ABSTRACT
Objective To systematically analyze the relevant scientific literature on the effect of nasopharyngeal carcinoma radiotherapy on thy?roid function and to elucidate the current research status in this field. Methods We searched the relevant literature published over a recent de?cade using the PubMed database,and parameters such as the number of published articles and high?frequency keywords were counted. Cluster analysis of the high?frequency keywords was performed using SPSS software. Results We identified 2928 references and 11 high?frequency key?words. The clustering analysis results identified five main aspects. Conclusion In recently published articles on the effect of nasopharyngeal carci?noma radiotherapy on thyroid function,the high?frequency keywords were clustered mainly in the following five categories:nasopharyngeal carcino?ma radiotherapy,complications of nasopharyngeal neoplasms and etiology of hypothyroidism,radiation dose and side effects,effect of radiation ther?apy on thyroid functional,nasopharyngeal neoplasm pathology,and secondary carcinoma.
ABSTRACT
The flushing pump which is applied to clean operative wound has no temperature controlling function up to now, and doctors have to prepare the flushing fluid that has previously been warmed. The flushing pump system with medical constant temperature designed in our laboratory can absorb flushing fluid at the room temperature, and then eject flushing fluid with the temperature in accordance with the requirements of operations at a controlled constant flow rate. The system combines flow rate control with temperature control functions. The flushing pump system includes flushing part, temperature controlling part, key inputting part, liquid crystal displaying part and exceptional situation monitoring part. The present paper introduces the design method and principle of each part of the system at first, and then gives the debug method of all the system parameters. Finally the paper discusses the performance of the system according to the result of the experiment.
Subject(s)
Equipment Design , Equipment and Supplies , TemperatureABSTRACT
In surgery operations, wound should be cleaned with warm sterilized saline solution. In order to reach rapidly warming the washing solution from the room temperature during the surgery, we designed a thermostatic medical infusion pump. The present paper mainly presents researches on the two temperature control methods in the standby mode and in the flushing mode of the system. In the standby mode, the traditional proportional-integral-derivative (PID) control algorithm was adopted. In the flushing mode, dynamic characteristics of the system was changed in real time, which made the thermostatic control process more complex, and the fitted control function combined with the PID control algorithm was adopted in this mode. The temperature control parameters were adjusted in real time according to the initial temperature and the flow rate of the washing solution to obtain a constant temperature of the washing solution, no matter how the initial temperature and the flow rate are changed. The experiment results showed that this kind of control system performed well with a high accuracy.
Subject(s)
Algorithms , Equipment Design , Infusion Pumps , Sodium Chloride , Solutions , TemperatureABSTRACT
Aims: To give researches on gastric interdigestive pressure activity, including gastrointestinal (GI) physiological motility recording method, data processing and analysis method, as well as to give reasonable interpretation on how to generate such gastric pressure activity. Study design: Basic application study. Place and Duration of Study: School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology (USST), between June 2010 and October 2011. Methodology: We introduced a telemetric method to get the gastric physiological pressure activity inside the GI tract and the general process for processing such gastric Migrating Motor Complex (MMC) pressure activity including the process of abnormal value removing, medians of five-three-Hanning (53H) weighted average smoothing, and the fluctuation frequency estimation. Results: Using the process of abnormal value removing, medians of five-three-Hanning (53H) weighted average smoothing, and the fluctuation frequency estimation, we well obtained gastric interdigestive pressure activity (MMC). Conclusion: The methods introduced in the paper including abnormal value removing, the 53H weighted average smoothing, and the fluctuation frequency estimation were helpful for researches on gastric interdigestive pressure activity.
