Raspberry ketones have important therapeutic properties such as anti-influenza and prevention of diabetes. In order to obtain raspberry ketone from Chlamydomonas reinhardtii, two enzymes catalyzing the last two steps of raspberry ketone synthesis, i.e. 4-coumaryl-CoA ligase (4CL) and polyketide synthase (PKS1), were fused using a glycine-serine-glycine (GSG) tripeptide linker to construct an expression vector pChla-4CL-PKS1. The fusion gene 4CL-PKS1 driven by a PSAD promoter was transformed into a wild-type (CC125) and a cell wall-deficient C. reinhardtii (CC425) by electroporation. The results showed the recombinant C. reinhardtii strain CC125 and CC425 with 4CL-PKS1 produced raspberry ketone at a level of 6.7 μg/g (fresh weight) and 5.9 μg/g (fresh weight), respectively, both were higher than that of the native raspberry ketone producing plants (2-4 μg/g).
Acyl Coenzyme A ; Butanones ; Chlamydomonas reinhardtii/genetics* ; Ligases ; Polyketide Synthases

Objective:To estimate the diagnostic value of anti-cyclic citrullinated peptide (CCP) antibody for juvenile idiopathic arthritis (JIA) by Meta-analysis.Methods:The databases, including Wanfang Database, VIP citation databases, China National Knowledge Infrastructure (CNKI), China Biology Medicine disc, Excerpta Medica Database (EMbase), PubMed, Cochrane Library, were searched for the studies related to the diagnostic value of anti-CCP antibody for JIA. After extracting literature data and assessing the articles by Quality Assessment of Diagnostic Accuracy Studies tool. The sources of heterogeneity, sensitivity (SEN), specificity (SPE), area under the curve (AUC) were calculated by the summary receiver operator characteristic (SROC) curve by Stata 12.0 software.Results:A total of 30 literature reports were included in this Meta-analysis. The heterogeneity and the threshold effect of the included articles were tested, a mixed effects model was selected to calculate the pooled weighted SEN [0.16, 95% CI(0.11, 0.22)], SPE [0.99, 95% CI(0.98, 0.99)] and AUC [0.86, 95% CI(0.83, 0.89)]. The sensitivity estimates were highly heterogeneous, which was partially explained by the higher sensitivity in the rheumatoid factor-positive polyarthritis (RF+PA) subtypes [0.65, 95% CI (0.51, 0.76)] than in the other subtypes. The ability of diagnostic differentiation between of JIA and healthy children was better than the diagnostic differentiation between JIA and other patients ( Z=7.9, P<0.01). Conclusion:Although anti-CCP antibody cannot be used as an early diagnostic indicator of JIA, it can provide a certain guiding role in the initial screening and early treatment of the disease. CCP has clinical value in the differential diagnosis of RF+PA subtypes.

Salidroside, as one of the main active ingredients of Rhodiala plant, has the effects of anti-hypoxia, anti-radiation, anti-fatigue, anti-tumor, hypoglycemia and improving immunity. With the increasing demand for salidroside and the decreasing of plant resources, microbial production of salidroside has attracted much attention due to its advantages of short period and easy controlling. At present, microbial production of salidroside is still at the basic research stage. In order to make it easier for researchers to understand the advances of microbial synthesis of salidroside, the biosynthesis pathways, uridine diphosphate glucosyltransferases, wild strain/natural enzymes and engineered strain/recombinant enzymes were reviewed.
Biosynthetic Pathways ; Glucosides ; metabolism ; Phenols ; metabolism

Objective: To observe the feasibility and clinical curative effect of anatomical reduction and fixation of lesser trochanter through posterior-lateral approach for the treatment of comminuted femoral intertrochanteric fractures in elderly patients. Methods: A total of 19 cases with comminuted femoral intertrochanteric fracture, aged 60 to 84 years were enrolled in this study.According to Evans-Jensen classification, 13 cases had type Ⅱ B and 6 cases had type Ⅲ femoral fracture.Through the posterior-lateral approach, the lesser trochanter was fully exposed, reduced and fixed by screw.The proximal and distal femur were well fixed with dynamic hip screw(DHS)or proximal femoral locking plate(PFLP). The lesser trochanter reduction was observed by intraoperative vision and postoperative X-ray.The function evaluation after hip surgery was performed by using Sanders hip function score. Results: All 19 patients obtained the anatomic reattachment in the lesser trochanter.All patients were followed up for an average of 18 months.After treatment, the Sanders hip function score were excellent in 7 patients(36.8%), fine in 12 cases(63.2%). And curative efficacy was good in all cases(100%). There was no significant difference in the curative effect between patients with different classification and gender. Conclusions Through the posterior-lateral approach, the surrounding structures of the lesser trochanter can be fully exposed, and the lesser trochanter can be anatomically repositioned.The posterior-lateral approach to anatomical reduction and fixation of lesser trochanter is an effective and feasible method for the treatment of comminuted intertrochanteric fracture in the elderly.

