Nonpronuclear- and monopronuclear-derived blastocysts do not impair subsequent perinatal and maternal outcomes.

CONTEXT: The routine clinical practice is to prioritize the transfer of blastocysts derived from 2PN embryos if they are available. For women who only have blastocysts resulting from 0PN and 1PN embryos, whether to transfer these embryos or discard them has been an ongoing debate over the years. OBJECTIVE: To investigate the perinatal and obstetric outcomes following the transfer of vitrified-warmed single blastocysts derived from 0PN and 1PN zygotes. DESIGN: Retrospective cohort study. SETTING: University-affiliated IVF center. PATIENT(S): This study included singletons born to women who had undergone 0PN and 1PN vitrified-warmed single blastocyst transfers, compared to those resulting from 2PN vitrified-warmed single blastocyst transfers from 2012 to 2020. INTERVENTIONS: None. MAIN OUTCOME MEASURE(S): Perinatal and obstetric outcomes. RESULT(S): A total of 7,284 women were included in the final analysis. Of these, 386, 316, and 6582 cycles resulted from 0PN-, 1PN-, and 2PN-derived blastocysts transfer, respectively. The rates of clinical pregnancy, miscarriage, and live birth were similar across the study cohorts in both unadjusted and adjusted analyses. When comparing the 0PN and 2PN groups, no differences were found in birth outcomes after adjusting for confounders. Similarly, maternal complications and mode of delivery were comparable between these two study cohorts. Birth parameters were also similar between the 1PN and 2PN blastocyst groups, except for more male births in the 1PN cohort. Furthermore, a comparison between the 1PN and 2PN groups did not reveal any significant differences in maternal outcomes. CONCLUSION: The current study showed that the transfer of 0PN and 1PN blastocysts did not compromise reproductive outcomes or increase maternal and perinatal complications. This information is valuable for clinicians to counsel couples effectively and guide them in making informed decisions.

Acquired Fanconi syndrome in mixed cryoglobulinemia patients: a single-center case series.

PURPOSE: Cryoglobulinemia is a pathological condition characterized by the presence of cryoglobulins in the blood, with cryoglobulinemic glomerulonephritis being the most frequent form of renal involvement. Fanconi syndrome presents as a generalized dysfunction of the proximal tubule, characterized by the presence of polyuria, phosphaturia, glycosuria, proteinuria, proximal renal tubular acidosis, and osteomalacia. We aimed to present five cases co-occurring with Fanconi syndrome and cryoglobulinemia. METHODS: We retrospectively summarized the cases of five patients with Fanconi syndrome and cryoglobulinemia at Peking Union Medical College Hospital from January 2012 to June 2022. The clinical features, diagnosis, treatment, and prognosis were systematically analyzed. RESULTS: All five patients exhibited typical features of Fanconi syndrome, and cryoglobulinemia was concurrently detected in all cases. These patients also exhibit positive anti-nuclear antibody spectrum and hyperglobulinemia, and IgM constitutes the predominant monoclonal component in cryoglobulins. In addition to supplemental treatment, timely immunosuppressive therapy may potentially benefit the long-term renal prognosis of patients with this condition. CONCLUSION: Our findings highlight the rare co-occurrence of Fanconi syndrome and cryoglobulinemia in clinical practice. Despite the lack of causal evidence, the coexistence of Fanconi syndrome and tubulointerstitial injury is also noteworthy in patients with cryoglobulinemia, underscoring the importance of thorough evaluation and tailored management in patients presenting with overlapping renal manifestations. Key Points • Patients with mixed cryoglobulinemia can clinically present with tubulointerstitial injury, specifically manifesting as Fanconi syndrome. • In addition to typical symptoms of Fanconi syndrome, these patients also exhibit positive anti-nuclear antibody spectrum and hyperglobulinemia, while IgM constitutes the monoclonal component in cryoglobulins. • Timely immunosuppressive therapy may improve long-term renal prognosis in these patients.

