ABSTRACT
Decoction is a classical dosage form of traditional Chinese medicines. In the process of decocting, various complex components produce physical interactions and chemical reactions, among which physical interactions include van der Waals force, hydrogen bond, electrostatic interaction, π-π stacking, etc., and chemical reactions include Maillard reaction, oxidation reaction, hydrolysis reaction, degradation reaction, polymerization reaction, etc. New substances and original ingredients from chemical reactions can be further activated. These effects form the basis of particle formation in the broth. The sizes of the particles in decoctions range from nanoscale to micron scale, mostly composed of polysaccharide, protein matrix, wrapped in water insoluble molecules, can increase the dispersion of insoluble components and the stability of unstable components, as well as reduce the volatile components and toxic components of volatile components, and ultimately achieve the purpose of efficient absorption and toxicity reduction. From the angle of physical change and chemical reaction in the process of decoction, this paper expounds the formation mechanism of particles in decoction, expounds the research method of particles, analyzes the components in particles and the interaction between components, and then explains the pharmacodynamic characteristics of traditional Chinese medicine decoction, which provides the foundation for the modernization of Chinese decoction.
ABSTRACT
As an edible eukaryotic microorganism, Saccharomyces cerevisiae has the characteristics of high safety, rapid proliferation, low cost, easy transformation, etc. It has been widely used to produce vaccines, antibodies, insulin, etc. Up to now, yeast components, such as cell wall and yeast microcapsules, have been widely used in the treatment of tumors, inflammatory virus infection, post-traumatic osteoarthritis and other diseases. Among them, the components of yeast cell membrane are relatively simple and stable, which are easy to be extracted on a large scale. Therefore, yeast cell membrane material was used to construct yeast membrane vesicle nanosystem, and its biomedical application was preliminarily explored. In this study, Saccharomyces cerevisiae membrane vesicle (SMV) was prepared by co-extrusion method, and the particle size and surface potential of SMV, drug loading and release characteristics, stability, cell safety, and in vitro therapeutic effect were investigated. The results showed that the average particle size of SMV was 185.1 nm. Curcumin and silica nanoparticles were effectively encapsulated by co-incubation and ultrasonic methods, and the characteristics of cell membrane proteins were maintained. Moreover, SMV had good stability and biocompatibility. In addition, SMV could be effectively uptaken by macrophages RAW 264.7, and curcumin loaded SMV could effectively eliminate reactive oxygen species (ROS). In conclusion, the yeast plasma membrane vesicles prepared in this study could effectively deliver curcumin drugs and encapsulate nanoparticles, and could be effectively absorbed by macrophages and effectively eliminate ROS, providing new ideas and new methods for biomedical applications of yeast membrane materials.
ABSTRACT
The shortage of donor corneal tissue worldwide has led to extensive research for alternate corneal equivalents utilizing tissue engineering methods. We conducted experiments using Poly D, L lactic acid polymer along with a copolymer (Eudragit) in varying concentrations to create a biodegradable scaffold suitable for in vitro growth of corneal epithelial stem cells. It was found that stable, spherical, and porous microparticles can be prepared by combining PDLLA and Eudragit RL100 polymers in the ratio of 90:10 and 70:30. The microparticles can then be fused to form scaffold membranes with porous architecture and good water retention capacity at room temperature using methanol, which can withstand handling during transplantation procedures. The scaffolds made using a 70:30 ratio were found to be suitable for the promotion of growth of laboratory corneal epithelial stem cell lines (SIRC cell lines). This innovation can pave way for further developments in corneal stem cell research and growth, thus providing for viable laboratory-derived corneal substitutes.
