Objective To investigate the retinal pigment epithelium cells(RPE) and makeit the base of clinical application for transplantation.Methods The cultured RPE cells labeled by BrdU was injected into the host's subretinal space by external transcleral.The structure and ultrastructure of the transplanted RPE cells were observed by microscope and transmission electron microscope.Results The transplanted cells had the normal structure as the host's cells.They attached to Bruch's membrane with basal infoldings and forming microvill on the surface of the cells.Conclusion The external transcleral is a practical method for RPE transplatation.The transplanted cells can vitalize and form some normal ultrastrcture.

Objective To compare imaging quality and radiation dose of different anode filter combination for full-field digital mammography.MethodsThe image of FLUKE NA 18-220 phantom were taken at full-field digital mammography (FFDM),system with Mo/Mo,Mo/Rh,Rh/Rh anode/filter combination by automatic exposure control,record the exposure factors and doses.The images on monitor with the best window width and window level were read by 4 independent radiologists,the images of specks groups,nylon fibers and masses was assessed by the 4 experienced readers at the criterion of American College of Radiology 1999 mammography quality control manual.Statistical analysis was performed using analysis of variance.ResultsThe nylon fibers scores of Mo/Mo,Mo/Rh and Rh/Rh were 5.50 ± 2.12,4.50 ± 1.85 and 4.38 ± 1.38 ; the specks groups scores of Mo/Mo,Mo/Rh and Rh/Rh were 5.38 ± 1.98,4.25 ± 1.56 and 4.38 ± 1.38;the masses scores of Mo/Mo,Mo/Rh and Rh/Rh were 5.38 ± 1.98,4.38 ±1.68 and 4.25 ± 1.56,the detection of specks groups,nylon fibers was not statistically significant ( F =4.56 and 4.32,P ＞ 0.05 ),but the detection of masses was statistically significant ( F =36.65,P ＜ 0.05).The radiation doses were different,the entrance surface dose (ESD) and average glandular dose (AGD) of Rh/Rh were (5.11 ± 1.89) and ( 1.08 ± 0.13 ) mGy,the ESD and AGD of Mo/Mo were (6.66 ± 2.33 )and ( 1.29 ± 0.38 ) mGy,the ESD and AGD of Mo/Rh anode/filter combination were (5.67 ± 2.02) and ( 1.29 ± 0.38) mGy.ConclusionsThe radiation dose of Rh/Rh and Mo/Rh anode/filter combination of FFDM were lower,and the imaging was clear,so Rh/Rh and Mo/Rh anode/filter combination of FFDM prefer to mostly patients,especially when the thinkness is large,Rh/Rh anode/filter combination is preferred,because the kV values was higher,the penetration of X-ray was stronger.The Mo/Mo anode/filter combination was used when the needs of high-resolution,because the detection of masses of it was better.

Objective To compare imaging quality and radiation dose on full-field digital mammography (FFDM) system and dual-sided reading CR mammography (DSCRM) system.Methods The TRM of ALVIM statistics phantom was exposed by FFDM system and (DSCRM) with the same radiation dose.The exposure parameters and the entrance surface dose (ESD) were recorded.Then,the phantom was exposed by DSCRM system with different mAs and same kV,and the ESD was recorded.The images obtained above were read by three radiologists on the monitor of diagnosis work station with the same window width and window level.The evaluation of the images were given using a five-level confidence scale.ROC curves were drawn and probability Pdet were calculated.Results The ROC analysis of microcalcifications and masses showed A(z) values of 0.730-0.925 and 0.670-0.945 for FFDM,and 0.632-0.815 and 0.575-0.785 for DSCRM when the radiation dose is 1.36 mGy.The radiation dose with DR system could be reduced compared with dual-sided reading CR system at the same probabilities Pdet,the ESD reduced from 1.63 mGy to 1.36 mGy by 19.8％,the AGD reduced from 0.65 mGy to 0.56 mGy by 16.0％.Conclusions The observable details with FFDM system were proved to be superior to DSCRM system at the same radiation dose.With the similar image quality,the radiation dose of FFDM system was less than the DSCRM system.

