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1.
Article in Chinese | WPRIM | ID: wpr-864039

ABSTRACT

Objective:To investigate the etiology, prognosis and prognostic factors of pediatric acute liver failure(PALF), in order to provide the basis for clinical treatment of PALF.Methods:The clinical data of children with PALF hospitalized at Hunan Children′s Hospital from May 2008 to May 2018 were collected, and the causes and prognosis were analyzed.According to the prognosis, the patients were divided into the death group and the survival group, whose biochemical indexes were then compared.After that, the statistical analysis of different data were carried out by using t-test, Wilcoxon test and χ2 test separately. Results:In 120 PALF cases, there were 68 males and 52 females, and there were 36 infants, 34 toddlers, 22 preschoolers and 28 school-age children.Twenty cases (16.7%) were caused by sepsis, 19 cases (15.8%) by genetic metabolic diseases, 18 cases (15.0%) by poisoning, 12 cases (10.0%) by viral infection, 6 cases (5.0%) by drugs, 1 case (0.8%) by bile polyp, and 1 case (0.8%) by tumor disease.Besides, the etiology of 43 cases (35.9%) was unknown.Among the cases with known etiologies, genetic metabolic and infectious diseases were the main cause of disease in infants, toddler patients were mostly caused by infectious diseases and drug/toxicants, and drug/toxicants and hereditary metabolic diseases were the dominant cause of disease in school-age children and preschoolers.Mortality rate of children with PALF was 50.0%.Among them, the mortality of Epstein-Barr virus-associated hemophagocytic syndrome, sepsis, Citrin deficiency and Tyrosinemia was higher than that of other diseases.Compared with the survival group, the total bilirubin (TB)[159.00(73.05, 274.00) μmol/L vs.62.75(2.65, 221.75)μmol/L], direct bilirubin(DB)[83.00(41.43, 160.00) μmol/L vs.38.74(10.98, 128.75) μmol/L], prothrombin time (PT)[39.60(24.93, 62.60) s vs.24.65(21.43, 29.83) s], international standardized ratio (INR)[3.40(2.30, 6.74) vs.2.09(1.85, 2.84)], and blood ammonia (NH 3) levels [109.50(85.25, 149.75) μmol/L vs.80.00(60.25, 102.75) μmol/L] in the death group were significantly increased, and the diffe-rences were statistically significant(all P<0.05); while the levels of albumin[(28.72±5.88) g/L vs.(33.69±4.96) g/L], alanine aminotransferase (ALT) [586.50(223.25, 1 082.00) U/L vs.1 434.00(615.00, 3 334.50) U/L]and aspartate aminotransferase (AST) [827.50(545.00, 2 024.00) U/L vs.1 663.50(821.00, 4 886.75) U/L]in the death group were significantly decreased, and the differences were statistically significant(all P<0.05). However, the blood glucose and cholesterol levels in both groups had no statistically significant difference. Conclusion:The mortality of children with PALF is high, and different age groups have different etiologies.The increase of TB, DB, PT, INR, NH3 and the ratio of hepatic encephalopathy, and the decrease of albumin, AST and ALT suggest poor prognosis.

2.
Chinese Journal of Biotechnology ; (12): 1450-1458, 2020.
Article in Chinese | WPRIM | ID: wpr-826831

ABSTRACT

Heparin and heparan sulfate are a class of glycosaminoglycans for clinical anticoagulation. Heparosan N-sulfate-glucuronate 5-epimerase (C5, EC 5.1.3.17) is a critical modifying enzyme in the synthesis of heparin and heparan sulfate, and catalyzes the inversion of carboxyl group at position 5 on D-glucuronic acid (D-GlcA) of N-sulfoheparosan to form L-iduronic acid (L-IdoA). In this study, the heparin C5 epimerase gene Glce from zebrafish was expressed and molecularly modified in Escherichia coli. After comparing three expression vectors of pET-20b (+), pET-28a (+) and pCold Ⅲ, C5 activity reached the highest ((1 873.61±5.42) U/L) with the vector pCold Ⅲ. Then we fused the solution-promoting label SET2 at the N-terminal for increasing the soluble expression of C5. As a result, the soluble protein expression was increased by 50% compared with the control, and the enzyme activity reached (2 409±6.43) U/L. Based on this, site-directed mutations near the substrate binding pocket were performed through rational design, the optimal mutant (V153R) enzyme activity and specific enzyme activity were (5 804±5.63) U/L and (145.1±2.33) U/mg, respectively 2.41-fold and 2.28-fold of the original enzyme. Modification and expression optimization of heparin C5 epimerase has laid the foundation for heparin enzymatic catalytic biosynthesis.


