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BACKGROUND:Studies have testified that nano-ultrasound contrast agents have a strong permeability, making it possible to image the targeted tissues outside blood vessels and overcome the limitation that micron contrast agents are only available for the blood pool imaging. OBJECTIVE:To construct the folate-modified nanoparticles targeting breast cancer as ultrasound contrast agents, as wel as to observe their ability to specifical y bind to cel s and imaging effect in vitro. METHODS:Both contrast agents, pegylated lactic acid-glycolic acid copolymer wrapping liquid fluorocarbon formed nanoparticles (mPP/PFOB) and folate modified pegylated lactic acid-glycolic acid wrapping liquid fluorocarbon formed nanoparticles (mPPF/PFOB), were constructed by phacoemulsification-evaporation method. (1)Biocompatibility detection:HFF-1 and MCF-7 cel s in the logarithmic phase were cultivated with various concentrations (0, 0.005, 0.01, 0.02, 0.05, 0.1, 0.2 and 1 g/L) of mPP/PFOB or mPPF/PFOB for 24 hours respectively, and then the cel viability was measured. (2)Targeting ability detection in vitro:HFF-1 and MCF-7 cel s in the logarithmic phase were divided into three groups. Cy5-labled mPP/PFOB and mPPF/PFOB were added into groups A and B, respectively;the cel s in group C were pretreated with folate for 2 hours, and sequential y Cy5-labled mPPF/PFOB was added into group C. Fluorescence intensity was detected by flow cytometry after 0.5 hours of culture. The distribution of contrast agents in cel s was observed using confocal microscopy after 20 minutes of culture. (3)Ultrasound imaging in vitro:there were three groups:saline was as group A;the suspension of saline and mPPF/PFOB nanoparticles was prepared as group B;MCF-7 cel s were resuspended with the mixture of saline and mPPF/PFOB nanoparticles to prepare the suspension of nanoparticles and cel s as group C. In each group, the suspension was added into latex gloves, that were then tightened and immersed in water. Final y, the ultrasound was use to detect the ultrasound imaging effect in vitro. RESULTS AND CONCLUSION:Neither nanoparticles were with significant cytotoxicity. The flow cytometry showed that the mean fluorescence intensity in MCF-7 cel s of group B was significantly higher than that of groups A and C. But there were no significant differences in the mean fluorescence intensity in HFF-1 cel s among the three groups. It was observed that mPPF/PFOB mainly gathered around the MCF-7 cel membrane, while mPP/PFOB randomly distributed in the cytoplasm. After mPPF/PFOB binding to MCF-7 cel s, they could enhance ultrasound echo in vitro. These findings indicate that the targeted nanoparticles mPPF/PFOB have good biocompatibility and can specifical y bind to breast cancer MCF-7 cel s in vitro and enhance the imaging capability.
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Sal-like 4 (SALL4) plays an important role in promoting the cellular proliferation and maintaining the pluripotency of embryonic stem cells and tumor cells.In fully differentiated cells,the expression of SALL4 is silenced or down-regulated.However,the expression of SALL4 is found to be restored or up-regulated in a variety of non-germ cell tumors.Besides,the expression of SALL4 is often associated with disease progression,treatment effect and prognosis.Therefore,examining the expression level of SALL4 will be of great importance in the diagnosis of disease and monitoring the disease progression for non-germ cell tumors.
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Pancreatic cancer is a highly malignant tumor. Animal models of pancreatic cancer included xenograft models, carcinogen induced models, genetically engineered models. Xenograft models are relatively reliable and feasible, but the growth pattern is different between serve immunodeficienct animals and human beings. Carcinogens induced models simulated the environmental factors to reconstruct the development of pancreatic cancer. But carcinogens could have other biological effects. Genetically engineered models could make the occurrence of pancreatic cancer at the molecular level. But it is difficult to control the transgenic product accurately. No model could meet all the needs of different experiments. It is important to choose a suitable animal model in different experiments.
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A microcapsule is a spherical, with a diameter that can be controlled in the range of 200 -1500 μm and biocompatible semipermeable membrane, which allows the bidirectional diffusion of nutrients,oxygen, secreted therapeutic product, and waste but prevents the penetration of high molecular weight substances from the microcapsule, such as antibodies and immunocytes. In comparison to monolayer culture and multicellular tumor spheroid model, orthotopic injection of microencapsulated tumor cells has uncomparable advantages in cell proliferation, mimicking the in vivo situation, making orthotopic tumor model and distant organ metastases model. Microencapsulated tumor assay has the potential of being widely used for in vitro anticancer drug screening and evaluation of the effects. This article mainly reviews the advantages of microencapsulated tumor assay and its application.
