RESUMEN
BACKGROUND: Polycystic ovary syndrome (PCOS) is the main cause of anovulatory infertility in women of reproductive age, and low-grade chronic inflammation plays a key role in the occurrence and development of PCOS. However, obesity, as a likely confounding factor, can affect the inflammatory state of PCOS patients. OBJECTIVE: The aim of this study was to comprehensively investigate intra-ovarian inflammatory states and their impact on embryo quality in PCOS patients with a normal BMI undergoing IVF treatment. METHODS: DIA-mass spectrometry-based proteomics and bioinformatic analysis were combined to comprehensively profile the protein expression of granulosa cells (GCs) from 5 normal-BMI PCOS patients and 5 controls. Thirty-four cytokines were further systematically detected in follicular fluid (FF) from 32 age- and BMI-matched normal-BMI patients using Luminex liquid chip suspension technology. Next, the differentially expressed cytokines were evaluated by enzyme-linked immunosorbent assay (ELISA) in 24 newly recruited subjects, and the relationship between these cytokines and embryo quality in PCOS patients was analysed. Finally, these cytokine levels were compared and evaluated in PCOS patients with different androgen levels. RESULTS: Proteomic analysis showed that the suppression of substance metabolism and steroid biosynthesis, more interestingly, resulted in an enhanced immune and inflammatory response in the GCs of normal-BMI PCOS patients and prompted the involvement of cytokines in this process. Luminex analysis further showed that FF macrophage inflammatory protein-1 beta (MIP-1ß) and stromal cell-derived factor-1 alpha (SDF-1α) levels were significantly increased in normal-BMI PCOS patients compared to controls (P = 0.005; P = 0.035, respectively), and the ELISA results were consistent with these findings. Besides, FF MIP-1ß showed an inverse correlation with the number of D3 good-quality embryos and the good-quality blastocyst rate in patients with PCOS (P = 0.006; P = 0.003, respectively), which remained significant after correction for multiple comparisons. Moreover, SDF-1α levels had no relationship with embryo development in PCOS patients. Additionally, SDF-1α levels were significantly lower in PCOS patients with high androgen levels than in controls (P = 0.031). CONCLUSIONS: Local ovarian inflammation was present in normal-BMI PCOS patients, affecting follicular development, and FF MIP-1ß may be a potential biomarker associated with embryo quality in normal-BMI PCOS patients.
Asunto(s)
Síndrome del Ovario Poliquístico , Humanos , Femenino , Síndrome del Ovario Poliquístico/metabolismo , Quimiocina CCL4/metabolismo , Quimiocina CXCL12/metabolismo , Proteómica , Andrógenos/metabolismo , Índice de Masa Corporal , Líquido Folicular/metabolismo , Citocinas/metabolismo , Inflamación/metabolismo , Fertilización In VitroRESUMEN
Due to the different physicochemical properties of lead (Pb) and arsenic (As), coupled remediation processes of contaminated soils containing Pb and As have always been a technical challenge. In the present study, a novel solidifying agent (BER) was synthesized using alkaline oxygen furnace slag (BOFS), modified electrolytic manganese residue (EMR) and red mud (RM). The solidifying agent was synergistically used with Fenton reagent for solidifying/stabilizing Pb- and As-contaminated soils. The experimental results indicated that the low stability of Ca-As(III) complexes, which serve as the primary fixation mechanism for As in BER-solidified soil, led to inadequate stabilization of As by BER. As a result, the leaching concentration of As in the BER-solidified soil reached 2.61 mg/L. The incorporation of Fenton reagent had minimal effect on the strength of the solidified soil and Pb stabilization, but it substantially improved As stabilization. The immobilization efficiency of the BER-Fenton for lead and arsenic reached 99% and 98%, respectively. The strength of solidified/stabilized soil using BER-Fenton was close to that of cement-based solidified/stabilized soil. Microscopic analysis showed that the incorporation of Fenton reagent increased the content of goethite and ettringite, which effectively filled the soil pores and enhanced the adsorption of Pb and As. The use of BER and Fenton reagent increased the content of As(V) to 88%, which promoted the complexation between As and Fe(III)/Mn(IV). This study provides a novel method for efficient remediation of heavy metal-contaminated sites, as well as a useful reference for ensuring the safe reuse of hazardous wastes.
