Breast Neoplasms ; metabolism ; pathology ; Cell Polarity ; Epithelial Cells ; metabolism ; pathology ; Epithelial-Mesenchymal Transition ; Eye Proteins ; metabolism ; Female ; Humans ; Membrane Proteins ; metabolism ; Nerve Tissue Proteins ; metabolism ; Receptors, Proteinase-Activated ; metabolism ; Tumor Suppressor Proteins ; metabolism

To observe the clinical results of innercapsular phacoemulsification with primary intraocular lens ( lOL ) implantation in the treatment of lens dislocation.METHODS: A total of 23 cases ( 23 eyes ) of lens dislocation ( lla and llb ) were underwent innercapsular phacoemulsification combined with primary lOL implantation. lOL implantation were underwent during operation, the complications of intraoperative and postoperative, postoperative vision, intraocular pressure ( lOP ) , corneal endothelial cell, lOL location were analyzed.RESULTS: The operations were successfully completed for all patients in accordance with the pre - surgery program; lens nucleus or its fragments did not crash into the vitreous cavity; 20 cases of corneal edema and 17 cases of lOP presented at the first day after surgery, the deviation or displacement of lOL and serious complications such as retinal detachment were not appeared. At the first week postoperation, the average lOP was 15. 81 ± 2. 10mmHg, with statistically significant differences when compared with the preoperative ( P<0. 01 ) , the visual acuity in all eases increased, with statistically significant differences when compared with the preoperative ( P < 0. 01 ). Corneal endothelial cell density and percentage of hexagonal cells decreased, the variation coefficient increased in first week of postoperative, with no statistically significant differences when compared with the preoperative (P>0. 05) CONCLUSlON: lnnercapsular phacoemulsification combined with primary lOL implantation in the treatment of the whole lens dislocation (‖a and ‖b ) can restore function in patients with diplopia and control lOP effectively, reduce corneal endothelial cell damage, which is an effective method to treat the whole traumatic lens dislocation.

AIM: To observe the inhibition of c-Met inhibitor on proliferation of lens epithelial cells (LECs).METHODS: Human's LECs were cultured and hepatocyte growth factor (HGF) and K252a were added to second passage of cells supplied with Dulbecco's modified eagle's medium (DMEM). MTT assay was used to examine the proliferation of LECs, and Western-blot was used to detect the expression change of Bcl-2 and Caspase-3.RESULTS: The photodensity (A) of HGF (50nmol/L) + K252a (30nmol/L) was not significantly different from that of DMEM control (P＞0.05). The expression of Bcl-2 and Caspase-3 were not significantly different from that in the control group.CONCLUSION: K252a, the inhibitor of c-Met, can effectively inhibit the proliferation of LECs.

Background Studies demonstrated that human amniotic epithelial cells (AECs) have some characteristics of embryonic stem cells and they were used to re-establish the surface of eyes. Human AECs may serve as new seed cells in tissue engineering for corneal epithelium reconstitution in the future. Objective The present study was to investigate the application value of human amniotic epithelium cells transfected by lentiviral vectormediated enhanced green fluorescent protein (EGFP) gene as new seed cell source for engineering the corneal surfacelayer. Methods Lentiviral vector carrying the objective gene EGFP was transfected into human amniotic epithelial cells (pLenti6/V5-DEST),and the transient expression of the transgene in the human amniotic epithelial cells was observed under the fluorescence microscope. Flow cytometry was used to detect the positive expression rates of EGFP in transfected cells. The transfected human amniotic epithelial cells were seeded onto the fresh corneal stromal surface of New Zealand white rabbit and cultured in vitro. The stem cell deficiency ( SCD ) models were established by cutting off the limbus of cornea in 20 eyes of New Zealand white rabbits, and the model rabbits were then divided into 2 groups randomly. The transplanted grafts carrying the pLenti6/V5-DEST-EGFP gene-transferred human amniotic epithelium cells were regarded as the pLenti6/V5-DEST-EGFP group, and the corneal stroma graft without any epithelial cell served as the control group. The opacity of stroma and corneal conjunctivalization and vascularization were observed daily. The rabbits' eyes were extracted one month after operation. The expression of EGFP in the cornea was detected under the fluorescence microscope, and the expression of CK8, CK18 and CK12 in cornea was detected by immunohistochemical staining. Results The shape of the transferred human amniotic epithelial cells resembled normal human amniotic epithelial cells. 48 hours after the transient transfection of EGFP presented with the highest expression level throughout the observation duration, with a positive expression rate of EGFP of 61.5% ,showing significant differences in comparison with that of 12 ( 5.24% ) , 24 ( 38.27% ) or 96 ( 39. 10% ) hours ( P ＜0. 05) post-transfection; but no obvious difference was found in the positive rate of transiently transfected EGFP between 48 hours and 72 hours ( 58.36% ) ( P＞0. 05 ). Six cornea grafts were clear in 1 month and two corneas were rejected during the observation period in the pLenti6/V5-DEST-EGFP group. A few new blood vessels were seen around the graft. Ten corneas of the control group became opaque and cloudy with new blood vessels growth around the grafts. Imunohistochemistry revealed the positive expressions of CK8, CK1 8 and CK12 in the corneal epithelial layer in the pLenti6/V5-DEST-EGFP group. However,the expression of CK12 was absent in the control group. Conclusion Human amniotic epithelium cells transfected with the pLenti6/V5-DEST-EGFP gene is a new and ideal feed cell type to reconstruct the corneal surface layer. Lentivirus is a relatively safe gene transfection vector.

