1.Ultrasensitive quantification of trace amines based on N-phosphorylation labeling chip 2D LC-QQQ/MS
Xiqing BIAN ; Yida ZHANG ; Na LI ; Menglin SHI ; Xiaolin CHEN ; Hui-Lu ZHANG ; Jie LIU ; Jian-Lin WU
Journal of Pharmaceutical Analysis 2023;13(3):315-322
Trace amines(TAs)are metabolically related to catecholamine and associated with cancer and neuro-logical disorders.Comprehensive measurement of TAs is essential for understanding pathological pro-cesses and providing proper drug intervention.However,the trace amounts and chemical instability of TAs challenge quantification.Here,diisopropyl phosphite coupled with chip two-dimensional(2D)liquid chromatography tandem triple-quadrupole mass spectrometry(LC-QQQ/MS)was developed to simul-taneously determine TAs and associated metabolites.The results showed that the sensitivities of TAs increased up to 5520 times compared with those using nonderivatized LC-QQQ/MS.This sensitive method was utilized to investigate their alterations in hepatoma cells after treatment with sorafenib.The significantly altered TAs and associated metabolites suggested that phenylalanine and tyrosine metabolic pathways were related to sorafenib treatment in Hep3B cells.This sensitive method has great potential to elucidate the mechanism and diagnose diseases considering that an increasing number of physiological functions of TAs have been discovered in recent decades.
3. Application of closed negative pressure drainage technique in repair of wound of hand trauma infection
Xiaoyan XIONG ; Zhaoguo ZHANG ; Chao LIU ; Xiqing CHENG
Chinese Journal of Primary Medicine and Pharmacy 2019;26(9):1042-1045
Objective:
To explore the application value of closed negative pressure drainage technique in wound healing of hand trauma.
Methods:
From August 2013 to October 2017, 80 patients with hand trauma in the Traditional Chinese Medicine Hospital of Cixi were divided into two groups according to the random principle, with 40 cases in each group.The control group was treated with conventional wound repair, and the observation group was treated with closed negative pressure drainage.The repair effect, healing, secondary operation, antibiotic use, hospitalization, histopathological score and patients' satisfaction were observed.
Results:
The total effective rate of the observation group (97.50%) was obviously higher than that of the control group(80.00%)(χ2=6.13,
4.Establishment and performance evaluation of light-initiated chemiluminescent assay for quantitation of prolactin in human serum
Jie REN ; Shenglin LIU ; Huiqiang LI
Chinese Journal of Clinical Laboratory Science 2019;37(7):495-498
Objective:
To establish an analytical method for serum prolactin (PRL) based on the photoinduced chemiluminescence technology, and evaluate its performance.
Methods:
A pair of PRL monoclonal antibodies were labeled with luminescent nanospheres and biotin respectively, and the double antibody sandwich detection system was formed with the serum prolactin and streptavidin-labeled photosensitive microspheres (universal photosensitive solution) under homogeneous conditions. The performance index and correlation of the detection system were evaluated.
Results:
The precision of intra-assay and within-day (coefficient of variation) of the developed assay were 4.60% and 5.25%, respectively. The functional sensitivity was 2.48 μIU/mL, and its reportable results were ranged from 2.48 to 4 240 μIU/mL. The recovery rates of different PRL calibrators (42.2, 424, 4 240 μIU/mL) added to human sera were ranged from 96.25% to 102.93%. There was no interference from bilirubin<20 mg/dL, hemoglobin<200 mg/dL, triglyceride<3 000 mg/dL and biotin<20 ng/mL. Also, the light-initiated chemiluminescent assay for PRL (PRL-LICA) correlated well with Beckman Unicel Dxi 800 Access 2.
Conclusions
LICA showed effective performance for detecting PRL in human serum, and it could meet the basic requirements of clinical diagnosis.