Acute Kidney Injury in Different Anticoagulation Strategies: A Large-Scale Pharmacoepidemiologic Study Using Real-World Data.

PURPOSE: Acute kidney injury (AKI) following anticoagulant application has received growing attention as a significant emerging complication of anticoagulation. Nevertheless, there remains a lack of real-world studies to compare the incidence, clinical features, and prognosis of AKI across different anticoagulant regimens. METHODS: Disproportionality analysis and Bayesian analysis were used to identify suspected AKI cases after different anticoagulant use within the Food and Drug Administration's Adverse Event Reporting System from January 2004 to March 2023. The time to onset, fatality, and hospitalization rates of anticoagulant-associated AKI were also investigated. RESULTS: We identified 9313 anticoagulant-associated AKIs, which appeared to influence mostly patients over 65 years old (65.37%). Lepirudin displayed a stronger AKI association than others, based on the highest reporting odds ratio (ROR = 6.66, 95% CI = 3.97-11.18), proportional reporting ratio (PRR = 6.08, χ2 = 69.12), and empirical Bayes geometric mean (EBGM = 6.08, the lower 90% one-sided CI = 3.95). Warfarin showed the slightest association with AKI in oral anticoagulants, lower than any direct oral anticoagulants excluding apixaban. Edoxaban exhibited the highest potential renal risk among direct oral anticoagulants, with an ROR of 3.32 (95% CI = 2.95-3.72). The median time to AKI onset was 36 (IQR 7-205) days following the initiation of anticoagulation therapy, and most AKI cases occurred within the first month. CONCLUSION: Particular attention should be directed toward monitoring renal function in individuals receiving anticoagulants, including warfarin and direct oral anticoagulants, as well as other anticoagulant agents. This diligence is particularly imperative within the first month after anticoagulant administration for individuals with a tendency for AKI.

New-onset metabolic syndrome is associated with accelerated renal function decline partially through elevated uric acid: an epidemiological cohort study.

Background: The burden of metabolic syndrome (MetS) continues to rise globally and is associated with complications of multiple organ systems. We aimed to identify the association between changes in MetS status and accelerated renal function progression through a regional epidemiological survey in China, thus discovering influence factors with treatable potential. Methods: This study was a population-based survey conducted in 2008 and 2014, assessing a representative sample of 5,225 individuals from rural areas of China. They were divided into four subgroups according to their MetS status in 2008 and 2014 (Never, Previously abnormal, New-onset, and Consistent). Multivariate logistic regression and stratification analysis evaluated the relationship between clinical factors and renal function decline under different MetS statuses. Smooth curve fitting further addressed the role of serum uric acid, illustrating the vital turning point of uric acid levels in the background of renal function deterioration. Results: Of all groups of MetS states, the new-onset MetS showed the most significant eGFR decline, with a 6.66 ± 8.21 mL/min/1.73 m2 decrease over 6 years. The population with newly-onset MetS showed a considerable risk increase in delta eGFR with a beta coefficient of 1.66 (95%CI=1.09-2.23) after necessary correction. In searching for the drivers, the strength of the association was significantly reduced after additional adjustment for uric acid levels (ß=0.91, 95%CI=0.35-1.45). Regarding the turning point, uric acid levels exceeding 426 µmol/L were more significantly associated with the stepped-up deterioration of kidney function for those with new-onset MetS. Conclusion: Metabolic syndrome demonstrated a solid correlation with the progression of renal function, particularly in those with newly-onset MetS status. In addition to the diagnostic components of MetS, hyperuricemia could be used as a marker to identify the high risk of accelerating eGFR decline early. Furthermore, we suggested a potential renal benefit for the newly-onset MetS population when maintaining their serum uric acid level below the criteria for asymptomatic hyperuricemia.

Association of metabolic-dysfunction associated steatotic liver disease with polycystic ovary syndrome.

