Integrating UPLC-QTOF-MS/MS and UPLC-DAD to evaluate the influence of sulfur-fumigated Paeoniae Radix Alba on the overall quality of three Si-Wu-Tang formulations.

INTRODUCTION: Sulfur-fumigation of Paeoniae Radix Alba (PRA) could induce the chemical transformation of its bioactive component paeoniflorin into a sulfur-containing derivative paeoniflorin sulfite, and thus alter the quality, bioactivities, pharmacokinetics, and toxicities of PRA. However, how sulfur-fumigated PRA (S-PRA) affects the quality of PRA-containing complex preparations has not been intensively evaluated. OBJECTIVES: We intend to evaluate the influence of S-PRA on the overall quality of three kinds of Si-Wu-Tang (SWT) formulations, i.e., decoction (SWT-D), granule (SWT-G), and mixture (SWT-M). MATERIAL AND METHODS: An UPLC-DAD multi-components quantification method was used to compare the transfer rates of paeoniflorin sulfite and other 10 bioactive components between S-PRA-containing and NS-PRA-containing SWT formulations. An UPLC-QTOF-MS/MS-based target metabolomics approach was applied to explore the differential sulfur-containing derivatives in S-PRA-containing SWT formulations. RESULTS: The transfer rates of paeoniflorin sulfite in three S-PRA-containing SWT formulations were all higher than 100%. Moreover, S-PRA also increased the transfer rate of 5-hydroxymethylfurfural, 1,2,3,4,6-O-pentagalloylglucose, whereas decreased that of paeoniflorin, albiflorin, and ferulic acid in three SWT formulations. Six pinane monoterpene glucoside sulfites originally identified in S-PRA, were also detectable in three S-PRA-containing SWT formulations. In addition, seven phenolic acid sulfites including (3Z)-6-sulfite-ligustilide, (3E)-6-sulfite-ligustilide, 6,8-disulfite-ligustilide, ferulic acid sulfite, neochlorogenic acid sulfite, chlorogenic acid sulfite, and angelicide sulfite (or isomer) were newly identified in these three S-PRA-containing formulations. CONCLUSION: S-PRA could differentially affect the transfer rate of paeoniflorin sulfite and other bioactive components during the preparation of three SWT formulations and subsequently the overall quality thereof.

Rationality of the ethanol precipitation process in modern preparation production of Zishui-Qinggan decoction evaluated by integrating UPLC-QTOF-MS/MS-based chemical profiling/serum pharmacochemistry and network pharmacology.

INTRODUCTION: Zishui-Qinggan decoction (ZQD) is a classical traditional Chinese medicine formula (TCMF) for alleviating menopausal symptoms (MPS) induced by endocrine therapy in breast cancer patients. In the production of TCMF modern preparations, ethanol precipitation (EP) is a commonly but not fully verified refining process. OBJECTIVES: Chemical profiling/serum pharmacochemistry and network pharmacology approaches were integrated for exploring the rationality of the EP process in the production of ZQD modern preparations. MATERIAL AND METHODS: Ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS) was applied to identify the chemical profiles and absorbed components of ZQD. Network pharmacology was used to identify targets and pathways related to MPS-relieving efficacy. RESULTS: The chemicals of ZQDs without/with EP process (referred to as ZQD-W and ZQD-W-P, respectively) were qualitatively similar with 89 and 87 components identified, respectively, but their relative contents were different; 51 components were detectable in the serum of rats orally administered with ZQD-W, whereas only 19 were detected in that administered with ZQD-W-P. Key targets, such as AKT1, and pathways, such as the PI3K-Akt signalling pathway, affected by ZQD-W and ZQD-W-P were similar, while the neuroactive ligand-receptor interaction pathway among others and the MAPK signalling pathway among others were specific pathways affected by ZQD-W and ZQD-W-P, respectively. The specifically absorbed components of ZQD-W could combine its specific key targets. CONCLUSION: The EP process quantitatively altered the chemical profiles of ZQD, subsequently affected the absorbed components of ZQD, and then affected the key targets and pathways of ZQD for relieving MPS. The EP process might result in variation of the MPS-relieving efficacy of ZQD, which deserves further in vivo verification.

Integrating serum pharmacochemistry and network pharmacology to identify chemical markers for quality control of Apocyni Veneti Folium.

