OBJECTIVE:To probe into a feasible work mode for special clinical pharmacist of hypertension.METHODS:Clinical pharmacists' works were summarized by considering the experience of clinical pharmacists and examples of pharmaceutical care in hypertension wards.RESULTS&CONCLUSIONS:Special pharmacists of hypertension should participate in clinical treatment team,check prescription every day and acquaintance with status quo of patients' medication so as to carry out pharmaceutical care suitability.It also should find out and resolve situation of ADR in time and master the characteristics of hypertension.

Enantioselective hydrolysis of 2-carboxyethyl-3-cyano-5-methylhexanoic acid (CNDE) is the key step in chemoenzymatic synthesis of pregabalin. We purified an intracellular carboxyl esterase from Morganella morganii ZJB-09203, which exhibited high enantioselectivity and activity towards CNDE. The carboxyl esterase was purified to electrophoretic homogeneity by ammonium sulfate fraction precipitation, Phenyl Sepharose 6 FF hydrophobic interaction chromatography, anion exchange with DEAE Sephadex A-50 and Bio-Scale CHT column. The purified enzyme was a monomer with molecular mass of 68 kDa determined by SDS-PAGE and gel chromatography. Substrate specificity of the enzyme towards p-nitrophenyl esters suggested that the purified enzyme was an esterase. The optimal reaction pH for CNDE hydrolysis was 9.0, and optimal temperature was 45 degrees C. The esterase was stable between pH 7.0 and 9.0, and at 40 degrees C. The enzyme activity was enhanced by Ca2+, Cu2+ and Mn2+, whereas strongly inhibited by Co2+, Fe3+, Ni2+ and EDTA. Meanwhile, we investigated the kinetic parameters of the esterase towards p-nitrophenyl esters and effect of CNDE concentration on conversion. The present study reported the esterase capable of stereospecific hydrolysis of CNDE for the first time. Our research will provide foundations for industrial production of Pregabalin using the new biocatalyst.
Chromatography, Gel ; Electrophoresis, Polyacrylamide Gel ; Esterases ; metabolism ; Hydrogen-Ion Concentration ; Kinetics ; Molecular Weight ; Morganella morganii ; enzymology ; Substrate Specificity ; Temperature

Anidulafungin is an effective antifungal medicine, which can inhibit activities of candida in vitro and in vivo. Echinocandin B (ECB) is the key precursor of Anidulafungin, thus the price and market prospect of Anidulafungin is directly due to the fermentation titer of ECB. In this study, Aspergillus nidulans was used for ECB fermentation, and the influence of adding microparticles on ECB fermentation was studied, such as talcum powder, Al2O3, and glass beads. The particle size and concentration were the key factors for mycelium morphology and ECB production, and ECB production could reach 1 262.9 mg/L and 1 344.1 mg/L by adding talcum powder of 20 g/L (d50 = 14.2 μm) and 7 glass beads (6 mm), an increase by 33.2% and 41.7%, respectively. The results indicated that the mycelium morphology of filamentous microorganisms and the product yield of fermentation could be improved by adding microparticles remarkably, and it provide an important method for the fermentative optimization of filamentous microorganisms.
Antifungal Agents ; metabolism ; Aspergillus nidulans ; metabolism ; Echinocandins ; biosynthesis ; Fermentation ; Fungal Proteins ; biosynthesis ; Industrial Microbiology ; methods

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3-Hydroxypropionic acid is an important building block to synthesize lots of industrially valuable chemicals. In this study, we firstly investigated the effects of cell, substrate and product concentrations on biosynthesis of 3-hydroxypropionic acid from 1,3-propanediol by Gluconobacter oxydans ZJB09112 in 50-mL shake flask containing 10 mL transformation liquid. To avoid the inhibition of substrate and product, we adopted fed-batch biotransformation and fed-batch biotransformation coupled with in situ product removal in 2-L bubble column reactor containing 1 L transformation liquid. The results show that high concentrations of substrate and product could inhibit the biotransformation by decreasing the initial reaction rate, and the optimal reaction conditions were as follows: cell concentration 6 g/L, pH 5.5. Fed-batch biotransformation in which the substrate concentration was maintained at 15-20 g/L could obtain product concentration of 60.8 g/L after 60 h, which gave a productivity of 1.0 g/(Lh) and a yield of 84.3%. Furthermore, fed-batch biotransformation coupled with in situ product removal could achieve the total product concentration of 76.3 g/L after 50 h, which gave a productivity of 1.5 g/(L x h) and a yield of 83.7%. The results obtained here may be useful for the application of G. oxydans in biocatalysis industry by using its characteristic of incomplete oxidation of alcohols.
Batch Cell Culture Techniques ; Biotransformation ; Gluconobacter oxydans ; metabolism ; Lactic Acid ; analogs & derivatives ; biosynthesis ; Oxidation-Reduction ; Propylene Glycols ; metabolism