Non-alcoholic fatty liver disease (NAFLD), which has a prevalence of over 25% in adults, encompasses a wide spectrum of liver diseases. Metabolic-dysfunction associated steatotic liver disease (MASLD), the new term for NAFLD, is characterized by steatotic liver disease accompanied by cardiometabolic criteria, showing a strong correlation with metabolic diseases. Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disease affecting 4-21% of women of reproductive age. Numerous studies have indicated that NAFLD and PCOS often occur together. However, as MASLD is a new term, there is still a lack of reports describing the effects of MASLD on the development of PCOS. In this review article, we have summarized the complex and multifaceted connections between MASLD and PCOS. Understanding the pathogenesis and treatment methods could not only guide the clinical prevention, diagnosis, and treatment of PCOS in patients with MASLD, but also increase the clinical attention of reproductive doctors to MASLD.

Paternal age does not jeopardize the live birth rate and perinatal outcomes after in vitro fertilization: an analysis based on 56,113 frozen embryo transfer cycles.

BACKGROUND: The global trend of delaying childbearing has led to an increasing number of couples seeking in vitro fertilization. The adverse effects of advanced maternal age on pregnancy and perinatal outcomes are well documented, regardless of the conception method. In addition, advanced paternal age may contribute to poor reproductive potential because of high levels of sperm DNA fragmentation. However, it remains challenging to guide older men regarding the effect of paternal age on pregnancy and birth outcomes in the field of assisted reproduction. OBJECTIVE: This study aimed to investigate the association of paternal age with live birth and perinatal outcomes following in vitro fertilization-frozen embryo transfer. STUDY DESIGN: A retrospective study was performed at a university-affiliated fertility center, involving women who were younger than 36 years and had undergone frozen embryo transfer from January 2011 to June 2021. Subjects were categorized into 6 groups based on paternal age: <25, 25 to 29, 30 to 34, 35 to 39, 40 to 44, and ≥45 years. A generalized estimating equation logistic regression model was used to account for the clustered nature of data and to adjust for confounders. Paternal age between 25 and 29 years served as the reference group in the logistic regression models. RESULTS: A total of 56,113 cycles who met the inclusion criteria were included in the final analysis. On unadjusted analyses, the reproductive outcome parameters showed a considerable decline with increasing male age. The live birth rate decreased from 47.9% for men aged 25 to 29 years to 40.3% among men aged ≥40 years. Similarly, the clinical pregnancy rate decreased from 54.4% in the reference group to 47.8% in the ≥40 years age group. Conversely, the miscarriage rate increased as male age increased, from 10.2% among men aged 25 to 29 years to 13.5% among men aged ≥45 years. However, the differences in the reproductive outcomes mentioned above were no longer significant in the multivariable models. Compared with the younger controls, advanced paternal age was not associated with a lower chance of live birth (males aged 40-44 years: adjusted odds ratio, 0.94; 95% confidence interval, 0.85-1.04; males aged ≥45 years: adjusted odds ratio, 0.93; 95% confidence interval, 0.79-1.10). In addition, the rates of clinical pregnancy (males aged 40-44 years: adjusted odds ratio, 0.95; 95% confidence interval, 0.85-1.05; males aged ≥45 years: adjusted odds ratio, 0.94; 95% confidence interval, 0.79-1.12) and miscarriage (males aged 40-44 years: adjusted odds ratio, 1.05; 95% confidence interval, 0.85-1.31; males aged ≥45 years: adjusted odds ratio, 1.07; 95% confidence interval, 0.77-1.50) were comparable between the reference and advanced paternal age groups. Furthermore, men in the youngest age group (<25 years) did not have worse pregnancy outcomes than those in the reference group. Regarding perinatal outcomes, there was no difference among the study cohorts in terms of preterm birth, low birthweight, macrosomia, small for gestational age, and large for gestational age, both in the unadjusted and confounder-adjusted models. CONCLUSION: This study did not demonstrate a significant association between paternal age and live birth and perinatal outcomes after in vitro fertilization-frozen embryo transfer when the female partners were younger than 36 years. With the global trend toward delaying childbirth, our findings provide useful information for counseling patients that increasing paternal age may not adversely affect pregnancy and perinatal outcomes in assisted reproduction.

