ABSTRACT
Objective To build the lentiviral vectors of pigment epithelial derived factor (PEDF) gene,and investigate their expression in human umbilical cord mesenchymal stem cells (hUCMSCs).Methods The PEDF lentiviral vectors (LV-PEDF) were built by DNA recombination and confirmed by DNA sequencing.hUCMSCs were transfected by LV-PEDF with MOI 10,30,50,respectively.The transfection efficiency was observed under fluorescence microscope.Cell immunofluorescence,immunocytochemistry and real-time PCR methods were used for detecting the expression of PEDF and VEGF.Results The PEDF cDNA was sub-cloned into pCDH-CMV-MCS-EF 1-copGFP vector successfully.DNA sequencing analysis confirmed that PEDF gene sequence was exactly the same with that reported in GenBank.pCDH-PEDF infected cells could show green fluorescence under fluorescence microscope.The transfection efficiency was 72.1％ in PEDF-MSCs.Immunofluorescence and immunochemical staining confirmed that PEDF protein was overexpressed in hUCMSCs.The relative expression of PEDF mRNA in experimental group and control group was (0.170±0.028) and (0.015 ± 0.007) respectively by RT-PCR,the difference was statistically significant (P＜0.00 1).The relative expression levels of VEGF mRNA in the two groups were (0.265 ± 0.022) and (0.285 ± 0.049),respectively,with no significant difference (P＞0.05).Conclusions We successfully built a lentivius vector carrying PEDF gene and obtained hUCMSCs with overexpressed PEDF.
ABSTRACT
Objective To evalute the indication and clinical application value of microwave abla-tion in spleen-preserving surgery in treament of patients with traumatic rupture.Methods The clinical data of 45 patients with traumatic splenic rupture were retrospectively analyzed.Spleen preserving surgery was feasible by preoperative CT evaluation and intraoperative observation.36 patients received simply mi-crowave ablation,and 9 cases received microwave ablation assisted partial splenectomy.Postoperative com-plications were observed,and the changes of platelet in peripheral blood were detected before and after op-eration in the 3 th,7th,14th day to evaluate the function of spleen.Results 15 patient was complicated with left pleural effusion,1 patient was complicated with splenic fossa fluid associated with infection.Dur-ing the follow-up of at least 2 months,No patients had postoperative bleeding,hemoglobinuria,gastrointes-tinal leak,deep vein thrombosis and pancreatic leak complications,no deaths.Seven days after microwave ablation spleen-preserving surgery,blood platelet count were significantly higher than the levels before the operation (P 0.05).The CT scan and the ultrasonic examina-tion that reviewed after operation in the 30-60th day showed good in spleen imaging in spleen-preserving patients,No patients had portal vein thrombosis.Conclusion Microwave ablation spleen-preserving sur-gery in the treatment of partial traumatic splenic rupture will be a safe and effective operation which could expand the spleen-preserving indication and be used in clinical practice.
ABSTRACT
Objective To Discuss the effect of surgical treatment from enteral and parenteral nutrition that in perioperative period of hepatolithiasis patients who taking hepatectom.Methods Retrospective analysis 55 cases who taking hepatectomy treatment in Fuling Central Hospital of Chongqing City from March 2011 to March 2015,all elective operations.According to the different nutrition support methods in perioperative period,randomly divided into enteral and parenteral group (n =25) and parenteral nutrition group (n =30).Results The postoperative complication rate and recovery time of bowel function in patients with enteral and parenteral nutrition,it was better than parenteral nutrition group (28 ％ vs 40％),[(4.50 ± 0.50) d vs (5.50 ± 1.00) d],the difference had statistical significance (P ＜ 0.05).Before and after operation in two groups of body mass index [(22.10 ± 1.80) vs (22.30 ± 1.70)],prealbumin impovement [(130.00 ± 45.00) mg/L vs (124.00 ± 55.00) mg/L],albumin impovement [(35.50 ± 2.72) g/L vs (36.50 ± 2.70) g/L],had no statistical significance (P ＞ 0.05).Conclusion Enteral and parenteral nutrition in perioperative can significantly improve the nutritional status of patients and the effect of surgical treatment.
