 4 weeks after operation) were compared among groups.</p><p><b>RESULTS</b>The baseline data among these groups were comparable in terms of age, gender, tumor TNM staging, pathological types, serum albumin level, and the rate of receiving neoadjuvant therapy(all P>0.05). There were no significant differences in operative time, blood loss, and postoperative hospital stay among groups(all P>0.05). No patients died during perioperative peried. Anastomotic leakage occurred in 2 cases, one from broad group and another from narrow group. The incidences of arrhythmia and postoperative pulmonary complications, including infection, atelectasis, pneumothorax, and pleural effusion were similar among groups (all P>0.05). The average amount of gastric juice drainage in narrow group was (98±57) ml/day, which was markedly lower than that in broad group [(157±62) ml/day, P=0.000] and in moderate group [(123±68) ml/day, P=0.008]. One week after operation, the overall incidence of DGE was 10.6%(30/282), the incidence of DGE in broad, moderate, narrow groups was 17.2%(16/93), 14.3%(10/70), and 3.4%(4/119) respectively, and broad and moderate groups had higher incidence as compared to narrow group (P=0.001 and P=0.006).</p><p><b>CONCLUSION</b>During the Ivor-Lewis operation, application of a narrow gastric conduit (width < 3.0 cm), which completely position the esophageal bed with fixation to the pericardium posterior wall, can significantly reduce the incidence of postoperative early DGE.</p>

Tissue and Induction Expression Profiles of Rice Phospholipid Hydroperoxide Glutathione Peroxidase at Protein Level / 生物化学与生物物理进展

Proteins arc the major molecules performing life activities, and their spatial and temporal expression profiles in organisms are very important for understanding their accurate functions. Phospholipid hydroperoxide glutathione pcroxidase (PHGPx) is a unique antioxidant enzyme that directly reduces lipid hydroperoxides in biomembranes and plays a vital role in defense against mernhrane peroxidation damage. The protein expression profiles of rice PHGPx (OsPHGPx) were investigated in different rice tissues and under various stress treatments by using Western blot analysis. The results showed that in mature rice plants, OsPHGPx was mainly distributed in leaves, especially flag leaves, and in rice seedlings, OsPHGPx was detected in shoots and leaves. Moreover, OsPHGPx expression in rice seedlings could be markedly induced by H2O2 and NaCI, but weakly influenced by several plant hormones. Time- and dose-dependent effects were observed in both H2O2 and NaCI treatments, and the strongest induction was observed when rice seedlings were treated with 0.5 mmol/L H2O2 for 12 h or 500 mmol/L NaCI for 24 h. Additionally, dimethylthiourea, a H2O2 trap, inhibited H2O2-enhanced expression of OsPHGPx, but did not impair the enhanced effect of NaCI, implying that NaCl-induced OsPHGPx expression was not mediated by H2O2. These results will contribute greatly to further study the exact physiological function of OsPHGPx in rice.

In vitro proliferation of mesenchymal stem cells on three-dimensional macroporous scaffolds of chitosan-gelatin-basic fibroblast growth factor composite / 中国组织工程研究

BACKGROUND: Stem cell differentiation potential is strongly correlated with culture condition. The alteration in scaffold material surface function, three dimensional (3D) structure, and addition of growth factors can control stem cell proliferation and differentiation.OBJECTIVE: To develop 3D macroporous scaffolds with optimal porosity and porous structure to provide a microenvironment that promotes the growth of multi-potent stem cells.DESIGN: Repetitive measurement.SETTING: Department of Anatomy, College of Basic Medicine, Guangzhou University of Chinese Medicine.MATERIALS: Healthy adult SD rats were provided by the Experimental Animal Center in Guangzhou University of Chinese Medicine. Chitosan and basic fibroblast growth factor (bFGF) were purchased from Sigma Corporation (St. Louis,MO).METHODS: The experiment was performed at the Department of Anatomy, College of Basic Medicine, Guangzhou University of Chinese Medicine from March 2003 to December 2006. Using a freeze-drying method, 3D macroporous scaffolds made of different ratios of chitosan-gelatin with bFGF were fabricated that could release bFGF with controlled porosity and porous structure. Bone marrow was obtained from the femur and tibia of SD rats, and mesenchymal stem cells (MSCs) were isolated, cultured and seeded on the scaffolds with bFGF. MSCs seeded on scaffolds with no bFGF served as control. The procedure during experiment was accorded with animal ethical requirements.MAIN OUTCOME MEASURES: 3D structure and release performance of the scaffolds were observed by ELISA and scanning electron microscope; the effect of 3D macroporous scaffolds that released bFGF on MSC growth and viability were observed by HE staining, MTT, cell counting and SEM.RESULTS: There was no significant difference in pore size between scaffolds with and without bFGF (P ＞ 0.05). Scaffolds with bFGF significantly improved MSC survival rate, promoted cell adhesion, proliferation, and viability compared with scaffolds without bFGF (P ＜ 0.05).CONCLUSION: The results suggest that 3D macroporous scaffolds with bFGF release improve MSC survival on scaffolds,and lay a foundation for its application in tissue engineering.