ABSTRACT
The present study aims to correlate the sample-to-cutoff ratios (S/CO) distributions of reactive results for HTLV-1/2 antibodies with the detection of proviral DNA in a population of blood donor candidates. It was carried out a retrospective data search of 632 HTLV-1/2 reactive samples, submitted to confirmatory testing from January 2015 to December 2019. Serological screening was performed by chemiluminescent microparticle immunoassay Architect rHTLV-I/II, whereas confirmatory testing was performed by in-house real-time polymerase chain reaction method. 496 out of 632 samples (78%) had undetectable HTLV-1/2 proviral DNA and 136 (22%) had detectable proviral DNA. HTLV infection was not confirmed in any individual for whom the S/CO ratio value was <4, and proviral DNA detection rates gradually escalated as S/CO ratio values increased. The sensitivity and predictive positive value found for the Architect rHTLV-I/II was 100% and 22%, respectively. The receiver operating characteristic (ROC) curve analysis showed that the optimal S/CO ratio value for predicting the presence of HTLV-1/2 was 18.11. High S/CO ratios were more associated with the detection of proviral DNA. The S/CO ratio value <4 suggests excluding true HTLV infection and the risk of blood transmission (AU).
O estudo tem como objetivo correlacionar às distribuições das razões sample-to-cutoff (S/CO) de resultados reagentes para anticorpos HTLV-1/2 com a detecção de DNA proviral em uma população de candidatos à doação de sangue. Realizou-se uma busca retrospectiva de dados de 632 amostras reagentes para HTLV-1/2 submetidas à testagem confirmatória entre janeiro de 2015 a dezembro de 2019. A triagem sorológica foi realizada pelo imunoensaio quimioluminescente de micropartículas Architect rHTLV-I/II, enquanto o teste confirmatório foi realizado pelo método de PCR em tempo real in-house. 496 de 632 amostras (78%) apresentaram DNA proviral indetectável e 136 (22%) apresentaram DNA proviral detectável. A infecção por HTLV não foi confirmada em nenhum indivíduo com valor de S/CO <4 e as taxas de detecção de DNA proviral escalonaram gradualmente à medida que as razões S/CO aumentaram. A sensibilidade e valor preditivo positivo encontrados para o Architect rHTLV-I/II foram 100% e 22%, respectivamente. Utilizando análise de curva ROC, o valor de razão S/CO ideal para predizer a presença de DNA proviral foi de 18,11. Razões S/CO elevadas foram mais associadas à detecção de DNA proviral. Em suma, o valor de S/CO <4 sugere a exclusão de infecção por HTLV e o risco de transmissão pelo sangue (AU).
Subject(s)
Blood Donors , Immunoassay , Human T-lymphotropic virus 1 , Human T-lymphotropic virus 2 , Real-Time Polymerase Chain Reaction , InfectionsABSTRACT
The dense extracellular matrix (ECM) of the tumor severely limits the deep penetration of nanomedicine and weakens its anti-tumor effect. Based on this, the yeast vesicle biomimetic nanomedicine with active deep penetration ability of tumor tissue was designed and developed for enhanced tumor therapy. Results of characterization showed that the yeast cell vesicles (YCV) displayed a spherical morphology with diameter of around 100 nm and was well dispersed. Then the chemotherapeutic drug doxorubicin (DOX) was selected as a model drug, and DOX was loaded into YCV to obtain YCV/DOX through electrostatic interaction, the encapsulation efficiencies of DOX were calculated as 82.5%. The drug release profile of YCV/DOX implied that DOX release showed a manner of pH-dependent, it may be that pH has affected the electrostatic effect of YCV and DOX. Compared with liposomes (Lipo), in vitro cell experiments showed that YCV from natural sources had stronger permeability in three-dimensional multicellular spheres. It is speculated that the mechanism may be good deformation capacity of YCV. A 4T1 xenograft tumor model was established to evaluate the therapeutic efficacy of YCV/DOX. The results suggested that YCV/DOX has stronger tumor tissue penetration ability and could effectively inhibit the tumor growth. All animal experiments were performed in line with national regulations and approved by the Animal Experiments Ethical Committee of Zhengzhou University. This study brings new ideas for the development of biomimetic nanomedicine to overcome the ECM of solid tumors.