ABSTRACT
Confronted with the gradual exhaustion of the earth's fossil energy resources and the grimmer environmental deterioration, the bio-based process to produce high-value added platform chemicals from renewable biomass is attracting growing interest. Escherichia coli has been chosen as a workhouse for the production of many valuable chemicals due to various advantages, such as clear genetic background, convenient to be genetically modified and good growth properties with low nutrient requirements. Rational strain development of E. coli achieved by metabolic engineering strategies has provided new processes for efficiently biotechnological production of various high-value chemical building blocks. This review focuses on recent progresses in metabolic engineering of E. coli that lead to efficient recombinant biocatalysts for production of high-value organic acids such as succinic acid, 3-hydroxypropanoic acid and glucaric acid as well as alcohols like glycerol and xylitol. Besides, this review also discusses several other platform chemicals, including 2,5-furan dicarboxylic acid, aspartic acid, glutamic acid, itaconic acid, levulinic acid, 3-hydroxy-gamma-butyrolactone and sorbitol, which have not been produced by E. coli until now.
Subject(s)
Acids , Metabolism , Alcohols , Metabolism , Biocatalysis , Escherichia coli , Genetics , Metabolism , Industrial Microbiology , Methods , Metabolic Engineering , Methods , Metabolic Networks and Pathways , Genetics , Organic Chemicals , MetabolismABSTRACT
BACKGROUND: To synthetically analyze the correlation between cytokeratin 19 and pancreas stem cells.DATA SOURCES:References about the correlation between cytokeratin 19 and pancreas stem cells were retrieved in Medline and Ovid database with the key words of "cytokeratin, pancreas, stem cells, marker, differentiation" in English from January 2000 to May 2006.STUDY SELECTION:Literatures about the correlation between cytokeratin 19 and pancreas stem cells were retrieved and those characterized by strong pertinency, published in the near future and selected from authoritative journals were selected in this paper.DATA EXTRACTION: Among 86 articles, 35 of them met the inclusion criteria and 51 were excluded.DATA SYNTHESIS:Pancreas stem cells are characterized by multi-directional differentiation.Cytokeratin 19 has a positive expression in pancreas stem cells and a molecular sign of pancreas stem cells. This provides a new way to further understand bionomics of pancreas stem cells,study separation,purification,transconformation and direction differentiation to islet cells, and treat diabetes mellitus with transplantation of pancreas stem cells.CONCLUSION: Expression of cytokeratin 19 is observed during the differentiation of pancreas stem cells and cytokeratin19 is a sign of pancreas stem cells.
ABSTRACT
BACKGROUND: Pancreatic stem cells can be differentiated into endocrine functioning cells under a suitable cultivation condition, suggesting that pancreatic stem cells can be used as a new source of islet in treating diabetes mellitus.OBJECTIVE: To discuss the expression of stem cell in different purities of pancreatic islets.DESING: A controlled observation based on cells.SETTING: Third People's Hospital of Mianyang City.MATERIALS: The experiment was conducted in the Center Laboratory (Key Laboratory of Chongqing City), the First Affiliated Hospital of Chongqing Medical University from March 2005 to March 2006. Thirty healthy male SD rats, of clean grade, aged 50 days were used in this study. During the experiment, the disposal of animals corresponded to Animal Ethical Standard. Cytokeratin (CK)-19 and β-actin were designed by Shanghai Invitrogen Biological Co., Ltd.Brdu reagent, mouse anti-rat Brdu monoclonal antibody, rabbit anti-mouse (RAM) CK-19 polyclonal antibody and Brdu immunohistochemical kit were from Boster Co., Ltd. (Wuhan); HistostainTM -DS double immunohistochemical staining kit was purchased from Beijing Zhongshan Company.METHODS: Rats were intraperitoneally injected with Brdu for labeling, and subjected to perfusion with Ⅴ-type collagenase via pancreatic ducts, with pancreas being excised, ripped, beaten up, digested and centrifuged to obtain pancreatic islet sediments. After being precipitated, the sediments were divided into 3 groups. Group A: The isolated sediments of islets were not purified; Group B: The sediments were added slowly with 25% Ficoll-400 and Hanks solution for purification; Group C: The sediments were slowly added with 25% Ficoll-400, 11% Ficoll-400 for purification in order.MAIN OUTCOME MEASURES: Expressions of Brdu and CK-19-positive cells in the islet samples were detected by immunohistochemical method, and mRNA expression of CK-19 in the different purities of islets was detected by reverse transcription-polymerase chain reaction.RESULTS: ① The purity of islets in the group A was the lowest. The purity of islets obtained by different purification techniques showed significances among the three groups (F =89.42, P ＜ 0.05). ②Positive expressions of Brdu and CK-19 in the group A were significantly higher than those in the group B and group C (F =18.64, 22.12, 38.61. P ＜0.01). ③Expression of CK-19 mRNA in the group A was significantly higher than that in other groups, with group C showing the lowest.CONCLUSION: Expressions of Brdu and CK-19-positive cells exist in different purities of islets, suggesting the existence of more stem cells in the low purify of islets.