INTRODUCTION: Apocyni Veneti Folium (AVF) is a commonly used traditional Chinese medicinal herb for the treatment of hypertension. Chemical markers are crucial for the quality control of herbal medicines; however, the therapeutic components of AVF remain to be well elucidated. OBJECTIVES: This study was intended to integrate serum pharmacochemistry and network pharmacology to identify chemical markers of AVF and establish an efficacy-related quality control method of AVF. MATERIAL AND METHODS: Ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS) was applied to identify the absorbed AVF constituents in rat serum. Network pharmacology was further used to identify anti-hypertension-related chemical markers. Subsequently, a quantitative method was established using UPLC with diode array detection (DAD) and applied for quality evaluation of commercial AVF samples. RESULTS: Thirteen prototype constituents were unequivocally or tentatively characterized in serum samples, among which quercetin, kaempferol, hyperoside, isoquercitrin, chlorogenic acid, cryptochlorogenic acid, and neochlorogenic acid were identified as dominant chemicals related to anti-hypertensive efficacy. The quantitative data showed that the total contents of seven marker components even showed 2-fold variation among 14 batches of commercial AVF samples with RSD values ranging from 12.15% to 75.61%. Hierarchical cluster analysis and heatmap analysis showed that 14 batches of commercial AVF samples could be divided into three main groups. CONCLUSION: The chemical markers obtained from this study could be applicable for efficacy-related quality control of AVF.

Quality consistency evaluation of commercial Prunellae Spica by integrating determination of secondary metabolites and saccharides.

INTRODUCTION: Prunellae Spica (PS) is a commonly used medicinal herb in China. Secondary metabolites and saccharides are major bioactive components of PS. However, holistic quality consistency of commercial PS is ambiguous due to lack of comprehensive evaluation methods and reliable quality control markers. OBJECTIVES: Integrating multiple chromatographic and chemometric methods to comprehensively evaluate the holistic quality of PS. MATERIAL AND METHODS: Ultrahigh-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-TQ-MS/MS) was applied to quantify 12 secondary metabolites of PS. High-performance liquid chromatography coupled with photodiode array/evaporative light scattering detection (HPLC-PDA/ELSD) and high-performance gel permeation chromatography (HPGPC) methods were used to characterise the saccharides. Multivariate statistical analysis was adopted to evaluate the quality consistency of commercial PS and explore the potential quality control markers. RESULTS: The contents of secondary metabolites and saccharides were significantly different among commercial PS. All samples could be classified into three groups with ferulic acid, protocatechualdehyde, gallic acid, ursolic acid/oleanolic acid, sucrose, p-coumaric acid, chlorogenic acid as the major contributing components responsible for the difference. The content of rosmarinic acid was correlated with that of betulinic acid, hyperposide, chlorogenic acid, rutin, caffeic acid, p-coumaric acid and glucose, whereas polysaccharides, ferulic acid, protocatechualdehyde and ursolic acid/oleanolic acid, quercetin, sucrose and majority monosaccharides were not. CONCLUSION: The holistic quality of commercial PS was inconsistent. Together with rosmarinic acid, ferulic acid, protocatechualdehyde, ursolic acid/oleanolic acid, polysaccharides and sucrose might be recommended as potential quality control markers for the holistic quality control of PS.

SubPhaser: a robust allopolyploid subgenome phasing method based on subgenome-specific k-mers.

With advanced sequencing technology, dozens of complex polyploid plant genomes have been characterized. However, for many polyploid species, their diploid ancestors are unknown or extinct, making it impossible to unravel the subgenomes and genome evolution directly. We developed a novel subgenome-phasing algorithm, SubPhaser, specifically designed for a neoallopolyploid or a homoploid hybrid. SubPhaser first searches for the subgenome-specific sequence (k-mer), then assigns homoeologous chromosomes into subgenomes, and further provides tools to annotate and investigate specific sequences. SubPhaser works well on neoallopolyploids and homoploid hybrids containing subgenome-specific sequences like wheat, but fails on autopolyploids lacking subgenome-specific sequences like alfalfa, indicating that SubPhaser can phase neoallopolyploid/homoploid hybrids with high accuracy, sensitivity and performance. This highly accurate, highly sensitive, ancestral data free chromosome phasing algorithm, SubPhaser, offers significant application value for subgenome phasing in neoallopolyploids and homoploid hybrids, and for the subsequent exploration of genome evolution and related genetic/epigenetic mechanisms.