ABSTRACT
Blood is an important component of living organism which is responsible for material transportation. The microenvironment of blood flow plays an important role in physiological and pathological processes of angiogenesis and cardiac development, erythrocytes aggregation and blood viscosity, tumor metastasis and atherosclerosis. Besides, micro-fluid environment significantly affects drug delivery, cell screening, and artificial organ design. Thus, the measurement and quantitative analysis of micro-fluid contribute to the biomedical engineering filed. Micro-particle imaging velocimetry (Micro-PIV) combines conventional PIV with microscopy technique. Correlation analysis is conducted in two groups of images captured by high speed camera at different time intervals, and the velocity profiles in micro-fluid environment are successfully measured. Compared with other velocity measurement methods, Micro-PIV has high temporal resolution and spatial resolution. The main setup of Micro-PIV and its principle analysis method were introduced in this review. Recent studies of Micro-PIV applications in biomedical engineering field were then summarized. Moreover, the drawbacks of Micro-PIV technique and prospect of its applications were discussed.
ABSTRACT
Objective:To clarify the characteristics of renal cortical microcirculation and its relationship with the expression of plasma endothelial microparticle (EMP) in septic rats, and to evaluate the effect of Xuebijing injection as an adjuvant therapy of antibiotics on septic AKI.Methods:The 8-10 weeks old specific pathogen free (SPF) male Sprague-Dawley (SD) rats were divided into sham operation group (Sham group), positive drug control group and Xuebijing group by the random number table method, with 10 rats in each group. The cecal ligation and puncture (CLP) with large ligation (ligated 75% of the cecum) was used to prepare a rat high-grade sepsis model; in the Sham group, the cecum was stretched without ligation or puncture. Due to the high mortality of CLP with large ligation, Xuebijing injection (4 mL/kg, 12 hours per time) and imipenem/cilastatin injection (90 mg/kg, 6 hours per time) were administered to the rats in the Xuebijing group via the tail vein immediately after the model was produced. Normal saline and imipenem/cilastatin were administered to the rats by the same methods in the positive drug control group. The rats in the Sham group were treated with the same volume of normal saline as any of the other two groups at the same frequency. At 48 hours after model reproduction, the mean arterial pressure (MAP) and blood lactic acid (Lac) of the rats were measured. The renal cortical microcirculation was monitored by using side stream dark-field imaging. Renal hypoxia signals were assessed by pimonidazole chloride immunohistochemistry. Plasma EMP levels were determined by using flow cytometry, and then the correlation between EMP and microcirculation parameters of renal cortex was analyzed. At the same time, the serum creatinine (SCr) was measured, and the renal injury score (Paller score) was used to evaluate the severity of renal tissue pathological damage.Results:Compared with the Sham group, perfused vessel density (PVD), microvascular flow index (MFI) and MAP in the positive drug control group and the Xuebijing group decreased significantly, the positive expression of hypoxia probe (pimonidazole) increased, Lac, EMP, Paller score and SCr increased significantly. However, compared with the positive drug control group, the renal cortical microcirculation in the Xuebijing group was improved significantly, PVD and MFI were increased significantly [PVD (mm/mm 2): 16.20±1.20 vs. 9.77±1.12, MFI: 2.46±0.05 vs. 1.85±0.15, both P < 0.05], Lac was reduced significantly (mmol/L: 4.81±1.23 vs. 6.08±1.09, P < 0.05), MAP increased slightly [mmHg (1 mmHg = 0.133 kPa): 84.00±2.00 vs. 80.00±2.00, P > 0.05], suggested that Xuebijing injection improved renal microcirculation perfusion in septic rats, and this effect did not depend on the change of MAP. The positive expression of pemonidazole in renal cortex of the Xuebijing group was significantly lower than that of the positive drug control group [(35.89±1.13)% vs. (44.93±1.37) %, P < 0.05], suggested that Xuebijing injection alleviated renal hypoxia. The plasma EMP levels of rats in the Xuebijing group were significantly lower than those in the positive drug control group (×10 6/L: 3.49±0.17 vs. 5.78±0.22, P < 0.05), and the EMP levels were significantly negatively correlated with PVD and MFI ( r values were -0.94 and -0.95, respectively, both P < 0.05), indicated that the increase of plasma EMP was highly correlated with renal microcirculation disorder, and Xuebijing injection inhibited the increase of plasma EMP levels. The Paller score in the Xuebijing group was significantly lower than that in the positive drug control group (46.90±3.84 vs. 62.70±3.05, P < 0.05), and the level of SCr was also significantly lower than that in the positive drug control group (μmol/L: 121.1±12.4 vs. 192.7±23.9, P < 0.05), which suggested that Xuebijing injection relieved kidney injury and improved renal function in septic rats. Conclusion:As an adjuvant therapy of antibiotics, Xuebijing injection could inhibit the expression of plasma EMP in rats with sepsis, improve renal cortex microcirculation, and reduce kidney injury.