ABSTRACT
Objective:To investigate the expression of ?-caten in and focal adhesion kinase(FAK)in colon carcinoma and to reveal their correlation with tumor clinicopathological features. Methods:The expression of ?-catenin and FAK in tumor tissue and normal tissue of 34 colon carcinoma patients were detected by immunohistochemistry and Western blot.Results:The expression of ?-catenin and FAK were detected both in tumor tissue and normal tissue,mostly in plasmalemma and endochylema.The expression of ?-catenin was significantly lower in tumor tissue than that in normal tissue(t=10.693,P
ABSTRACT
Background and purpose:The theory of cancer stem cell offers us a new thought about tumors, more and more kinds of cancer stem cell were isolated and indentif ied from corresponding cancer tissue. Our aim was to investigate the content of side population cells in SW480 human colorectal cancer ce11 line and to enrich cancer stem- like cells in SW480 through serum-free medium (SFM) culture. Methods:The percentage of side population cells in human colorectal cancer cell line SW480 was detected with ? ow cytometry. SW480 cell line was cultivated in serum- free medium(SFM) supplemented with growth factors and the cancer stem-like cells reforming into ? oating spheres were isolated. The isolated cancer stem-like cells were identifi ed by limited-dilution assay, differentiation assay, self- renewal assay, and alternative cultivation assay. Results:The percentage of SP cells was 1.2% in SW480,In the absence of serum, a minority (0.54%-0.62%) of cancer stem-like cells in SW480 cells survived, proliferated and formed into the suspended tumor cell spheres. SW480 cancer stem-like cells possessed proliferative, self-renewal and differentiation potential, which were responsible for the ? oating tumor clone. Serum addition into SFM resulted in the proliferation of cancer stem-like cells; after several generations and alternated cultivation in SSM and SFM, cancer stem-like cells maintained their characteristics. Conclusions:SW480 cell line contains a tiny minority of SP cells with stem cell properties.The cancer stem-like cells in SW480 line can be maintained in SFM using a floating culture method.
ABSTRACT
Objective To investigate the changes of immunity in patients with hepatocellular carcinoma (HCC) treated by high intensity focused ultrasound (HIFU). Methods HIFU system was used to treat 21 patients with mid-advanced HCC. The blood tests of cellular and humoral immunity related indexes were made before and on the 3rd, 7th, 14th, 21st day following the sonication. The histological changes of the target lesions were observed by light and electron microscope for the cases undergoing second stage operation. Results Light and electron microscope all revealed that HIFU could lead to irreversible changes in the tumor cells of the target lesions. There were not significant differences between the values of CD 3, CD 4, CD 8, CD 16, IgG, IgA, IgM,etc. before and after the ablation. Conclusions HIFU could destroy the tissues of HCC effectively. HIFU could not change the immunity of the patients with mid-advanced HCC remarkably. It may be ideal to unite immunotherapy and the other methods to get a sound clinical prognosis for HCC patients.
ABSTRACT
0.05).Conclusions RFA treatment of PHC patients does not lead to hematogenous tumor dissemination.
ABSTRACT
4.0 cm,the extension living time and elevation survival rate were better in palliative surgery operation than RFA treatment.However,RFA treatment has lower complications rate and higher postoperation quality of living.