Subject(s)
Animals , Carbohydrate Epimerases , Chemistry , Genetics , Escherichia coli , Gene Expression , Heparin , Metabolism , Heparitin Sulfate , Metabolism , Iduronic Acid , Metabolism , Zebrafish Proteins , Chemistry , Genetics
3.
Chinese Journal of Biotechnology ; (12): 1689-1698, 2020.
Article in Chinese | WPRIM | ID: wpr-826808

ABSTRACT

Enterokinase is a class of serine proteases that specifically recognize the cleavage DDDDK sequences. Therefore, enterokinase has been widely used as a tool enzyme in the field of biomedicine. Currently, the expression level of enterokinase in Pichia pastoris is low, which hinders related practical applications. In this study, the effects of six different signal peptides SP1, SP2, SP3, SP4, SP7 and SP8 on the secretory expression of enterokinase in Pichia pastoris were studied. Compared with α-factor, SP1 significantly increased the secretory expression of enterokinase (from 6.8 mg/L to 14.3 mg/L), and the enterokinase activity increased from (2 390±212) U/mL to (4 995±378) U/mL in shaking flask cultures. On this basis, the enterokinase activity was further enhanced to (7 219±489) U/mL by co-expressing the endogenous protein Kex2. Moreover, the activity that the mutant strain with N-terminal fusion of three amino acids of WLR was increased to (15 145±920) U/mL with a high specific activity of (1 174 600±53 100) U/mg. The efficient secretory expression of enterokinase laid a foundation for its applications in near future.

4.
Chinese Journal of Biotechnology ; (12): 567-576, 2019.
Article in Chinese | WPRIM | ID: wpr-771352

ABSTRACT

Ethyl carbamate (EC), a carcinogenic and teratogenic chemical that is widely distributed in various alcoholic beverages, has attracted much attention. Microbial enzymatic degradation of EC in rice wine is always efficient and attractive. In this review, we summarize the research progress and problems of microbial enzymatic elimination of EC in rice wine from three aspects: the mechanisms of EC formation in rice wine, the research progress of acid urease, and the research progress of urethanase. Then, we propose the corresponding strategies to solve the problems: screening new urethanase with satisfied enzyme properties, food-grade expression and directed evolution of the bifunctional Fe³⁺-dependent acid urease and acid urease used in combination with urethanase to eliminate both urea and EC in rice wine.


Subject(s)
Oryza , Urea , Urease , Urethane , Wine
5.
Chinese Journal of Biotechnology ; (12): 805-815, 2019.
Article in Chinese | WPRIM | ID: wpr-771329

ABSTRACT

Hyaluronic acid (HA) is widely used in many fields, such as medicine, cosmetics and food. The bioactivity of HA depends on its molecular weight (Mw). Owing to the important physiological activities and special physiological functions, HA oligosaccharides have important application prospects in medicine fields. Streptococcus zooepidemicus has wide applications in commercial production of HA, due to its short fermentation cycle and strong production intensity. In order to efficiently synthesize HA oligosaccharides and solve the dissolved oxygen in the fermentation process, in this study, we overexpressed HA synthase (HasA) and introduced and optimized the leech hyaluronidase LHAase in Streptococcus zooepidemicus WSH-24. As a result, HA oligosaccharides were efficiently produced with improved dissolved oxygen. After 24 h, HA oligosaccharides production intensity reached to 294.2 mg/(L·h), and the concentration accumulated to 0.97 g/L in flask cultures, which was 1.82 times of the wild strain. Impressively, HA oligosaccharides were increased to 7.06 g/L in 3 L fermentor. The constructed Streptococcus zooepidemicus strain for producing HA oligosaccharides would have broad application prospects.