ABSTRACT
BACKGROUND:Currently,there is not a standard method for in vitro culture and large scale amplification of umbilical cord blood mesenchymal stem cells(UCB-MSCs).OBJECTIVE:To investigate the isolation,purification and culture of UCB-MSCs in vitro,and to detect its surface marker variation.METHODS:The monocytes were harvested from UCB using 1.077 g/cm3 lymphocytes separating solution and density gradient centrifugation,followed by incubation in an incubator containing 5%CO2 at 37℃.The cell morphological changes were observed at different time points and the expression of surface marker was detected using flow cytometry.RESULTS AND CONCLUSION:The monocytes isolated from the UCB grew initially into numerous hematopoietic cell clones,most of which were granulocyte/macrophage colony-forming units and burst forming unit-erithroid,increasing by(37.1±2.3)and (10.4±1.7),respectively.Switzerland staining showed most of them were granulocyte clones(80,1±85.2)%,next was erythroid clones(14.2±1.8)%.At 7 days after culture,some shuttle fibroblast-like cells and fiat osteogenic-like cell spread the whole plastic well.At 14 days after culture,flow cytometry showed CD38+ cells accounted for 1.64%,and CD34+/CD38+ cells accounted for 1,71%,and CD34+/CD38- were 0.55%.PI+ and Annexin-V+ cells accounted for 0.05% and 0.18% respectively.At 21 days after culture,CD38+,CD34+/CD38+ and CD34+/CD38- cells were 74.32%,1.61%,and 0.24%.The results reveled that UCB-MSCs can be isolated and cultured in vitro.
ABSTRACT
The experiment was performed at Basic Medical College and First Affiliated Hospital, Zhengzhou University from September 2004 to December 2005. Totally 60 Kunming mice were divided into 5 groups randomly: ①blank control group (n =15) and simple radiation group (n =15). The mice were given 0.2 mL sterile saline by intraperitoneal injection. ②antineoplastic polypeptide from Buthus Martensii Venom (APBMV) group (n =10) and APBMV plus radiation group (n =10) received 0.2 mL APBMV according to prepared concentration by intraperitoneal injection. ③Katsutoxin extract Ⅲ plus radiation group (n =10) received 0.2 mL katsutoxin extract Ⅲ by intraperitoneal injection every other 5.5 hours for 7 days. After 24 hours from the last injection, the mice were endured 60Co g ray radiation (80 cm, 7.5 Gy irradiation dose, 0.27 Gy/min dose rate). Then katsutoxin extract Ⅲ was given same as above for 7 days. Then bone marrow was extracted to be cultured to colony-forming unit-granulocyte and monocyte (CFU-GM). The findings showed that colony amount of APBMV plus radiation group and katsutoxin extract Ⅲ plus radiation group was obviously more than that of simple radiation group [(32?5),(27?3),(2?1)pieces/well,P
ABSTRACT
BACKGROUND:Human mesenchymal stem cells (hMSCs) become aging and even die after several passages. Some investigations have shown that telomere has a close correlation with life span of the cells. Whether the ectopic expression of human telomerase reverse transcriptase (hTERT) could induce the activity of the telomerase, maintain the length of telomere, and finally prolong the life cycle of MSCs without losing their multipotent differentiation capacity is still uncertain.OBJECTIVE: To observe the influence of the ectopic expression of hTERT on the telomerase activity and cell life cycles of hMSCs.DESIGN: Repetitive measurement trails.SETTING: Research Center of Stem Cell, Zhengzhou University Medical College.MATERIALS: The experiment was conducted in the Research Center of Stem Cell, Zhengzhou University Medical College from October 2003 to December 2005. hMSCs were obtained from 20 healthy donators from the Department of Pediatric Surgery and Outpatient, the Third and First Affiliated Hospitals of Zhengzhou University. Enhanced green fluorescent protein plasmid (pEGFP-C1) and pEGFP-hTERT were provided by Dr. Chantal Autexier of Canada. DH5α strain provided by Dr. Hou Wei-hong, the Key Molecular Medical Laboratory of Zhengzhou University Medical College.METHODS.: Under sterile condition, 2 mL bone marrow of sternum of healthy donors were harvested, and prepared after centrifugalization,dilution and passage.