RESUMEN
STUDY QUESTION: Does the transfer of single low-grade blastocysts result in acceptable reproductive and perinatal outcomes compared to the transfer of single good-grade blastocysts? SUMMARY ANSWER: The transfer of single low-grade blastocysts resulted in a reduced live birth rate of around 30% (14% for very low-grade blastocysts) compared to 44% for single good-grade blastocysts, but does not lead to more adverse perinatal outcomes. WHAT IS KNOWN ALREADY: It is known that low-grade blastocysts can result in live births. However, the current studies are limited by relatively small sample sizes and single-centre designs. Furthermore, evidence on perinatal outcomes after transferring low-grade blastocysts is limited. STUDY DESIGN, SIZE, DURATION: We conducted a multi-centre, multi-national retrospective cohort study of 10 018 women undergoing 10 964 single blastocyst transfer cycles between 2009 and 2020 from 14 clinics across Australia, China, and New Zealand. PARTICIPANTS/MATERIALS, SETTING, METHODS: Blastocysts were graded individually based on assessment of the morphology and development of the inner cell mass (ICM) and trophectoderm (TE), and were grouped into three quality categories: good- (AB, AB, or BA), moderate- (BB), and low-grade (grade C for ICM or TE) blastocysts. CC blastocysts were individually grouped as very low-grade blastocysts. Logistic regression with generalized estimating equation was used to analyse the association between blastocyst quality and live birth as well as other reproductive outcomes. Binomial, multinomial logistic, or linear regression was used to investigate the association between blastocyst quality and perinatal outcomes. Odds ratio (OR), adjusted OR (aOR), adjusted regression coefficient, and their 95% CIs are presented. Statistical significance was set at P < 0.05. MAIN RESULTS AND THE ROLE OF CHANCE: There were 4386 good-grade blastocysts, 3735 moderate-grade blastocysts, and 2843 low-grade blastocysts were included in the analysis, for which the live birth rates were 44.4%, 38.6%, and 30.2%, respectively. Compared to good-grade blastocysts, the live birth rate of low-grade blastocysts was significantly lower (aOR of 0.48 (0.41-0.55)). Very low-grade blastocysts were associated with an even lower live birth rate (aOR 0.30 (0.18-0.52)) and their absolute live birth rate was 13.7%. There were 4132 singleton live births included in the analysis of perinatal outcomes. Compared with good-grade blastocysts, low-grade blastocysts had comparable preterm birth rates (<37 weeks, aOR 1.00 (0.65-1.54)), birthweight Z-scores (adjusted regression coefficient 0.02 (0.09-0.14)), and rates of very low birth weight (<1500 g, aOR 0.84 (0.22-3.25)), low birth weight (1500-2500 g, aOR 0.96 (0.56-1.65)), high birth weight (>4500 g, aOR 0.93 (0.37-2.32)), small for gestational age (aOR 1.63 (0.91-2.93)), and large for gestational age (aOR 1.28 (0.97-1.70)). LIMITATIONS, REASONS FOR CAUTION: Due to the nature of the retrospective design, residual confounding could not be excluded. In addition, the number of events for some perinatal outcomes was small. Between-operator and between-laboratory variations in blastocyst assessment were difficult to control. WIDER IMPLICATIONS OF THE FINDINGS: Patients undergoing IVF should be informed that low-grade blastocysts result in a lower live birth rate, however they do not increase the risk of adverse perinatal outcomes. Further research should focus on the criteria for embryos that should not be transferred and on the follow-up of long-term outcomes of offspring. STUDY FUNDING/COMPETING INTEREST(S): H.Z. is supported by a Monash Research Scholarship. B.W.J.M. is supported by a NHMRC Investigator grant (GNT1176437). R.W. is supported by an NHMRC Emerging Leadership Investigator grant (2009767). B.W.J.M. reports consultancy, travel support, and research funding from Merck. The other authors do not have competing interests to disclose. TRIAL REGISTRATION NUMBER: N/A.