ABSTRACT
OBJECTIVE: To construct a novel daunorubicin-loaded microparticles (DNR-MPs) drug delivery system, and study its proliferation inhibiton effect on leukemia cells. METHODS: Both DNR-MPs-IDL(DNR microparticles with intracellular drug loading method) and DNR-MPs-EDL (DNR microparticles with extracellular drug loading method) were prepared from HL-60 cells, and the average particle size, Zeta potential and drug loading efficiency of DNR-MPs were measured. The uptake of DNR-MPs by HL-60 cells was analyzed by confocal laser scanning microscopy (CLSM) and flow cytometry (FCM). The inhibition effect of DNR-MPs on the proliferation of HL-60 cells was evaluated by MTT assay. RESULTS: There was no significant difference in the average particle size and Zeta potential of the two DNR-MPs. The drug loading efficiency of DNR-MPs-EDL was higher compared with DNR-MPs-IDL. The uptake results showed that the two DNR-MPs significantly increased the drug uptake compared with free DNR (P < 0.05). DNR-MPs- IDL showed a higher drug uptake by the cells than DNR-MPs-EDL did (P < 0.05). They also exhibited an enhanced inhibition effect on the proliferation of HL-60 cells compared with DNR (P < 0.05). CONCLUSION: The different ways of preparation of drug-loaded microparticles can cause varying anti-tumor effect. Microparticles can significantly augment the anti-leukemia efficacy of DNR, indicating a promising drug carrier for the therapy of leukemia.
ABSTRACT
Microvesicles (MVs, also known as microparticles) are small vesicles that originate from plasma membrane of almost all eukaryotic cells during apoptosis or activation. MVs can serve as extracellular vehicles to transport bioactive molecules from their parental cells to recipient target cells, thereby serving as novel mediators for intercellular communication. Importantly, more and more evidence indicates that MVs could play important roles in early pathogenesis and subsequent progression of cardiovascular and metabolic diseases. Elevated plasma concentrations of MVs, originating from red blood cells, leukocytes, platelets, or other organs and tissues, have been reported in various cardiometabolic diseases. Circulating MVs could serve as potential biomarkers for disease diagnosis or therapeutic monitoring. In this review, we summarized recently-published studies in the field and discussed the role of MVs in the pathogenesis of cardiometabolic diseases. The emerging values of MVs that serve as biomarker for non-invasive diagnosis and prognosis, as well as their roles as novel therapeutic targets in cardiometabolic diseases, were also described.
Subject(s)
Humans , Biomarkers , Metabolism , Cardiovascular Diseases , Blood , Diagnosis , Therapeutics , Cell Communication , Cell-Derived Microparticles , Metabolism , Metabolic Diseases , Blood , Diagnosis , TherapeuticsABSTRACT
Background@#Microparticles (MPs) are small extracellular plasma membrane particles shed by activated and apoptotic cells, which are involved in the development of atherosclerosis. Our previous study found that microRNA (miR)-19b encapsulated within endothelial MPs (EMPs) may contribute to the upregulation of circulating miR-19b in unstable angina patients. Hypoxia is involved in atherosclerosis as a critical pathological stimulus. However, it still remains unclear whether the increase of miR-19b levels in EMPs is related to hypoxia and if the effect of miR-19b - wrapped within EMPs - stimulates hypoxia on vascular endothelial cells. This study aimed to explore the changes of miR-19b in EMPs induced by hypoxia as well as their effects on endothelial cells.@*Methods@#Human umbilical vein endothelial cells (HUVECs) were cultured in vitro and arranged to harvest EMPs in two parts: the first part consisted of EMP and EMP and the second part included EMP, EMP, and EMP. Cell migration was detected by scratch migration and transwell chamber migration. Angiogenesis was assessed by tube formation assays. Furthermore, we predicted the target gene of miR-19b by bioinformatics analysis, and luciferase assay was used to verify the targeted gene of miR-19b. Data were analyzed by one-way analysis of variance. Student's t-test was used when two groups were compared.@*Results@#Compared with EMP- and EMP-inhibited migration of cells by scratch migration assay (80.77 ± 1.10 vs. 28.37 ± 1.40, P < 0. 001) and transwell chamber migration assay (83.00 ± 3.46 vs. 235.00 ± 16.52, P < 0.01), the number of tube formations was markedly reduced by 70% in the EMP group (P < 0.001) in vitro analysis of HUVECs. Meanwhile, a strong inhibition of migration and tube formation of HUVECs in the presence of miR-19b-enriched EMP was observed. This effect might be due to the delivery of miR-19b in EMPs. Transforming growth factor-β2 (TGFβ2) was predicted to be one of the target genes of miR-19b, and we further confirmed that TGFβ2 was a direct target gene of miR-19b using the luciferase assay. The expression of TGFβ2 in HUVECs was inhibited by treatment with EMP and EMP.@*Conclusions@#MiR-19b in EMPs induced by hypoxia could reduce endothelial cell migration and angiogenesis by downregulating TGFβ2 expression, which may have inhibited the progression of atherosclerosis.