① Transfection of pEGFP-hTERT into hMSCs and the screening and amplification of resistance cloning:The 5th passage cells were seeded in a 24-well plate,and transfected by pEGFP-hTERT with lipofectamine method.The cells were divided into four groups including untransfected group,lipofectamine group,pEGFP-C1 group and pEGFP-hTERT group. Resistance cloning screen and amplification was performed by G418. ②hTERT mRNA expression and detection of telomerase activity:RT-PCR and PCR-ELISA were used to detect the hTERT mRNA expressions of the fifth passage hMSCs transfected with pEGFP-hTERT, and pEGFP-C1, the untransfected tenth passage hMSCs and K562 cells (positive control), and the telomerase activity of the fifth and thirtieth passage hMSCs transfected with pEGFP-hTERT,the fifth pEGFP-C1-transfected cells and the tenth passage untransfected cells. ③Karyotype analysis of hTERT-transfected MSCs: Chromosome analysis was performed by conventional Giemsa staining.④Inducement of differentiation from telomerase-positive MSCs into neuron-like cells and RE-PCR identification:The transfected MSCs were cultured in a medium containing epidermal growth factor and basic fibroblast growth factor, which could induce the cells differentiate into neuron-like cells. The culture solution was changed every 3 days, and the changes in cell growth condition and morphologic characteristics were observed under an inverted microscope. The microtubule associate protein (MAP2) and neurofliament subunit M (NF-M) were identified by RT-PCR.MAIN OUTCOME MEASURES:①hMSCs transfection with different kathion liposomes and the screening and amplification of resistance cloning; ②hTERT mRNA expressions of each group and detection of telomerase activity; ③Karyotype analysis of pEGFP-hTERT-transfected MSCs; ④ Induction of differentiation from telomerase-positive MSCs into neuron-like cells and RE-PCR identification.RESULTS: ①With the decrease of G418 concentration, the cells in the untransfected and lipofectamine groups died, and stably EGFP expressed MSCs were obtained; after G418 screening, there was a cell clone undergone 35 passages and continued to proliferate, whose appearance and growth characteristics were similar to the untransfected MSCs observed under inverted microscope. ②The fifth passage pEGFP-C1-transfected hMSCs and tenth passage untransfected hMSCs remained telomerase-negative, but the K562 and fifth passage hTERT-transfected cells showed positive telomerase activity. ③The telomerase activity of the fifth and thirtieth passage hTERT-transfected cells was positive. ④The hTERT-MSCs at passage 10, 20 and 30 had 23 pairs of chromosomes, and two X chromosomes. So they were still normal diploid with normal chromosome appearance and number. ⑤Many hTERT-transfected MSCs had the typical appearance of neuron-like cells. RT-PCR analysis showed that th expressions of MAP2 and NF-M were increased.CONCLUSION:Ectopic expression of the hTERT gene is found in hMSCs,and can induce the telomerase activity of hMSCs.The ectopic expression of the hTERT gene in hMSCs could extend the life spans of cells and maintain their multipotent differentiation capacity.