Asunto(s)
Nacimiento Prematuro , Embarazo , Humanos , Recién Nacido , Femenino , Estudios Retrospectivos , Transferencia de Embrión/métodos , Nacimiento Vivo , Peso al Nacer , BlastocistoRESUMEN
PURPOSE: Poor sleep and mood may be predisposing factors for acne. We aimed to investigate the associations between acne and sleep quality, circadian preferences, and mood. METHODS: This case-control study recruited patients with acne and age- and sex-matched healthy controls. We used the Investigator's Global Assessment to evaluate acne severity and various validated questionnaires to measure sleep quality, daytime sleepiness, sleep apnea, circadian preference, and mood symptoms. RESULTS: A total of 81 patients with acne (age: 21.6 ± 5.0 years, 52% female) and 76 controls were recruited. Compared to controls, patients had a higher score on the Pittsburgh Sleep Quality Index (5.2 ± 2.6 vs. 4.1 ± 2.3, p = 0.008) and State-Trait Anxiety Inventory (State: 44.6 ± 9.7 vs. 40.6 ± 6.6, p = 0.003; Trait: 47.9 ± 8.2 vs. 45.3 ± 6.2, p = 0.03), and a lower score on a reduced version of the Morningness and Eveningness Questionnaire (13.9 ± 2.6 vs. 14.7 ± 2.3, p = 0.05) and Epworth Sleepiness Scale (7.4 ± 3.4 vs. 8.6 ± 3.6, p = 0.04). Acne severity was associated with sleep quality (ß = 0.33), eveningness (ß = 0.34), depression (ß = 0.66), and anxiety (State: ß = 1.73; Trait: ß = 1.21), even when adjusted for education level and family history of acne. CONCLUSION: Acne is highly associated with poor sleep and mood. Dermatologists are advised to attend closely to the psychological impact of acne. Improvements in sleep and mood may benefit the treatment of acne.
Asunto(s)
Acné Vulgar , Trastornos del Inicio y del Mantenimiento del Sueño , Humanos , Femenino , Adolescente , Adulto Joven , Adulto , Masculino , Calidad del Sueño , Estudios de Casos y Controles , Afecto , Sueño , Encuestas y Cuestionarios , Ritmo CircadianoRESUMEN
A novel solvent terminated microextraction method based on a natural deep eutectic solvent (L-menthol and lactic acid at a molar ratio of 1:2) coupled with high-performance liquid chromatography was proposed, which was utilized for the separation and enrichment of bisdemethoxycurcumin, demethoxycurcumin and curcumin in Curcumae Longae Rhizoma and turmeric tea. The effects of independent parameters on extraction efficiency were optimized by single-factor analysis. Subsequently, four predominated parameters affecting the extraction procedure, including extractant volume, salt concentration, demulsifier consumption, and demulsification time, were further evaluated by a central composite design. Under the optimized conditions, the linear ranges of calibration curves were 0.005-0.5 µg/mL for bisdemethoxycurcumin, 0.004-0.4 µg/mL for demethoxycurcumin, and 0.0045-0.45 µg/mL for curcumin, respectively. In addition, the developed method provided low detection limits (0.1-0.4 ng/mL) and high enrichment factors (279-350). Its intra-day and inter-day precision were carried out by relative standard deviation ranging from 2.2 to 9.2%. Finally, the applicability of this method was assessed by the analysis of Curcumae Longae Rhizoma and turmeric tea samples. The results showed that these samples were detected successfully and the spiked recoveries over the range of 85.3-108.9% with relative standard deviations of 1.6-8.9% were attained, indicating its high relative recoveries with good precision in real sample analysis.
Asunto(s)
Curcumina , Microextracción en Fase Líquida , Cromatografía Líquida de Alta Presión , Curcuma/química , Disolventes Eutécticos Profundos , Diarilheptanoides , Límite de Detección , Microextracción en Fase Líquida/métodos , Solventes/química , Té/químicaRESUMEN
BACKGROUND: Long-term dietary fat intake is thought to affect metabolism and pregnancy of polycystic ovary syndrome (PCOS) patients, and the type of fatty acids one consumes plays an important role. Previous studies mostly used questionnaires to analyze the type and proportion of fatty acids. METHODS: This prospective study included 91 PCOS patients. Serum fatty acids were measured by the gas chromatograph-mass spectrometry method before ovulation induction. We compared the fatty acids between the pregnancy group and the nonpregnancy group and explored the influence of the fatty acids on live births and pregnancy loss. RESULTS: Nervonic acid was lower in the pregnancy group than in the nonpregnancy group (0.25% vs. 0.30%, p = .017). The following trans-fatty acids were significantly lower in the pregnancy group than in the nonpregnancy group: trans-10-heptadecenoic acid, trans-vaccenic acid, trans-11-eicosenoic acid, and brassidic acid. The level of polyunsaturated fatty acids in the live birth group was significantly higher than the pregnancy loss group (16.95% vs. 15.10%, p = .039). Among individual PUFAs, the levels of linoleic acid (p = .043), docosapentaenoic acid (p = .024), alpha-linolenic acid (p = .042), and eicosapentaenoic acid (p = .035) were higher in the live birth group than in the pregnancy loss group. After adjusting for infertility duration, age, and body mass index, our findings suggested an inverse association between pregnancy and nervonic acid, trans-10-heptadecenoic acid, trans-vaccenic acid, trans-11-eicosenoic acid, and brassidic acid and pregnancy. CONCLUSIONS: Our findings indicate that polyunsaturated fatty acids are associated with live birth in PCOS patients. Serum trans-fatty acids and nervonic acid might be risk factors for nonpregnancy. The mechanism of the influence of different fatty acids on pregnancy and live birth merits further exploration.