Subject(s)
Humans , Cell Hypoxia , Genetics , Physiology , Cell Movement , Genetics , Physiology , Endothelial Cells , Metabolism , Human Umbilical Vein Endothelial Cells , Metabolism , MicroRNAs , Genetics , Metabolism , Neovascularization, Physiologic , Genetics , Physiology , Transforming Growth Factor beta2 , Genetics , MetabolismABSTRACT
The microparticle is a kind of small particle formed in the process of cell activation and apoptosis.Currently,the mi-croparticle is taken as the main medium of inflammatory reaction.However,it may also play a role of atherosclerotic plaque formation and subsequent onset,development and formation of acute coronary syndrome.Furthermore,the microparticle participates in the atherosclerotic plaque formation and rupture,coagulation startup and thrombus formation.Through the study of microparticle,the coronary atherosclerotic heart disease pathogenesis can be deeply understood.
ABSTRACT
Objective To establish a liquid chromatography-tandem mass chromatography ( LC-MS/MS) method for determination of vancomycin concentration in human serum and compare its methodological performance with chemiluminescence microparticle immuno-assay (CMIA). Methods The proteins in serum sample were precipitated by methanol and then vancomycin was eluted and separated on Agilent Poroshell 120EC C18 column (2.1 mm×50 mm, 2.7 μm) with mobile phase of methanol and water in a gradient percentage. Both phases of methanol and water contained 0.1% formic acid. The flow rate was 0.5 mL/min. Norvancomycin was applied as internal standard. Electrospray ionization source was applied and operated in positive ion mode of multiple reaction monitoring (MRM). A total of 112 serum samples collected from the patients taking vancomycin in our hospital, and the results were compared with those of CMIA. Results The method exhibited good linearty within the concentration range of 1 to 100 μg/mL (r2＝ 0.9964).The intra-and inter-day accuracy and precision were all satisfactory for the requirements of quantitative drug testing. No matrix effect and carry-over effect were observed. The results of Wilcoxon symbol rank test showed a difference with statistical significance was found between the serum con-centrations of vancomycin determined by LC-MS/MS and CMIA (P＜0.05), but strong positive correlation and good linear regression were shown between the two methods (Y＝1.06X－0.37, r＝0.986). Conclusion LC-MS/MS should be a cost-saving method superior to CMIA. Its results highly correlated with those of CMIA method. The advantages of LC-MS/MS on accuracy and precision obviously outperformed those of CMIA. Since all the results of LC-MS/MS are comparable to CMIA, this method should be promising to use in determining concentration of vancomycin in serum samples with high clinical value.
