Nowadays,small peptides are always expressed in the form of fusion protein.The expression product contains many superfluous amino acids which can affect the biological functions of small peptides even expressed by GST fusion protein expression system.SUMO protease can cut SUMO fusion protein expressed by fusion expression system without any amino acid residues left on target protein thus become a hot topic in this field.Recombinant His-UlP1/pET3c/BL21(DE3)engineering strain was constructed by genetic engineering technology and the expression conditions were optimized in shake flaks.The process of high density fermentation was explored and different purification conditions were detected by chromatography.The results showed that SUMO protease could be expressed well after inducing the engineering strain by IPTG of 1.0mmol/L at 30℃ for 6 hours.The expression level of the strain in fermentation pots could reach 24.3% analyzed by SDS-PAGE.The purity of SUMO protease was more than 98% after further purification by cation exchange chromatography.The yield was 355mg SUMO protease per liter fermentation liquid.Western blot analysis demonstrated that there were immune reactions between IlP1 and 6?His antibodies,so it has established a good foundation for large-scale industralazation in the future.

Hydrolytic amino acids were extracted by acid hydrolysis method, then derivatized with phenyl isothiocyanate (PITC). And the samples were analysed by HPLC on an Ultimate Prime C18 (4.6 mm x 250 mm, 5 microm) column with gradient elution of 0.1 mol x L(-1) sodium acetate buffer solution (adjusted to pH 6. 5)-acetonitrile (93:7) (A) and acetonitrile-water (8:2) (B) at a flow rate of 1.0 mL x min(-1). Column temperature was 40 degrees C and the detected wavelength was 254 nm. Amino acids derivative solution remained stable in 36 hours. The response was linear for 16 amino acids with a correlation coefficient r > 0.999 5. The average recoveries were 98.01% -101.8%. The method is reliable with good accuracy and repeatability, which is useful for the determination of amino acids in Galli Gigerii Endothelium Corneum.
Amino Acids ; analysis ; Animals ; Chickens ; Chromatography, High Pressure Liquid ; Chromatography, Reverse-Phase ; Endothelium ; chemistry ; Gizzard, Avian ; chemistry

Objective To investigate the effect of total flavonoids of Herba Epimodii (HEF) on the expression of core binding factor?1 (Cbfal) in the bone of ovariectomized rats.Methods Fifty-four female SD rats were allocated into 6 groups (9 in each): sham operation group,ovariectomized (OVX) group,OVX followed by three kinds of HEF doses(40,80 and 160 mg?kg~(-1)?d~(-1))and nilestriol (0.1 mg?kg~(-1)?week~(-1))for 12 weeks respectively.Bone mineral density (BMD) of whole body was determined by DEXA.The levels of osteocalcin (BGP) and estradiol (E_2) in serum were measured by radioimmunologic method.All rats were then sacrificed,and total RNA were directly isolated from the skull.Relative quantification of Cbfal mRNA expression was detected by real time quantitative PCR.Results Serum E_2 and BMD of whole body in the OVX group were lower than those in sham group significantly after 12 weeks (both P0.05).Relative quantification of Cbfal mRNA expression in the OVX group was significantly lower than that in sham group after 12 weeks (P

