Objective:To examine the clinical efficacy of endovascular treatment on symptomatic occlusion of intracranial vertebral artery (ICVA) in early non-acute stage.Methods:Nine consecutive patients who presented with aggressive ischemic events in the early non-acute stage of ICVA occlusion from January 2014 to December 2019 and received endovascular treatment at Department of Neurosurgery, Peking University First Hospital were retrospectively reviewed.There were 7 males and 2 females, aged 63.4 years old(range: 52 to 72 years).The average preoperative modified Rankin scale(mRS) was 4.3(range: 4 to 5), the National Institute of Health stroke scale(NIHSS) was 12.3(range: 8 to 18). Among them, 2 patients received a single stage endovascular treatment, and the other 7 patients received staged endovascular treatment.The strategy of staged treatment was as follows: firstly, the occlusion part was passed through by a micro-guidewire and dilated with balloons to maintain the blood flow above Thrombolysis In Cerebral Infarction grade 2b. Then, the intravascular large load thrombus was eliminated by the fibrinolytic system and strengthened antiplatelet drugs. After that, a second stage of angioplasty with stenting was performed on the severe residual stenosis part.The complications and the recanalization rate were collected, and the National NIHSS and mRS after endovascular treatment and in follow-up period were recorded.Results:In the 2 cases received single stage endovascular treatment, although revascularization was achieved lastly, one patient suffered embolus translocation and the other suffered re-occlusion after mechanical thrombectomy during the operation, respectively.Technical success was achieved in 6 of the 7 patients received staged endovascular treatment.On discharge, the average NIHSS scores was 5.7(range: 3 to 4) of the patients. Three months after operation,the average mRS was 1.6(range:0 to 3) and it was 0.9(range: 0 to 2) at the latest follow-up, which were better than preoperative status.Conclusions:Staged endovascular treatment might be a safe, efficient, viable option in carefully selected patients with symptomatic ICVA occlusion in early non-acute stage. It needs to be confirmed by further investigation, preferably in a large controlled setting.

Objective:To examine the clinical efficacy of endovascular treatment on symptomatic occlusion of intracranial vertebral artery (ICVA) in early non-acute stage.Methods:Nine consecutive patients who presented with aggressive ischemic events in the early non-acute stage of ICVA occlusion from January 2014 to December 2019 and received endovascular treatment at Department of Neurosurgery, Peking University First Hospital were retrospectively reviewed.There were 7 males and 2 females, aged 63.4 years old(range: 52 to 72 years).The average preoperative modified Rankin scale(mRS) was 4.3(range: 4 to 5), the National Institute of Health stroke scale(NIHSS) was 12.3(range: 8 to 18). Among them, 2 patients received a single stage endovascular treatment, and the other 7 patients received staged endovascular treatment.The strategy of staged treatment was as follows: firstly, the occlusion part was passed through by a micro-guidewire and dilated with balloons to maintain the blood flow above Thrombolysis In Cerebral Infarction grade 2b. Then, the intravascular large load thrombus was eliminated by the fibrinolytic system and strengthened antiplatelet drugs. After that, a second stage of angioplasty with stenting was performed on the severe residual stenosis part.The complications and the recanalization rate were collected, and the National NIHSS and mRS after endovascular treatment and in follow-up period were recorded.Results:In the 2 cases received single stage endovascular treatment, although revascularization was achieved lastly, one patient suffered embolus translocation and the other suffered re-occlusion after mechanical thrombectomy during the operation, respectively.Technical success was achieved in 6 of the 7 patients received staged endovascular treatment.On discharge, the average NIHSS scores was 5.7(range: 3 to 4) of the patients. Three months after operation,the average mRS was 1.6(range:0 to 3) and it was 0.9(range: 0 to 2) at the latest follow-up, which were better than preoperative status.Conclusions:Staged endovascular treatment might be a safe, efficient, viable option in carefully selected patients with symptomatic ICVA occlusion in early non-acute stage. It needs to be confirmed by further investigation, preferably in a large controlled setting.