Asunto(s)
Aborto Espontáneo , Síndrome del Ovario Poliquístico , Ácidos Grasos trans , Embarazo , Humanos , Femenino , Síndrome del Ovario Poliquístico/complicaciones , Estudios Prospectivos , Ácidos Grasos , Ácidos Erucicos , Tasa de Natalidad , Inducción de la Ovulación/métodos , Aborto Espontáneo/etiologíaRESUMEN
BACKGROUND: Variations in the oral microbiota have been significantly correlated with the progress of autoimmune diseases, such as immunoglobulin A nephropathy and Henoch-Schönlein purpura (HSP). However, there is no report outlining the character of tongue coating microbiota variations in children with Henoch-Schönlein purpura nephritis (HSPN). METHOD: A total of 20 children with HSPN and 14 healthy controls were recruited for this research. Tongue coating samples of two groups were collected for 16S rRNA gene sequencing. The diversity, principal component analysis (PCA), nonmetric multidimensional scaling (NMDS), partial least squares discriminant analysis (PLS-DA), and linear discriminant analysis (LDA) effect size (LEfSe) were performed. Microbial function was assessed using the PICRUST. RESULTS: The ACE and Chao indices were greatly lower in the HSPN group than in the HG (P = 0.001). The Shannon and Simpson indices were dramatically reduced in children with HSPN compared with those in the healthy controls (P = 0.005). Bacteroidales, Selenomonadales, Lactobacillales, Fusobacteriales, Neisseriales, and Actinomycetales composed more than 80% of all sequences, while Bacteroidales was the most generous order in both groups. PCA, NMDS and PLS-DA showed a marked difference between the control and HSPN groups. LEfSe analysis showed alteration of tongue coating microbiota in the HSPN group. There were 30 metabolic functions significantly differed between the two groups. CONCLUSIONS: Children with HSPN have substantially various tongue coating microbiota compared to healthy controls. Even though this research does not indicate causality, it is beneficial to enhance the possibility for coming microbial-based treatments to enhance the clinical effects of HSPN in children.
Asunto(s)
Vasculitis por IgA , Microbiota , Nefritis , Niño , Humanos , ARN Ribosómico 16S/genética , LenguaRESUMEN
In this study, a vortex-assisted hydrophobic deep eutectic solvent dispersive liquid-phase microextraction was developed and used for the extraction and preconcentration of six main active compounds in Zi-Cao-Cheng-Qi decoction. The deep eutectic solvent, prepared by mixing tetrabutylammonium chloride and hexanoic acid at a molar ratio of 1:1, was added to the sample solution containing the analytes. In the absence of disperser, the extractant was rapidly dispersed into fine droplets by the aid of vortex and adequately contacted with the analytes. Some key parameters affecting the approach including extraction solvent type and volume, sample phase pH, extraction time, centrifugation time, and salt concentration were investigated and optimized. Under the optimum conditions, enrichment factors of the target analytes were in the range of 3-330. The calibration graphs were linear with a correlation coefficient (r) ≥ 0.9929. The detection limits were 0.3-0.9 ng/mL, and the satisfactory precisions (relative standard deviations, 0.5-8.9%) and accuracies (relative recoveries, 91.1-102.2%) were also obtained. The developed method was rapid (only 2 min), eco-friendly, effective, and easy to operate. And it has been successfully applied to simultaneous extraction, enrichment, and determination of the main active compounds in a traditional Chinese medicinal formula coupled with high-performance liquid chromatography.