Objective To study the role of hematopoietic growth factor(HGF)of placenta chorionic villus in fetal hematopoiesis during embryo ontogeny by observation of the appearance time and the content changes with the fetal growth, which was compared with HGF in cord blood. Methods Thirty embryo villus (2 g each) and 30 cord blood (2 mL each) were collected separately from early pregnant stage(6- 8 weeks), middle pregnant stage(16-22 weeks)and late pregnant stage (37-42 weeks). The levels of HGF were detected by enzyme - linked immunosorbent assay. Results HGF were produced on the early pregnant stage and the content of FL-T3,IL-3 increased gradually.There were significantly differences at different stages(P

Traditional herbal medicines, Panax ginseng, Panax quinquefolium and Panax notoginseng, attract our attention for their extensive and powerful pharmacological activities. Ginsenosides are the main active constituents of these medicinal herbs. The related glycosyltransferases involved in ginsenoside biosynthesis are the key enzymes which catalyze the last important step. Modification of ginsenoside aglycones by glycosyltransferases produces the complexity and diversity of ginsenosides, which have more extensive pharmacological activity. At present, ginsenoside aglycones and compound K have been obtained by synthetic biology. As the last step of ginsenoside biosynthesis, glycosylation of ginsenoside aglycones has been studied intensively in recent years. This review summarizes the basic strategies and research advances in studies on glycosyltransferases involved in ginsenoside biosynthesis, which is expected to lay the theoretical foundation for the in-depth research of biosynthetic pathway of ginsenosides and their production by synthetic biology.
Biosynthetic Pathways ; Ginsenosides ; biosynthesis ; Glycosyltransferases ; metabolism ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Synthetic Biology

Pichia pastoris is one of the most important systems used in the field of molecular biology for the expression of recombinant proteins. The system has advantages of high expression, high stability, high secretion, easy high-density fermentation and low cost. Many factors affect the expression of recombinant protein, such as gene copy number, codon usage preference, type of promoter, molecular chaperones, glycosylation, signal peptide and fermentation process. In this review, research advances of the above aspects are summarized, which lay a foundation for improving the expression of recombinant proteins in P. pastoris.
Fermentation ; Gene Dosage ; Glycosylation ; Molecular Chaperones ; Pichia ; metabolism ; Promoter Regions, Genetic ; Protein Sorting Signals ; Recombinant Proteins ; biosynthesis