Subject(s)
Bioreactors , Fermentation , Hyaluronan Synthases , Genetics , Metabolism , Hyaluronic Acid , Genetics , Metabolism , Industrial Microbiology , Oligosaccharides , Genetics , Metabolism , Streptococcus equi , Genetics , Metabolism
6.
Article in Chinese | WPRIM | ID: wpr-751772

ABSTRACT

Objective To study the protein binding rates ofgardenioside,paeoniflorin and naringin in Weiyanling granules in bovine serum albumin,rat plasma and human plasma.Methods The equilibrium dialysis method was carried to determine the binding-rates of three components in Weiyanling granules in bovine serum albumin,rat plasma and human plasma.The HPLC method was used to determine the mass concentration of three components in weiyanling granules in inner and outer dialysis and to calculate the plasma protein-binding rates.Results The linear relationship,precision,extraction recovery and stability of gardenoside,paeoniflorin and naringin all conformed to the methodological requirements.Within the low,middle and high concentration (5.084,10.04 and 20.08 g/L) of Weiyanling granules,the protein-binding rates of gardenioside and bovine serum albumin were 23.77% ± 13.39%,10.75% ± 1.34%,5.82% ± 6.53%,respectively;the protein-binding rates of paeoniflorin were 51.81% ± 6.53%,30.12% ± 6.61%,21.39% ± 9.45%,respectively;the protein-binding rates of naringin were 70.21% ± 3.31%,67.61% ± 1.31%,56.00% ± 3.46%,respectively;the protein-binding rates of gardenioside and rat plasma were 71.56% ± 3.00 %,43.56% ± 12.22%,40.63% ± 4.73%,respectively;the protein-binding rates ofpaeoniflorin and rat plasma were 55.72% ± 1.60%,47.59% ± 6.33%,40.07% ± 0.72%,respectively;the protein-binding rates of naringin and rat plasma were 85.85% ± 3.53%,85.38% ± 0.99%,79.99% ± 2.57%,respectively;the protein-binding rates of gardenioside and human plasma were 13.56% ± 2.40%,3.02% ± 2.41%,5.82% ± 2.08%,respectively;the protein-binding rates of paeoniflorin and human plasma were 9.49% ± 5.23%,5.01% ± 1.10%,3.98% ± 1.54%,respectively;the protein-binding rates of naringin and human plasma were 25.04% ± 2.61%,26.09% ± 13.82%,20.20% ± 2.24%,respectively.Conclusions Within 5.084,10.04 and 20.08 g/L,the protein binding-rate of gardenioside,paeoniflorin and naringin were shown asfollow,rat plasma >bovine serum albumin > human plasma.There were significant differences in species.

7.
Article in Chinese | WPRIM | ID: wpr-789171

ABSTRACT

Objective To establish a method for the simultaneous content determination ofquercetin,kaempferol and pinocembrin in Penthorum Chinense Pursh.Methods The HPLC analysis was carried out on Alltima C18 (250 mm × 4.6 mm,5 μm) with a mixture of methanol-0.2% phosphate (45:55) as the mobile phase.The determination wavelength was set at 270 nm with the flow rate of 1.0 ml/min.The column temperature was set at 30 ℃.Results The quercetin,kaempferol and pinocembrin showed good linearity in the range of 0.316-10.120 μg (r=0.999 9),0.012-0.397 μg (r=0.999 6) and 0.063-2.016 μg (r=0.999 8) respectively.The average recovery rates of quercetin,kaempferol and pinocembrin were 99.34% (RSD=1.51%),99.76% (RSD=l.96%) and 98.34% (RSD=1.56%) respectively.Conclusions The method is accurate and sensitive,which can be used for the content determination of quercetin,kaempferol and pinocembrin in Penthorum Chinense Pursh.

8.
Chinese Journal of Biotechnology ; (12): 1222-1233, 2019.
Article in Chinese | WPRIM | ID: wpr-771806

ABSTRACT

Sulfated compounds are widely present in cytoplasm, on cell surface, and in extracellular matrix. These compounds play important roles in cell development, differentiation, immune response, detoxication, and cell signal transduction. 3-Phosphoadenosine-5-phosphosulfate (PAPS) is the universal sulfate group donor for the biosynthesis of sulfated compounds. Up to now, the synthesis of PAPS is still too expensive for industrial applications. This review focuses on the recent progress of PAPS production and summaries the application of PAPS, particularly in the production of glucosinolate, heparin, condroitin sulfate, and oxamniquine production.


