Survivin expresssion in embryo spleen, embryo kidney, embryo liver and in many cancer tissues was determined by RT-PCR, while not in the health liver tissue. Construct the engineered Escherischia coli expressing human survivin and identify the expressed human survivin by Westen-blot. The combination activity of Survivin and RhSmac was determined in vitro. L929 cells transferred with Survivin can survive longer than which transferred with BSA.

Objective: To clone the cDNA of human sPLA2-IIA,construct the engineered Escherischia coli expressing human sPLA2-IIA and identify the expressed human sPLA2-IIA. Methods: Total RNAs were purified from human fetal spleen. The cDNA of human sPLA2-IIA was cloned by RT-PCR and inserted into plasmid pET32a(+) between NcoI and EcoRI sites for expressing the recombinant human sPLA2-IIA in Escherischia coli BL21(DE3). The recombinants were screened by SDS-PAGE. The engineered Escherischia coli expressing trxA-human sPLA2-IIA fusion protein was established. The expressed human sPLA2-IIA exists in the form of inclusion body and accounts for about 25% of the total proteins of Escherischia coli BL21(DE3). Conclusion: the engineered E. coli methods are suitable for preparing plenty of human sPLA2-IIA which has laid base for the large-scale expression,purification and basic studies of human sPLA2-IIA.

To analyze the regularity in combined medication with Xiyanping injection (Xiyanping for short) in the real world by as- sociation rules. Totally 5 822 patients using Xiyanping injection was collected from the 18 Class III Grade I hospitals nationwide to study the combined medication information of the patient with lung infection and make the analysis by using association rules and Apriori. According to the results, major drugs combined with Xiyanping in treatment of lung infection included compound amino acid, inosine, coenzyme A, cytidine triphosphate, vitamin C. Common drugs combined with Xiyanping can be divided into 5 categories: nutrition support therapy (vitamin C, compound amino acid) , coenzymes (coenzyme A, cytidine triphosphate, inosine), expectorants and antiasthmatics (ambroxol, salbutamol, doxofylline), hormones (dexamethasone, budesonide), antibiotics (mainly cefminox). The main combined medicines mostly conformed to the regularity for drugs treating lung infection. In addition, there were two most common medical combination models: the model for Xiyanping combined a single medicine is Xiyanping + nutrition support therapy, while the model for Xiyanping combined two or more than two medicines is Xiyanping + nutrition support therapy + coenzyme. Pharmacologically, Xiyanping is mostly combined with western medicines with similar pharmacological effects to substitute or supplement the antibiotic effect in treating lung infection. However, further studies shall be conducted for the safety and rationality of the combined medication based on clinical practices, in order to provide reference for clinical medication.
Adult ; Anti-Bacterial Agents ; administration & dosage ; Ascorbic Acid ; administration & dosage ; Cephamycins ; administration & dosage ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hospital Information Systems ; Humans ; Lung Diseases ; drug therapy ; Male ; Middle Aged ; Young Adult

OBJECTIVETo study the changes of amino acids in cerebral spinal fluid (CSF) in children with spastic or athetotic cerebral palsy (CP) by examining CSF levels of glutamic acid (Glu), gamma-aminobutyric acid (GABA) and aspartate (ASP).

METHODSCSF samples were obtained from 13 children with spastic CP, from 14 children with athetotic CP, and from 10 children without central nervous system and infectious diseases (control group). CSF levels of Glu, GABA and ASP were determined by high-performance liquid chromatography.

RESULTSCSF levels of GABA, ASP and Glu in the control group were 13.04+/-2.19, 10.21+/-0.45 and 8.41+/-2.26 micromol/L, respectively. Compared with the control group, CSF GABA levels in the spastic and the athetotic CP groups (8.02+/-2.03 and 10.01+/-2.68 micromol/L respectively) significantly decreased (P<0.01), whereas CSF levels of Glu (20.99+/-8.15 and 28.77+/-17.62 micromol/L respectively) and Asp (13.53+/-3.93 and 14.02+/-2.88 micromol/L respectively) in the spastic and the athetotic CP groups significantly increased (P<0.01). There were statistical differences in the GABA level between the spastic and the athetotic CP groups (P<0.05). In children with spastic CPCSF Glu level was positively correlated to muscle tension.

CONCLUSIONSCSF excitatory amino acid levels increased, while CSF inhibitory amino acid levels decreased in children with CP. There were differences for CSF amino acid levels in different types of CP. The changes of amino acid levels may contribute to the pathogenesis of CP.