5.Comparative analysis of the efficacy of endovascular treatment and surgical clipping for intracranial ruptured aneurysms: a single-center retrospective case series study
Dajun LIU ; Fen'gen LI ; Xiqing JIANG
International Journal of Cerebrovascular Diseases 2018;26(9):677-683
Objective To investigate the safety and effectiveness of endovascular treatment and surgical clipping for the treatment of intracranial ruptured aneurysms. Methods From January 2012 to December 2017, patients with ruptured intracranial aneurysm treated at the Department of Neurosurgery, the Second People's Hospital of Taizhou were enrolled retrospectively. The demographics, baseline clinical data,outcomes, and complications were compared between the endovascular treatment group and the surgical clip group. Results A total of 220 patients were enrolled, they aged 55. 1 ±11. 8 years. There were 117 patients in the endovascular treatment group and 103 in the surgical clipping group. There were no significant differences in perioperative complications (26. 2% vs. 19. 6%; χ2 = 1. 340, P = 0. 247), in-hospital mortality (6. 0%vs. 4. 9%; χ2 = 0. 135, P = 0. 713), and good outcomes at discharge (85. 5% vs. 81. 6%; χ2 =0. 614, P = 0. 433) between the two groups. Multivariate logistic regression analysis showed that age (odds ratio [OR] 1. 072, 95% confidence interval [CI] 1. 025-1. 124; P < 0. 001), smoking (OR 6. 325, 95% CI 2. 367-16. 901; P < 0. 001 ), and high World Federation of Neurosurgery Societies (WFNS) grade (OR 5. 218, 95% CI 1. 881-14. 449; P < 0. 001) had significant independent correlation with the poor clinical outcome at discharge. The imaging follow-up data in 155 aneurysms (81 in the endovascular treatment group and 74 in the surgical clipping group) were available. The follow-up time was 14. 3 ± 6. 9 months (range, 6-36 months); 20 aneurysms (12. 9%) had recurrence. There was no significant difference in the recurrence rate of the endovascular treatment group and surgical clipping group (17. 3% vs. 8. 1%; χ2 =2. 900, P = 0. 089). The clinical follow-up data of 188 patients (95 in the endovascular treatment group and 93 in the surgical clipping group) were available. The follow-up time was 15. 5 ± 6. 8 months (range, 6- 36 months). There was no significant difference in the good outcome rate between the endovascular treatment group and surgical clipping group (95. 8% vs. 90. 3%; χ2 = 2. 182, P = 0. 140 ). Multivariate logistic analysis showed that smoking (OR 4. 872, 95% CI 1. 719-13. 872; P < 0. 001 ) and high WFNS grade (OR 3. 512, 95% CI 1. 446-8. 583; P < 0. 001) were the independent risk factor for long-term poor outcome. Conclusion The efficacy and safety of surgical clipping for ruptured intracranial aneurysms were comparable to endovascular treatment. Age, smoking, and WFNS grade were the important factors affecting the outcomes of patients.
6.Distribution and antimicrobial resistance profile of the clinical bacterial strains isolated from Beijing Children's Hospital from 2009 to 2015
Fang DONG ; Yan WANG ; Xiqing LIU ; Jinghui ZHEN ; Hongyan ZHENG ; Mei CHANG ; Yang ZHEN ; Qingxiu ZHANG ; Wenqi SONG
Chinese Journal of Infection and Chemotherapy 2017;17(1):61-70
Objective To investigate the distribution and antimicrobial resistance profile of the common pathogens isolated during the period from 2009 to 2015.Methods All the bacterial strains isolated from pediatric inpatients in Beijing Children's Hospital during the period from 2009 to 2015 were analyzed. Antimicrobial susceptibility was determined by disk diffusion method and Phoenix 100 Automated Microbiology System. Results were analyzed according to the guidelines of CLSI (2014) using WHONET 5.6 software.Results The total strains were 26630. The most common gram-positive