1,3-Dihydroxyacetone is widely used in cosmetics, medicines and food products. We reviewed the recent progress in metabolic pathways, key enzymes, as well as metabolic engineering for microbial production of 1,3-dihydroxyacetone. We addressed the research trend to increase yield of 1,3-dihydroxyacetone by improving the activity of glycerol dehydrogenase with genetic engineering, and regulating of fermentation process based on metabolic characteristic of the strain.
Dihydroxyacetone ; biosynthesis ; Fermentation ; Genetic Engineering ; methods ; Gluconobacter oxydans ; genetics ; metabolism ; Industrial Microbiology ; methods ; Metabolic Engineering ; methods ; Sugar Alcohol Dehydrogenases ; metabolism

There is growing interest in biodiesel and this results in the accumulation of glycerol. The exploitation and application of glycerol has attracted more and more attention. In the current study, glycerol was biotransformed to produce 3-hydroxypropionaldehyde by genetic engineering bacteria. It is known that 3-hydroxypopionaldehyde has been widely used as an important intermediate for chemicals, effective antimicrobial agent, and fix agent for tissues. A pair of primers was designed on the basis of the sequence of both NH2-terminus and the amino acid sequence of glycerol dehydratase reported by NCBI, and a fragment about 1.6 kb was obtained by PCR amplification using the total genome DNA of Lactobacillus reuteri as template, then the fragment was cloned to the pMD18-T vector and sequenced. Two specific primers were designed according to the obtained sequence, and a fragment with length of 1674 bp was amplified using PCR with these two specific primers. Consequently, the resulting products were digested with EcoR I and Hind III and ligated using T4 DNA ligase to the pET28b vector digested with the same enzymes. The recombinant plasmid, named pET28b-dhaB, was transformed into E. coli BL21. The positive clones were induced with IPTG and the expression products were further analyzed by SDS-PAGE, indicating that protein with a molecule weight of around 65 kD was obtained. Furthermore, the glycerol dehydratase activity was evaluated and compared with the wild type strain as well.
Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Glyceraldehyde ; analogs & derivatives ; chemistry ; metabolism ; Hydro-Lyases ; biosynthesis ; genetics ; Lactobacillus reuteri ; enzymology ; genetics ; Propane ; chemistry ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics

Objective To describe the general characteristics of patients with acute chest pain in order to analyze factors associated with patients’utilization of emergency medical services (EMS).Methods A total of 747 eligible patients with acute chest pain admitted to emergency department of Qilu Hospital were consecutively enrolled from October 2014 to April 2015.Clinical data including demographic features, mode of arrival,past medical history,risk factors,symptoms and signs were collected prospectively by using standardized case report form.Univariate and multivariate analyses were carried out to investigate the association between the decision to use EMS and related factors including demographic features,past medical history,risk factors,symptoms and signs.Results Of the total 747 eligible patients,414 (55.4%)were male ,and the mean age was (57.2 ± 15.8)year;333 (44.6%)were female,and the mean age was (61.7 ±14.9)year.Of them,171 (22.9%)patients used EMS,and 143 chest pain patients with more than 75 years old were more inclined to use EMS (P <0.01),whereas 152 patients in 65 -75 years age group accounted for the lowest proportion of using EMS.Men were more inclined to use EMS than women (P <0.05),and 483 patients with typical chest pain used more EMS than patients with atypical chest pain (P <0.05);Of them,356 patients with a history of hypertension and 54 patients with a history of cerebral infarction were more inclined to use EMS (P <0.05 and P <0.01,respectively).Multivariate logistic regression analysis showed that male,older than 75 years,history of cerebral infarction were independent factors associated with EMS use (P <0.05).Conclusions This study indicated that only less than one-third of emergency department visits with acute chest pain decide to use EMS when symptoms occurred. Factors including male,older than 75 years,and a history of cerebral infarction were associated with more use of EMS.In order to promote patient asking for EMS timely,more work should be done.