ABSTRACT
BACKGROUND: Stem cells are relatively primitive cells possessing the capabilities of self-renewal, high proliferation and multi-potential differentiation in vivo under certain conditions. Pancreatic stem cells and umbilical cord blood mesenchymal stem cells (MSCs) may serve therapeutic purpose clinically, but they are still difficult to culture in vitro at present.OBJECTIVE: To explore the method for isolation, purification and culture of pancreatic stem cells and umbilical cord blood MSCs in vitro and observe their morphological changes during culture in vitro.DESIGN: Completely randomized experiment with repeated measurement.SETTING: Stem Cell Research Center, Teaching and Research Division of Physiology, Medical School of Zhengzhou University.MATERIALS: This experiment was conducted in the Stem Cell Research Center, Teaching and Research Division of Physiology, Medical College of Zhengzhou University, between April 2004 and January 2005. Ten to fifteen newborn SD rats (1-3 days) were selected for culture in vitro of pancreatic stem cells, and fresh umbilical cord blood was collected from healthy woman (24-35 years old, with informed consent) at full-term delivery for culture in vitro of umbilical blood SMCs.METHODS: The abdomen of the newborn SD rat was opened under aseptic condition to obtain the pancreas, which was cut into small tissue blocks and digested with type-V collagenase for islet isolation. The isolated islets were purified in continuous roller-bottle culture. Umbilical cord blood was freshly collected for isolating the monocytes by means of density gradient centrifugation in lymphocyte separation medium (with density of 1.077 g/cm3). The islet cells and umbilical cord blood monocytes were cultured in the incubator at 37 ℃ with 5% CO2. The morphological changes of the cells were observed at designed time points and flow cytometry was used to determine the expression of cell surface molecules.MAIN OUTCOME MEASURES: The isolation and culture of pancreatic stem cells and umbilical cord blood MSCs, and their morphological changes during culture in vitro.RESULTS: During culture in vitro, the fusiform islet progenitor cells showed adherent polar growth and continuous proliferation, which covered the whole bottom of the flask after 12-14 days and could be subcultured for passages. However round cells appeared after removal of the growth factor and serum in the culture medium. The monocytes isolated from the umbilical cord blood grew initially into numerous hematopoietic cell clones, most of which proved to be granulocyte clones by Switzerland staining. Seven days later, flat flask wall-adhering epithelial cells and long fusiform fibroblasts were observed mixed with a number of osteoclasts. As the cell culture was prolonged, the cell number increased steadily.CONCLUSION: Pancreatic stem cells and umbilical cord blood SMCs can be cultured in vitro for further experiments.
ABSTRACT
AIM: To observe the inhibitory effects of polypeptide from Buthus martensii venom (PBMV) on human tumor cell lines and the influence of PBMV on cell cycle migration, expression of tumor-suppressor gene p53 and the membrane potential of CNE-2Z cells. METHODS: MTT colorimetric method, the colony formation and mitosis index, flow cytometry assay and intracellular recording with glass microelectrodes were used. RESULTS: PBMV had obvious cytotoxicity on several tumor cell lines. The cells grow was inhibited by PBMV, colony formation rate and mitotic index of tumor cells were reduced and the number of polykaryocyte was decreased. CNE-2Z cells in S phase were reduced evidently after they were treated with PBMV (P
ABSTRACT
AIM: To construct a recombinant retrovirus vector carrying hTERT for establishing UCBMSCs with hTERT(hTERT-MSCs) to overcome their limited life span and detecting whether telomerized UCBMSCs line maintained long-term self-renewal and differentiation capacity.METHODS: The whole cDNA was generated by PCR amplifications from the plasmid pEGFP-hTERT-C1.The hTERT segments were subcloned into pLNCX2.The target cells were infected with these retroviral particles.The stably transfected cells were selected by neomycin and expanded life span which were designated hTERT-MSCs was observed.The expression of hTERT in mRNA level was detected by RT-PCR and the telomerase activity was measured by TRAP(PCR)-ELISA assay.The hTERT-MSCs were induced with 5-azacytidine to cardiac muscle cells and the specific marker of myocardiocyte was detected.RESULTS: The constructed plasmids were digested with restriction endonucleases(BglⅡand NotⅠ).Two characteristic segments including 6.1 kb and 3.6 kb were obtained.The hTERT-MSCs expressed hTERT in mRNA level.The telomerase activity of hTERT-MSCs was positive.The growth kinetics of hTERT-MSCs was higher than those in UCBMSCs.The hTERT-MSCs were induced to myocardiocyte.CONCLUSION: The hTERT recombinant retrovirus vector has been successfully constructed.The hTERT gene activates the telomerase and prolongs the life-span of cells.No effect of hTERT gene on some type of differentiation potential of MSCs is present.