Subject(s)
Cell Differentiation , Chondroitin Sulfates , Phosphoadenosine Phosphosulfate , Metabolism , Sulfates
9.
Chinese Medical Journal ; (24): 1666-1673, 2018.
Article in English | WPRIM | ID: wpr-688061

ABSTRACT

<p><b>Background</b>One of the main aims of the updated Prostate Imaging Reporting and Data System Version 2 (PI-RADS v2) is to diminish variation in the interpretation and reporting of prostate imaging, especially among readers with varied experience levels. This study aimed to retrospectively analyze diagnostic consistency and accuracy for prostate disease among six radiologists with different experience levels from a single center and to evaluate the diagnostic performance of PI-RADS v2 scores in the detection of clinically significant prostate cancer (PCa).</p><p><b>Methods</b>From December 2014 to March 2016, 84 PCa patients and 99 benign prostatic shyperplasia patients who underwent 3.0T multiparametric magnetic resonance imaging before biopsy were included in our study. All patients received evaluation according to the PI-RADS v2 scale (1-5 scores) from six blinded readers (with 6 months and 2, 3, 4, 5, or 17 years of experience, respectively, the last reader was a reviewer/contributor for the PI-RADS v2). The correlation among the readers' scores and the Gleason score (GS) was determined with the Kendall test. Intra-/inter-observer agreement was evaluated using κ statistics, while receiver operating characteristic curve and area under the curve analyses were performed to evaluate the diagnostic performance of the scores.</p><p><b>Results</b>Based on the PI-RADS v2, the median κ score and standard error among all possible pairs of readers were 0.506 and 0.043, respectively; the average correlation between the six readers' scores and the GS was positive, exhibiting weak-to-moderate strength (r = 0.391, P = 0.006). The AUC values of the six radiologists were 0.883, 0.924, 0.927, 0.932, 0.929, and 0.947, respectively.</p><p><b>Conclusion</b>The inter-reader agreement for the PI-RADS v2 among the six readers with different experience is weak to moderate. Different experience levels affect the interpretation of MRI images.</p>

10.
Article in Chinese | WPRIM | ID: wpr-805901

ABSTRACT

Objective@#To investigate the clinical features of children with infectious mononucleosis (IM), to compare the difference of therapeutic effects between general treatment to antiviral therapy for IM.@*Methods@#This prospective study analyzed the clinical data and laboratory test results of 201 cases with IM in our hospital from January 1, 2016 to December 31, 2016. The follow-up period was 6-12 months. The patients were divided into two groups according to the order of admission. The clinical symptoms and laboratory test results of the two groups were observed after hospitalization.@*Results@#Of the total of 201 patients, male to female is 1.72∶1; Age: 8 months to 13 years 6 months (average 4.8±2.8 years), The disease frequently occurred in summer and autumn, accounted for 64.18%.The major clinical manifestations was fever (97.51%), angina (79.10%), enlarged of lymph node(68.66%), eyelid swelling (67.16%), hepatomegaly (53.73%) and splenomegaly (46.77%). There was no statistical difference in duration of fever, improved angina time, lymph nodes(liver, spleen) enlargement recovery time, heterotypic lymphocytes normalization time, lymphocyte function normalization time.There was significant difference in reducing the serum/plasma or total blood EBV-DNA in the short term between antiviral group and general treatment group (P<0.01), but for the long-term reduction of EBV-DNA, there was no significant difference (P>0.05). The low affinity EBV-CA-IgG was converted to high affinity EBV-CA-IgG in 89.41% of patients after 3 months and in 98.68% of patients after 6 months. With the recovery of the disease, the positive rate of EBV-CA-IgM and EBV-EA-IgG decreased. But the positive rate of EBV-NA-IgG increased. Serum/plasma EBV-DNA was completely overcast after 3 months, but after 12 months, the patient’s total blood EBV-DNA positive rate was still 71.11%.@*Conclusions@#In general IM patients, there was no significant difference between antiviral therapy and general treatment, so antiviral therapy is not generally recommended. EBV DNA in the whole blood of patients with IM will continue to be positive more than 6-12 months, indicating that to observe changes of IM should not use whole blood samples for EBV DNA testing.