Chromosomal integration of heterologous genes or pathways is preferred over the use of episomal plasmids for its inherently stability and thus more desirable in the industrial setting. However, the position of integration of heterologous genes in the genome influences the expression levels. In combination of high throughput transformation of the Yeast Knock-out Collection (YKO) and FACS analysis, the position effect on heterologous reporter gene gfp was identified across the whole genome in yeast. In total 428 high-expressed sites and 444 low-expressed sites were spotted, providing massive data to analyze patterns and reasons for region dependency of gene expression on the genome-wide scale.
Gene Expression Regulation, Fungal ; Gene Knock-In Techniques ; Genes, Reporter ; Genome, Fungal ; Saccharomyces cerevisiae ; genetics

Ketogulonigenium vulgare is an acid-producing strain in the process of two-step vitamin C fermentation. L-sorbosone dehydrogenase (SNDH) is one of the key enzymes during the biosynthesis of 2-keto-L-gulonic acid (2-KGA), the precursor of vitamin C. However, the catalytic mechanism of SNDH is unclear. According to the whole genome sequencing of K. vulgare, two genes encoding sorbosone dehydrogenases, one derived from the chromosome (named as sndhg) and one from plasmid (named as sndhp), were introduced into an industrial strain K. vulgare. The overexpression of gene sndhg had hardly effect on 2-KGA production, and the overexpression of gene sndhp produced an obvious byproduct in the fermentation broth. Combinational expression of sndhg/sndhp with pqqA (obtaining sndhg-pqqA and sndhp-pqqA modules) in K. vulgare resulted in the similar fermentation phenotype to two previous strains. After serial sub-cultivation of co-cultured Bacillus endophyticus with each engineered K. vulgare for 50 d, the conversion rate of 2-KGA increased by 15.4%, 179%, 0.65% and 125% compared with that of the parental K. vulgare with B. endophyticus. This study shows that adaptive evolution of microbial consortium is an effective strategy to increase the fitness between functional modules and chassis, thus quickly getting better strains for production of 2-KGA.
Aldehyde Oxidoreductases ; genetics ; metabolism ; Ascorbic Acid ; Bacillus ; Bacterial Proteins ; genetics ; metabolism ; Coculture Techniques ; Fermentation ; Industrial Microbiology ; Microorganisms, Genetically-Modified ; Rhodobacteraceae ; enzymology ; genetics ; Sugar Acids ; metabolism

In order to study the inherent difference among terpenes producing yeasts from the point of metabolomics, we selected taxadiene producing yeasts as the model system. The changes of cellular metabolites during fermentation log phase of artificial functional yeasts were determined using metabolomics methods. The results represented that compared to W303-1A as a blank control, the metabolites in glycolysis, tricarboxylic acid cycle (TCA) cycle and several amino acids were influenced. And due to the changes of metabolites, the growth of cells was inhibited to a certain extent. Among the metabolites identified, citric acid content in taxadiene producing yeasts changed the most, the decreasing amplitude reached 90% or more. Therefore, citric acid can be a marker metabolite for the future study of artificial functional yeasts. The metabolomics analysis of taxadiene producing yeasts can provide more information in further studies on optimization of terpenes production in heterologous chassis.
Alkenes ; metabolism ; Amino Acids ; metabolism ; Citric Acid ; analysis ; Citric Acid Cycle ; Diterpenes ; metabolism ; Fermentation ; Glycolysis ; Metabolome ; Metabolomics ; Yeasts ; metabolism

The key challenge to generate engineered cells by synthetic biology for producing 7-dehydrocholesterol (7-DHC) in a high titer is the match between functional module and chassis. Our study focused on solving this problem by combining different promoters and yeast chassis to increase 7-DHC production. To optimize the chassis in order to accumulate zymosterol, the substrate for 7-DHC synthesis, we overexpressed truncated HMG-CoA reductase (tHmglp) and squalene epoxidase (Erglp), both are key genes of yeast endogenous zymosterol biosynthetic pathway. In addition, we knocked out C-24 methyl transferase (Erg6p) and C-22 dehydrogenase (Erg5p) to inhibit the conversion of zymosterol to ergosterol. By introducing heterologous C-24 reductase under three promoters with different strengths, namely TDH3p, PGK1p and TDH1p, we constructed functional modules of diverse activities. Nine engineeredcells were generated based on the combination of these three modules and three chassis. The result shows that the engineered cell composed of functional module regulated by TDH3p and chassis SyBE_000956 had the highest 7-DHC production, indicating a better match than others. This study provides evidences for importance of match and empirical support for rational design of subsequent researches.
Cholesterol ; metabolism ; Cytochrome P-450 Enzyme System ; genetics ; Dehydrocholesterols ; metabolism ; Gene Knockout Techniques ; Hydroxymethylglutaryl CoA Reductases ; metabolism ; Industrial Microbiology ; Methyltransferases ; genetics ; Promoter Regions, Genetic ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; genetics ; Synthetic Biology