ABSTRACT
AIM: To observe the growth inhibiting effect of antineoplastic polypeptide from Buthus Martensii venom(APBMV)on liver neoplasm METHODS: MTT calorimetric method, trypin blue exclusion,colony formation and H 22 -bearing mouse model RESULTS: The ability to metabolize MTT of SMMC-7721 cells was lower than control distinctly after the cells were treated by APBMV The IC 50 of APBMV was 11 3 ?g/mL The growth of SMMC-7721 cells was inhibited obviously by APBMV and the dose-response relationship was clear, the IC 50 were 15 87 ?g/mL,13 05 ?g/mL and 8 70 ?g/mL respectively The colony formation rate of SMMC-7721 cells was also decreased evidently compared with control when treated concentrations of APBMV were higher than 8 ?g/mL Tumor growth of H 22 -bearing mice was inhibited by APBMV evidently, the growth inhibiting rate was reached 37 31%( P
ABSTRACT
AIM: To explore the regulatory effects of cytokines such as EGF, bFGF on expression of neural-specific molecules in mononuclear cells (MNCs) cultured in H-DMEM medium. METHODS: The umbilical cord blood samples were collected from health puerperal natural delivery. The mononuclear cells were isolated by centrifugation over Lymphoprep and planted in T-75 flasks containing H-DMEM medium with or without addition of EGF, bFGF or EGF plus bFGF at a final concentration of 20 ?g/L, respectively. Phenotypic changes were monitored by inverse phase-contrast microscopy. Tau and MAP2 mRNA were determined by reverse-transcript polymerase chain reaction (RT-PCR). Tau and MAP2-positive cell were determined by immunocytochemistry. RESULTS: The expression of tau protein mRNA was negative in uncultured cells, but MAP2 mRNA was positive. In cultured cells, tau protein mRNA expressed positively, MAP2 mRNA expression was upregulated by EGF+bFGF, EGF or bFGF compared with control group (no cytokines). The upregulatory capability of EGF+bFGF to MAP2 mRNA expression was stronger than that of EGF or bFGF alone. The same upregulatory tendency was noted in tau mRNA expression. In the group of control, bFGF, EGF, EGF+bFGF, the rate of MAP2-positive cells was 14.4%, 19.6%, 25.6%, 33.5%, respectively. Tau protein-positive cells were 13.5%, 15.3%, 21.4%, 29.8%, respectively. Under inverse microscopy, the freshly isolated MNCs were small and round, after culturing, the cells became larger with some big, long cytodenrites in the EGF+bFGF group, with 1 or 2 threadlike cytodenrites in the EGF group, or with some short multi-dendron like-astrocyte in the bFGF group and control group, but the number of astrocyte-like cells in the control group was less than that in bFGF group. CONCLUSION: MNCs derived from human umbilical cord blood cells express some neural specific molecules and are upregulated by cytokines, especially EGF and bFGF, which have the synergetic action. [
ABSTRACT
Objective To investigate the effect of excessive all-trans retinoic acid (atRA) on mouse embryonic palatal fusion and the mechanism. Method Palatal shelves from embryonic D 13 embryonic mice were cultured in BGJb medium and treated with vehicle control only or 5 ?mol/L atRA for 72 h. Palatal fusion was examined by hemagglutinin esterase. Apoptosis and laminin were detected by TUNEL and immunohistochemistry, respectively. The level of Smad2 phosphorylation (pSmad2) was analyzed by Western blot. Results atRA led to failure of palatal fusion and inhibited the migration and apoptosis of medial edge epithelial cells (MEE) and degradation of basal lamina within, compared with control palatal shelves in cultures. Additionally, apoptosis was detected in mesenchyme of atRA-treated palatal shelves. Further experiment revealed that pSmad2 was abrogated by atRA. Conclusion atRA induced failure of palatal fusion through inhibition of apoptosis of the MEE cell and degradation of basal lamina within medial edge epithelial seam. Inhibition of pSmad2 may account for the failure of palatal fusion by atRA.