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With the development of cell separation technique, hepatocyte transplantation becomes a hot topic; however, the application is limited by donor deficiency and immunological rejection. Microencapsulated hepatocytes contribute to the promotion and application for liver cell transplantation, for which provide a large amount of high activity and good function of liver cells, in this paper, liver cell microencapsulation technology and its progress in applications were reviewed, providing prospective way for large-scale and high-active culture in vitro and long-term cryopreservation.
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BACKGROUND: Contacting with blood, most of polymer materials lead to different extents of blood coagulation, which limits their clinical application. Therefore, developing polymer materials with excellent anticoagulant property has become a key to clinical study of bioartificial liver materials.OBJECTIVE: To in vitro detect the blood dompatibility of polyacrylamide grafted polypropylene (PP) membrane (PP-g-AAm), a novel artificial liver reactor material.METHODS: Prior to and after modification, hemolytic test, prothrombin time and activated partial thromboplastin time tests of PP membrane were performed; blood platelet CD62P and CD63 expression rates were determined by flow cytometry, and platelet adhesion on PP and PP-g-AAm membranes by scanning electron microscopy.RESULTS AND CONCLUSION: The hemolysis ratio of PP and PP-g-AAm membranes was 1.32% and 1.46%, respectively.Compared with PP-g-AAm membrane, prothrombin time and activated partial thromboplastin time of PP membrane weremarkedly shorter (P < 0.05). CD62P and CD63 expression rates in the PP-g-AAm membrane were significantly lower than PP membrane (P < 0.05). Scanning electron microscopy results revealed that there were obvious changes of platelets adhering to these two membranes, but platelets adhering to PP-g-AAm membrane were fewer than PP membrane. These results indicate that PP-g-AAm membrane exhibits good blood compatibility.
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BACKGROUND: Microcarrier culture technology has become a new and large scale cell culture technology. It has been mainly used in the amplification research of tissue engineering seed cells. Microcarder possesses the advantage of larger surface area and plays an essential role in microcarrier culture technology.OBJECTIVE: To sum up the biomaterials and methods of microcarrier preparation, and provide theoretical foundation for the study of microcarrier culture technology and tissue engineering.METHODS: Articles were retrieved from PubMed, Wanfang, and VIP databases with the key words of "micrecarrier, biomaterials cell culture, tissue englneering" in both English and Chinese between 1967/2009 and 1990/2009, respectively. Inclusion criteria:study addressing microcarrier materials, preparation, and performance; study of microcerrier cell culture; animal experiments and clinical applications. A total of 34 articles were originally retrieved based on their titles and abstracts.RESULTS AND CONCLUSION: Although a lot of studies have reported research and preparation of microcarrier, clinical application remains still difficult. Recently, varying materials will be made into novel compound materials by new technology,which can adjust mechanics and biodegredation of microcarder via surface modification.
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Objective To evaluate reoperations for benign bile duct strictures after a prewousRoux-en-Y hepaticojejunostomy.Methods Clinical date of 28 patients with previous reconstruction of Roux-en-Y hepaticojejunostomy for benign bile duct strictures were retrospectively analyzed.For data staftstical analysis t-test and stepwise logistic regression analysis were used.Results Reoperative surgery was performed for residual biliary stones with bile duct stricture in 10 cases(35.7%),simple anastomotic stricture of hepaticojejunostomy in 11 cases(39.3%),remained biliary stricture after initial rear in 6 cases (21.4%).anastomotic leakage with duodenal leakage in one case(3.6%).Mode of reoperation:18 cases (64.3%)underwent hepatic lobectomy with Roux-en-Y hepaticojejunostomy,liver splitting approach to Roux-en-Y hepaticojejunostomy in 5 cases(17.9%),right hemihepatectomy in one case(3.6%),resection of anastomotic stenosis involved segment and Roux-en-Y hepaticojejunostomy in one case(3.6%),abdominal drainage and duodenum fistulization and jejunum ostomy in one case(3.6%),choledocholithotomy with T-tube drainage in 2 cases(7.1%);Thirteen patients(46.4%)developed postoperative complications.Conclusion Biliary tract stenosis remains the main cause for reoperation in patients undergoing a faeled reconstruction.Wide and patent biliary tract drainage and reconstruction somenmes necessitate a hepatic lobectomy.
