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

AIM: To evaluate the effects of 23G vs 20G pars plana vitrectomy ( PPV ) combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation for macular epiretinal membrane with cataract. ·METHODS: Totally 45 eyes of 45 patients with macular epiretinal membrane and cataract were enrolled in this retrospective non-randomized controlled clinical study. All eyes were treated with PPV combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation. There were 20 eyes in 23G PPV group, and 25 eyes in 20G PPV group. The best corrected visual acuity ( BCVA ) , intraocular pressure (IOP), counting of corneal endothelial cells ( CEC) and central retinal thickness ( CRT ) were examined before surgery. BCVA results were converted to the logarithm of the minimum angle of resolution ( LogMAR ) visual acuity. All operations were performed by the same doctor. Operation time for vitrectomy and membrane peeling, average ultrasound energy ( AVE) and effective phacoemulsification time ( EPT ) were recorded. BCVA and CRT were observed postoperatively at 30d and 90d, counting of CEC was observed postoperatively at 90d. IOP was observed postoperatively at 1d and 7d. ·RESULTS:The mean operation time for vitrectomy were 12. 57± 1. 35min in 23G group and 17. 30 ± 1. 19min in 20G group. The difference was statistically significant ( t =-12. 488, P<0. 01). There were no statistical significances in operation time for membrane peeling, AVE and EPT between 23G and 20G groups ( t=-0. 68,-1. 186,-0. 737, P=0. 500, 0. 242,0. 465). On 1d after surgery, IOP in 23G group was lower than that in 20G group, the difference was statistically significant (t= -2. 345, P=0. 024). The BCVA and CRT of the two groups both improved after operations. There were no statistically significant differences between two groups in terms of IOP, BCVA, and CRT ( F = 0. 465, 1. 895, 0. 689; P = 0. 499, 0. 176, 0. 411). IOP, BCVA and CRT were significant statistical different in different time-point within each group ( F=291. 245, 103. 06, 665. 402, P<0. 01 ). Different surgical methods of 23G and 20G had interactive effects on IOP with different time points ( F = 13. 245, P<0. 01 ), but different surgeries had no interactive effects on BCVA and CRT with different time points (F=1. 212, 2. 293;P=0. 283, 0. 129). The counting CEC in 23G group was more than that in 20G group postoperatively at 90d, the difference was statistically significant (t=2. 049, P=0. 048). ·CONCLUSION: The 23G PPV combined with internal limiting membrane peeling, phacoemulsification, intraocular lens implantation for macular epiretinal membrane with cataract is effective. Compared with 20G PPV, 23G PPV has advantages in operation time for vitrectomy and counting CEC. But lower IOP is likely in 23G PPV on 1d after surgery

OBJECTIVETo evaluate the effectiveness of manual therapy and traction for lumbar disc herniation and analyze the current status of this kind of randomized clinical trial (RCT).

METHODSDatabase of CNKI, VIP, WANFANG, PubMed and OVID were searched. Some relevant journals were manually retrieved. A total of 2 874 literatures on manual therapy and traction for lumbar disc herniation were collected, of which 17 articles met the inclusion criteria. The Jadad score scale was used to evaluate the quality,and RevMan5.0 was used for meta-analysis of literatures.

RESULTSThe results of the meta-analysis of all trials involved were as followed:the combined effect of the effective rate was RR = 1.10, 95% CI [1.06, 1.14], the combined effect of the cure rate was RR = 1.36, 95% CI [1.21,1.52], the combined effect of the VAS was RR = 1.37, 95% CI [1.28, 1.45], the combined effect of the JOA was RR = 4.75, 95% CI [4.40, 5.09].

CONCLUSIONThe overall quality of the current RCT researches about manual therapy for lumbar disc herniation was lower,and did not support the conclusion that manual therapy was more effective than traction for lumbar disc herniation.

Humans ; Intervertebral Disc Displacement ; surgery ; therapy ; Lumbar Vertebrae ; surgery ; Musculoskeletal Manipulations ; methods ; Randomized Controlled Trials as Topic ; Traction ; methods

The uhrastructure and atrial natriuretic peptide(ANP)mRNA in myocardium of streptozotocin-induced diabetic rats were studied along with the effect of rosiglitazone.It was found that the myocardial ultrastructure was demaged in diabetic rats.Rosiglitazone may ameliorate the lesion in myocardium,and lower the ANP mRNA in diabetic rats(1.14?0.05 at 6 weeks and 1.12?0.09 at 10 weeks vs 0.97?0.14,both P

OBJECTIVETo investigate the allele frequency distribution, gene diversity and haplotype diversity of 12 Y-specific short tandem repeats (STR) in Guangzhou Han population and evaluate their forensic application.

METHODSTwelve Y-STR loci in 401 unrelated male Han individuals in Guangzhou were amplified with PowerPlex(R) Y System, and the PCR products were detected with 3100 Genetic Analyzer.

