The natural isoflavones biosynthetic pathway is only limited in legumes plant. To study the isoflavone in bacteria by metabolic engineering requires transformation of multi-gene of the whole pathway into the host strain to resembling the expression and metabolism of the genes. The multi-gene transformation and expression strategy become necessary because of this. This article talks about the multi-gene transformation strategy using one or many vectors, taking the five genes of isoflavonoid biosynthetic pathway to E. coli. The recombinant bacteria carry five genes with two vectors, the whole Isoflavonoid biosynthetic pathway was constructed into E. coli. Fermented with L-tyrosine as substrate and IPTG as an inducer, the recombinant bacteria can produce a new isoflavone related metabolite showed on HPLC analysis profile.
Amino Acid Sequence ; Base Sequence ; Biosynthetic Pathways ; genetics ; Enzymes ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Engineering ; methods ; Isoflavones ; biosynthesis ; metabolism ; Molecular Sequence Data ; Oxygenases ; biosynthesis ; genetics ; Soybeans ; chemistry ; Transformation, Bacterial

OBJECTIVETo investigate the clinical features of invasive candidiasis in children and the risk factors for Candida bloodstream infection.

METHODSA retrospective study was performed on 134 children with invasive candidiasis and hospitalized in 5 tertiary hospitals in Urumqi, China, between January 2010 and December 2015. The Candida species distribution was investigated. The clinical data were compared between the patients with and without Candida bloodstream infection. The risk factors for Candida bloodstream infection were investigated using multivariate logistic regression analysis.

RESULTSA total of 134 Candida strains were isolated from 134 children with invasive candidiasis, and non-albicans Candida (NAC) accounted for 53.0%. The incidence of invasive candidiasis in the PICU and other pediatric wards were 41.8% and 48.5% respectively. Sixty-eight patients (50.7%) had Candida bloodstream infection, and 45 patients (33.6%) had Candida urinary tract infection. There were significant differences in age, rate of use of broad-spectrum antibiotics, and incidence rates of chronic renal insufficiency, heart failure, urinary catheterization, and NAC infection between the patients with and without Candida bloodstream infection (P<0.05). The multivariate logistic regression analysis showed that younger age (1-24 months) (OR=6.027) and NAC infection (OR=1.020) were the independent risk factors for Candida bloodstream infection.

CONCLUSIONSThe incidence of invasive candidiasis is similar between the PICU and other pediatric wards. NAC is the most common species of invasive candidiasis. Candida bloodstream infection is the most common invasive infection. Younger age (1-24 months) and NAC infection are the risk factors for Candida bloodstream infection.

Adolescent ; Age Factors ; Candidiasis ; drug therapy ; microbiology ; Child ; Child, Preschool ; Female ; Fungemia ; etiology ; Humans ; Infant ; Infant, Newborn ; Logistic Models ; Male ; Retrospective Studies ; Risk Factors

OBJECTIVETo investigate the time course of calpain activity changes in rat neurons following fluid percussion injury (FPI) under normothermia (37 degrees celsius;) and mild hypothermia (32-/+0.5) degrees celsius;.

METHODSIn vitro cultured rat neurons were subjected to FPI followed by application of mild hypothermia for intervention at different time points, and the changes in intraneuronal calpain activity following FPI and the interventional effect of mild hypothermia on calpain activity were evaluated by UV-spectrophotometry at different time points.

RESULTSRemarkable changes occurred in calpain activity in the neurons following FPI at 37 degrees celsius;, and mild hypothermia produced obvious interventional effect on calpain activity in close relation to the timing of intervention initiation.

CONCLUSIONIntraneuronal calpain activity changes following FPI are involved in the pathological process of cellular injury, and mild hypothermia might offer protection against traumatic brain injury to some extent by regulating calpain activity. The interventional effect of mild hypothermia is associated with the timing of the intervention initiation.

Animals ; Calpain ; metabolism ; Female ; Hypothermia, Induced ; Neurons ; metabolism ; pathology ; Percussion ; Pregnancy ; Rats ; Rats, Wistar ; Time Factors

Objective To investigate time course changes of intraneuronal calpain activity following fluid percussion injury(FPI)under normothermia(37℃)and mild hypothermia(32+0.5)℃.Methods Rats' neurons produced by external culture and FPI models prepared, mild hypothermia was applied for intervention at different time points and the changes in intraneuronal calpain activity following FPI and the intervention effect of mild hypothermia on calpain activity during different time courses were detected by UV-spectrophotometry. Results Remarkable changes occurred to calpain activity following FPI at 37℃. While the intervention effect of mild hypothermia on calpain activity was different, closely related to the starting time point of intervention. Conclusion The changes of intraneuronal calpain activity following FPI join the pathological process of cellular trauma; mild hypothermia might protect patients with traumatic brain injury(TBI)to some extent through the regulation of calpain activity. And, the intervention effect of mild hypothermia has some correlation with the starting time point of intervention.