Integrating Multi-Type Component Determination and Anti-Oxidant/-Inflammatory Assay to Evaluate the Impact of Pre-Molting Washing on the Quality and Bioactivity of Cicadae Periostracum.

Cicadae Periostracum (CP) is a traditional Chinese medicinal herb derived from the slough that is molted from the nymph of the insect Cryptotympana pustulata Fabricius. Washing with water to remove residual silt is a primary processing method of CP that is recommended by the Chinese Pharmacopoeia, but how washing methods affect the quality and bioactivity of CP is unknown. In this study, the quality and bioactivity of non-washed CP (CP-NW), post-molting-washed CP (CP-WAT), and pre-molting-washed CP (CP-WBT) were comparatively investigated. The quality of these CP samples was evaluated in terms of the UPLC-QTOF-MS/MS-based chemical profiling and semi-quantification of 39 N-acetyldopamine oligomers (belonging to six chemical types), the HPLC-UV-based quantification of 17 amino acids, the ICP-MS-based quantification of four heavy metals, and the contents of ash; the bioactivities of the samples were compared regarding their anti-oxidant and anti-inflammatory activities. It was found that, compared with CP-NW, both CP-WBT and CP-WAT had significantly lower contents of ash and heavy metals. Moreover, compared with CP-WAT, CP-WBT contained lower levels of total ash, acid-insoluble ash, and heavy metals and higher contents of N-acetyldopamine oligomers and amino acids. It also had enhanced anti-oxidant and anti-inflammatory activities. A Spearman's correlation analysis found that the contents of N-acetyldopamine oligomers and free amino acids were positively correlated with the anti-oxidant/-inflammatory activities of CP. All these results suggest that pre-molting washing can not only remove the residual silt but can also avoid the loss of the bioactive components and assure higher bioactivities. It is concluded that pre-molting washing could enhance the quality and bioactivity of CP and should be a superior alternative method for the primary processing of qualified CP.

Poor cardiovascular health status among Chinese women.

BACKGROUND: Systematic investigation and analysis of cardiovascular health status (CVHS) of Chinese women is rare. This study aimed to assess CVHS and atherosclerotic cardiovascular disease (ASCVD) burden in the Chinese women physicians (CWP) and community-based non-physician cohort (NPC). METHODS: In this prospective, multicenter, observational study, CVHS using the American Heart Association (AHA) defined 7 metrics (such as smoking and fasting glucose) and ASCVD risk factors including hypertension, hyperlipidemia and type-2 diabetes were evaluated in CWP compared with NPC. RESULTS: Of 5832 CWP with a mean age of 44 ± 7 years, only 1.2% achieved the ideal CVHS and 90.1% showed at least 1 of the 7 AHA CVHS metrics at a poor level. Total CVHS score was significantly decreased and ASCVD risk burden was increased in postmenopausal subjects in CWP although ideal CVHS was not significantly influenced by menopause. Compared to 2596 NPC, fewer CWP had ≥ 2 risk factors (8% vs. 27%, P < 0.001); CWP scored significantly higher on healthy factors, a composite of total cholesterol, blood pressure, fasting glucose (P < 0.001), but, poorly on healthy behaviors (P < 0.001), specifically in the physical activity component; CWP also showed significantly higher levels of awareness and rates of treatment for hypertension and hyperlipidemia, but, not for type-2 diabetes. CONCLUSION: Chinese women's cardiovascular health is far from ideal and risk intervention is sub-optimal. Women physicians had lower ASCVD burden, scored higher in healthy factors, but, took part in less physical activity than the non-physician cohort. These results call for population-specific early and improved risk intervention.

In-depth transcriptome characterization uncovers distinct gene family expansions for Cupressus gigantea important to this long-lived species' adaptability to environmental cues.