Objective To investigate the impact of polyclonal neural cell adhesion molecule antibody (P-NCAM-Ab) on the potency of botulinum toxin A (BTX-A).Methods Ninety male Sprague-Dawley rats were randomly divided into 3 equal groups:a normal control group,a BTX-A group and a P-NCAM-Ab group.The rats in the normal control group were injected with 100 μl of saline solution in their right gastrocnemius,while those in the BTX-A and P-NCAM-Ab groups were injected with 100 μl of BTX-A (0.5 U).In addition,the rats in the P-NCAM-Ab group were also injected with 100 μl of P-NCAM-Ab (the dosage was 20 U) at the same site on the 3rd day after the BTX-A injection.The rats' gastrocnemius muscle strength was evaluated with a self-made system for evaluating neuromuscular function before and after the toxin injection,on the 3rd day,as well as 1,2,4,6,8,10 and 12 weeks after the BTX-A injection.Any wet weight changes in the muscles were observed,and immunochemistry methods were employed to observe any structural changes in the motor endplates and nerve fibers at the different time points.Results After the saline injection,the average gastrocnemius muscle strength of the control group increased with time,while strength in the BTX-A and P-NCAM-Ab groups demonstrated a decrease in strength followed by a gradual increase.The average gastrocnemius muscle strength of the rats in the BTX-A and P-NCAM-Ab groups was significantly lower than that of the control group at all time points.Compared with the BTX-A group,the muscle strength of the P-NCAM-Ab group rats decreased further.Strength recovery in the BTX-A and P-NCAM-Ab groups was significantly slower than in the control group.The wet weight percentage in the BTX-A and P-NCAM-Ab groups at first decreased and then recovered with time.After the BTX-A injection,the average wet weight percentage of the P-NCAM-Ab group rats was significantly lower than that of the BTX-A group after 3 days,and 1,2 and 4 weeks.Karnovsky-Roots AchE staining showed that the motor endplates' color in the BTX-A and P-NCAM-Ab groups deepened gradually,though the color of the P-NCAM-Ab group was lighter than that of the BTX-A group at each time point.The mean optical density of the motor endplates' positive reaction area increased with time in both groups,but the P-NCAM-Ab group was lower than that of the BTX-A group at 1,2,4,8 and 12 weeks.Counting the nerve fibers dyed by gold chloride showed similar trends with both experimental groups significantly different from the control group.Conclusion P-NCAM-Ab can increase the potency of BTX-A and prolong its action.

Nitriles are an important type of synthetic intermediates in the production of fine chemicals because of their easy preparations and versatile transformations. The traditional chemical conversion of nitriles to carboxylic acids and amides is feasible but it requires relatively harsh conditions of heat, acid or alkali. Nitrile converting enzymes (nitrilase, nitrile hydratase and amidase) which are used as biocatalyst for the production of fine chemicals have attracted substantial interest because of their ability to convert readily available nitriles into the corresponding higher value amides or acids under mild conditions with excellent chemo-, regio- and stereo-selectivities. Many nitrile converting enzymes have been explored and widely used for the production of fine chemicals. In this paper, various examples of biocatalytic synthesis of pharmaceuticals and their intermediates, agrochemicals and their intermediates, food and feed additives, and other fine chemicals are presented. In the near future, an increasing number of novel nitrile converting enzymes will be screened and their potential in the production of useful fine chemicals will be further exploited.
Amides ; metabolism ; Amidohydrolases ; metabolism ; Aminohydrolases ; metabolism ; Carboxylic Acids ; metabolism ; Chemical Industry ; methods ; Hydro-Lyases ; metabolism ; Nitriles ; chemistry