Linalool is an important monoterpene, and widely used in food, pharmaceutical and cosmetic industry. The low concentration in plants and the difficulties in extraction restrict its large scale production. Saccharomyces cerevisiae can provide the monoterpene precursor, geranyl diphosphate (GPP) through its endogenous isoprenoid pathway. Therefore, it could be used as the host for monoterpene production. However, the weak metabolic flux through the isoprenoid pathway leads to the insufficient supply of GPP, and results in low monoterpene productivity. In order to increase the metabolic flux, we constructed the integrated expression plasmid pRS305-tHMG1 and free expression plasmid pYLIS-IDI1 to enhance the expression levels of isopentenyl diphosphate isomerase (IDI1) and a truncated 3-hydroxyl-3-methylglutaryl-CoA reductase gene (tHMG1). The two plasmids were separately transformed into S. cerevisiae CEN.PK2-1C, resulting in strains LS01 and LS02. The plasmid pYLIS-IDI1 was further transformed into strain LS01, resulting in strain LS03. GC-MS analysis showed that the linalool concentration was increased by 1.3 times and reached (127.71 +/- 7.68) microg/L. In conclusion, enhancement of the supply of GPP precursors through the regulation of isoprenoid pathway could increase the linalool production in S. cerevisiae.
Biosynthetic Pathways ; genetics ; Butadienes ; metabolism ; Hemiterpenes ; metabolism ; Monoterpenes ; metabolism ; Pentanes ; metabolism ; Saccharomyces cerevisiae ; genetics ; metabolism

Alpha-ketogluratate is one of the key intermediates in the TCA cycle, playing an important role in the connection of carbon and nitrogen metabolism. This article aims at stating recent research progress in the production of alpha-ketoglutarate by microbial fermentation. First, a large group of microbes have been screened to accumulate alpha-ketoglutarate including prokaryotes and eukaryotes. Second, physiological characterization of over-accumulation of alpha-ketoglutarate is caused by thiamine defect and nitrogen starvation. Third, the process of fermentation was controlled and optimized by the manipulation of pH, dissolved oxygen and cofactors. Fourth, many metabolic engineering strategies were also presented for alpha-ketoglutarate production focusing on regeneration of cofactor and manipulation of the pathway. Last, we discussed the limitation of current progress and proposed the future research needs for microbial production of alpha-ketoglutarate.
Bacteria ; growth & development ; metabolism ; Fermentation ; Fungi ; growth & development ; metabolism ; Industrial Microbiology ; methods ; Ketoglutaric Acids ; metabolism ; Metabolic Engineering ; Yarrowia ; growth & development ; metabolism

PURPOSE: Methylenetetrahydrofolate reductase (MTHFR) has been implicated in lung cancer risk and response to platinum-based chemotherapy in advanced non-small cell lung cancer (NSCLC). However, the results are controversial. We performed meta-analysis to investigate the effect of MTHFR C677T polymorphism on lung cancer risk and response to platinum-based chemotherapy in advanced NSCLC. MATERIALS AND METHODS: The databases of PubMed, Ovid, Wanfang and Chinese Biomedicine were searched for eligible studies. Nineteen studies on MTHFR C677T polymorphism and lung cancer risk and three articles on C677T polymorphism and response to platinum-based chemotherapy in advanced NSCLC, were identified. RESULTS: The results indicated that the allelic contrast, homozygous contrast and recessive model of the MTHFR C677T polymorphism were associated significantly with increased lung cancer risk. In the subgroup analysis, the C677T polymorphism was significantly correlated with an increased risk of NSCLC, with the exception of the recessive model. The dominant model and the variant T allele showed a significant association with lung cancer susceptibility of ever smokers. Male TT homozygote carriers had a higher susceptibility, but the allelic contrast and homozygote model had a protective effect in females. No relationship was observed for SCLC in any comparison model. In addition, MTHFR 677TT homozygote carriers had a better response to platinum-based chemotherapy in advanced NSCLC in the recessive model. CONCLUSION: The MTHFR C677T polymorphism might be a genetic marker for lung cancer risk or response to platinum-based chemotherapy in advanced NSCLC. However, our results require further verification.
Carcinoma, Non-Small-Cell Lung/*drug therapy/enzymology/genetics ; Genetic Predisposition to Disease ; Humans ; Lung Neoplasms/drug therapy/*enzymology/*genetics ; Methylenetetrahydrofolate Reductase (NADPH2)/*genetics ; Platinum/*therapeutic use ; Polymorphism, Genetic/genetics