BACKGROUND: Cupressus gigantea, a rare and endangered tree species with remarkable medicinal value, is endemic to the Tibetan Plateau. Yet, little is known about the underlying genetics of the unique ecological adaptability of this extremely long-lived conifer with a large genome size. Here, we present its first de novo and multi-tissue transcriptome in-depth characterization. RESULTS: We performed Illumina paired-end sequencing and RNA libraries assembly derived from terminal buds, male and female strobili, biennial leaves, and cambium tissues taken from adult C. gigantea. In total, large-scale high-quality reads were assembled into 101,092 unigenes, with an average sequence length of 1029 bp, and 6848 unigenes (6.77%) were mapped against the KEGG databases to identify 292 pathways. A core set of 41,373 genes belonging to 2412 orthologous gene families shared between C. gigantea and nine other plants was revealed. In addition, we identified 2515 small to larger-size gene families containing in total 9223 genes specific to C. gigantea, and enriched for gene ontologies relating to biotic interactions. We identified an important terpene synthases gene family expansion with its 121 putative members. CONCLUSIONS: This study presents the first comprehensive transcriptome characterization of C. gigantea. Our results will facilitate functional genomic studies to support genetic improvement and conservation programs for this endangered conifer.

miRNAS in cardiovascular diseases: potential biomarkers, therapeutic targets and challenges.

Cardiovascular diseases (CVD) are the leading cause of morbidity and mortality in the world. Although considerable progress has been made in the diagnosis, treatment and prognosis of CVD, there is still a critical need for novel diagnostic biomarkers and new therapeutic interventions to decrease the incidence of this disease. Recently, there is increasing evidence that circulating miRNAs (miRNAs), i.e. endogenous, stable, single-stranded, short, non-coding RNAs, can be used as diagnostic biomarkers for CVD. Furthermore, miRNAs represent potential novel therapeutic targets for several cardiovascular disorders. In this review we provides an overview of the effects of several CVD; including heart failure, acute myocardial infarction, arrhythmias and pulmonary hypertension; on levels of circulating miRNAs. In addition, the use of miRNA as therapeutic targets is also discussed, as well as challenges and recommendations in their use in the diagnosis of CVD.

Prenatal phthalate exposure and placental size and shape at birth: A birth cohort study.

OBJECTIVE: There is concern over the potential placental effects of prenatal phthalate exposure, and the potential adverse effects of prenatal phthalate exposure require further study; however, few data are available in humans. We investigated the associations between phthalate exposure in each trimester and both placental size and shape at birth. METHODS: We measured the urinary concentrations of phthalate metabolites among 2725 pregnant women in the Ma'anshan Birth Cohort. Before collecting urine samples from each of the three trimesters, the pregnant women were interviewed via questionnaires. Placental information was obtained from hospital records. We estimated the sex-specific associations between urinary phthalate concentrations in each trimester and both placental size and shape at birth using adjusted multiple regression. A linear mixed model was used for the repeated measures analysis with subject-specific random intercepts and slopes for gestational age at sample collection to test the effect of phthalate levels on placental size and shape and to estimate the effect sizes. RESULTS: Overall, placental breadth increased by 0.148cm (95% CI: 0.078, 0.218) with each 1 ln-concentration increase in MBP in the first trimester. The difference between placental length and breadth (length-breadth) decreased by 0.086cm (95% CI: -0.159, -0.012) and 0.149cm (95% CI: -0.221, -0.076) with each 1 ln-concentration increase in MMP and MBP, respectively, in the first trimester. In the second trimester, placental thickness increased by 0.017cm (95% CI: 0.006, 0.027), 0.020cm (95% CI: 0.004, 0.036), 0.028cm (95% CI: 0.007, 0.048), and 0.035cm (95% CI: 0.018, 0.053) with each 1 ln-concentration increase in MMP, MBP, MEOHP, and MEHHP, respectively. In the third trimester, placental thickness increased by 0.037cm (95% CI: 0.019, 0.056) and 0.019cm (95% CI: 0, 0.037) with each 1 ln-concentration increase in MBP and MEHP, respectively. Multiple linear regression for each offspring sex indicated that prenatal phthalate exposure increased placental thickness in both the first and second trimesters in males, whereas the corresponding relationship was close to null in females. Linear mixed models (LMMs) yielded similar results. CONCLUSION: Our results suggest the presence of associations between prenatal phthalate exposure and placental size and shape. Exposure to certain phthalates may cause the placenta to become thicker and more circular. Associations appeared stronger for the subsample representing male offspring than those for the subsample representing female offspring. Given the few studies on this topic, additional research is warranted.