isolates were Streptococcus pneumoniae,Staphylococcusaureusand coagulase-negative Staphylococcus (CNS), while the most frequently isolated gram-negative microorganisms were Klebsiella spp.,Pseudomonas aeruginosa and Escherichia coli. The prevalence of S. pneumoniae was up to 25.7 % (4101/15973) in all respiratory tract specimens. About 50.2 % of the S. pneumoniae isolates were not susceptible to penicillin. The prevalence of methicillin-resistant strains was 20.6 % in S. aureus (MRSA) and 87.8 % in coagulase negative Staphylococcus (MRCNS) on average. The prevalence of MRSA increased from 11.1 % in 2009 to 29.8 % in 2015. No S. pneumoniae or staphylococcal strains were found resistant to vancomycin or linezolid. The Enterococcus strains were still highly susceptible to vancomycin and linezolid. Overall 0.3 % of the Enterococcus faecium isolates were resistant to vancomycin. The extended-spectrum beta-lactamases (ESBLs) producing strains accounted for 71.4 % -78.1 % of E. coli and 65.1 % - 76.9 % of K. pneumoniae isolates. The carbapenem-resistant E. coli and K. pneumoniae were reported for the first time in 2010, but in 2014, the strains resistant to carbapenems had increased to more than 7 % in E. coli, and higher than 20 % in K. pneumoniae. In 2015, up to 27.7 % and 25.7 % of P. aeruginosa isolates were resistant to imipenem and meropenem, respectively, and 59.9 % of the A. baumannii isolates were resistant to imipenem and meropenem. Beta-lactamase was positive in 46.3 % of the H. influenzae isolates. Conclusions MRSA and the carbapenem-resistant strains of E. coli,K. pneumoniae and A. baumannii are still on the rise in pediatric inpatients, which poses a serious threat to clinical practice and implies the importance of strengthening infection control.
7.Effects of coupling meglumine cyclic adenylate and the human granulocyte colony-ostimulating factor on rat with diastolic heart failure
Liu LIAN ; Zhang JINGUO ; Xiqing WEI
Chinese Journal of Geriatrics 2017;36(1):74-79
Objectives To explore the clinical effect of coupling meglumine cyclic adenylate (MCA)and the human granulocyte colony-ostimulating factor (G-CSF)on rat with diastolic heart failure(DHF).Methods Totally 60 rats of DHF model were evenly divided into 4 groups according to random number:Control group(n=15,control),Model group(n=15,DHF model),MCA group(n =15,treated with MCA)and MCA+GCSF group(n=15,treated with MCA plus G-CSF).MCA group were administered by intragastric injection of MCA 30 mg/kg/d for 15 d,MCA+G-CSF group were administered by intragastric injection of MCA 30 mg/kg/d and plus G-CSF 100 μg/kg/d for 15 d,while Control group and Model group were given same volume of saline solution.BIOPAC SYSTEM was used to analyze the model establishment.The mRNA levels of GATA-4 and Cx43 were measured by RT-PCR.The protein expressions of GATA-4,Cx43,cTNI and c-kit were measured with western blotting.ELISA and flow cytometry were used to detect cAMP and differentiation rate of bone marrow mesenchymal stem cells (BMSCs),respectively.Results Compared with MCA group,the denaturation degree of myocardial tissues in DHF rat was significantly improved than in MCA+G-CSF group.Moreover,the level of GATA-4 (1.62 ± 0.09),Cx43 (1.02 ± 0.07),cTNI (1.42 ± 0.12),c-kit (0.65±0.02),cAMP(283.67± 18.09)nmol/L and BMSCs cell differentiation rate(38.62 ± 1.52)% in MCA + GCSF group were significantly promoted (all P< 0.05)than in MCA group,GATA-4 (0.82±0.07),Cx43 (0.52±0.05),cTNI(0.86 ± 0.13),c-kit (0.48 ± 0.03),cAMP(198.83 ± 16.03) nmol/L and BMSCs cell differentiation rate (19.82 ± 0.89)%.Conclusions The combination of MCA with G-CSF is significantly improved DHF than single MAC treatment,which may regulate BMSCs differentiation though cAMP/PKA signaling pathways.
