The article thinks that the environment of embedded users' clinical scientific research is the key to the subject service.It explains the background and causes of embedded clinical scientific research and analyses the problems exist in subject service of library in hospital.At last,it comes up with some effective measures in solving the subject service of embedded clinical scientific research.

Penicillin expandase, also known as deacetoxycephalosporin C synthase (DAOCS), is an essential enzyme involved in cephalosporin C biosynthesis. To evaluate the catalytic behaviors of penicillin expandase under high penicillin G concentration and to identify mutants suitable for industrial applications, the specific activities of wild-type DAOCS and several mutants with increased activities toward penicillin G were determined by HPLC under high penicillin G concentrations. Their specific activity profiles were compared with theoretical predictions by different catalytic dynamics models. We evaluated the specific activities of wild-type DAOCS and previous reported high-activity mutants H4, H5, H6 and H7 at concentrations ranging from 5.6 to 500 mmol/L penicillin G. The specific activities of wild-type DAOCS and mutant H4 increased as penicillin G concentration increased, but decreased when concentrations of substrate go above 200 mmol/L. Other mutants H5, H6 and H7 showed more complex behaviors under high concentration of penicillin G. Among all tested enzymes, mutant H6 showed the highest activity when concentration of penicillin G is above 100 mmol/L. Our results revealed that the substrate inhibition to wild-type DAOCS' by penicillin G is noncompetitive. Other DAOCS mutants showed more complex trends in their specific activities at high concentration of penicillin G (>100 mmol/L), indicating more complex substrate inhibition mechanism might exist. The substrate inhibition and activity of DAOCS mutants at high penicillin G concentration provide important insight to help select proper mutants for industrial application.
Catalysis ; Intramolecular Transferases ; genetics ; Mutation ; Penicillin G ; pharmacology ; Penicillin-Binding Proteins ; genetics ; Streptomyces ; enzymology ; genetics

Objective To compared analgesic effect of sufentanil and fentanyl in surgery patients during mechanical ventilation, and to explore the rational dosage of analgesic and sedative drugs. Methods A prospective randomized controlled trial was conducted. 600 postoperative critically ill patients underwent mechanical ventilation for 12-72 hours admitted to Department of Critical Care Medicine of the Second Affiliated Hospital of Kunming Medical University from April 2013 to March 2015 were enrolled. They were randomly divided into two groups, sufentanil and fentanyl was used for analgesia respectively, and 300 patients in each group. The initiate dosage of sufentanil and fentanil was 5 μg/h and 50 μg/h, and the dosage was adjusted. A postoperative pain score (Prince-Henry score) of 0-1, and Richmond agitation-sedation scale (RASS) score -1-0 were targeted. 1 mg/kg of propofol was used if patient could not fall in sleep or felt anxious after loading dose of sufentanil (5 μg) or fentanil (50 μg) for 5 minutes. The use of analgesic drugs, the proportion and dosage of propofol was observed in the two groups, and adverse reactions were recorded. Results The mean dose of sufentanil for analgesia was (0.07±0.02) μg·kg-1·h-1, and the mean dose of fentanyl was (0.67±0.12) μg·kg-1·h-1. The patients in the two groups received propofol 40 to 60 mg/h in night, and the use proportion of propofol in sufentanil group was slightly less than that in fentanyl group (25.7% vs. 28.3%), but the difference was not statistically significant (P > 0.05). It was found by subgroup age analysis that, the mean analgesic dose of sufentanil or fentanyl in patients over 80 years old was lower than that in 70-79 years, 60-69 years and < 60 years groups but without statistical significance. There were 11 cases (3.7%) and 21 cases (7.0%) patients suffered from respiratory depression in sufentanil group and fentanyl group, respectively, without statistical significance (P = 0.069). The hemodynamics of patients in two groups was stable during analgesia, and no accidental extubation due to restlessness was found. Conclusions A smaller dose of sufentanil for postoperative patients underwent mechanical ventilation with satisfactory analgesia was (0.07±0.02) μg·kg-1·h-1, but need to be added with 40-60 mg/h and a small dose of propofol to improve anxiety and sleep. The proportion of patients needing propofol addition was slightly lower than that of fentanyl.

Objective To investigate the ipratropium bromide combined with Ambroxol Aerosol Inhalation on preventive effect of pulmonary complications after thoracotomy. Method 115 patients undergoing thoracic operation were randomly divided into 2 groups, 2 groups were given routine nursing before operation, the control group on the basis of Mucosolvan atomization inhalation, the observation group in the conventional nursing combined with ipratropium bromide and atomization inhalation of ambroxol. Results the curative effect of therapeutic group was signiifcantly better than the control group (P<0.05); lung infection in the observation group was significantly lower than that of the control group (P<0.05). Conclusion ipratropium bromide and Mucosolvan aerosol inhalation signiifcantly on the prevention of pulmonary complications after thoracotomy and effect, will help the patient restore.