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Objective To evaluate the quality of life and influencing factors in patients with breast cancer after breast-conserving surgery, so as to provide theoretical support for preoperative health education and postoperative rehabilitative intervention for these patients. Methods 120 breast conserving surgery (BSC) and 140 modified radical mastectomy (MRM) patients were assessed with Functional Assessment of Cancer Therapy-Breast (FACT-B V4.0) one week after operation. Life quality was evaluated between the two groups, and the influencing factors were also analyzed. Results Postoperative breast cancer patients after breast conserving surgery were with good breast complementary attention, lower emotion state. The other scores of quality of life had no statistical difference. Influencing factors on the quality of life of breast cancer patients were anxiety state, relationship of family members, position of surgery and character type. Conclusions It suggests that nursing specialists should make out corresponding, scientific and reasonable nursing intervention schemes based on the characteristics of different groups of people to ease psychological burden, and elevate the quality of life effectively for postoperative breast conserving surgery patients.
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OBJECTIVE:To create an in vitro harvesting method of culturing a large number of adult bone marrow MSCs(BMSCs) DESIGN,TIME AND SETTlNG:The randomized, controlled study was performed at the Shanghai Institute of Digestive Surgery (Key Laboratory of Education Committee of Shanghai City),as well as Department of General Surgery and Organ Transplantation Center,Ruijin Hospital,Medical College.Shanghai Jiao Tong University from September 2005 to April 2006.MATERIALS:Bone marrow samples were collected from normal persons.who did bone marrow examination at the Department of Hematology,Ruijin Hospital,Medical College.Shanghai Jiao Tong University.Donors were volunteers who signed the informed consent.METHODS:Human BMSCs were harvested using Pemoll gradient centrifugation and adherence method.and then incubated in microcarrier cytodex3.Common monolayer polystyrene was incubated as controls.Cell phenotype and proliferative activity were tested utilizing flow cytometry and MTT.MAIN OUTCOME MEASURES:Collection.incubation,morphology of human BMSCs.and prolireration and cell cycle of human BMSCs on the cytodex 3 were measured.RESULlTS:Flow cytometry detection showed that the surface marker of human BMSCs on the cytodex3 was ldentical to that on the common monolayer polystyrene;BMSCs were positive for CD29,CD44 and CD105.but negative for CD14,CD34,CD45,VLA-1 and HLA-DR.MTT detection demonstrated that human BMSCs were in the adaptive phase at days 1-3.and entered logarithmic phase frOm day 3.No significant difference was detected in human BMSCs on the monolayer polystyrene and cytodex3(P＞0.05).On the monolayer polystyrene,human BMSCs entered degenerating stage from day 6,whereas on the cytodex3,human BMSCs were still in the logarithmic growth phase at day 9(P＜0.05).Flow cytometry detection confirmed that the cell cycle of human BMSCs was the same both on the monolayer polystyrene and cytodex3 (P＞0.05). CONCLUSION:Using cytodex3 culture technique,a large amount of human BMSCs can be obtained,and the proliferative activity of these BMSCs is good.
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BACKGROUND: Presently, there is not an optimal cryopreservation protocol of the microcapsules, which has restrained the application of the microcapsules. OBJECTIVE: To investigate the characteristics of ice crystal and the morphology of alginate-chitosan-alginate (ACA) microcapsules cryopreserved at different solutions and different cooling rates, and to explore the optimal cryopreservationprotocol for ACA microcapsules. DESIGN, TIME AND SETTING: An observational study was performed at the Laboratory of Cryomicroecope in Shanghai University of Science and Technology (China) from February to April in 2008. MATERIALS: The high-voltage pulsing microcapsule shaping device was used to prepare ACA microcapsules.METHODS: The ACA microcapsules were preserved at different cooling rates (1 ℃/minute, 10 ℃/minute, 30 ℃/minute and 100 ℃/minute) by the cryomicroscopy system and then rawarmed at 50 ℃/minute. The protocols were repeated after the supplement of 10% dimethyl sulphoxide. MAIN OUTCOME MEASURES: The growth of ice crystals and the morphology of ACA microcapsules were checked at different cooling rates and in different solutions. The changes of forms and the rates of damage were checked after the microcapsules were rewarmed.RESULTS: The ice crystals grew into big crystals at the freezing process when the cooling rate was low than 10 ℃/minute and cryoprotector was not used. The growth of ice crystals would result in the distortion of microcapsules. It also could reduce the cryodamage of the microcapsules. The size of the ice crystals would grow down when raising the cooling rate and using thecryoprotector. The post-thaw ACA microcapsules were intact when dimethyl sulphoxide was used at a concentration of 10% and the cooling rate was higher than 30 ℃/minute (P ＜ 0.05). CONCLUSION: Mechanical damage occurs mainly during the growing of ice crystals at the time of microcapsules cryopreservation process. The growth of the ice could be restrained effectively by raising the cooling rate and using the cryoprotector.