OBJECTIVE:To provide reference for TCM in the treatment of coronary heart disease (CHD) unstable angina and new drug research and development.METHODS:TCM inheritance auxiliary platform software was adopted to build prescription database of TCM in the treatment of CHD unstable angina.The software function of clinical collection,platform management,information management,knowledge retrieval,statistics,data analysis were adopted to analyze prescription database in respects of four properties,five tastes,channel tropism distribution,frequency statistics,rules of prescriptions and potential new prescriptions,etc.RESULTS:Through analyzing 178 prescriptions of CHD unstable angina,152 ingredients were involved.Top 3 in the list of distribution frequency of four properties were warm (617),cold (474) and mild (288).Top 3 in the list of distribution frequency of five tastes were bitter (750),sweet (732) and pungent (497).Top 3 in the list of distribution frequency of channel tropism were liver (860),heart (573) and spleen (549).A total of 40 ingredients were used more than 10 times,and top 5 were Ligusticum chuanxiong,Salvia miltiorrhiza,Astragalus membranaceus,Angelica sinensis and Carthamus tinctorious.According to the frequency of the drug combination,top 5 in descending order were L.chuanxiong-S.miltiorrhiza,L.chuanxiong-A.sinensis,L.chuanxiong-A.membranaceus,C.tinctorious-L.chuanxiong and S.miltiorrhiza-A.membranaceus.A total of 8 new prescriptions were obtained after evolution.CONCLUSIONS:Main prescriptions for CHD unstable angina included in CNKI are mild drug property,sweetness and bitterness in taste.Main selected ingredients can promote blood circulation,remove blood stasis and relieve pain,relieve restlessness and tranquilize the mind.It embodies the CHD unstable angina treatment principles of "treat both the incidental and fundamental aspects,tonification and purgation in combination".

The chalcone synthase (CHS) superfamily of the type III polyketide synthases (PKSs) generates backbones of a variety of plant secondary metabolites. Benzalacetone synthase (BAS) catalyzes a condensation reaction of decarboxylation between the substrates of 4-coumaric coenzyme A and malonyl coenzyme A to generate benzylidene acetone, whose derivatives are series of compounds with various biological activities. A BAS gene Pcpks2 and a bifunctional CHS/BAS PcPKSI were isolated from medicinal plant P. cuspidatum. Crystallographic and structure-based mutagenesis studies indicate that the functional diversity of the CHS-superfamily enzymes is principally derived from small modifications of the active site architecture. In order to obtain an understanding of the biosynthesis of polyketides in P. cuspidatum, which has been poorly described, as well as of its activation mechanism, PcPKS2 was overexpressed in Escherichia coli as a C-terminally poly-His-tagged fusion protein, purified to homogeneity and crystallized, which is helpful for the clarification of the catalytic mechanism of the enzyme and lays the foundation for its genetic engineering manipulation.
Butanones ; Catalytic Domain ; Crystallization ; Fallopia japonica ; enzymology ; Polyketide Synthases ; genetics ; metabolism

Glycosyltransferases (GTs) catalyze the transfer of a sugar residue of an activated sugar donor to an acceptor molecule. Many families 1 GTs utilize an uridine diphosphate (UDP) activated sugar as donor in the glycosylation reaction, and most of these belong to a group of GTs referred to as the UGTs. The relationship between the degree of amino acid sequence identity and substrate specificity of the plant UGTs is highly complicated, and the prediction of substrate specificity based on phylogenetic analyses need to be improved by more biochemical characterization. This review summarizes the three dimensional structures of plant UGTs published in the Protein Data Bank (PDB), including the detailed substrate interactions with the sugar and receptor binding pockets and mutational analyses of some critical amino acids. It will be helpful for biochemical characterization the substrate specificity of the individual UGT, and lay the foundation for the enzymatic and genetic manipulation of plant UGTs in the future.
Amino Acid Sequence ; Glycosylation ; Glycosyltransferases ; chemistry ; Phylogeny ; Plant Proteins ; chemistry ; Plants ; enzymology ; Protein Structure, Tertiary ; Substrate Specificity ; Uridine Diphosphate ; chemistry

The aim of this paper is to compare the cytotoxicity and cellular uptake efficiency of three kinds of poly(b-benzyl-L-amino) block-poly(ethylene glycol) nanoparticles (PXA-PEG-NPs) using Calu-3 cells, and select one as a nasal drug delivery vector for curcumin (Cur). Poly(gamma-benzyl-L-glutamate) block-poly(ethylene glycol) nanoparticles (PBLG-PEG-NPs), poly(gamma-benzyl-L-lysine) block-poly(ethyleneglycol) nanoparticles (PZLL-PEG-NPs) and poly(gamma-benzyl-L-aspartate) block-poly(ethylene glycol) nanoparticles (PBLA-PEG-NPs) were prepared by emulsion-solvent evaporation method. MTT assays were used to evaluate the cytotoxicity of PXA-PEG-NPs against Calu-3 cells. The cellular uptake of nanoparticles was visualized by an inverted fluorescence microscope and quantified by a flow cytometer. The results indicated that even at high concentration of 2 mg x mL(-1) the three nanoparticles had no cytotoxicity on Calu-3 cells. Compared to the curcumin solution, the three curcumin-loaded PXA-PEG-NPs showed significantly higher cellular uptake efficiency on Calu-3 cells (at equal concentration of curcumin with 5 microg x mL(-1) Cur solution), PBLG-PEG-NPs group was the highest. The cellular uptake increased with incubation time, and has positive correlation with nanoparticle concentration. In brief, PXA-PEG-NPs are conducive to delivery Cur into cells, and PBLG-PEG-NPs might be provided as a good nasal drug delivery carrier.