Lanosterol synthase is encoded by the erg7 gene and catalyzes the cyclization of 2, 3-oxidosqualene, which is a rate-limiting step of the inherent mevalonate (MVA)/ergosterol metabolic pathway in Saccharomyces cerevisiae. The intermediate 2, 3-oxidosqualene is also the precursor of triterpenoids. Therefore, the cyclization of 2, 3-oxidosqualene is the key branch point of ergosterol and triterpenoids biosynthesis. Down-regulation of 2, 3-oxidosqualene metabolic flux to ergosterol in S. cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway reconstructed by the synthetic biology approach. To construct erg7 knockout cassette harboring the loxP-Marker-loxP element, long primers were designed, which were homologous to the sequences of both erg7 ORF and plasmid pUG66. The cassette was transformed into diploid wild strain INVSc1 by LiAc/SS Carrier DNA/PEG method and then erg7 gene haploid deficient mutant was obtained by homologous recombination. The results of semiquantitative PCR and real-time quantitative PCR revealed that erg7 expression level in erg7 gene haploid deficient mutant is one time lower than that in wild strain. The results of TLC and HPLC showed that the ergosterol content in deficient mutant decreased to 42% of that in wild strain.
Chromatography, High Pressure Liquid ; DNA Primers ; Down-Regulation ; Ergosterol ; metabolism ; Haploidy ; Intramolecular Transferases ; genetics ; Polymerase Chain Reaction ; Saccharomyces cerevisiae ; genetics ; Squalene ; analogs & derivatives ; metabolism

OBJECTIVETo compare the difference in clinical efficacy on children attention deficit hyperactivity disorder (ADHD) between the combined therapy of scalp acupuncture and EGG biofeedback and the simple EEG biofeedback therapy so as to search the better therapeutic method for ADHD.

METHODSOne hundred patients were randomized into an observation group and a control group, 50 cases in each one. In the control group, the simple EEG biofeedback therapy was adopted. In the observation group, on the basis of biofeedback therapy, scalp acupuncture was added and applied to Dingzhongxian (MS 5), Dingpangyixian (MS 8), Baihui (GV 20), Sishencong (EX-HN 1), etc. The ten treatments made one session. After four sessions of treatment, FIQ value in Wechsler intelligence scale, CIH score in Conners children behavior questionnaire, the ratio of 0 wave and p wave in EEG, FRCQ and FAQ in the integrated visual and auditory continuous performance test (IVA-CPT) and clinical comprehensive efficacy were observed before and after treatment in the two groups separately.

RESULTSThree cases were dropped out in the observation group and 2 cases were out in the control group. In the two groups, FIQ, FRCQ and FAQ were all increased after treatment (P < 0.01, P < 0.05); the increases in the observation group were much more significant than those in the control group after treatment (all P < 0.05). In the two groups, CIH score and the ratio of 0 wave and p wave were all reduced after treatment (P < 0.01, P < 0.05); the reduction in the observation group were much more apparent as compared with those in the control group (both P< 0.05). The total effective rate was 91.5% (43/47) in the observation group and better than 83. 3% (40/48, P < 0.01) in the control group.

CONCLUSIONThe combined therapy of scalp acupuncture and EEG biofeedback achieves the superior efficacy on children ADHD as compared with the simple biofeedback therapy. This combined therapy rapidly relieves the essential symptoms of ADHD and improves EEG waveform in children patients. Importantly, this therapy obtains and consolidates the significant efficacy.

Acupuncture Therapy ; Adolescent ; Attention Deficit Disorder with Hyperactivity ; psychology ; therapy ; Biofeedback, Psychology ; Child ; Combined Modality Therapy ; Electroencephalography ; Female ; Humans ; Male ; Scalp

AIM:To investigate the feasibility of using human amniotic epithelial cells(HAECs) sheets,which are cultured by using a biodegradable fibrin sealant to re-establish corneal surface layer.METHODS:Human amniotic membrane(AM) harvested at the time of elective caesarean section was digested by collagenase and trypsin respectively to obtain HAECs.The HAECs were planted on Millicell culture dishes coated with biodegradable fibrin glue,and cultured by using the air-lifting cultivation.The HAECs sheets were investigated morphologically by Hematoxylin-Eosin staining,scanning electron microscopy(SEM) and were identified by cytokeratin immunohistochemistry.RESULTS:HAECs sheets were transparent and successfully cultured by using a biodegradable fibrin sealant.Most of the cultured cells were round or polygon and typical slabstone-like appearance.Many microvilli were observed on cell surfaces by SEM.Cytokeratin staining was positive.HAECs had stratificated growth tendency and became corneal-like stratificating epithelial cells.CONCLUSION:A commercially available fibrin sealant was an effective means of tissue engineering to create a carrier-free,transplantable HAECs sheets.The sheets were potential graft to re-establish corneal surface layer.