Objective To explore the bioactive constituents of Gentiana rhodantha for its serum pharmacochemistry research.Methods The blood migrating constituents of Gentiana rhodantha were determined by comparing the HPLC fingerprints of the aqueous extracts,drug contained serum and blank serum.Results Twenty compounds were detected in drug contained serum,four among which were original constituents of Gentiana rhodantha,the rest might be metabolites of the original ingredients.Conclusion These twenty constituents absorbed in blood could be the bioactive components of Gentiana rhodantha.Further studies will be useful to clarify the bioactive constituents and action mechanisms of Gentiana rhodantha.

Objective To compare treatment response according to the PERCIST1.0,RECIST1.1,EORTC,and WHO criteria in patients with colorectal liver metastases (CLM) who received neoadjuvant chemotherapy.Methods A total of 41 CLM patients (27 males,average age 68.48 years;14 females,average age 62.43 years) from January 2010 to September 2013 were included in this retrospective study.PET/CT scan was performed before chemotherapy and after 4-6 cycles′ chemotherapy.The baseline and the sequential follow-up 18F-FDG PET/CT of each patient were evaluated according to the PERCIST1.0,RECIST1.1,EORTC,and WHO criteria.The response was categorized into 4 levels including CR,PR,SD,PD.PET/CT images were used for both metabolic and anatomic evaluation.The concurrent diagnostic CT or MRI images (performed within 1 week of PET/CT) were also utilized when needed.The agreements of criteria were analyzed using Kappa test.The response rate (RR) and disease control rate (DCR) were compared using χ2 test.Results The RR and DCR according to the PERCIST1.0,EORTC and RECIST1.1 criteria were 31.71％(13/41) and 63.41％(26/41),31.71％(13/41) and 60.98％(25/41),17.07％(7/41) and 68.29％(28/41),respectively.The general comparison of PERCIST1.0 and RECIST1.1,EORTC and RECIST1.1 criteria showed good agreements (κ values: 0.711,0.689).Significant difference was not found in the DCR(χ2=2.000,P>0.05) but found in the RR(χ2=6.000,P<0.05) between PERCIST1.0 and RECIST1.1.Difference of DCR between EORTC and RECIST1.1 was not significant(χ2=3.000,P>0.05),while the RR had significant difference(χ2=6.000,P<0.05).The RR and DCR according to WHO criterion were 12.20％(5/41) and 70.73％(29/41),which had a good consistency with those according to PERCIST1.0 criteria (κ=0.629).Significant statistical difference was not found in the DCR(χ2=3.000,P>0.05) but found in the RR(χ2=8.000,P<0.05) between PERCIST1.0 and WHO criteria.Conclusions In evaluating CLM treatment response,anatomical criteria and metabolic criteria have a good consistency.But metabolic criteria are more sensitive for RR evaluating.

Objective: To investigate the safety and efficacy of pingyangmycin (bleomycin A5) sclerotherapy for early peripheral arteriovenous malformations (AVM). Methods: Thirteen cases of early-stage [Schobinger clinical stage (Ⅰ/Ⅱ)] peripheral AVM patients (11 stage Ⅰ and 2 stage Ⅱ patients) aged between 3 months and 51 years were selected between January 2012 and May 2015. Pingyangmycin sclerotherapy injections were administered with B-scan ultrasonography or digital subtraction angiography positioning. All patients underwent relevant supplementary examinations before and after the procedure, and clinical evaluation was performed based on the improvement of the clinical symptoms of the patient and re-examination of lesion by imaging. Results: A total of 88 injections were administered to the 13 patients with an average of 6.7 injections per patient, 3-6 years of follow-up. Based on clinical evaluation, 7 patients were generally cured, 3 patients had significant improvement, 2 patients had partial improvement, and 1 patient had no improvement. Seven patients had different levels of postoperative swelling that resolved on its own. No serious complications occurred. Conclusions Pingyangmycin sclerotherapy for treatment of early peripheral AVM was effective and had few complications. It can control further progression of pathological changes.

3-Deoxy-D-manno-octulosonate 8-phosphate synthase (KDO8PS) is the key enzyme to synthesize eight-carbon sugar in plant and gram-negative bacterial cell wall. To analyze the polymerization and characterization in plant KDO8PS, the candidate gene was cloned from fresh Phyllostachys edulis seedling by RT-PCR. The open reading frame of PeKDO8PS is 876 bp deduced into 291 amino acid residues. The target protein was overexpressed in Escherichia coli induced by IPTG and then lager amount of fusion protein was purified through two-step methods with affinity chromatography and size-exclusion chromatography (SEC). SEC analysis shows that PeKDO8PS protein existed mainly in the form of dime in solution. Glutaraldehyde cross-linking experiments confirmed that the enzyme could form dimers. Further we identified that KDO8PS at high concentration two dimers could form tetramer in aqueous solution by analytical ultracentrifuge (AUC) analysis. The pH of the catalytic reaction was between 4.0 and 9.0, the optimum pH value was 8, the thermal stability range was between 25 and 65 ℃, and the optimum temperature was about 55 ℃. The enzyme activity was inhibited by some metal ions at lower concentrations, especially in the presence of Fe3+metal ion and activated by metal protease inhibitor EDTA at low concentration.