RESULTSThe data of allele distribution and gene diversity of the 12 Y-STR loci were obtained from these individuals. A total of 398 haplotypes were observed and the overall haplotype diversity for the 12 Y-STR loci was 0.99997. The DNA samples of 13 different species of animals were amplified and no specific products were observed. Different tissues from the same individual exhibited the same Y-STR haplotype.

CONCLUSIONThe 12 Y-STR loci exhibit high genetic polymorphism and are of important value in understanding of the human origin, individual identification for mixed male/female DNA samples and paternity identification.

China ; Chromosomes, Human, Y ; genetics ; Forensic Medicine ; methods ; Gene Frequency ; Genetic Variation ; Genotype ; Haplotypes ; Humans ; Male ; Microsatellite Repeats ; genetics ; Polymorphism, Genetic ; Population Groups

The cyclization of 2,3-oxidosqualene is the key branch point of ergosterol and triterpenoid biosynthesis. Downregulation of 2,3-oxidosqualene metabolic flux to ergosterol in Saccharomyces cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway. In our study, primers were designed according to erg7 gene sequence of S. cerevisiae. Three fragments including 5' long fragment, 5' short fragment and erg7 coding region fragment were amplified by PCR. 5' long fragment consists of the promoter and a part of erg7 coding region sequence. 5' short fragment consists of a part of promoter and a part of erg7 coding region sequence. These fragments were inserted reversely into pESC-URA to construct antisense expression plasmids. The recombinant plasmids were transformed into S. cerevisiae INVSc1 and recombinant strains were screened on the nutritional deficient medium SD-URA. The erg7 expression level of recombinant strains, which harbored antisense expression plasmid of erg7 coding region, was similar to that of INVScl by semi-quantitative PCR detection. But erg7 expression level of recombinant strains, which harbored 5' long antisense fragment and 5' short antisense fragment, was significantly lower than that of the control. The results of TLC and HPLC showed that the ergosterol content of recombinant strains, which harbored 5' long antisense fragment, decreased obviously. The ergosterol contents of the others were almost equal to that of INVSc1. Lanosterol synthase gene expression was downregulated by antisense RNA technology in S. cerevisiae, which lays a foundation for reconstructing triterpenoid metabolic pathway in S. cerevisiae by synthetic biology technology.
DNA Primers ; Down-Regulation ; Gene Expression ; Intramolecular Transferases ; genetics ; metabolism ; Plasmids ; Polymerase Chain Reaction ; RNA, Antisense ; Saccharomyces cerevisiae ; enzymology ; genetics ; Squalene ; analogs & derivatives ; metabolism ; Transformation, Genetic

OBJECTIVETo investigate the therapeutic effect and the possible mechanism of Bushen Huayu Qutan Recipe (BHQR, a Chinese recipe formulated according to the therapeutic principle of nourishing Shen, resolving stasis and dispelling phlegm) on polycystic ovarian syndrome (PCOS).

METHODSBHQR was used, both internally and externally, to treat 46 patients with PCOS, and combined with diformin if necessary. Six successive months treatment was regarded as one course. The clinical symptoms and signs, including obesity, polytrichia, acne and acanthosis, were observed and scored before and after treatment; and serum levels of hormones and insulin were measured as well.

RESULTSAfter treatment, the scores of symptoms and signs of patients were significantly lowered (P<0.05 for obesity and P <0.01 for others), significant lowering was also found in serum level of testosterone (P<0.05), fasting serum insulin (P<0.01) and the values of insulin and insulin area under curve (P<0.05) 30 min and 60 min after glucose load.

CONCLUSIONBHQR could not only significantly relieve the symptoms and signs of patients with PCOS, but also regulate well the ovarian function.

Adolescent ; Adult ; Capsules ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Insulin ; blood ; Metformin ; therapeutic use ; Phytotherapy ; Polycystic Ovary Syndrome ; drug therapy ; pathology ; Testosterone ; blood ; Treatment Outcome ; Young Adult

Objective: To investigate the chemical constituents from the herbs of Corydalis edulis. Methods: The compounds were isolated and purified by various column chromatographies, and their structures were identified by physiochemical properties and spectroscopic data. Results: Fifteen compounds were isolated from the n-BuOH layer of EtOH extract of the herbs of C. edulis. and identified as 6, 7-methylenedioxydihydroisocoumarin (1), oxohydrastinine (2), 6, 7-dimethoyisoquinolin-1 (2H)-one (3), 6, 7-methylenedioxy-1 (2H)-isoquinolinone (4), (+)-syringaresinol (5), (+)-medioresinol (6), salidroside (7), isosalicin (8), icariside D2 (9), salicin (10), icariside F2 (11), icariside B2 (12), nicotinamide (13), pyrocatechol (14), and 5-hydroxyl-2-hydroxymethylpyridine (15), respectively. Conclusion: Compound 1 is a new natural product. Compounds 3, 5-12, 14, and 15 were isolated from Corydalis species for the first time. Compounds 2, 4, and 13 were firstly obtained from C. edulis.