Photocatalytic reduction of CO2 based on a CeO2 photocatalyst loaded with imidazole fabricated N-doped graphene and Cu(ii) as cocatalysts.

Cocatalysts are vital for improving photocatalytic activity. Incorporating nitrogen atoms on a graphene frame using an imidazole cycle resulted in a new N-doped graphene (denoted as ING). Cerium(iv) oxide (CeO2) nanoparticles were dispersed on ING sheets, producing an ING/CeO2 hybrid material. The ING/CeO2 hybrid material was characterized using X-ray diffraction, transmission electron microscopy, Raman spectroscopy, ultraviolet-visible diffuse reflectance spectroscopy and surface photovoltage spectroscopy. Copper(ii) ions [Cu(ii)] were adsorbed on the ING/CeO2 hybrid material to directly form Cu(ii)/ING/CeO2, which could capture the photogenerated electrons to reduce carbon dioxide (CO2) to methanol (CH3OH) under incident light irradiation. The results showed that the yield from reducing CO2 to CH3OH during the photocatalytic process using Cu(ii)/ING/CeO2 as the photocatalyst approached 385.8 µmol g-1 cat. h-1, whereas the yield was only 3.57 µmol g-1 cat. h-1 using ING/CeO2 as the photocatalyst. This shows that the Cu(ii) ions play a vital role during photocatalytic reduction of CO2 by forming copper(i) ions [Cu(i)]. The percentage of ING in the ING/CeO2 hybrid material was investigated, and the results indicated that 3.6% of ING achieved an optimal yield of CH3OH during the photo-reduction process. The simultaneous roles of Cu(ii) ions and ING sheets demonstrate a synergistic strategy for improving the photocatalytic CO2 reduction.

Vitamin D and risk of preterm birth: Up-to-date meta-analysis of randomized controlled trials and observational studies.

AIM: We performed a meta-analysis of randomized controlled trials (RCT) and observational studies to answer the two following questions: (i) whether low maternal circulating 25 hydroxyvitamin D (25-OHD) is associated with an increased risk of preterm birth (PTB) or spontaneous PTB (sPTB); and (ii) whether vitamin D supplementation alone during pregnancy can reduce the risk of PTB. METHODS: Literature search was carried out using Pubmed, Web of Science and Embase databases up to June 2016. Pooled OR or relative risk (RR) with 95%CI were computed using fixed or random effects models depending on the size of heterogeneity. Subgroup analysis was used to explore potential sources of between-study heterogeneity. Publication bias was evaluated using Egger's test and Begg's test. RESULTS: Twenty-four articles (six RCT and 18 observational studies) were identified. Maternal circulating 25-OHD deficiency (pooled OR, 1.25; 95%CI: 1.13-1.38) rather than insufficiency (pooled OR, 1.09; 95%CI: 0.89-1.35) was associated with an increased risk of PTB, and vitamin D supplementation alone during pregnancy could reduce the risk of PTB (pooled RR, 0.57; 95%CI: 0.36-0.91). This was also the case for the sPTB subgroup (circulating 25-OHD <50 vs >50 nmol/L; pooled OR, 1.45; 95%CI: 1.20-1.75). CONCLUSIONS: Maternal circulating 25-OHD deficiency could increase PTB risk and vitamin D supplementation alone during pregnancy could reduce PTB risk. Extrapolation of the results, however, must be done with caution, and there is urgent need for larger, better-designed RCT to confirm this effect.

Spectroscopic and Molecular Modeling Studies on Binding of Fleroxacin with Human Serum Albumin.