11.
Chinese Journal of Biotechnology ; (12): 1772-1783, 2018.
Article in Chinese | WPRIM | ID: wpr-776291

ABSTRACT

Glucaric acid (GA), a top value-added chemical from biomass, has been widely used for prevention and control of diseases and the production of polymer materials. In GA biosynthesis pathway, the conversion of inositol to glucuronic acid that catalyzed by myo-inositol oxygenase is the limiting step. It is necessary to improve MIOX activity. In the present study, we constructed a high-throughput screening system through combing the concentration of GA with the green fluorescent protein fluorescence intensity. By applying this screening system, three positive variants (K59V/R60A, R171S and D276A) screened from the mutant library. In comparison, the recombinant strain Escherichia coli BL21(DE3)/MU-R171S accumulated more GA, 136.5% of that of the parent strain.


Subject(s)
Biosensing Techniques , Biosynthetic Pathways , Escherichia coli , Glucaric Acid , Chemistry , Inositol Oxygenase , Chemistry
12.
Chinese Journal of Biotechnology ; (12): 1784-1793, 2018.
Article in Chinese | WPRIM | ID: wpr-776290

ABSTRACT

Heparin is a very important anticoagulant drug. Currently, heparin is mainly extracted from porcine mucosa. However, animal-derived heparin shows low anticoagulant activity due to the low proportion of the anticoagulant active unit, the GlcNS6S-GlcA-GlcNS6S3S-Ido2S-GlcNS6S pentasaccharide. In this study we proposed an enzymatic strategy to sulfate the animal-sourced heparin to increase the proportion of anticoagulant pentasaccharide and the anticoagulant activity. First, three sulfotransferases HS2ST, HS6ST, and HS3ST were expressed tentatively in Escherichia coli and Pichia pastoris. After measuring the sulfotransferase activity, we confirmed P. pastoris GS115 is the better host for sulfotransferases production. Then, the maltose binding protein (MBP) and thioredoxin (TrxA) were fused separately to the N-terminal of sulfotransferases to increase enzyme solubility. As a result, the yields of HS2ST and HS6ST were increased to (839±14) U/L and (792±23) U/L, respectively. Subsequent sulfation of the animal-sourced heparin with the recombinant HS2ST, HS6ST and HS3ST increased the anticoagulant activity from (76±2) IU/mg to (189±17) IU/mg.


Subject(s)
Animals , Escherichia coli , Heparin , Chemistry , Oligosaccharides , Chemistry , Pichia , Sulfotransferases , Swine
13.
Article in Chinese | WPRIM | ID: wpr-299306

ABSTRACT

<p><b>OBJECTIVE</b>To construct a cell model of 4.1R gene knockout in murine macrophage cell line RAW264.7 using CRISPR/Cas9 technique.</p><p><b>METHODS</b>Three high?grade small?guide RNAs (sgRNAs) that could specifically identify 4.1R gene were synthesized and inserted into lentiCRISPRv2 plasmid. RAW264.7 cells were infected with sgRNA?Cas9 lentivirus from 293T cells transfected with the recombinant sgRNA?lentiCRISPRv2 plasmid, and the positive cells were screened using puromycin and the monoclonal cells were obtained. The expression of 4.1R protein in the monoclonal cells was measured by Western blotting, and the mutation site was confirmed by sequence analysis. Result A 4.1R gene knockout RAW264.7 cell line was obtained, which showed a 19?bp deletion mutation in the 4.1R gene sequence and obviously enhanced proliferation.</p><p><b>CONCLUSION</b>We successfully constructed a 4.1R gene knockout macrophage cell line using CRISPR/Cas9 technique, which may facilitate further investigation of the function of 4.1R in macrophages.</p>

14.
Chinese Journal of Biotechnology ; (12): 228-236, 2017.
Article in Chinese | WPRIM | ID: wpr-310595

ABSTRACT

Glucaric acid, a high value-added organic acid, is widely used in food, pharmaceutical and chemical industries. For microbial production of glucaric acid in Saccharomyces cerevisiae, we constructed a synthetic glucaric acid biosynthetic pathway by coexpressing the genes encoding myo-inositol oxygenase from mice and uronate dehydrogenase from Pseudomonas putida. Moreover, myo-inositol-1-phosphate synthase was identified as a rate-limiting enzyme in glucaric acid pathway and was upregulated, resulting in the production of glucaric acid of (107.51±10.87) mg/L, a 2.8-fold increase compared to the parent strain. Then, by repressing the activity of phosphofructokinase, the concentration of glucaric acid further increased to (230.22±10.75) mg/L. The strategy could be further used to construct cell factories for glucaric acid production.