Asunto(s)
Disolventes Eutécticos Profundos/química , Medicamentos Herbarios Chinos/química , Microextracción en Fase Líquida/métodos , Cromatografía Líquida de Alta Presión/métodos , Interacciones Hidrofóbicas e Hidrofílicas , Límite de DetecciónRESUMEN
BACKGROUND: Propionic acidemia (PA) is a severe monogenic disorder characterized by a deficiency of the mitochondrial protein propionyl-CoA carboxylase (PCC) enzyme, which is caused by mutations in the PCCA or PCCB gene. Preconception carrier screening could provide couples with meaningful information for their reproductive options; however, it is not widely performed in China. CASE PRESENTATION: This report describes a case of dizygotic twin siblings conceived by in vitro fertilization (IVF) and diagnosed with propionic acidemia (PA). Their parents had no history of PA. Tandem mass spectrometry and urine gas chromatography/mass spectrometry (GC/MS) of the twin siblings revealed markedly elevated propionyl carnitine (C3), C3/C2, and 3-hydroxypropionate in the plasma and urine. Whole-exome sequencing was performed for the twin siblings. A homozygous missense mutation, c.2002G > A (p.Gly668Arg) in PCCA, was identified in the twin siblings. Sanger sequencing confirmed the homozygous mutation in the twin siblings and identified their parents as heterozygous carriers of the c.2002G > A mutation in PCCA. Both neonates in this case died. This is an emotionally and financially devastating outcome that could have been avoided with genetic carrier screening before conception. If couples are screened before IVF and found to be silent carriers, then reproductive options (such as preimplantation genetic diagnosis or prenatal diagnosis) can be offered to achieve a healthy newborn. CONCLUSION: This case is a reminder to infertile couples seeking IVF that it is beneficial to clarify whether they are silent carriers before undergoing IVF.
Asunto(s)
Ligasas de Carbono-Carbono/genética , Metilmalonil-CoA Descarboxilasa/genética , Mutación , Acidemia Propiónica/genética , Adulto , China , Femenino , Fertilización In Vitro , Humanos , Recién Nacido , Masculino , Padres , Embarazo , Hermanos , Gemelos DicigóticosRESUMEN
Self-assembled supramolecular dispersive liquid-phase microextraction combined with high-performance liquid chromatography was developed and introduced for simultaneous extraction and determination of the trace level rhein, chrysophanol, and physcion in Rhubarb. Compared with conventional dispersive liquid-phase microextraction, the proposed method used a self-assembled ternary supramolecular consisting of the mixed extraction solvent (heptanol and nonanol) and dispersant (acetone) to achieve high enrichment factors of target analytes. Several factors affecting performance were investigated and optimized, including the mixed extraction solvent, type and volume of the dispersant, the pH of sample phase, salt concentration, shaking time, volume of sample phase, centrifugation time, and rate. Meanwhile, the method mechanism of self-assembled supramolecular dispersive liquid-phase microextraction was analyzed and described. Under the optimized extraction conditions, the enrichment factors of rhein, chrysophanol, and physcion were 116.5, 325.9, and 356.1, respectively. Good linearities (r ≥ 0.9952) for all analytes, low limits of detection (less than 0.04 ng/mL), satisfactory precisions (0.1-8.9%), and accuracies (recoveries, 88.2-104.1%) were achieved. The experimental results showed that the approach was simple, fast, with short extraction time, high enrichment factors, good linearities, and low limits of detection.
Asunto(s)
Antraquinonas/análisis , Microextracción en Fase Líquida , Rheum/química , Cromatografía Líquida de Alta Presión , Concentración de Iones de Hidrógeno , Sustancias Macromoleculares/análisisRESUMEN
Two proposed syringe membrane filter solid phase microextraction and syringe membrane filter liquid/solid phase microextraction, coupled separately with high performance liquid chromatography, were developed for simultaneous enrichment and determination of the trace level of flavonoids in traditional Chinese medicine. In syringe membrane filter solid phase microextraction, the membrane of syringe membrane filter was served as a solid adsorption film to adsorb target analytes. And in syringe membrane filter liquid/solid phase microextraction, the membrane of syringe membrane filter was used as not only an adsorption phase, but also as a holder of extraction solvent to realize liquid-solid synergistic extraction. The simple operation, rapid extraction, and little or no organic solvent consumption make the two approaches very interesting. To evaluate the two proposed approaches, the crucial parameters affecting the enrichment factors of target analytes were investigated and optimized, and the two microextractions were intercompared. Moreover, their microextraction mechanisms were analyzed and described. Under the optimized conditions, both the new approaches achieved good linearities, accuracies, precisions, and low limits of detection, and the two methods were successfully applied for concentration of the flavonoids in traditional Chinese medicines.