Obstetric and perinatal outcomes after embryos cultured in one-step versus sequential culture media systems in vitrified-warmed single blastocyst transfer cycles.

RESEARCH QUESTION: Does type of culture medium used influence obstetric and perinatal outcomes after vitrified-warmed single blastocyst transfers? DESIGN: Retrospective cohort study involving singletons after vitrified-warmed single blastocyst embryo transfers, using embryos cultured in either Irvine Continuous Single Culture medium (CSC) or Vitrolife G5TM PLUS medium culture system between 2013 and 2020. RESULTS: A total of 2475 women who had singleton deliveries were included for final analysis: 1478 had embryos cultured in CSC and 997 had embryos cultured in G5TM PLUS medium. Birth outcomes, including preterm birth, mean birth weight, gestational age- and sex-adjusted birth weight (Z-scores), rates of large-for-gestational-age, small-for-gestational-age, low birth weight and macrosomia, and the distribution of newborn gender did not differ significantly between groups in crude and adjusted analyses. Women whose embryos were cultured in G5TM PLUS frequently suffered from pregnancy-induced hypertensive disorders compared with those who had embryos cultured in CSC (4.7% versus 3.0%; P = 0.031). This difference was no longer significant after adjusting for several key confounders (adjusted odds ratio 1.49, 95% CI 0.94 to 2.38, P = 0.087). Other obstetric complications, including gestational diabetes mellitus, preterm premature rupture of membranes, abnormal placentation, postpartum haemorrhage and the mode of delivery were all similar between the two groups. CONCLUSIONS: The present study adds new information to the current evidence by suggesting that the embryo culture medium does not affect birth outcomes and obstetric complications when comparison is limited to Irvine CSC and Vitrolife G5TM PLUS in vitrified-warmed single blastocyst transfer cycles.

A Collaborative Learning Model for Skin Lesion Segmentation and Classification.

The automatic segmentation and classification of skin lesions are two essential tasks in computer-aided skin cancer diagnosis. Segmentation aims to detect the location and boundary of the skin lesion area, while classification is used to evaluate the type of skin lesion. The location and contour information of lesions provided by segmentation is essential for the classification of skin lesions, while the skin disease classification helps generate target localization maps to assist the segmentation task. Although the segmentation and classification are studied independently in most cases, we find meaningful information can be explored using the correlation of dermatological segmentation and classification tasks, especially when the sample data are insufficient. In this paper, we propose a collaborative learning deep convolutional neural networks (CL-DCNN) model based on the teacher-student learning method for dermatological segmentation and classification. To generate high-quality pseudo-labels, we provide a self-training method. The segmentation network is selectively retrained through classification network screening pseudo-labels. Specially, we obtain high-quality pseudo-labels for the segmentation network by providing a reliability measure method. We also employ class activation maps to improve the location ability of the segmentation network. Furthermore, we provide the lesion contour information by using the lesion segmentation masks to improve the recognition ability of the classification network. Experiments are carried on the ISIC 2017 and ISIC Archive datasets. The CL-DCNN model achieved a Jaccard of 79.1% on the skin lesion segmentation task and an average AUC of 93.7% on the skin disease classification task, which is superior to the advanced skin lesion segmentation methods and classification methods.

Contributions of NR1H3 genetic polymorphisms to susceptibility and effects of narrowband UVB phototherapy to nonsegmental vitiligo.