Objective:To improve the quality of 18F-fluorodeoxyglucose (FDG) PET images at different acquisition times through deep learning (DL) PET image reconstruction methods. Methods:A total of 45 patients (20 males, 25 females; age (52.0±13.6) years) with malignant tumors and PET/CT scans from September 2020 to October 2020 in the Department of Nuclear Medicine of the First Hospital of Shanxi Medical University were included in this retrospective study. The short acquisition time 30 s/bed PET images from the raw list mode were selected as the input of DL model. DL image reconstruction model, based on the Unet algorithm, was trained to output imitated PET images with full dose standard acquisition time (3 min). The image quality evaluation and quantitative analysis were carried out for four groups of images: DL images, 30 s, 90 s, and 120 s images, respectively. The quality of PET images in four groups was evaluated using the five-point method. Liver background activities, lesions quantification parameters (maximum standardized uptake value (SUV max), mean standardized uptake value (SUV mean), standard deviation (SD), signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR)), and first-order texture features (skewness, kurtosis, uniformity, entropy) were measured. Kappa test, χ2 test and one-way analysis of variance (least significant difference t test) were used for data analysis. Results:The image quality scores between four groups were highly consistent ( Kappa=0.799, P<0.001). The number of patients with scores≥3 in DL, 30 s, 90 s and 120 s groups were 6, 4, 7 and 8, respectively ( χ2=125.47, P<0.001). The liver SD of DL group was significantly lower than that of 30 s group (0.26±0.07 vs 0.43±0.11; F=3.58, t=-7.91, P<0.05). The liver SNR of DL group was higher than that of 30 s group (11.04±4.36 vs 5.41±1.41; F=10.22, t=5.40, P<0.05). The liver SD and SNR of DL group were similar to those of 90 s group (0.39±0.16, 8.46±3.34; t values: -0.87 and 2.17, both P>0.05). In 18 tumor lesions with high uptake, SNR and CNR of DL group were significantly higher than those of 30 s group (60.21±29.26 vs 38.38±16.54, 22.26±15.85 vs 15.41±9.51; F values: 13.09 and 7.05; t values: 5.20 and 4.04, both P<0.001). There were statistically significant differences among four groups in the first-order texture features ( F values: 4.30-9.65, all P<0.05), but there was no significant difference between DL group and 120 s group ( t values: from -1.25 to 0.15, all P>0.05). Conclusion:DL reconstruction model can improve the quality of short-frame PET images, which meets the needs of clinical diagnosis, efficacy evaluation and radiomics research.

Arabinose-5-phosphate isomerase (KdsD) is the first key limiting enzyme in the biosynthesis of 3-deoxy-D-manno-octulosonate (KDO). KdsD gene was cloned into prokaryotic expression vector pET-HTT by seamless DNA cloning method and the amount of soluble recombinant protein was expressed in a soluble form in E. coli BL21 (DE3) after induction of Isopropyl β-D-1-thiogalactopyranoside (IPTG). The target protein was separated and purified by Ni-NTA affinity chromatography and size exclusion chromatography, and its purity was more than 85%. Size exclusion chromatography showed that KdsD protein existed in three forms: polymers, dimmers, and monomers in water solution, different from microbial KdsD enzyme with the four polymers in water solution. Further, the purified protein was identified through Western blotting and MALDI-TOF MASS technology. The results of activity assay showed that the optimum pH and temperature of AtKdsD isomerase activities were 8.0 and 37 ℃, respectively. The enzyme was activated by metal protease inhibitor EDTA (5 mmol/L) and inhibited by some metal ions at lower concentration, especially with Co²⁺ and Cd²⁺ metal ion. Furthermore, when D-arabinose-5-phosphate (A5P) was used as substrate, Km and Vmax of AtKdsD values were 0.16 mmol/L, 0.18 mmol/L·min. The affinity of AtKdsD was higher than KdsD in E. coli combined with substrate. Above results have laid a foundation for the KdsD protein structure and function for its potential industrial application.
Aldose-Ketose Isomerases ; biosynthesis ; Arabidopsis ; enzymology ; Arabidopsis Proteins ; biosynthesis ; Cloning, Molecular ; Escherichia coli ; metabolism ; Metals ; Pentosephosphates ; Recombinant Proteins ; biosynthesis