Asunto(s)
Flavonoides/análisis , Microextracción en Fase Líquida , Microextracción en Fase Sólida , Jeringas , Cromatografía Líquida de Alta Presión , Microextracción en Fase Líquida/instrumentación , Medicina Tradicional China , Estructura Molecular , Microextracción en Fase Sólida/instrumentaciónRESUMEN
BACKGROUND: Fertilization failure after intracytoplasmic sperm injection continues to affect couples and the etiology is not well-understood. CASE PRESENTATION: We characterized a couple with 2-year history of primary unexplained infertility. Three different assisted reproduction attempts (IVF + rescue ICSI, ICSI and ICSI-AOA) showed repeated fertilization failure for MII oocyte retrieval after controlled ovarian hyperstimulation. After whole-exome sequencing and sanger sequencing of the couple and their family members, variant pathogenicity was assessed using SIFT, PolyPhen2, Mutation Taster, and Human Splicing Finder software. We identified novel compound heterozygous mutations, c.1535 + 3A > G and c.946C > T (p. Leu316Phe), in WEE2 in the female proband. Trios analysis of the variations revealed an autosomal recessive pattern. c.1535 + 3A > G in WEE2 was predicted to break the wild-type donor site and affect splicing, and the missense mutation c.946C > T (p. Leu316Phe) of WEE2 was predicted to be pathogenic. CONCLUSION: A novel compound heterozygous mutation in WEE2 was identified in an infertile female who experienced repeated fertilization failure even after ICSI-AOA. These novel mutations in WEE2 provided genetic evidence for fertilization failure.
Asunto(s)
Proteínas de Ciclo Celular , Infertilidad Femenina , Proteínas Tirosina Quinasas , Adulto , Proteínas de Ciclo Celular/genética , Femenino , Humanos , Infertilidad Femenina/genética , Infertilidad Femenina/terapia , Mutación , Proteínas Tirosina Quinasas/genética , Inyecciones de Esperma Intracitoplasmáticas , Insuficiencia del TratamientoRESUMEN
A three-phase hollow-fiber liquid-phase microextraction based on deep eutectic solvent as acceptor phase was developed and coupled with high-performance capillary electrophoresis for the simultaneous extraction, enrichment, and determination of main active compounds (hesperidin, honokiol, shikonin, magnolol, emodin, and ß,ß'-dimethylacrylshikonin) in a traditional Chinese medicinal formula. In this procedure, two hollow fibers, impregnated with n-heptanol/n-nonanol (7:3, v/v) mixture in wall pores as the extraction phase and a combination (9:1, v/v) of methyltrioctylammonium chloride/glycerol (1:3, n/n) and methanol in lumen as the acceptor phase, were immersed in the aqueous sample phase. The target analytes in the sample solution were first extracted through the organic phase, and further back-extracted to the acceptor phase during the stirring process. Important extraction parameters such as types and composition of extraction solvent and deep eutectic solvent, sample phase pH, stirring rate, and extraction time were investigated and optimized. Under the optimal conditions, detection limits were 0.3-0.8 ng/mL with enrichment factors of 6-114 for the analytes and linearities of 0.001-13 µg/mL (r2 ≥ 0.9901). The developed method was successfully applied to the simultaneous extraction and concentration of the main active compounds in a formula of Zi-Cao-Cheng-Qi decoction with the major advantages of convenience, effectiveness, and environmentally friendliness.
Asunto(s)
Medicamentos Herbarios Chinos/análisis , Microextracción en Fase Líquida , Medicina Tradicional China , Concentración de Iones de Hidrógeno , Cloruro de Sodio/química , Solventes/químicaRESUMEN
This study proposed a new ballpoint connector-protected salt-oil-salt liquid phase microextraction for extraction and enrichment of trace rhein and chrysophanol in rhubarb prior to determination of the analytes by high performance liquid chromatography. In this study, a handy ballpoint connector (between ballpoint tip and ink chamber) was used as extraction device, in which its cavity was filled with n-octanol, and the bare n-octanol in its two opening ends was covered with a thin layer of sodium chloride film. The design subtly assembled salt film onto ballpoint connector for extraction and enrichment, which greatly improved the enrichment factors of the target analytes. Moreover, the novel procedure and its extraction mechanism were described and analyzed, and several crucial parameters reflecting the extraction effect were investigated and optimized. Under optimum conditions, high enrichment factors (247 and 127), good linearities with r ≥ 0.9998, limits of detection (0.6-1.1 ng/mL), relative standard deviations of intra- and interday (2.2-8.8% and 4.3-8.9%), and average recoveries (97.6-98.1%), were obtained, respectively. The proposed method can not only eliminate the negative effects from viscosity and ion strength at high salt concentration of sample phase, but also make salting-out effect be focused on small area so as to maximize the extraction effect.