Vitiligo is the most common depigmenting disorder to which both genetic and environmental factors contribute. The aim of the current work was to evaluate the relationship between polymorphisms of the gene nuclear receptor subfamily 1 Group H member 3 (NR1H3) and the risk of vitiligo and phototherapy effects in the Chinese Han population. Two independent samples were enrolled to form the discovery set (comprised of 1668 nonsegmental vitiligo [NSV] patients and 2542 controls) and the validation set (comprised of 745 NSV patients and 1492 controls). A total of 13 tag single nucleotide polymorphisms (SNPs) were genotyped in the samples from the discovery stage. SNPs that achieved nominal significance were validated in another independent sample set. The serum level of NR1H3 protein was assayed using enzyme-linked immunosorbent assay kits in the validation set. Genetic association analysis was carried out at allelic and genotypic levels. The therapeutic effects of significant SNPs were examined in the validation set. The SNP rs3758672 was significantly associated with NSV. The A allele was correlated with NSV risk and poorer therapeutic effects. The A allele was strongly correlated with the increased level of serum NR1H3 in both controls and patients. In summary, SNP rs3758672 in NR1H3 was significantly associated with both disease susceptibility and individualized therapeutic effects of NSV in study participants with Han Chinese ancestry.

Clustering-based fusion for medical information retrieval.

Medicine is a fast-moving field, and the number of medical publications has increased rapidly over recent years. How to find relevant information from this vast pool of research effectively and efficiently has therefore become highly challenges. Previous studies have demonstrated that data fusion can improve search performance if properly utilized. However, in most cases effectiveness is the only concern and efficiency is not considered. A fusion-based system is by nature more complicated and expensive computationally than other retrieval models such as BM25, because many component retrieval systems and an extra layer of fusion are required. The number of component retrieval systems involved is an important indicator of complexity of the fusion-based system. We aim to select the optimal k-subset of component retrieval systems for any given number k, to optimize both fusion performance and reduce the cost of data fusion. A clustering-based approach is proposed. First all the candidates are divided into clusters by the Chameleon clustering algorithm, then representatives from every cluster are chosen by Sequential Forward Selection for fusion. Evaluated with two datasets from TREC, the proposed method performs more effectively than the other baseline methods including the state-of-the-art subset selection method significantly. When either of the two typical fusion methods is used, an improvement rate of over 10% is observed for both measures Mean Average Precision and Recall-level Precision, and an improvement rate of over 5% is observed for both measures Precision at 10 document level and Mean Reciprocal Rank.

Design and Optimization of Piezoelectric Cantilever Beam Vibration Energy Harvester.

Piezoelectric cantilever beams are commonly utilized to harvest energy from environmental vibrations due to their simple structures. This paper optimizes a single crystal trapezoidal hollow structure piezoelectric cantilever beam vibration energy harvester with a copper substrate to achieve high energy density at a low frequency. Finite element analysis (FEA) is adopted to optimize the size of the copper substrate at first, and the piezoelectric energy harvester (PEH) is further optimized with a trapezoidal hollow structure under the optimal size of the copper substrate. The developed PEH with a trapezoidal hollow structure (La = 20 mm, Lb = 15 mm, and Lh = 40 mm), with a copper substrate of 80 mm × 33 mm × 0.2 mm, can obtain the best output performance. Under the condition of 1 g acceleration, the resonance frequency and peak voltage output were 23.29 Hz and 40.4 V, respectively. Compared with the unhollowed PEH, the developed trapezoidal hollow structure PEH can reduce its resonant frequency by 12.18% and increase output voltage by 34.67%, while also supplying a power density of 7.24 mW/cm3. This study verified the feasibility of the optimized design through simulation and experimental comparison.

Multilayer structured CNF/rGO aerogels and rGO film composites for efficient electromagnetic interference shielding.

The development of efficient electromagnetic interference (EMI) shielding materials with high electromagnetic waves (EMWs) absorption capacity is of great significance to alleviate secondary EMWs pollution. Herein, multilayered composites were prepared by stacking cellulose nanofibril (CNF)/reduced graphene oxide (rGO) aerogels and rGO film together. The porous aerogels and the dense film serve as the EMWs absorbing layer and reflecting layer, respectively. When the EMWs enter the multilayer composites, they go through the process of absorption-reflection-reabsorption, resulting in a high EMI shielding effectiveness (SE) of ~32 dB. Furthermore, both experimental and theoretical analyses were adopted to explore the effect of arrangement order of CNF/rGO aerogels on EMI shielding performance. The results indicate that composites with progressively higher graphene content exhibit a higher EMWs absorption capacity at the same total EMI SE. This work offers a feasible design for improving EMWs absorption without affecting the overall EMI shielding performance of the material.