Amino Acids ; cerebrospinal fluid ; Cerebral Palsy ; cerebrospinal fluid ; physiopathology ; Child, Preschool ; Chromatography, High Pressure Liquid ; Female ; Humans ; Male ; Muscle Tonus

Objective: To investigate the coagulation function in patients with acute-on-chronic liver failure (ACLF) patients using thromboelastography (TEG), and to comprehensively and dynamically evaluate patients’ bleeding and coagulation status. Methods: The clinical data of ACLF patients were collected, and TEG was used to evaluate whole blood clotting kinetics in these patients. Routine biochemical parameters were measured, and complications were evaluated. The t-test was used for comparison of continuous data, the chi-square test was used for comparison of categorical data, and the Pearson correlation coefficient was used for correlation analysis. P < 0.05 was considered statistically significant. Results: A total of 60 patients (39 male and 21 female patients) were enrolled, with a mean age of 47.20±16.20 years. The TEG results showed that all patients had normal thrombokinetics, but TEG parameters were correlated with coagulation function, markers of systemic inflammatory response syndrome, laboratory markers, and prognosis. The patients with a higher R value had higher risks of infection (6.23±2.91 vs 4.74±1.12, P = 0.009), hepatorenal syndrome (5.64±2.54 vs 3.21±1.43 P < 0.01), and bleeding (6.71±3.51 vs 4.80±1.63, P = 0.01), and the patients with a lower K value (0.72±1.36 vs 1.64±1.43, P = 0.02), an increased α-angle (63.33°±10.02° vs 56.62°±12.13°, P = 0.03), and an increased MA (56.83±11.07 vs 50.40±10.81, P = 0.03) had increased risks of hepatic encephalopathy. Conclusion ACLF patients have low coagulation function, and TEG truly reflects the "rebalance" status of this low level. Abnormal TEG parameters suggest increased risk of complications in these patients, indicating a poor prognosis.

In this work,a new pyrylium derivatization-assisted liquid chromatography-mass spectrometry(LC-MS)method was developed for metabolite profiling of the glutathione anabolic pathway(GAP)in cancer tissues and cells.The pyrylium salt of 6,7-dimethoxy-3-methyl isochromenylium tetrafluoroborate(DMMIC)was used to label the amino group of metabolites,and a reductant of dithiothreitol(DTT)was employed to stabilize the thiol group.By combining DMMIC derivatization with LC-MS,it was feasible to quantify the 13 main metabolites on the GAP in complex biological samples,which had good linearity(R2=0.9981-0.9999),precision(interday precision of 1.6％-19.0％and intraday precision of 1.4％-19.8％)and accuracy(83.4％-115.7％).Moreover,the recovery assessments in tissues(82.5％-107.3％)and in cells(98.1％-118.9％)with GSH-13C2,15N,and Cys-15N demonstrated the reliability of the method in detecting tissues and cells.Following a methodological evaluation,the method was applied successfully to investigate difference in the GAP between the carcinoma and para-carcinoma tissues of esophageal squamous cell carcinoma(ESCC)and the effect of p-hydroxycinnamaldehyde(CMSP)on the GAP in KYSE-150 esophageal cancer cells.The results demonstrate that the developed method provides a promising new tool to elucidate the roles of GAP in physiological and pathological processes,which can contribute to research on drugs and diseases.

Objective To investigate the antibacterial resistance in nationwide's tietiary hospitals and understand the trend of antimicrobial resistance.Methods All the clinical isolates were collected from 18 hospitals and the minimal inhibitory concentrations (MICs) were tested using agar dilution method recommended by Clinical and Laboratory Standards Institute (CLSI) in central laboratory.The susceptibilities of isolates to antimicrobial agents were determined by using CLSI or European Committee on Antimicrobial Susceptibility Testing (EUCAST) 2017 guideline.Results A total of 4333 pathogenic isolates from 18 tertiary hospitals in 18 cities nationwide over the period from July 2015 to June 2016 were studied.Based on the MIC results,Escherichia coli and Klebsiella pneumoniae showed extended spectrum β-lactamase (ESBLs) phenotype rates of 59.4％ and 27.5％,respectively;decreased by 7 to 10 percentage points comparing the last time.Carbapenems,amikacin,moxalactam,β-lactam/β-lactamase inhibitor combinations,tigecycline,and fosfomycin displayed desirable antibacterial activity against Enterbacteriaceae,but a significant increasing of carbapenems resistance Klebsiella pneumoniae were noted.For non-fermenting Gram-negative isolates,resistance rate of Pseudomonas aeruginosa and Acinetobacter baumannnii to imipennnem were 29.5％ and 69.8％ and multidrug-resistant (MDR) detection rate were 35.6％ and 78.3％,extensively drug-resistant (XDR) were 10.2％ and 72.5％,respectively.Klebsiella pneumoniae isolated from children were more resistant to β-lactam than those from adults and the old people,so bacterial resistance in children is an important problem in China.Conclusion Though the decline of ESBLs detection rate,carbapenem non-susceptible Klebsiella pneumoniae rates continued to increase,which should be paid more attention.