ABSTRACT
Objective To investigate the changes in serum level of microparticles (EMPs) in mice with ventilator-induced lung injury (VILI), and explore its significance in VILI. Methods Forty-eight grade SPF male C57BL/6J mice were randomly divided into two groups, with 24 mice in each group: the mice in mechanical ventilation (MV) group were given high tidal volume (VT 30 mL/kg) MV for 4 hours after tracheal intubation, and those in spontaneous breathing group were spontaneously breathed for 4 hours. The apical blood of 12 mice in each group were collected, and serum levels of interleukins (IL-1β, IL-6) and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA), and serum EMPs levels were determined by flow cytometer. The correlations between EMPs and IL-1β, IL-6, and TNF-α were analyzed by linear regression analysis. The lung tissues of other 12 mice in each group were harvested, and wet/dry weight (W/D) ratio was assessed. After hematoxylin-eosin (HE) staining, the morphological changes in lung tissue were observed under light microscope. After double staining of uranium acetate and lead citrate, the ultrastructural changes in lung tissue were observed with electron microscope. Results Compared with spontaneous breathing group, the levels of lung W/D ratio in MV group was significantly increased (5.47±0.14 vs. 4.34±0.11), the levels of IL-1β, IL-6, TNF-α and EMPs were also significantly increased [IL-1β (ng/L): 42.4±4.4 vs. 7.7±3.6, IL-6 (ng/L): 1 239.5±66.3 vs. 21.7±4.6, TNF-α (ng/L):237.6±25.8 vs. 37.1±19.1, EMPs (cells/μL): 28.6±1.8 vs. 5.9±1.8, all P < 0.01]. It was shown by correlation analysis that EMPs were positively related with IL-1β, IL-6, and TNF-α (r value was 0.968, 0.932, 0.945, respectively, all P = 0.000). It was shown by fitting linear regression analysis that when EMPs increased by 1 cell/μL, IL-1β increased by 2.4 ng/L [95% confidence interval (95%CI) = 1.9-2.8, P < 0.001], IL-6 increased by 34.5 ng/L (95%CI = 25.1-44.0, P < 0.001), and TNF-α increased by 13.6 ng/L (95%CI = 10.3-16.9,P < 0.001). It was shown by light microscope that the structure of lung tissue and alveolar of mice in spontaneous breathing group appeared normal, while the shrinks of alveolar and disappearance of alveolar architecture were found in MV group. It was shown by electron microscopy that alveolar wall edema and thickening and broken alveolar septa were found in MV group, by contrast, the structure of alveolar was normal in spontaneous breathing group. Conclusion 30 mL/kg VT ventilation for 4 hours could induce VILI with increase in EMPs, suggesting EMPs closely related to VILI, and EMPs level may be putative biomarker of VILI.
ABSTRACT
Abstract Gastroretentive floating microparticles were developed and evaluated for the controlled metronidazole delivery for treatment of gastric disease. Floating microparticles, varying in proportions of chitosan and hydroxypropyl methylcellulose or ethylcellulose, were obtained by spray drying. Floating microparticles were characterized by physicochemical and in vitro studies, according to their floating ability and drug delivery. Microparticles presented mean diameter from 1.05 to 2.20 µm. The infrared spectroscopy confirmed the drug encapsulation and showed no chemical linkage between microparticles components. X-ray diffraction showed changes in the drug`s solid state, from crystalline to amorphous, indicating partial drug encapsulation, due to the presence of some crystalline peaks of metronidazole in microparticles. All microparticles floated immediately in contact of simulated gastric fluid and both floating and drug release profiles were dependent of microparticles composition. Microparticles samples constituted by chitosan and hydroxypropyl methylcellulose revealed the best relationship between floating duration and drug release, remaining floating during the occurrence of the drug release, ideal condition for the floating gastroretentive systems.
Subject(s)
Solid Waste Grinding , Drug Liberation , Metronidazole/administration & dosage , Chitosan/pharmacokinetics , Hypromellose DerivativesABSTRACT
Objective To detect the reactive samples of enzyme-linked immunosorbent assay (ELISA1) by chemiluminescence microparticle immunoassay (CMIA),and to analyze the application value of CMIA in HCV infection validation of blood donors.Methods Nucleic acid 3-item combined testing (NAT),another ELISA2,HCV antibody supplementary test(Western Blot test,WB) and CMIA test supplemented in blood samples of 102 ELISA1 anti-HCV reactive blood donors were retrospectively analysed.Results Among 102 blood donors of anti-HCV positive,32 cases (31.37%,32/102) were HCV RNA reactive samples,50 cases (49.02%,50/102) were ELISA2/WB reactive simultaneously.With CMIA NAT results as the reference standard,CMIA was poorly correlated with HCV RNA (Spearman correlation coefficient rs=0.395,P<0.01),and the consistency between them was weak by Kappa test (Kappa=0.270,P<0.01).With ELISA2/WB detection results as the reference standard,CMIA was highly correlated with the results(Spearman correlation coefficient rs=0.713,P<0.01),and which showed high consistency by Kappa test (Kappa=0.674,P<0.01).Conclusion CMIA as a detection method of protein label after HCV infection has great value in the HCV infection confirmation in low-risk population.