Fleroxacin (FLRX) is a new member of the class of fluoroquinolones, its effects on human serum albumin (HSA) and the mechanism of action are poorly understood, Especially, the secondary structural alterations of HSA induced by FLRX and the inner filter effect, which resulted in a spurious decrease in the observed fluorescence intensity and affected the binding parameters calculated from it are not considered. In this paper, binding of FLRX to HSA has been studied using multi-spectroscopy and molecular modeling methods. Fluorescence spectra revealed that the observed fluorescence quenching of HSA by FLRX was due to a 1∶1 complex formation by a static quenching process with a constant of 105 L·mol-1. The thermodynamic parameters (ΔH and ΔS) were calculated to be -107.99 kJ·mol-1 and -240.99 J·mol-1·K-1 via the Van't Hoff equation, which indicated that hydrogen bond and van der Waals force were the dominant intermolecular force. From the synchronous fluorescence, FT-IR and three dimensional fluorescence spectra, it was evident that the binding of FLRX to HSA induced a conformational change in the protein, and the alterations of secondary structure were quantitatively calculated by the evidence from FTIR spectra with reductions of α-helices of about 18.3%, decreases of ß-sheet structures of about 9.6%, and increases of ß-turn structures of about 18.0%. Site marker competitive experiments showed that phenylbutazone and FLRX shared a common binding site Ⅰ corresponding to the subdomain Ⅱ A of HSA. The binding details between FLRX and HSA were further confirmed by molecular docking studies, which revealed that FLRX was bound at subdomain Ⅱ A through multiple interactions, such as hydrogen bond, hydrophobic and van der Waals, etc. The accurate and full basic data in the work is beneficial to clarify the binding mechanism of FLRX with HSA and is helpful for understanding its effect on protein function during the blood transportation process.

Ultrasound-Assisted Extraction May Not Be a Better Alternative Approach than Conventional Boiling for Extracting Polysaccharides from Herbal Medicines.

In clinical practice polysaccharides from herbal medicines are conventionally prepared by boiling water extraction (BWE), while ultrasound-assisted extraction (UAE) has often been used instead employed in laboratory research due to its strong extraction ability and efficiency. However, if and how the polysaccharides obtained by UAE and BWE are comparable, and hence whether the UAE-based research is instructive for the actual usage of herbal polysaccharides still requires further evaluation. To address this issue, here we chemically analyzed and compared the UAE- and BWE-obtained polysaccharides from three herbal medicines, i.e., Ginseng Radix, Astragali Radix and Dendrobii Officinalis Caulis. Then, the spike recovery of two series of standard dextran and pullulan by UAE and BWE was tested. The results showed that the polysaccharides from the herbal medicines by UAE were quantitatively and qualitatively different with those by BWE. The powerful extraction ability and polysaccharide degradation caused by ultrasound collectively contributed to these differences. It was then revealed that not only the UAE conditions but also the polysaccharide structures could affect the extraction ability and polysaccharide degradation. Given these, we highly recommended that the effects of UAE on polysaccharides from herbal medicines should be first carefully considered before employing it in relevant chemical and pharmacological analysis.

The Levels of 21 Elements and Inter-element Interactions in Scalp Hair of Women at Childbearing Age in A Rural Area, Inner Mongolia.

Women of reproductive age are at an increased risk of minerals deficiencies, especially for women in recent years. The objective of this study was to investigate the concentrations of 21 elements (Ca, Mg, Cu, Zn, Fe, Mn, Cr, Ti, B, Co, Mo, Si, V, Ni, Cd, Al, Pb, Ba, Sr, Sn and Se) in scalp hair of apparently healthy women at childbearing age of Xinghe County, Ulanqab city of Inner Mongolia. The hair samples were determined by inductively coupled plasma-atomic emission spectrometry (ICP-AES) and atomic fluorescence spectrometry (AFS), respectively. The results revealed that the concentration levels of 21 elements were clearly differed from the mean levels reported in the literatures. Micronutrient deficiencies and apparent excess of toxic elements are present in female of childbearing age in studied area. The higher Mg and Al levels may be due to the largest bentonite deposit in North China lay in the study area. The soils in the studied areas had been detected to have low total Se concentration which may account for that 98% participants had Se concent below 0.2 µg·g-1. Inter-element interactions were studied by evaluation of correlation coefficients between two elements, as well as by multiple regression analysis. These elements form divalent ions and their sources are similar were significantly inter-correlated. The strongest relations found between the elements in the hair were as follows: Mg and Ca, Cr and B, Si and Pb, Mn and Pb, Fe and Mn. No correlation was found between Cd and Pb, which is different with other reports. This could be explained by the rarely industrial exposure to these elements. And kinds of geographical environment, dietary habits and other factors in different regions impact human trace element metabolism. Multiple regression analysis presented the results: Ca=f(Mg, Se) (Se are negatively correlated, ß<0), Fe=f(Cu, Mn, Ti), Zn=f(Ca, Se, Fe, Ni) (Fe, Ni are negatively correlated, ß<0), Cu=f(Pb, Fe, Cd), Al=f(Mg, Pb), Se=f(Zn, Cr, Ca) (Ca are negatively correlated, ß<0). These relations can be useful to study the relationships among different elements inside an organism of women. Cluster analysis (CA) was used for further classifying the different sources of elements on the basis of the similarities of their chemical properties. The significantly correlated subcluster of Mn-Pb-Si-Al proved that Pb exposure came from a nature source. The results will provide a reliable basis for improving macro- and microelements status of women in childbearing age in rural area. Meanwhile，this study may help to make more effective strategies to improve the reproductive-aged women's health and the pregnancy outcome.