Objective To investigate the gram-positive coccus resistance in nationwide's tietiary hospitals and understand the trend of antimicrobial resistance.Methods All the clinical isolates were collected from 18 hospitals and the minimal inhibitory concentrations (MICs) were tested using agar/broth dilution method recommended by Clinical and Laboratory Standards Institute (CLSI) in central laboratory.The susceptibilities of isolates to antimicrobial agents were determined by using CLSI or European Committee on Antimicrobial Susceptibility Testing (EUCAST) 2017 guideline.Results A total of 2301 Gram-positive cocci isolated from 18 hospitals in 18 cities nationwide were studied.Based on the MIC results,the prevalence of methicillin resistant Stapylococcus aureus (MRSA) and methicillin resistant Stapylococcus epidermidis (MRSE) were 39.9％ and 86.6％ respectively.No vancomycin insensitive Staphylococcus was detected.Staphylococcus aureus were 100％ susceptibile to linezolid and teicoplanin,but resistant or insensitive for drugs other than vancomycin were observed among Coagulase Negative Staphylococci (CoNS).Antibiotic resistance rate of Enterococcus faecalis and Enterococcusfaecium to ampicillin were 4.5％ and 85.1％.The detectation rate of vancomycin resistant Enterococcus(VRE) was 2.1％.Nonsusceptibility rate of Enterococcus faecalis to linezolid was 7.8％,showing slight increase than last time.The prevalence of penicillin nonsusceptible Streptococcus pneumoniae (PNSSP) was 6.6％ based on non-meningitis and parenteral administration criterion;while for cases of oral penicillin,the rate was 70.0％,was as flat as last time.There were no significant differenees of resistance rates of Stapylococcus aureus,Stapylococcus epidermidis Enterococcus faecalis and Enterococcus faecium among various groups such as different department,age,or specimen source.Conclusion Compared with past surveillance result,VRE detection ratio was steady,while MRSA detection ratio decreased.The emergence of resistance and non-susceptible strains to new antibiotics such as linezolid,tigecycline and daptomycin should be payed more attention.

Objective This study aimed to investigate the potential relationship between urinary metals copper (Cu), arsenic (As), strontium (Sr), barium (Ba), iron (Fe), lead (Pb) and manganese (Mn) and grip strength. Methods We used linear regression models, quantile g-computation and Bayesian kernel machine regression (BKMR) to assess the relationship between metals and grip strength.Results In the multimetal linear regression, Cu (β=-2.119), As (β=-1.318), Sr (β=-2.480), Ba (β=0.781), Fe (β= 1.130) and Mn (β=-0.404) were significantly correlated with grip strength (P < 0.05). The results of the quantile g-computation showed that the risk of occurrence of grip strength reduction was -1.007 (95％ confidence interval:-1.362, -0.652; P < 0.001) when each quartile of the mixture of the seven metals was increased. Bayesian kernel function regression model analysis showed that mixtures of the seven metals had a negative overall effect on grip strength, with Cu, As and Sr being negatively associated with grip strength levels. In the total population, potential interactions were observed between As and Mn and between Cu and Mn (Pinteractions of 0.003 and 0.018, respectively).Conclusion In summary, this study suggests that combined exposure to metal mixtures is negatively associated with grip strength. Cu, Sr and As were negatively correlated with grip strength levels, and there were potential interactions between As and Mn and between Cu and Mn.

To study the binding capacity of active ingredients of Daidai lipid-lowering flavonoid extract and plasma protein,investigate the ways to improve the traditional formula for calculating protein binding rates based on ultrafiltration,and increase the stability and reliability of the experimental results. UPLC-MS/MS was used to establish a quantitative analysis method for simultaneous determination of active ingredients( neohesperidin and narngin) in ultrafiltrate. The protein binding rates were calculated by the traditional ultrafiltration formula. The correction factors( F) were introduced later,and the binding rates calculated with the correction factors were compared with those without the correction factors. The binding capacity of the extract and plasma protein was evaluated. The quantitative analysis method established by UPLC-MS/MS had a good specificity. The standard curve and linear range,method accuracy,precision and lower limit of quantitation all met the requirements. The method met the requirement for quantitative detection of the active ingredients in ultrafiltrate after the rat plasma was filtrated in the ultrafiltration tube. Under the experimental conditions,the binding rates of both active ingredients( neohesperidin and narngin) were higher than 90%. The active ingredients and rat plasma protein were bound in a concentration-dependent manner,with statistically significant differences( P<0. 01). There was no statistically significant difference between the protein binding abilities of the two active ingredients with rat plasma protein. Therefore,the active ingredients of Daidai lipid-lowering flavonoid extract had a relatively strong binding strength with rat plasma protein,and they were bound in a concentration-dependent manner. Additionally,when calculating protein binding rates by the traditional ultrafiltration formula,the correction factors could be introduced to effectively reflect the errors of multiple ingredient groups in traditional Chinese medicine extracts.This correction method could provide a reference thinking and practical reference for the improvement of the determination method of the traditional Chinese medicine plasma protein binding ability based on ultrafiltration.
Animals ; Blood Proteins ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacology ; Flavonoids ; pharmacology ; Hypolipidemic Agents ; pharmacology ; Lipids ; Rats ; Reproducibility of Results ; Tandem Mass Spectrometry