Gentiana straminea Maxim. polysaccharide decolored via high-throughput graphene-based column and its anti-inflammatory activity.

Gentiana straminea Maxim. exhibits various biological activities. However, the purification and functions of polysaccharides in Gentiana straminea Maxim. have never been reported. Herein, by proposing a flexible 3D graphene-based decoloration column (3DD column), Gentiana straminea Maxim. polysaccharide (GMP) was high-throughput obtained and its anti-inflammatory activity was investigated. Benefiting from the large macroporous network of 3D NH2-graphene oxide hydrogel with selective adsorption towards pigments, the 3DD column exhibits high decoloration ratio (96.41%). In addition, the 3DD column provides superior practical functionality as compared to the traditional approaches, which are time-consuming and need toxic solvents, and exhibiting widespread-application for the purification of polysaccharide from other common plant species. More importantly, the decolored GMP as a natural product has promising anti-inflammatory activity on RAW264.7 cells without negative impact on cell viability. Overall, this work reveals a new functional polysaccharides and provides a flexible approach for polysaccharide decoloration, exhibiting a promising prospect for natural polysaccharides in practical application of pharmaceutical.

Multiscale nanocelluloses hybrid aerogels for thermal insulation: The study on mechanical and thermal properties.

Herein, multiscale nanocelluloses (NCs) were prepared and blended with fumed silica and organosilane to fabricate freeze-dried aerogels in various combinations. The cooperation of multiscale raw materials optimized the porous structures of aerogels, thus improving the thermal insulation properties. The use of NCs with different characteristics endowed the resultant aerogels with distinct mechanical performance. The addition of high-aspect-ratio NCs in the composite aerogels was essential for improving the bendability. Furthermore, lower-aspect-ratio NCs helped to resist the compression deformation of the cross-linked aerogels. The functional groups on NCs made a difference in the thermal stability of the as-prepared aerogels. However, after treating at 150-350 ℃, the aerogels could maintain structural integrity and high elastic recovery rate, possessing ultralow density (7.2 kg/m3) and thermal conductivity (25.4 mW m-1 K-1). The outstanding thermal properties and controllable mechanical performance make these aerogels potential candidates in different fields such as textile and building industries.

Metabolomic analysis of simvastatin and fenofibrate intervention in high-lipid diet-induced hyperlipidemia rats.

AIM: To investigate the metabolite changes caused by simvastatin or fenofibrate intervention in diet-induced hyperlipidemia rats using a GC-MS-based metabolomic profiling approach. METHODS: SD rats were fed with high-lipid diet for 4 weeks to induce hyperlipidemia, then the rats were fed with normal diet, and orally administered with simvastatin (10 mg·kg(-1)·d(-1)) or fenofibrate (150 mg·kg(-1)·d(-1)) for 2 weeks. Blood samples were collected once a week, and potential biomarkers were examined using commercial assay kits and a metabolomic approach. The metabolomics data were analyzed using a multivariate statistical technique and a principal component analysis (PCA). RESULTS: Oral administration of simvastatin or fenofibrate significantly decreased the plasma levels of total cholesterol (TC) and low-density lipoprotein (LDL) cholesterol and increased the plasma level of high-density lipoprotein (HDL) cholesterol in the hyperlipidemia rats. Plasma samples were scattered in the PCA scores plots in response to the diet and to the drugs administered. The main metabolites changed in the hyperlipidemia rats were cholesterol, creatinine, linoleic acid, ß-hydroxybutyric acid, tyrosine, isoleucine and ornithine. The plasma level of creatinine was significantly lower in the simvastatin-treated rats than in the fenofibrate-treated rats. The plasma tyrosine concentration was declined following intake of high-lipid diet, which was reversed by fenobrate, but not by simvastatin. CONCLUSION: A series of potential biomarkers including tyrosine, creatinine, linoleic acid, ß-hydroxybutyric acid and ornithine have been identified by metabolomic profiling, which may be used to identify the metabolic changes during hyperlipidemia progression.