OBJECTIVE: To eludicate the risk factors of mechanical ventilation and prolonged mechanical ventilation in patients with severe multiple injuries. METHODS: Consecutive patients with severe multiple injures who were treated in Peking University People's Hospital Trauma Medical Center between December 2016 and December 2019 were enrolled in this restropective chart-review study. According to mechanical ventilation and ventilatory time, the patients were divided into mechanical ventilation (MV) group and non-mechanical ventilation (NMV) groups, prolonged mechanical ventilation (PMV) group and shortened mechanical ventilation (SMV) groups. Clinical data such as gender, age, base excess, mechanism of injury, Glasgow Coma Scale (GCS), abbreviated injury scale (AIS) and injury severity score (ISS) were collected. To indentify the risk factors of mechanical ventilation and prolonged mecha-nical ventilation, univariate and multivariate Logistic analyses were carried out. RESULTS: In the present study, 112 patients (82 male, 30 female) with severe multiple injuries having a median age of 52 (range: 16-89 years) and a median ISS of 34 (range: 16-66) were enrolled. The primary mechanism of injury was traffic accident injury and falling injury. In the study, 62 and 50 patients were assigned to MV and NMV groups, respectively. Logistic analysis showed that GCS (OR=0.72, 95%CI: 0.53-0.92, P=0.03), base excess (OR=0.56, 95%CI: 0.37-0.88, P=0.002) and multiple rib fracture (OR=1.72, 95%CI: 1.60-2.80, P=0.012) were independent significant risk factors for mechanical ventilation after severe multiple injuries. Within the mechanical ventilation group, 38 and 24 patients were assigned to PMV and SMVgroups, respectively. Compared with the SMV group, the PMV group had a higher ISS and higher rate of severe head trauma. The length of hospital stay of PMV group was longer than that of SMV groups. Meanwhile, the incidence of tracheotomy in PMV group was high. CONCLUSIONS GCS, base excess and rib fracture might be independent risk factors for mechanical ventilation. Higher ISS and lower GCS might prolong the ventilatory time and the length of hospital stay. Meanwhile, the incidence of tracheotomy was high in PMV group because of the longer ventilatory time and poor consciousness.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Injury Severity Score ; Male ; Middle Aged ; Multiple Trauma ; Respiration, Artificial ; Retrospective Studies ; Risk Factors ; Young Adult

ObjectiveTo explore the effect of community education and service of mental health on improvement of people's diathesis.Methods483 subjects from 6 social strata were investigated with the questionnaire to find out the community resident's idea on mental health and need for psychological service.ResultsThere were significant differences in the concepts of criteria of mental health among people in different social strata and ages (P<0.001),and requirements for community education and service of mental health varied from one to another. Forms and contents of community education and service were multiplicate.ConclusionThe community education and service of mental health should be developed widely for improving people's diathesis.

OBJECTIVETo investigate the triterpenoids chemical constituents in rhizome of Alisma gramineum.

METHODSilicon gel chromatography and HPLC techniques were employed for isolation and purification of the constituents, and the structures were elucidated by various spectral analyses such as NMR, MS and IR.

RESULT5 protostane-type tetracyclic triterpenoids were isolated and identified as 16,23-oxidoalisol B(1), 11-deoxyalisol C(2), alisol F(3), 16beta-methoxyalisol B acetate (4), 16beta-hydroxyalisol B acetate (5).

CONCLUSIONAll of these compounds were found from A. gramineum for the first time.

Alisma ; chemistry ; Cholestanones ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Triterpenes ; chemistry ; isolation & purification

Objective To explore the clinical value of serum procalcitonin (PCT)-based antibiotic therapy in the second-classedexacerbations of chronic obstructive pulmonary disease (AECOPD). Methods 240 patients diagnosised as AECOPD were randomized to the PCT group and the control group. Serum PCT levels of patients from the PCT group were measured 1 h after hospitalized and the third, fifth, eighth day respectively. When PCT < 0.1 μg / L, patients will stop taking antibiotics and initiated while PCT≥0.1 μg / L. Antibiotic treatment in the control group was based on guidelines of COPD diagnosis and treatment. Results Duration of antibiotic therapy and hospitalization were respectively 5.6 ± 1.4 and 8.2 ± 1.1 days in the PCT group, 9.2 ± 2.2 and 11.4 ± 2.5 days in the control group (both P < 0.05). Mean costs of hospitalization expensesand antibiotic therapy were 5700 ± 201 and 1650 ± 189) yuan in the PCT group, 6210 ± 220 and 2350 ± 210 yuan in the control group (both P < 0.05). The clinical effective rate, times of exacerbation, one-year ΔFEV1, the 1-year hospitalization rate and time to next exacerbation all showed no significant differences between the two groups. Conclusion PCT-guided antibiotic treatment reduces antibiotic use inthe second-classed acute exacerbations patients.