15.
Chongqing Medicine ; (36): 3050-3052,3056, 2017.
Article in Chinese | WPRIM | ID: wpr-608783

ABSTRACT

Objective To access the diagnostic value of Prostate Imaging Reporting and Data System (PIRADS version 2)for prostate cancer (PCa) in prostate specific antigen (PSA) grey zone(4-10 ng/mL).Methods Treatment naive PCa and BPH patients with an increase of PSA 4-10 ng/mL from 2200 patients underwent prostate MRI from 2012 to 2016 were included,multiparameter magnetic resonance imaging (mp-MRI) prior biopsy or prostatectomy and clinical data were obtained,mp-MRI were retrospectively analyzed quantitatively by a radiology expert with 15 years experience in urogenital system imaging diagnosis and a doctor with 5 years experience in radiology diagnosis blind to the pathology results according to PIRADS v2,PIRADS v2 score and lesion zone were recorded respectively,in case of disagreement,dicision was made through discuss.TRUS guided biopsy or prostatectomy pathology serves as gold reference.Diagnostic value of PIRADS v2 for PSA grey zone PCa was calculated by receiver operating characteristic (ROC) curve,logistic regression analysis was used to access the risk factors of PCa.Results 15 PCa and 30 BPH patients were in eluded.There was no significant difference between these two groups in age,tPSA,fPSA,f/tPSA,prostate volume and PSA density.The area under ROC curve of PIRADS v2 in diagnosing PCa was 0.932[95 % CI 0.822-0.984],P<0.01.Using a cutoff PIRADS>4,the diagnosis sensitivity was 88.89 %,specificity 87.10 %,and positive predictive value 80 %,negative predictive value 93.10%,respectively.Logistic regression analysis showed that PIRADS v2 score was an independent risk factor for predicting PCa,with a hazard ratio 17.847[3.745-85.078],P<0.01.There was a positive correlation between PIRADS v2score and gleason score,r=0.585,P=0.022.Conclusion PIRADS v2 has a significantly high diagnosis value in diagnosing PSA grey zone PCa and a good correlation with pathology results.

16.
China Pharmacy ; (12): 745-748, 2017.
Article in Chinese | WPRIM | ID: wpr-507688

ABSTRACT

OBJECTIVE:To investigate the effects of fluvastatin combined with aspirin on the related indicators of patients with diabetic lower extremity arteriosclerosis occlusive desease(LEAOD). METHODS:80 patients with diabetic LEAOD were ran-domly divided into control group(40 cases)and observation group(40 cases). Based on convertional treatment,control group was given Rosuvastatin calcium tablet 5 mg+Aspirin enteric-coated tablet 100 mg every evening;observation group was given Fluvas-tatin sodium capsule 40 mg+Aspirin enteric-coated tablet 100 mg. They were treated for 3 months. Fibrinogen,plasma D-dimer, blood viscosity, cholesterol (TC), triglyceride (TG), glycosylated hemoglobin (HbAlc), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), carotid intima-media thickness, compliance situation of TC, TG, HbAlc,LDL-C and HDL-C,cases of carotid intima thickening,plaque formation,stenosis and occlusion before and after treat-ment were observed,and the incidence of adverse reactions was recorded. RESULTS:After treatment,fibrinogen,plasma D-di-mer,blood viscosity,TC,TG,HbAlc,LDL-C and carotid intima-media thickness in 2 groups were significantly lower than before treatment,and observation group was significantly lower than control group;HDL-C in 2 groups was significantly higher than be-fore,and observation group was significantly higher than control group,with statistical significance (P0.05). CONCLUSIONS:Fluvastatin combined with aspirin can significantly improve the pa-tients 'blood viscosity,blood lipid,vascular endothelial function in the treatment of diabetic LEAOD,it can reduce the thrombotic disease risk,and does not increase the incidence of adverse reactions.