Changes in expression and distribution of attractin in the testes of rats at different developmental stages.

Attractin (Atrn), an autosomal recessive gene, is widely expressed in the body and displays multiple physiological and pathological functions in different types of tissues. The objective of this study was to localize Atrn protein and mRNA in the testis and epididymis of rats at different stages of maturation. Testis and epididymidis samples were obtained from the following 5 groups of Sprague Dawley (SD) rats in different developmental stages: newborn (8 h after birth), prepubertal (5 days), pubertal (20 days), postpubertal (50 days) and mature (70 days). Tissues were fixed and prepared for indirect immunofluorescence, immunohistochemistry, in situ hybridization, confocal laser scanning microscopy and western blot assays. A polyclonal antiserum against mouse Atrn and oligonucleotide riboprobes were used in the above assays. At the different stages of maturation, Atrn protein and mRNA were both widely expressed in the rat testis, including Leydig cells, primitive spermatogonia, primary spermatocytes, spermatids, Sertoli and peritubular myoid cells. Staining of the Atrn protein was mainly located on the cell membrane and in the cell cytoplasm while Atrn mRNA was distributed in both the nucleus and cytoplasm. No immunopositive staining was detected in spermatozoa and epididymides. In the epididymis, comprised of the caput, corpus and cauda, there was no definitive immunopositive staining within the efferent ductules or epididymal ducts. Taken together, Atrn protein and mRNA are both expressed widely in the rat testis at different stages of maturation, which suggests that Atrn protein is involved and plays an important role in the development of the reproductive system. In addition, the rat testis has the ability to synthesize Atrn protein throughout sexual development.

Simultaneous determination of Eleutheroside B and Eleutheroside E in rat plasma by high performance liquid chromatography-electrospray ionization mass spectrometry and its application in a pharmacokinetic study.

Eleutheroside B and Eleutheroside E, two kinds of the major bioactive saponins of Eleutherococcus senticosus, play a pivotal role in biologic activity. In this study, a specific and sensitive high performance liquid chromatography-electrospray ionization-tandem mass spectrometry method (HPLC-MS/MS) was developed and validated for simultaneous determination of Eleutheroside B and Eleutheroside E in rat plasma. The analytes were extracted from rat plasma via a simple protein precipitation procedure with methanol and polygonin was used as internal standard. Chromatographic separation was achieved on a C18 column using a gradient elution program with acetonitrile and water containing 0.1% ammonium hydroxide solution as the mobile phase, with a flow rate of 0.2mL/min. The detection was performed on a triple-quadrupole tandem mass spectrometer by multiple reactions monitoring (MRM) mode in a negative ion mode via electrospray ionization (ESI). The transition monitored were m/z 371 [M-H](-)→209 for Eleutheroside B, m/z 741[M-H](-)→579 for Eleutheroside E and m/z 389[M-H](-)→277 for internal standard. Linear calibration curves were obtained in the concentration range of 1-2000ng/mL for both (Eleutheroside B and Eleutheroside E), with a lower limit of quantification of 1ng/mL. Extraction recovery was over 80% in plasma. The intra- and inter-day precision (RSD) values were below 12% and accuracy (RE) was -2.80 to 5.70% at three QC levels for both. The assay was successfully applied to study pharmacokinetics behavior in rats after oral and intravenous administration of the single substances (Eleutheroside B and Eleutheroside E). And further research was performed by comparing the difference in pharmacokinetic behavior between the single substances and an aqueous extract of E. senticosus after oral administration. Significant difference in pharmacokinetic characteristics between the single substances and an aqueous extract was found in rat, which would be beneficial for the pre-clinical research and clinical use of E. senticosus.

Rapid analysis of tadalafil in human blood plasma and seminal plasma by liquid chromatography/tandem mass spectrometry.