Objective To provide the candidate antigens for immunological diagnosis by analyzing the expression of nu -cleoprotein ( NP) of Ebola virus. Methods BioSun software was used to predict the NP epitopes. The bridging-PCR was used to synthesize the NP gene. The pBVIL1 vector was used to clone and express the NP gene. Results The 360-739 aa of NP was confirmed to be the dominant antigen by BioSun software. The recombinant NP dominant antigen was expressed in E.coli with molecular weight of 58 ×103.The specificity of ELISA based on recombinant NP was 99.24% (130/131) in negative samples. Conclusions The dominant NP antigen can be potentially used for developing Ebola virus diagnostic reagent.

In this study the chemical constituents of the higher polar sustances from Desmodium caudatum were investigated.The compounds were isolated by using column chromatographies over silicagel, polyamide, ODS, Sephadex LH-20, and preparative HPLC. The structures of these compounds were identified on the basis of NMR and MS spectra. Thirteen compounds were obtained and their structures were identified as vanillin(1), loliolide(2), indole-3-carboxaldehyde(3), salicylic acid(4), swertisin(5), saccharumoside C(6), isosinensin (7), kaempferol 3-O-β-D-glucopyranoside-7-O-α-L-rhamnopyranoside (8), isovitexin (9), vitexin (10), nothofagin(11), resveratroloside (12), and 2"-α-rhamnopyranosyl-7-O-methylvitexin (13). Except for compound 5, the remaining compounds were isolated from D. caudatum for the first time. Compounds 2, 3, 6-8, 11-13 were separated from the genus Desmodium for the first time.
Apigenin ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Fabaceae ; chemistry ; Molecular Structure ; Saponins ; chemistry ; isolation & purification ; Spectrometry, Mass, Electrospray Ionization

To study relevant risk factors of Shenmai injection induced adverse reactions by using Logistic model and ROC curve, and made the prediction for the occurrence of relevant adverse reactions/events. Case data of patients treated with Shenmai injection were collected by using the prospective, multi-center, large-sample, nested-case control method, in order to analyze the risk factors of Shenmai injection-induced adverse reactions/events, establish the logistic model and draw the receiver operating characteristic (ROC) curve for risk factors. During the study, 7632 patients (including 3 477 males and 4 155 females) were included, and eight of them suffered adverse reactions/events. Based on a multi-factor Logistic model analysis, the age (> or = 50 years) (OR = 5.061, 95% CI: 2.197-7.924; P = 0.001), the total number of medication days (OR = -1.020, 95% CI: -l.652 - 0.388; P = 0.002) and the single dose (OR = 0.245, 95% CI: 0.127-0.364; P = 0.000) were significant independent risk factors for Shenmai injection-induced adverse reactions/events. According to the results, ROC curves were drawn with age (> or = 50 years), the total number of days of inedication and single dose; The area under ROC curves the joint predictor (0.9753, 95% CI: 0.9443-1.000, P < 0.005) was larger than that of the other three single indexes, with a higher risk prediction value. The independent risk factors for Shenmai injection-induced adverse reactions/events included the age (> or = 50 years), the total number of days of medication and single dose. In clinical practice, the age (> or = 50 years), the total number of days of medication and the medication dose can be substituted in the joint predictor calculation formula (P = 1 / [1 + e(-(-21.58 + 5.061 x Xage - 1.020 x Xd + 0.245 x X(mL)] to predict the potential adverse reactions of patients and adjust the dosage regimen.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Drug-Related Side Effects and Adverse Reactions ; epidemiology ; etiology ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Female ; Humans ; Infant ; Logistic Models ; Male ; Middle Aged ; Prospective Studies ; ROC Curve ; Risk Factors ; Young Adult