Asunto(s)
Antraquinonas/análisis , Microextracción en Fase Líquida , Aceites/química , Rheum/química , Cloruro de Sodio/química , Cromatografía Líquida de Alta PresiónRESUMEN
A novel three-phase hollow fiber liquid-phase microextraction was developed based on reverse micelle as extraction solvent and acceptor phase, and compared with conventional two-phase hollow fiber liquid-phase microextraction. Both procedures were used in the extraction and concentration of four cinnamic acids (caffeic acid, p-hydroxycinnamic acid, ferulic acid, and cinnamic acid) in traditional Chinese medicines prior to high-performance liquid chromatography analysis. Parameters affecting the two procedures were investigated and optimized to obtain the optimum enrichment factors. The mechanism of the developed procedure was explored and elucidated by comparison with conventional two-phase hollow fiber liquid-phase microextraction. Under the optimized conditions, the analytes' enrichment factors were between 50 and 118 for the proposed procedure, and 31-96 for conventional two-phase mode. Satisfactory linear ranges (r2 ≥ 0.99), detection limits (0.1-0.6 ng/mL), precisions (<9.2%), and accuracies (recoveries: 80-123.1%) were observed for the two procedures. The results showed that the enrichment capacity of the proposed procedure for the cinnamic acids is better than that of conventional two-phase procedure, and both are eco-friendly, simple, and effective for the enrichment and detection of cinnamic acids in traditional Chinese medicines.
Asunto(s)
Cinamatos/análisis , Medicamentos Herbarios Chinos/análisis , Microextracción en Fase Líquida , Medicina Tradicional China , Micelas , Estructura MolecularRESUMEN
PURPOSE: To perform a preliminary exploration of a new embryo rank in clinical practice by combining the embryo chromosome copy number and mitochondrial copy number analysis of DNA extracted from embryo culture medium and blastocoel fluid. METHOD: Eighty-three ICSI embryos from day 2 or day 3 were cultured to day 5 or day 6. Thirty-two blastocysts of 3 cc or above were obtained. Culture medium and blastocoel fluid were collected at 24 h before blastocyst formation. The genomic DNA and mitochondrial DNA (mtDNA) from the culture medium combined with blastocoel fluid and the whole blastocyst were amplified and sequenced by MALBAC-NGS. We compared the chromosomal information generated by the new protocol from the culture medium and the information employed by the whole embryo method. A multivariable linear regression was performed to study the impact of the blastocyst morphological score, chromosomal abnormality, embryo mtDNA copy number, and female age on the culture medium mtDNA copy number. RESULTS: (1) The DNA from 31 blastocysts was successfully amplified, and the successful amplification rate was 96.9% (31/32). The success rate of the amplification of genomic DNA extracted from the culture medium was 87.5% (28/32). (2) There were 18 blastocysts in which the less invasive method and the whole embryo method revealed the same results. The consistency rate was 66.7% (18/27). (3) The culture medium mitochondrial DNA copy number (MCN) had a significantly positive correlation with the blastocyst mitochondrial DNA copy number (P = 0.001), female age (P = 0.012), and blastocyst score (P = 0.014), but there was no obvious correlation with blastocyst chromosome (P = 0.138). CONCLUSIONS: The preliminary exploration result of the less invasive approach for having an embryo rank was not satisfying, which still awaits further long-term evaluation.
Asunto(s)
Aneuploidia , Cromosomas/genética , Técnicas de Cultivo de Embriones/métodos , Diagnóstico Preimplantación , Adulto , Medios de Cultivo , ADN Mitocondrial/genética , Implantación del Embrión/genética , Transferencia de Embrión/métodos , Embrión de Mamíferos , Desarrollo Embrionario/genética , Femenino , Fertilización In Vitro , Pruebas Genéticas/métodos , Humanos , EmbarazoRESUMEN
A fast, simple, and efficient salt-assisted dispersive liquid-liquid microextraction coupled with high-performance liquid chromatography was developed and introduced for the simultaneous enrichment, extraction, and determination of the trace levels of matrine alkaloids (sophoridine, matrine, and sophocarpine) in Sophorae Flavescentis Radix and Composite Kushen injection. Compared with conventional dispersive liquid-liquid microextraction, the proposed method, with added salt but without dispersant and centrifuging, makes the operation simpler, greener, and leads to a higher enrichment factor. The crucial parameters affecting the enrichment factors of target analytes, such as type and volume of extraction solvent, pH of sample phase, salt concentration, volume of sample phase, and extraction time, were investigated and optimized, meanwhile, the extraction mechanism of the method was analyzed and described. Under the optimized conditions, the enrichment factors of the three matrine alkaloids were 150, 178, and 227, respectively. Good linearities (r2 ≥ 0.9992) for all analytes, low limits of detection (less than 0.08 ng/mL), satisfactory precisions (2.1-12.3%), and accuracies (recoveries, 99.3-103.9%) were achieved. The experimental results showed that the approach is a simple, fast, green, eco-friendly, and sensitive method and can be used for the preconcentration and determination of matrine alkaloids in traditional Chinese medicines and their preparations.