17.
China Pharmacy ; (12): 4995-4998, 2017.
Article in Chinese | WPRIM | ID: wpr-664365

ABSTRACT

OBJECTIVE:To investigate the role orientation of pharmacists in China,and to provide legislative suggestions for the formulation of pharmacist law in China. METHODS:Through literature study,the career and evolution process of pharmacists at home and abroad were traced. The obstacles of the role change of pharmacists in China were analyzed to put forward the enlight-enment to the legislation of pharmacist law in China. RESULTS & CONCLUSIONS:Pharmacists in the world had generally en-tered the stage of"taking medication safety of patients as the center"pharmaceutical care,but pharmacists in China were still in the transition stage from"taking drugs as the center"traditional pharmacy to"taking safe medication as the center"clinical phar-macy. The role cognition of pharmacists needed to be improved. Pharmaceutical care should be taken as the role orientation,and the scope of pharmacist law adjustment should be defined;the qualification admittance threshold of pharmacists should be im-proved with the goal of pharmaceutical care-oriented talents;Taking pharmaceutical care as the content,the rights and obligations of pharmacists should be defined. The legislation of pharmacists should be conducted in China to speed up the transformation of pharmacist's role into pharmaceutical care.

18.
Chinese Journal of Biotechnology ; (12): 1091-1100, 2017.
Article in Chinese | WPRIM | ID: wpr-242247

ABSTRACT

Terminators as regulatory signals are typically placed behind the last coding sequence to block the transcription of DNA to RNA and release the transcript. In the present study, the hairpin and the U-rich sequence of the bacteriophage λto terminator were first modified to investigate their effects on termination efficiency and mRNA stability in Bacillus subtilis 168. Compared with the native λto terminator, the terminator variants M3, M11 and M12 showed higher termination efficiency values. Moreover, the variantsM3, M4 and M11 showed significant positive effects on the mRNA stability of the upstream gfp gene. Additionally, insertion of RNase site also increased the mRNA stability. The results of this study suggested that the composition of the hairpin loop is not required for effective intrinsic termination in B. subtilis. Our results also showed that the terminator could also be used as a potential tool for increasing mRNA stability and the corresponding enzyme production in B. subtilis.

19.
Chinese Journal of Biotechnology ; (12): 936-945, 2017.
Article in Chinese | WPRIM | ID: wpr-242217

ABSTRACT

Heparosan is the start point for chemoenzymatic synthesis of heparin and it is of great significance to efficiently synthesize heparosan in microorganisms. The effects of overexpressing key enzyme genes of the UDP-glucuronic acid (UDP-GlcUA) pathway (pgcA, gtaB and tuaD) or the UDP-N-acetyl-glucosamine (UDP-GlcNAc) pathway (glmS, glmM and glmU) on the heparosan production and molecular mass were analyzed in the constructed heparosan-producing Bacillus subtilis ((1.71±0.08) g/L). On this basis, heparosan production was increased to (2.89±0.11) g/L with the molecular mass of (75.90±1.18) kDa through co-overexpressing the tuaD, gtaB, glmU, glmM and glmS genes in shake flask cultivation. In the 3 L fed-batch fermentation, heparosan production was improved to (7.25±0.36) g/L with the molecular mass of (46.66±2.71) kDa, providing the potential for heparosan industrial production.

20.
Article in Chinese | WPRIM | ID: wpr-337399

ABSTRACT

Ethyl carbamate (EC) as a potential carcinogen commonly exists in traditional fermented foods. It is important eliminate urea that is the precursors of EC in many fermented foods, including Chinese Rice wine. On the basis of achieving high-level overexpression of food-grade ethanol-resistant acid urease, we studied the hydrolysis of urea and EC with the recombinant acid urease. Recombinant acid urease showed degraded urea in both the simulated system with ethanol and Chinese Rice wine (60 mg/L of urea was completely degraded within 25 h), indicating that the recombinant enzyme is suitable for the elimination of urea in Chinese Rice wine. Although recombinant acid urease also has degradation catalytic activity on EC, no obvious degradation of EC was observed. Further investigation results showed that the Km value for urea and EC of the recombinant acid urease was 0.7147 mmol/L and 41.32 mmol/L, respectively. The results provided theoretical foundation for realizing simultaneous degradation of urea and EC.


Subject(s)
Oryza , Recombinant Proteins , Metabolism , Urea , Chemistry , Urease , Metabolism , Urethane , Chemistry , Wine
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