ABSTRACT
Objective To establish a high performance liquid chromatography (HPLC) method for determining phenytoin concentration in epilepsy patients' plasma,and compare this method with chemiluminescence microparticle immunoassay (CMIA),and to evaluate the consistency of the two methods.Methods HPLC and CMIA methods were applied to determine the plasma concentration of phenytoin in 60 epileptic patients,respectively.The difference of results was analyzed by two-side paired t-test,and then the correlation and consistency of the two methods were investigated with Passing-Bablok regression and Bland-Altman method.Results There was no significant difference between the results of the two methods (P ＞0.05).The regression equation of the determination results by HPLC (Y) and CMIA (X) was Y=0.992 9X +0.143 7 (R2 =0.992 6,n =60),which indicated the correlation of the two methods was good.Bland-Altman analysis showed that the consistency of the two methods for determining was good.Conclusion HPLC and CMIA method in monitoring plasma concentration of phenytoin have good correlation and consistency.Both methods can be used for therapeutic drug monitoring of phenytoin.
ABSTRACT
BACKGROUND: The syphilis diagnostic algorithms applied in different countries vary significantly depending on the local syphilis epidemiology and other considerations, including the expected workload, the need for automation in the laboratory and budget factors. This study was performed to investigate the efficacy of traditional and reverse syphilis diagnostic algorithms during general health checkups. METHODS: In total, 1,000 blood specimens were obtained from 908 men and 92 women during their regular health checkups. Traditional screening and reverse screening were applied to the same specimens using automatic rapid plasma regain (RPR) and Treponema pallidum latex agglutination (TPLA) tests, respectively. Specimens that were reverse algorithm (TPLA) reactive, were subjected to a second treponemal test performed by using the chemiluminescent microparticle immunoassay (CMIA). RESULTS: Of the 1,000 specimens tested, 68 (6.8%) were reactive by reverse screening (TPLA) compared with 11 (1.1%) by traditional screening (RPR). The traditional algorithm failed to detect 48 specimens [TPLA(+)/RPR(−)/CMIA(+)]. The median TPLA cutoff index (COI) was higher in CMIA-reactive cases than in CMIA-nonreactive cases (90.5 vs 12.5 U). CONCLUSIONS: The reverse screening algorithm could detect the subjects with possible latent syphilis who were not detected by the traditional algorithm. Those individuals could be provided with opportunities for evaluating syphilis during their health checkups. The COI values of the initial TPLA test may be helpful in excluding false-positive TPLA test results in the reverse algorithm.
Subject(s)
Female , Humans , Male , Agglutination , Automation , Budgets , Epidemiology , Immunoassay , Latex , Mass Screening , Plasma , Syphilis , Syphilis, Latent , Treponema pallidumABSTRACT
Objective To investigate the incidence of prozone phenomenon of rapid plasma reagin (RPR) test in syphilis serolog-ic testing and its correlation with the intensity of chemiluminescent microparticle immunoassay (CMIA) results.Methods A total of 101493 patients in our hospital from January 2014 to December 2015 were performed syphilis serologic testing by CMIA ,RPR and treponema pallidum particle agglutination (TPPA).The incidence rate of prozone phenomenon in RPR testing was evaluated.Its in-fluencing factors were investigated by using the Logistic regression.Results Among 101493 serum samples ,2180 cases were posi-tive by CMIA and 767 cases were positive by RPR ,the incidence rate of prozone phenomenon was 3.3% (26/767)in RPR.The Lo-gistic regression results indicated that the incidence of prozone phenomenon was significantly correlated with CMIA S /CO values and RPR titer ,but had no correlation with sex ,age and seasonality.Conclusion Although the incidence of prozone phenomenon is low in syphilis serologic testing ,but it is enough important.The patients with higher S/CO value in CMIA test have a higher inci-dence rate of RPR prozone phenomenon.