Asunto(s)
Alcaloides/síntesis química , Alcaloides/aislamiento & purificación , Microextracción en Fase Líquida , Extractos Vegetales/síntesis química , Extractos Vegetales/aislamiento & purificación , Sophora/química , Alcaloides/química , Concentración de Iones de Hidrógeno , Medicina Tradicional China , Extractos Vegetales/química , Sales (Química)/químicaRESUMEN
A sodium dodecyl sulfate sensitized switchable solvent liquid-phase microextraction method was developed and applied to the preconcentration of active alkaloids in Rhizoma coptidis followed by high performance liquid chromatography determination. Before extraction, nonionic triethylamine was converted to its cationic form in the presence of carbon dioxide. Then, the ionic solvent carrying target analytes was once more reverted to its nonionic form by adding sodium hydroxide, as well as phase separation and analytes enrichment were realized simultaneously. Several parameters affecting the approach, such as concentration of sodium dodecyl sulfate, extraction solvent volume, sodium hydroxide concentration, sample phase pH, injection solvent type, and extraction time, were investigated and optimized. The possible microextraction mechanism of double micelle supramolecular inclusion was explored. Under the optimum conditions, the enrichment factors of four protoberberine alkaloids were from 101.8 to 152.0. The linear ranges (with r2 ≥ 0.990) were 0.032-4.23, 0.031-4.33, 0.0026-10.04, and 0.0013-4.13 µg/mL for epiberberine, coptisine, palmatine, and berberine, respectively. The detection limits were in the range of 0.16-0.32 ng/mL. Satisfactory accuracies (recoveries 98.8-104.6%) and precisions (RSDs 1.9-10.9%) were also obtained. The results showed that the approach is rapid, effective, eco-friendly, and easy-to-handle for the enrichment and detection of active alkaloids in Rhizoma coptidis.
Asunto(s)
Alcaloides de Berberina/aislamiento & purificación , Medicamentos Herbarios Chinos/química , Microextracción en Fase Líquida , Dodecil Sulfato de Sodio/química , Alcaloides de Berberina/química , Solventes/químicaRESUMEN
A sensitivity-improved ultrasonic sensor is proposed and demonstrated experimentally in this present study. The device is comprised only a fiber-optic microcavity that is formed by discharging a short section of hollow core fiber (HCF). The key to ensuring the success of the sensor relies on the preprocessing of hydrogen loading for HCF. When discharging the HCF, the hydrogen is heated up during the formation of the air bubble, which enlarges the bubble diameter, smoothens its surfaces simultaneously and decreases Young's modulus of the material of the bubble. Ultimately, this results in the probe being highly sensitive to ultrasound with a SNR of 69.28 dB. Once the compact air cavity is formed between the end face of the leading-in fiber and the top wall of the bubble, a well-defined interference spectrum is achieved based on the Fabryâ»Perot interference. By using spectral side-band filtering technology, we detect the ultrasonic waves reflected by the seismic physical model (SMF) and then reconstruct its three-dimensional image.
RESUMEN
A novel kind of fiber optic ultrasonic sensor based on matching fiber Bragg gratings (FBGs) is proposed and demonstrated. The sensors consist of a pair of matching FBGs fixed to a special bracket. The bracket plays a role in stretching and squeezing the FBGs, with the pushâ»pull effect efficiently coupling the ultrasonic signal to the sensor, thus, improving the sensor’s sensitivity. Side-band filtering technology-based intensity interrogation was used to detect ultrasounds in water. With the synergic effect of the matching FBGs, the sensor performed with a high signal-to-noise ratio (56.9 dB at 300 KHz, 53 dB at 1 MHz and 31.8 dB at 5 MHz) and the observed ultrasonic sinusoidal signals were undistorted and distinguishable in the time domain.