Distribution of Mineral Elements and Influencing Factors in Scalp Hair of Childbearing Age Women in the County of Xinghe, Inner Mongolia of China.

The distribution and influencing factors on age, reproductive history and dietary habits in scalp hair of rural childbearing female in the county of XingHe, Inner Mongolia of China were studied. 21 mineral elements including essential and toxic elements (Al, B, Ba, Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Mo, Ni, Pb, Se, Si, Sn, Sr, Ti, V and Zn ) in scalp hair samples from 180 females of childbearing age were measured with inductively coupled plasma-atomic emission spectrometry (ICP-AES) and atomic fluorescence spectrometry (AFS), respectively. The results demonstrated that the content of the most mineral elements (B, Cd, Co, Cr, Cu, Fe, Mg, Mn, Mo, Ni, Pb, Se, Si, Sn, Sr, Ti, V and Zn) existed in a descending order in the hair of young age group (18～29 years) to the elder age group (40～45 years), while elements of Ca, Mg, Mn and Pb were found to have the lowest content in the middle age group (30～39 years). Women having two children were more likely to have the lowest content of Ca, Mg, Mn, Mo, Ni, Pb, Sn, Sr and Zn due to the increased numbers of pregnancies, the statistically correlations had been proved among Ca, Pb and Sn. In addition, dietary habits also can affect the level of the mineral elements in hair, for example, consuming suancai food frequently can cause lower level of Zn, Fe, Cu, Mn, Sr, Mo and Pb, but higher Se, the frequent intake of meat resulted in increased content of Zn and Se. The individuals that declared intaking vegetables regularly had more Si in their hair. Also, daily intake of fruit leads to higher level of Mn, Ni and Ti. This study will provide basic and useful information when addressing reproductive and women health challenges in the rural areas where poor dietary habits are prevailing.

Studies on the Interaction of Perfluorononanoic Acid with Human Serum Albumin by Multi-Spectroscopic, Molecular Docking and Isothermal Titration Calorimetry Techniques.

Perfluorononanoic acid (PFNA) is the third most frequently detected in serum among all perfluoroalkyl acids (PFAAs) which is a kind of toxic emerging environmental contaminant. The influence of PFNA on the conformation and even function of human serum albumin (HSA) is still just at the beginning of research. The attempt of this paper was to completely elucidate the interaction mechanism of PFNA with HSA by means of multi-spectroscopic, molecular docking and isothermal titration calorimetry (ITC) techniques. The inner filter effect of all fluorescence data in the paper was eliminated to get accurate binding parameters. The results showed that the fluorescence of HSA was quenched by PFNA through a combined quenching procedure of dynamic and static quenching. Through site marker competitive experiments, subdomain IIA of HSA had been assigned to possess the high-affinity binding site of PFNA. Furthermore, molecular docking reconfirmed that PFNA was bound in subdomain IIA mainly through polar force, hydrophobic interaction and halogen-bond, and the calculated free energy was -26.54 kJ·mol(-1) which indicated that the PFNA molecule exhibited large binding affinity towards HSA. The thermodynamic characterizations of two different classes of binding sites by ITC displayed that the first class with a higher affinity constant was dominated by an enthalpic contribution due to electrostatic interactions and halogen-bond, whereas the second class with a lower affinity constant was preponderated by hydrophobic interaction. The three-dimensional fluorescence revealed that the conformation of HSA was changed and the hydrophobicity of the Trp and Tyr residues microenvironment increased after formation of PFNA-HSA complex. The alterations of the protein secondary structure were quantitatively calculated from circular dichroism (CD) spectroscopy with reduction of α-helix content about 14.3%, ß-sheet 5.3%, ß-turn 3.5%, and augment in random content from 14.4% to 37.5%. Above results revealed that the binding of PFNA with HSA can alter the secondary structure of HSA, further probably affecting HSA physiological function. The results can provide insights with the binding mechanism of PFNA with HSA and salient biophysical and biochemical clues on elucidating the transport and distribution of PFNA in vivo.