Objective To analyze the chemical components of the ethanol-soluble portion of Liuwei Dihuang Decoction,and to explore its mechanism in kidney-Yin deficiency of mice.Methods Bioactive components were analyzed by HPLC,the method was used at Hypersile C1s column(4.6 mm × 250 mm,5 μm),the mobile phase consisted of methanol-water (27 ∶ 73),the detective wavelength was 236 nm,the flow rate was 1.0 mL · min-1,the column temperature was 25 ℃.The mice were divided randomly into three groups (n =10):blank group,model group,experimental group.The mice were gavaged with oral administration of corresponding medicines (9.75 g · kg-1)for nine weeks.At time 6 d after administration,in addition to that the normal group was given distilled water,the other groups were gavaged with hydrocortisone injection per day according to 50 mg · kg-1 body weight for 4 d to establish the kidney Yin deficiency mice model.The contents of cyclic adenosine monophosphate (cAMP),cyclic guanosine monophosphate (cGMP) were determined by ELISA and the follicle-stimulating hormone (FSH),estradiol (E2),testosterone (T) were determined by radioimmunoassay after administration.Results The linear range of morroniside,sweroside,paeoniflorin and loganin were 0.24-2.40 μg (r=0.999 6),0.14-1.38 μg(r =0.999 1),0.12-1.20 μg(r =0.999 1),0.24-2.40 μg(r =0.999 4) respectively.The recovery and relative standard deviation of them were (100.22 ± 1.80) ％,(100.83 ± 1.94) ％,(102.40 ± 1.47) ％,(101.40 ± 1.50) ％,and RSD were 1.80％,1.92％,1.44％,1.50％.The average content of morroniside,sweroside,paeoniflorin and loganin in Liuwei Dihuang Decoction water extract-alcohol soluble parts were calculate in terms of material were 0.20％,0.02％,0.06％,0.13％.The plasma level of cAMP in model group and experimental group were (8.20 ±0.63),(6.90 ±0.15) nmol · L-1 with significant different (P ＜0.01).The serum contents of FSH,E2,T in model group and experimental group were(0.54 ±0.10),(0.88 ±0.04)mU · mL-1;(13.93 ± 0.29),(15.48 ± 0.43) pg · mL-1;(2.23 ± 0.14),(5.63 ± 0.48) ng · mL-1 with significant different (P ＜ 0.01).Conclusion It is found that ethanol-soluble portion might be one of the bioactive component of Liuwei Dihaung Decoction to improve kidney Yin deficiency,and it mainly contains loganin,morroniside,sweroside and paeoniflorin.Its function mechanism of tonifying kidney may be related to the regulation of cAMP content and the levels of hormones in the HPG axis.

We determined a component-target-disease network for Carthamus tinctorius L. and the key compounds, identified by topological analysis, were related to vasculitis, coronary heart and cerebrovascular disease. Based on these compounds, the chromatographic fingerprint of Carthamus tinctorius L. was established. Firstly, 132 compounds were obtained from TCMID and TCMSP databases. Their targets were predicted in the PharmMapp and HemMapper databases. CardioGenBase, Therapeutic Target Database and DisGeNET databases were used to collect targets of vasculitis, coronary heart disease and cerebrovascular disease. The corresponding relationships between component and target protein were established by mapping. Finally, the "component-target-disease" network was built with Cytoscape software. The core network and key nodes were analyzed with the Cytohubba plug-in. The results showed that the 24 key compounds were alpha-tocopherol, adenosine, quinone chalcone pigments such as hydroxysafflor yellow A, safflower yellow, quercetin, kaempferol and other flavonoids, organic acids such as stearic acid, linolenic acid, coumaric acid and cinnamic acid. This resulting chromatographic fingerprint of Carthamus tinctorius L. showed good consistency, and the core chemical compounds obtained by topological analysis of the network of "component-target-disease", could be used as quality control markers. Our research provides a new approach for the identification of quality control indicators in Chinese medicinal materials.

Paecilomyces hepiali is a new species of fungus isolated from a field collection of Ophiocordyceps sinensis from Baima snow mountain, Diqing Tibetan Autonomous Prefecture, Yunnan Province by the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences. The specimen was identified and named as Paecilomyces hepiali by Qing-Tao Chen, the professor of the Institute of Microbiology, Chinese Academy of Sciences (Paecilomyces hepiali) (2008), who identified a dried culture of living strain 82-2 as the holotype. Until now, the holotype (the voucher specimen) was deposited in the Herbarium of the Institute of Chinese Materia Medica (HICMM), China Academy of Chinese Medical Sciences, Beijing. The P. hepiali neotype designated by the paper "Neotypification of P. hepiali (Hypocreales)" published in TAXON 64 (1) by Yao Yi-Jian et al. in February 2015 is untenable.