BACKGROUND AND PURPOSE: The wnt/β-catenin signaling pathway plays a critical role in embryonic development and adult-tissue homeostasis. Recent investigations implicate the importance of wnt/β-catenin signaling in normal wound healing and its sustained activation being associated with fibrogenesis. We investigated the immunolocalization and activation of wnt/β-catenin in polymyositis (PM), dermatomyositis (DM), and Duchenne muscular dystrophy (DMD). METHODS: Immunofluorescence staining and Western blot analysis of β-catenin were performed in muscle specimens from 6 PM, 8 DM, and 6 DMD subjects. The β-catenin/Tcf4 DNA-binding activity in muscle was studied using an electrophoretic mobility shift assay (EMSA), and serum wnt/β-catenin/Tcf transcriptional activity was measured using a luciferase reporter gene assay. RESULTS: Immunoreactivity for β-catenin was found in the cytoplasm and nuclei of muscle fibers in PM, DM, and DMD. The protein level of β-catenin was elevated, and EMSA analysis confirmed the activation of wnt/β-catenin signaling. The transcriptional activities of β-catenin/Tcf in the circulation were increased in patients with PM, DM, and DMD, especially in those with interstitial lung disease, and these transcriptional activities decreased when PM or DM patients exhibited obvious clinical improvements. CONCLUSIONS: Our findings indicate that wnt/β-catenin signaling is activated in PM, DM, and DMD. Its activation in muscle tissue and the circulation may play a role in modulating muscle regeneration and be at least partly involved in the process of muscle and pulmonary fibrosis.
Blotting, Western ; Cytoplasm ; Dermatomyositis* ; Electrophoretic Mobility Shift Assay ; Embryonic Development ; Female ; Fluorescent Antibody Technique ; Genes, Reporter ; Homeostasis ; Humans ; Luciferases ; Lung Diseases, Interstitial ; Muscular Dystrophy, Duchenne* ; Polymyositis* ; Pregnancy ; Pulmonary Fibrosis ; Regeneration ; Wound Healing

This study aimed to further enhance 2-keto-L-gulonic acid (2-KLG) production efficiency. A strategy for enhancing Ketogulonigenium vulgare growth and 2-KLG production by improving B. megaterium growth with sucrose was developed based on the time course of osmolality during 2-KLG industrial scale fermentation and effects of osmolality on cells growth and 2-KLG production. Results showed that the accumulation of 2-KLG and the feeding of alkaline matter led to an osmolality rise of 832 mOsmol/kg in the culture broth. High osmotic stress (1 250 mOsmol/kg) made the growth ofB. megaterium and K. vulgare decreased 15.4% and 31.7%, respectively, and consequently the titer and productivity of 2-KLG reduced 67.5% and 69.3%, respectively. When supplement sucrose under high osmotic condition (1 250 mOsmol/kg), B. megaterium growth was significantly improved, with the result that 2-KLG production was increased 87%. Furthermore, by applying this sucrose addition strategy further to batch fermentation in 3 L fermentor, the productivity of 2-KLG increased 10.4%, and the duration of fermentation declined 10.8%. The results presented here provide a potential strategy for enhancing the target metabolites produced by mixed strains at environmental stress.
Bacillus megaterium ; genetics ; growth & development ; metabolism ; Fermentation ; Industrial Microbiology ; Osmosis ; Rhodobacteraceae ; genetics ; growth & development ; metabolism ; Stress, Physiological ; Sucrose ; pharmacology ; Sugar Acids ; metabolism

The aim of this study was to improve the 2-keto-L-gulonic acid (2-KLG) production efficiency by Ketogulonicigenium vulgare and Bacillus megaterium by using multi-stage pH control strategy. The effect of pH on the cell growths and 2-KLG production showed that the optimum pH for K. vulgare and B. megaterium cell growth were 6.0 and 8.0, respectively, while the optimum pH for 2-KLG production was 7.0. Based on the above results, we developed a three-stage pH control strategy: the pH was kept at 8.0 during the first 8 h, then decreased to 6.0 for the following 12 h, and maintained at 7.0 to the end of fermentation. With this strategy, the titer, productivity of 2-KLG and L-sorbose consumption rate were achieved at 77.3 g/L, 1.38 g/(L x h) and 1.42 g/(L x h), respectively, which were 9.7%, 33.2% and 25.7% higher than the corresponding values of the single pH (pH 7.0) control model.
Bacillus megaterium ; growth & development ; metabolism ; Culture Media ; chemistry ; Fermentation ; Hydrogen-Ion Concentration ; Rhodobacteraceae ; growth & development ; metabolism ; Sorbose ; metabolism ; Sugar Acids ; metabolism

Objective To evaluate the efficacy of biofeedback combined with swallowing function training at different times on post-stroke dysphagia. Methods 124 patients with post-stroke dysphagia were randomly divided into observation group and control group. The observation group was given biofeedback combined with swallowing function training for 2 weeks and 4 weeks of treatment. The control group was given routine swallowing function training for 2 weeks and 4 weeks of treatment. Kubota drinking water test was used to evaluate the swallowing function before treatment, 2 weeks and 4 weeks after treatment in both groups. Results There was no statistically significant difference in the scores of swallowing function between the two groups before treatment (P=0. 401). After 2 weeks of treatment, there was also no statistically significant difference in the evaluation of swallowing function between the observation group and the control group (P=0. 138). After 4 weeks of treatment, the evaluation of swallowing function in the observation group was significantly better than that in the control group(P=0. 003). Conclusion Biofeedback combined with swallowing function training is better than the routine swallowing function training. After 4 weeks of treatment, the efficacy of biofeedback combined with swallowing function training is better than the biofeedback combined with swallowing function training after 2 weeks, indicating that there is a correlation between the treatment effect and the length of treatment time.