Objective To investigate the expression of microRNA -216a(miR -216a)in patients suf-fering with breast cancer ,and identify the function and mechanism of miR -216 a on autophagy .Methods The expression of miR-216 a in 30 tumor tissues and paired normal tissue of breast cancer were detected by qRT -PCR.Inhibiton of miR-216a in MCF-7 cell lines was done by transfection of miR -216a inhibitor ( AMO-216a).Cell viability was detected by MTT assay .The level of Beclin 1 was detected by western blot .Results The level of miR-216 a was significant elevated in tumor tissue .Cell viability was markedly decreased owing to inhibition of miR-216a in MCF-7 cells.The level of Beclin 1 was significantly increased by transfection of miR-216a inhibitor in MCF-7cells(P<0.05).Conclusion The level of miR-216a is increased in breast canc-er tissue, and it might at least in part promote breast cancer via downregulating Beclin 1 and affecting autophagy .

Head and Neck Neoplasms ; genetics ; MicroRNAs

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.

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

OBJECTIVEThe study aimed to test if Paridis Rhizoma total saponins (PRTS) could induce apoptosis of human gastric cancer cell MKN-45.

METHODBased on the previous researches, PRTS was set by different concentrations to treat human gastric cancer cell for 12 h (5, 10, 20 mg x L(-1)). Fluorescent staining methods were adopted to observe apoptotic morphological changes of MKN-45. The apoptosis rates were analyzed by flow cytometry with Annexin V-FITC/PI staining. The enzymatic activities of caspase-3 and caspase-8 were measured by ELISA. The protein levels of Fas and FasL were detected by Western blotting.

RESULTUnder a fluorescence microscope, MKN-45 treated by PRTS was seen typical apoptotic morphological features. PRTS significantly increased the rate of apoptosis. Compared with the control group, there exsited significant differences in apoptosis rate of PRTS concentration of 20 mg x L(-1) (P < 0.01); besides, the enzymatic activities of caspase-3 and caspase-8 were promoted obviously after the effect of PRTS on MKN-45 cells for 12 h (P < 0.01). The protein levels of Fas and FasL in the MKN-45 were upgraded significantly.

CONCLUSIONPRTS can induce apoptosis of human gastric cancer cell MKN-45 , which is concerned with caspase-3 and caspase-8 and upgraded Fas and FasL.

Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Caspase 8 ; genetics ; metabolism ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Fas Ligand Protein ; metabolism ; Humans ; Magnoliopsida ; chemistry ; Rhizome ; chemistry ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Stomach Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; fas Receptor ; metabolism

Aged ; Female ; Fibroma ; pathology ; Fibroma, Ossifying ; pathology ; Gingival Neoplasms ; pathology ; Humans

AIM: To investigate the effect of Qingrehuayuziyin Fang on lymphocyte apoptosis and expression of Fas and FasL in peripheral blood in the patients with systemic lupus erythematosus(SLE).METHODS: Eighty patients with SLE were selected and divided randomly into two groups,40 in the therapy group and 40 in control group respectively.Patients were get the treatment of Qingrehuayuziyin Fang combined hormone(prednisone) as therapy group and get hormone treating only as control group.After treating three months,lymphocyte were collected to be detected the rate of apoptosis by flow cytometer.Meanwhile,the expression of Fas and FasL were analyzed by RT-PCR.RESULTS: After three months treating,the rate of lymphocyte apoptosis was decreased significantly in therapy group compared with that in control group(P

5 days.Blood gas analysis and blood pressure were determined at admitted day.Meanwhile,peripheral white blood cells at d1,3,5,and blood glucose were measured every day,respectively.GCS at d1,3,5 and hyperglycemia scorce(HS) were evaluated.Results Of the 82 studied patients,36 cases died.Univariate analysis showed that hypotension,lower GCS,higher peripheral white blood cells and HS were the independent death risk factors(Pa0.05).In multivariate logistic regression,the factors significantly associated with an increase in mortality were hypotension,lower GCS and higher HS.Conclusion Lower GCS,higher HS and hypotension are associated with poor outcome of children with severe trauma brain injury.