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OBJECTIVE: To analyze the research literatures related to bioartificial liver, and to make a conclusion concerning the development of bio-artificial liver.DATA SOURCES: Using bioartificial liver, liver cell, hepatocyte culture and bioreactor as search terms, searching Ovid, Springer Link database, and China National Knowledge Infrastructure, Vip Information database and Wanfang Date (1990.09-2008.09). Literatures search was limited to English and Chinese languages.DATA SELECTION: Researches regarding liver cells of bioartificial liver, reactors and auxiliary equipment was included, and the studies about immune and animal infection studies of bioartificial liver were excluded.MAIN OUTCOME MEASURES: ①The source, quantity and culturing of bio-artificial liver hepatocytes. ②Bioreactor type, nature and type of films. ③Composition of oxygen and temperature control devices of bioartificial liver.RESULTS: Totally 3898 documents seized initially in the searching by computer, according to inclusion and exclusion criteria, 29 were analyzed. Bioartificial liver was a hybrid device in which can culture hepatocytes in vitro, when the patient's blood flows through the device, material exchange with the cultured hepatocytes through semi-permeable membrane or direct contacting can take place, which can perform the same roles of detoxification, synthesis, biological transformation and other functions as real liver cells, so as to achieve the purpose of support and treatment. Bioartificial liver can also be involved in metabolism of the three major nutritive substances, as well as secretion of hepatocyte growth promo ting substances. So it is an effective alternative to the real liver as the function of detoxification and synthesis, and can fills the essential gap between the transplantation and acute liver failure.CONCLUSION: Although the bioartificial liver research has made significant progress, it still faces the problems such as limited liver cells sources, long-term maintenance of liver cell activity and function, and further optimization of the reactor design.
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Hepatocyte transplantation may be a viable alternative treatment to liver transplantation for acute/chronic liver failure and metabolic liver disorders. Hepatocyte transplantation is an effective treatment to support liver function around liver transplantation due to its relatively easy manipulation and mild wound. In recent two decades, hepatocyte transplantation have been applied in clinical treatment and showed some effect in acute/chronic liver failure and metabolic liver disorders. Here, we sum up the status of clinical hepatocyte transplantation, discuss its value in clinical application and some challenges need to resolve.
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BACKGROUND: Hepatocyte/polymer interface with good biocompatibility is the key factor in bioreactor design andconstruction, however, bioreactor used in the clinical practice currently is not an ideal one.OBJECTIVE: To establish human hepatocyte compatible polypropylene interface and to lay a foundation for establishingbioartificial liver reactor with polypropylene hollow fiber.DESIGN, TIME AND SETTING: The comparative observation, cell compatibility experiment was performed betweenFebruary and October 2003 at Shanghai Jiao Tong University, Shanghai, China.MATERIALS: Polypropylene Photochemical graft polymerization modification technique was used to graft hydrophilicacrylamide groups on the surface of polypropylene membrane by chemical bonds to form modified polypropylenemembrane.METHODS: L02 human hepatoeytes were seeded on polypropylene membrane, modified polypropylene membrane andpolystyrene membrane, and polystyrene membrane was used as normal control.MAIN OUTCOME MEASURES: Static water contact angle of polypropylene membrane before and after graftmodification; morphology, adherent rate and proliferation activity of L02 human hepatocytes on different material surfaces.RESULTS: Static water contact angle after polypropylene membrane graft modification was smaller than that before graftmodification (P < 0.05). The adherent rate of L02 human hepatocytes on the surface of modified polypropylene membranewas 0, and the proliferation activity of them, which grew as spherical aggregates, was markedly higher than that of cells onpolystyrene membrane and polypropylene membrane without graft modification.CONCLUSION: Grafting polyacrylamide on the surface of polypropylene can establish good interface of L02 humanhepatocytes/polypropylene and form hepatocyte spherical aggregates through simple static culture.