ABSTRACT
Objective To explore the clinical value of five methods commonly used for the detection of clinical syphilis antibody.Methods A total of 160 confirmed syphilis cases were chosen as the experimental group while 200 non-syphilis cases were set as the control group.Serum specimens were detected by methods as Treponema pallidum particle agglutination assay (TPPA),chemiluminescent microparticle immune assay (CMIA),enzyme linked immunosorbent assay (ELISA),emulsion method (TP-AD) and toluidine red unheated serum test (TRUST).Sensitivity and specificity were evaluated on five methods.Titers of syphilis antibody in different stages and pre/post on treament among syphilis patients were compared and analyzed under the five methods.Results The sensitivity vs.specificity of TPPA,CMIA,ELISA,TP-AD and TRUST appeared as 100.00％ vs.99.50％,99.38％ vs.99.00％,98.12％ vs.99.00％,94.38％ vs.94.50％ and 85.62％ vs.95.50％,respectively.Among the patients at primary or latent stages,the syphilis antibody positive rate detected by TRUST appeared lower than that detected by ELISA,TPPA,CMIA or TP-AD,and the differences were statistically significant (P＜0.01).There were no statistical differences in the syphilis antibody positive rate of syphilis patients in the secondary or tertiary stages detected by five methods (P＞0.05).In each stage of the syphilis patients,the syphilis antibody positive rate detected by ELISA or of CMIA combined with TRUST both reached 100.00％.Before and after treatment in 121 cases of confirmed syphilis,there was statistically significant difference in the syphilis antibody positive rate detected by TRUST method (P＜0.05).There was no statistical significance in the syphilis antibody positive rate detected by other four methods (P＞0.05).Conclusions The sensitivity and specificity of TPPA,CMIA and ELISA methods were better.Methods as ELISA or as CMIA combined with TRUST both appeared reliable for syphilis screening in every stage of the disease.TRUST was suitable for the determination of active stage syphilis and monitoring the effects after treatment.
ABSTRACT
Objective To explore the clinical value of five methods commonly used for the detection of clinical syphilis antibody.Methods A total of 160 confirmed syphilis cases were chosen as the experimental group while 200 non-syphilis cases were set as the control group.Serum specimens were detected by methods as Treponema pallidum particle agglutination assay (TPPA),chemiluminescent microparticle immune assay (CMIA),enzyme linked immunosorbent assay (ELISA),emulsion method (TP-AD) and toluidine red unheated serum test (TRUST).Sensitivity and specificity were evaluated on five methods.Titers of syphilis antibody in different stages and pre/post on treament among syphilis patients were compared and analyzed under the five methods.Results The sensitivity vs.specificity of TPPA,CMIA,ELISA,TP-AD and TRUST appeared as 100.00％ vs.99.50％,99.38％ vs.99.00％,98.12％ vs.99.00％,94.38％ vs.94.50％ and 85.62％ vs.95.50％,respectively.Among the patients at primary or latent stages,the syphilis antibody positive rate detected by TRUST appeared lower than that detected by ELISA,TPPA,CMIA or TP-AD,and the differences were statistically significant (P＜0.01).There were no statistical differences in the syphilis antibody positive rate of syphilis patients in the secondary or tertiary stages detected by five methods (P＞0.05).In each stage of the syphilis patients,the syphilis antibody positive rate detected by ELISA or of CMIA combined with TRUST both reached 100.00％.Before and after treatment in 121 cases of confirmed syphilis,there was statistically significant difference in the syphilis antibody positive rate detected by TRUST method (P＜0.05).There was no statistical significance in the syphilis antibody positive rate detected by other four methods (P＞0.05).Conclusions The sensitivity and specificity of TPPA,CMIA and ELISA methods were better.Methods as ELISA or as CMIA combined with TRUST both appeared reliable for syphilis screening in every stage of the disease.TRUST was suitable for the determination of active stage syphilis and monitoring the effects after treatment.