A selective, sensitive and rapid reversed-phased liquid chromatography with tandem mass spectrometry (LC-MS/MS) method was developed and validated for quantitation of tadalafil in human blood plasma and seminal plasma. Sample preparation was extracted with methyl tert-butyl ether under alkaline conditions and tadalafil was separated by HPLC using a C18 column and an isocratic mobile phase consisting of methanol and 2 mM ammonium acetate containing 0.05% formic acid in water (52:48, v/v) running at a flow rate of 0.2 mL/min for 5 min. The detection was carried out on a triple-quadrupole tandem mass spectrometer by multiple reactions monitoring (MRM) mode via electrospray ionization (ESI) and domperidone as internal standard. Calibration curves were linear over tadalafil concentration ranges of 2-1000 ng/mL in plasma and 2-100 ng/mL in seminal plasma with the lower limits of quantitation (LLOQ) of 2 ng/mL for both. The intra-day and inter-day accuracy and precision in two matrixes at all QC samples were within the generally accepted criteria for bio-analytical method that precision (RSD) values were below 15% and accuracy (RE) was 1.44-8.32% for plasma and -3.12% to 7.71% for seminal plasma. Finally, the validated method was applicable to clinical pharmacokinetic study of tadalafil in healthy volunteers following oral administration and to further investigate the correlation of tadalafil distribution in human blood plasma and seminal plasma, and tadalafil potential influence in sperm quality.

Simultaneous determination of oridonin, ponicidin and rosmarinic acid from Herba Isodi Rubescentis extract by LC-MS-MS in rat plasma.

An analytical method was developed for the simultaneous quantification of oridonin, ponicidin and rosmarinic acid in rat plasma using liquid chromatography-tandem mass spectrometry with electrospray ionization in positive and negative ion modes of operation. Multiple reaction monitoring was used for quantification using the precursor to product ion pairs of m/z 359.100 → 160.9 (rosmarinic acid), m/z 150.1 → 106.9 (acetaminophen as internal standard 1), m/z 365.1 → 347.3 (oridonin), m/z 363.3 → 345.2 (ponicidin) and m/z 258.3 → 201.0 (dextrorphan as internal standard 2). Optimum chromatographic separation was achieved with a BDS Hypersil C18 column (100 × 2.1 mm, 2.4 µm) and a mobile phase consisting of acetonitrile-5mM ammonium acetate containing 0.1% acetic acid (25:75, v/v). The analytes were extracted from rat plasma using a single liquid-liquid extraction technique and the calibration curve was linear within the concentration range of 2-1,000.0 ng/mL for three analytes. The extraction recovery was above 70%, the intra-day and inter-day precision of the quality control (QC) samples was less than 12.45% and the accuracy of the QC samples was 96.99-105.79% of the nominal values. The method was sensitive and reliable and was successfully applied to a pharmacokinetic and bioavailability study of Herba Isodi Rubescentis extract in rats.

Development and validation of a sensitive liquid chromatography/tandem mass spectrometry method for the determination of raddeanin A in rat plasma and its application to a pharmacokinetic study.

A simple and sensitive high-performance liquid chromatography-electro-spray ionization tandem mass spectrometry (LC-ESI-MS/MS) method was developed and validated to determine raddeanin A in rat plasma. After precipitation of rat plasma samples with methanol, chromatographic separation was achieved on a BDS Hypersil C18 column (100×2.1mm, 2.4µm) using the mobile phase consisted of acetonitrile and 2mM ammonium acetate with 0.05% formic acid (60:40, v/v). The detection was performed in a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode using negative ionization. The transition monitored were m/z 895.6→455.0 for raddeanin A and m/z 359.3→329.0 for IS, respectively. The method was linear over the concentration range of 2-1000ng/mL for raddeanin A. The intra-day and inter-day assay variations were <9.46%, and the accuracy values were between -2.04% and -6.52% relative error. The extraction recovery of raddeanin A was more than 70%, and the relative matrix effect ranges from 108.52% to 112.36%. The validated method has been successfully applied to determine the pharmacokinetic profile of raddeanin A in rat plasma following oral and intravenous administration.