Prokaryotic Expression and Purification of a Cotton Dehydration Responsive Element Binding Protein GhDBP1 and Its DNA Binding Activity / 生物化学与生物物理进展

Cotton (Gossypium hirsutum) is one of the most important economic crops in the world. Its growth and productivity were affected by environment stresses such as drought, cold and high salinity. Thus, the enhanced stress tolerance in this plant is of great importance. As the dehydration responsive element (DRE) binding protein (DBP) plays an important role in the regulation of plant resistance to environmental stresses and is quite useful for generating transgenic plants tolerant to these stresses, isolation and functional analysis of DBPs in cotton are important to cotton production. In the previous work, a DBP gene from cotton, named as GhDBP1, was isolated and its expression patterns in cotton plants was demonstrated at the transcriptional level. Here, the expression,purification and DNA binding activity of GhDBP1 were reported. The entire coding region of the GhDBP1 gene was inserted into an expression vector, pET28a, and transformed into Escherichia coli BL21 (DE3). The fusion protein was successfully expressed under IPTG induction and the purified recombinant protein was obtained by Ni-NTA affinity chromatography. Non-radioactive electrophoretic mobility shift assay revealed that the purified GhDBP1 protein was able to form a specific complex with the previously characterized DRE element. In addition, the computer modeling of the DNA-binding domain of GhDBP1 were performed using SWISS-MODEL software. The main-chain structures and the folding patterns of the DNA-binding domain of GhDBP1 were similar to the known structure of the DNA-binding domain of the Arabidopsis thaliana GCC box-binding protein AtERF1. These results indicate that GhDBP1 is a DRE-binding transcription factor and might use the structure similar to that of AtERF1 to interact with DRE sequence.

Molecular Characterization of Two Rice cDNAs Encoding GDP-Mannose-3', 5'-Epimerase and Their Expression Patterns / 生物化学与生物物理进展

GDP-mannose-3', 5' -epimerase (GME), which converts GDP-mannose into GDP-L-galactose, is essential for the biosynthesis of L-ascorbic acid in higher plants. The molecular characterization of two GME genes from rice has been reported. Firstly,both cDNAs were isolated from the rice mature leaves using RT-PCR technique. By comparing their sequences with homologues from other plants, it was found that GME genes were highly conserved among plant species, though phylogenetic study showed that all known GMEs could be divided into two distinct groups corresponding to monocots and dicots. Secondly, the genomic organization of rice OsGME genes was investigated, and a similarity of splice patterns was revealed. Finally, the expression patterns of the two cDNAs have been studied in various tissues and under different stress conditions by semiquantitative RT-PCR assay. The results showed that the OsGME1 transcript was up-regulated in response to cold stress, and gibberellin might regulate L-ascorbic acid levels by affecting transcription of both OsGME genes.

Anti-atherosclerotic Effect of Jiawei Zexie Tang in Rats / 中国药房

OBJECTIVE: To study the antiatherosclerotic effect of Jiawei zexie tang(GWZXT) and the possible mechanisms.METHODS: 48 healthy male SD rats were randomized to 5 groups,i.e normal control,model group,simvastatin group,and high dosage and low dosage GWZXT groups.Artherosclerosis rat model was established by feeding high fat forage and the corresponding drugs were administered intragastrically.Changes in serum levels of APoAⅠ,ApoB,Zn,Cu,Ca and Mg and the ultrastructure were detected.RESULTS: In high dosage GWZXT-treated group compared with model group,the Zn,Ca and ApoB levels decreased significantly(P

Radish Phospholipid Hydroperoxide Glutathione Peroxidase Gene Structure and Upstream Regulatory Sequence Analysis / 生物化学与生物物理进展

A novel radish RsPHGPx cDNA, which encodes a functional phospholipid hydroperoxide glutathioneperoxidase (PHGPx) protein, was identified in the previous work. In the study genomic organization and the upstream regulatory sequence analysis of this gene was presented. Southern blot analysis showed that RsPHGPx gene existed in radish genome in manner of single copy. Moreover, a 3.3 kb genomic DNA fragment of RsPHGPx gene was isolated by combination of common PCR and genome-walking method. Sequence analysis on this genomic fragment demonstrated that RsPHGPx gene consists of seven exons separated by six introns, and suggested that a short 5'-flanking sequence immediately before the exon 1 should be the putative RsPHGPx promoter region, which is proceeded by the upstream neighboring biotin synthase gene. Cis-acting elements search showed that the putative promoter contains elements responsive to hormones (eg. E-Box and W-Box), abiotic stresses (eg. MYB and MYC binding sites), and light (Box Ⅱ and Ⅰ-Box), etc. Northern blot analysis indicated that the expression of RsPHGPx was subjected to up-regulation of chilling and down-regulation of ABA and successive illumination (in etiolated seedlings), implying the regulatory roles of some predicted elements. However the up-regulation effect of herbicide paraquat, which can induce oxidative stress, suggested the presence of some unknown elements in the promoter region. This is the first report on gene structure and upstream regulatory sequence analysis in reported plant PHGPx genes, which will be a prerequisite to understand regulatory mechanism of PHGPx gene expression in plants.

Micropreparation of a Native PHGPx Protein From Radish Seedlings by Immunoaffinity Chromatography / 生物化学与生物物理进展

Radish phospholipid hydroperoxide glutathione peroxidase (RsPHGPx) was identified as a mitochondrion-targeting PHGPx in previous work. To determine its cleavage site of the targeting peptide, the immunoaffinity chromatography (IAC) purification approach was carried out to isolate the native RsPHGPx protein.Polyclonal antibodies directed against recombinant RsPHGPx were raised in rabbit. Monospecific anti-RsPHGPx antibodies were isolated by means of affinity chromatography using the recombinant RsPHGPx as affinity ligand, and employed in assembling an IAC column. A single-step, highly specific and easy-to-use protocol was developed for purification of the active RsPHGPx protein through the assembled IAC column. Using this approach, a specific protein of the expected molecular size was obtained from the mitochondrial fraction of radish seedlings. Western blot analysis showed that it could be specifically recognized by anti-RsPHGPx antibodies, and an enzyme activity assay indicated that it exhibited significant PHGPx activity, suggesting that the purified protein should be the desired native RsPHGPx. These results will lead to clarification of the targeting peptide and the active mature protein of RsPHGPx and will be helpful to further probe the intracellular localization mechanism and biological fun ction of this plant PHGPx.