The purpose of this research is to construct a kind of 3D-Scaffold with galactose-carrying polysaccharide for improving the function of hepatocytes in vitro. Galactose moieties were covalently coupled with hyaluronic acid through ethylenediamine. Galactosylated hyaluronic acid/chitosan scaffolds were prepared by lyophilization. The characteristics of the scaffolds such as morphology, hydrophilicity, and mechanical properties were investigated. The results indicated that the porosity and the pore size of the scaffolds made in -20 degrees C were useful used for culturing hepatocytes. And, the incorporating of hyaluronic acid in chitosan network improved the hydrophilicity and mechanical properties of the scaffolds. Rat primary hepatocytes growing in the scaffolds observed by phase-contrast microscope showed the multicellular spheroid morphologies. Therefore, galactosylated hyaluronic acid/chitosan scaffolds could be used as a promising scaffold for liver tissue engineering.
Animals ; Cells, Cultured ; Chitosan ; chemistry ; pharmacology ; Galactose ; chemistry ; pharmacology ; Hepatocytes ; physiology ; ultrastructure ; Hyaluronic Acid ; chemistry ; pharmacology ; Liver ; physiology ; ultrastructure ; Porosity ; Rats ; Tissue Engineering ; Tissue Scaffolds ; chemistry

Paclitaxel-loaded methoxy poly (ethylene glycol )-b-poly (L-lactic acid) diblock copolymer nanoparticles (PMT) were prepared by a self-emulsification/solvent evaporation method. The PMT morphology, size and its distribution, and drug release in vitro were investigated by DLS, UV, TEM and HPLC. The results indicate that PMT show a spherical morphology with inner core and outer shell. The diameter (nm) of PMT increases with the increase of the drug-loaded amount. The initial burst release is not observed, the drug releasing rate in vitro is lower, and the accumulated release increases with the increase of replacement amout of the pH7. 4 medium. This study develops a new formulation for paclitaxel and provides an experimental basis for the intravenous administration of paclitaxel.
Antineoplastic Agents, Phytogenic ; administration & dosage ; Delayed-Action Preparations ; Drug Carriers ; administration & dosage ; chemistry ; Humans ; Injections, Intravenous ; Nanoparticles ; Paclitaxel ; administration & dosage ; Polyesters ; administration & dosage ; chemistry ; Polyethylene Glycols ; administration & dosage ; chemistry ; Polymers ; administration & dosage ; chemistry

To study the alkaloid constituents in the seed of Sophora alopecuroides L. and isolation of lehmannine by transhydrogenation of sophocarbine to matrine present in the mixture. Methods　Various processes of exchange, ion exchange and column chromatograph with very similar physical and chemical properties was successfully separated by transhydrogenation. Results　5 alkaloids were obtained and identified by elementary analysis, IR, MS, 1HNMR and 13CNMR as oxymatrine, oxysophoridine, matrine, sophoridine and lehmannine (12, 13-dehydromatrine). Conclusion　Lehmannine was isolated from the seed of S. alopecuroides for the first time.

To summarize the application of proteomics to modernization research of Chinese materia medica (CMM) and offer the reference for modernization of CMM and new medicine preparation exploitation. Based on the references and our project group studies in proteomic of drug biosynthesis pathway and metabolization;the main content,technology strategies and important application of the proteomics were introduced significantly;and the further development of protemics was analyzed. The proteomics will become one of absolutely necessary tackles in the modernization research of CMM.