8.Preparation and Quality Control of Thermo-sensitive Paclitaxel Liposomes
Zhiping LI ; Xiqing ZHAO ; Shiqing ZHAO ; Hui ZHANG ; Yan LIU ; Ruolei XIAO
Herald of Medicine 2014;(11):1486-1490
Objective To prepare long-circulating temperature-sensitive liposomes with paclitaxel( LTSLP ),develop methods for determination of paclitaxel,related substances and monostearoyl phosphatidylcholine( MSPC ) in LTSLP,and the haemolysis of LTSLP in vitro. Methods HPLC-UV methods for paclitaxel content and related substances and HPLC-CAD method for MSPC in LTSLP were established and validated. Spectrophotometric method was used to determine hemolysis in vitro. Results There was a good linear relationship between peak area and concentration within the range of 60. 39-181. 17μg·mL-1 . Recovery and precision of the method for determination of paclitaxel content met the requirements. Specificity,sensitivity,and system suitability for related substances were consistent with requirements. There was a good linear relationship between peak area and concentration within the range of 1. 5-50. 0μg·mL-1 for the determination of MSPC with good specificity,sensitivity and recovery. Paclitaxel contents in three batches of self-prepared LTSLP were between 90. 0% and 110. 0%,single related substances were below 0. 5% and total impurities were below 2. 0%. There was almost no hemolysis in vitro. Conclusion The methods for determining paclitaxel content,related substances and haemolysis can be used to assess the quality of LTSLP. Self-produced LTSLP consistently meet the quality standards.
9.β-amyloid protein up-regulates the expression of the receptor for advanced glycation end products by increasing ROS production.
Weina KONG ; Jia ZHANG ; Weijuan GAO ; Qingtao LIU ; Liming ZHOU ; Xiqing CHAI
Journal of Southern Medical University 2013;33(8):1132-1136
OBJECTIVETo investigate the mechanism of β-amyloid protein (Aβ) in regulating the expression of the receptor for advanced glycation end products (RAGE).
METHODSAβ1-40 was injected into the bilateral hippocampus of rats, and 3 weeks later, the levels of reactive oxygen species (ROS) production were detected by flow cytometry. The expressions of RAGE, reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (gp9l(phox) and p47(phox)), nuclear factor-κB (NF-κB), and inhibitor of κB (IκB) were measured by Western blotting.
RESULTSInjection of Aβ1-40 caused a significant increase in the expressions of RAGE, gp9l(phox), p47(phox), phospho-p47(phox), phospho-IκBα, NF-κB and phospho-NF-κB in rat hippocampus but decreased the level of IκBα. Aβ1-40 injection also resulted in a significantly increased content of ROS in the hippocampus of the rats.
CONCLUSIONAβ up-regulates the expression of RAGE in rat hippocampus via NADPH/ ROS/NF-κB signaling pathway.
Amyloid beta-Peptides ; adverse effects ; Animals ; Hippocampus ; metabolism ; Male ; Membrane Glycoproteins ; metabolism ; NADP ; metabolism ; NADPH Oxidase 2 ; NADPH Oxidases ; metabolism ; NF-kappa B ; metabolism ; Oxidative Stress ; Peptide Fragments ; adverse effects ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Signal Transduction ; Up-Regulation
10.Effects of simvastatin on the expression of NADPH oxidase subunits mRNA in high glucose-induced injury of cardiomyocytes in neonatal rat
Xiqing WEI ; Hongyong TAN ; Jinguo ZHANG ; Hongsheng ZHANG ; Yu CHEN ; Lingai HU ; Zhihua LIU
Chinese Journal of Endocrinology and Metabolism 2012;28(5):419-422
Ventricular myocytes from hearts of the neonatal SD rats were treated with 10-7,10-6,and 10-5 mol/L simvastatin for 72 hours under high glucose condition. Compared with control group,the viability of cadiomyocyte was significantly lower in high glucose group (P<0.01 ).The activity of lactate dehydrogenase,the relative expressions of NADPH oxidase subunits p22phox,p47phox mRNA,and reactive oxygen species level in the high glucose group were higher than those of control group ( all P<0.05).lndexes mentioned above were significantly improved by simvastatin treatment in a dose-dependent manner.These results suggest that simvastatin ameliorates high glucose-induced injury of cardiomyocytes via increasing the expression of NADPH oxidase mRNA.

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