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BACKGROUND: Membrane materials of bioreactor have exchange of substance and good physiochemical characteristics as well as good biocompatibility.OBJECTIVE: To evaluate the biocompatibility of interface of human hepatocyte/microporous polypropylene, i.e. graft modified microporous polypropylene semipermeable ultrafiltration membrane (MPP).DESIGN, TIME AND SETTING: Animal observation was performed at the Organ Transplantation Center, Ruijin Hospital of Shanghai Jiao Tong University Medical School and Polymers Institute of Zhejiang University between September 2005 and October 2007.MATERIALS: The microporous polypropylene ultrafiltration plane thin membranes, 0.2 μm diameter, M<,r> 50 000-100 000 molecular blockage, were used. Photochemical graft polymerization modification technique was adopted to graft hydrophilic acrylamide group through chemical bonds on MPP surface and succeeded in constructing an interface of human hepatocyte/microporous polypropylene, i.e. bioreactor membrane of bioartificial liver, graft modified MPP.METHODS: The biocompatibility of modified MPP was evaluated by hemolysis test, cytotoxicity test, acute systemic toxicity test, pyrogen test, skin sensitization and percutaneous stimulation test according to the requirements and biological evaluation criteria of medical device of ISO10993-1:1992.MAIN OUTCOME MEAURES: The experimental results of hemolysis, cytotoxicity, general acute toxicity, pyrogen, skin sensitization and percutaneous stimulation of modified MPP.RESULTS: The hemolytic rate of modified MPP was 1.90% (<5%), which showed that modified MPP did not lead to hemolysis. The extract solution of modified MPP exhibited no significant inhibition on the proliferative activity of L929 cells. At 24, 48 and 72 hours after MPP injection, no mice death, significant changes in body mass, or acute systemic toxicity were observed, such as ptosis, dyspnea, eyanosis, abdominal stimulation, diarrhea, decreased movement or tremor. In rabbit pyrogen test, the body temperature changed in a range from -0.2 to 0.4, which was consistent with the evaluation criteria of biomedical materials without pyrogen. Only one case was found with very slight erythema in skin sensitization test; its integral was 1 and primary stimulation index was 0.25 (<0.4), and the primary stimulation index of percutaneous stimulation test was 0.2; the average primary stimulation index was 0.068, indicating that modified MPP had no skin irritation.CONCLUSION: Modified MPP has no haemolytieus, cytotoxicity, pyrogenicity or skin sensitization, suggesting good biocompatibility by photochemical graft acrylamide on the surface of MPP.
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Objective To study the clinical characteristics and diagnosis of the solid-psendopapillary tumor of pancreas (SPT).Methods The clinical data of 40 SPT from January 1996 to January 2008 were retrospectively analyzed. The average age was (32.9 + 13.6 )years. The average clinical course was (8.6±0.1) months.Clinical symptoms usually included distensible pains and secret anguish in abdomen (60.0%).No jaundice appeared in any case.Results The surgical resection was favorable for the treatment of SPT,which had excellent prognosis.No tumor recurrence were found in those following-up patients. Grossly,the cut surface showed areas of solid and papillary tissue,cystic degeneration,hemorrhage,and necrosis.Pathological features included a combination of solid and cystic components with pseudopapillae formation and degenerative regions without glands.Conclusions SPT has its uniquely clinical and pathological characteristics.Its main diagnosed points are helpful for clinical doctors to make timely diagnosis and reduce the rate of misdiagnosis and mistreatment.
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AIM: To evaluate the blood compatibility of a new bioartificial reactor membranous material (propylene-acidamide grafted poly propylene membrane, PP-g-AAm) in vitro. METHODS: Contacted PP-g-AAm membrane and PP (polypropylene) memb rane with platelet-rich plasma in a swing bed, 37 ℃, to simulate the conditions in vivo, and another group of PRP without any membranes was set as control group. ELISA was used to study the expression of ?-thromboglobulin, and flow cy tometry was used to study CD62P and CD63 expressio n of the activated blood platelets after contacting the two kinds of membranes w ith PRP. Scanning electrical microscopy was used to study the configuration and numbers of platelet cells adhered on the membranes. RESULTS: After contacting with PRP 30 min, ?-TG expression show ed marked difference between the two kinds of material groups and the control gr oup (P