Investigation of the Interaction between Perfluorododecanoic Acid and Human Serum Albumin by Multi-Spectroscopic and Molecular Modeling Techniques.

Perfluorododecanoic acid (PFDoA) is the most toxic emerging environmental contaminant among the 8～12 carbon chain perfluoroalkyl acids (PFAAs). A large amount of knowledge in the field of environmental PFAAs has been accumulated so far, while we are still just at the beginning of research into the interaction between PFDoA and human serum albumin (HSA). The goal of this study was to comprehensively determinate the binding mechanism of PFDoA with HSA by using fluorescence quenching technique in combination with molecular modeling and circular dichroism (CD) spectroscopy under the simulative physiological conditions. The quenching of HSA fluorescence by PFDoA was found to be a result of the combination of dynamic quenching and the formation of PFDoA-HSA complex. The calculated binding distance (r=3.65 nm) indicated that the non-radioactive energy transfer came into being in the interaction between PFDoA (acceptor) and HSA (donor). By performing displacement measurements, the specific binding of PFDoA in the vicinity of site I of HSA was clarified. Furthermore, the binding details between PFDoA and HSA were further confirmed by molecular docking studies, which revealed that PFDoA was bound at subdomain IIA by multiple interactions, such as the interaction between O1 of PFDoA with Arg 257 and Ser 287 predominately through polar force. And the best calculated docking energy is -25.87 kJ·mol-1, this high negative value indicated that the PFDoA molecule exhibited large binding affinity towards HSA. The effects of PFDoA on the conformation of HSA were analyzed by synchronous fluorescence spectra and three-dimensional fluorescence spectra, and the results exhibited that the hydrophobicity of the microenvironment around tryptophan residue was increased and the conformation of HSA was altered after binding PFDoA. The CD spectra quantitatively calculated the protein secondary structure, which suggested a loss of helical stability after the PFDoA-HSA complex formation. The binding research presented in this paper enriches our knowledge of the interaction dynamics of perfluoroalkyl acids to the HSA and reveals the chemical essence of the interaction between biomacromolecule and ligand.

Characterization of the Interaction between Acetylisovaleryltylosin Tartrat and Bovine Serum Albumin without or with Zn2+ and Cu2+ by Spectroscopic Analysis.

Acetylisovaleryltylosin tartrate (ATLL) is a new macrolide veterinary antibiotic, it is necessary to study the binding of ATLL to protein, which will directly correlate with the efficiency in vivo. Bovine serum albumin (BSA) is structure homologous with human serum albumin (HSA), and is commonly chosen as a model to investigate drug-protein interaction.There are many metal ions in plasma, as yet, the studies on mainly focus on single metal ion. In this study, the multiple systems formed by ATLL and BSA without or with Zn2+ and Cu2+ have been studied by mult-spectroscopy. The results showed that, the fluorescence of BSA was quenched by ATLL through a static quenching mechanism. The effective quenching constant (Ka) of ATLL to BSA decreased with Zn2+ and increased with Cu2+. Thermodynamic parameters revealed that hydrogen bonds and hydrophobic forces played significant roles in the binding of ATLL to BSA. The polarity of tryptophan and tyrosine residues changed when adding ATLL with or without Zn2+ and Cu2+. FT-IR spectra showed that ATLL changed α-helix and ß-sheet of BSA into ß-turn and random structure. The UV-Vis spectra indicated that the effects of Zn2+ on ATLL binding to BSA may cause by a competition binding, and Cu2+ possibly formed Cu2+-ATLL complex via metal ion bridge. All the knowledge obtained in this work will be helpful to understand the transport mechanism of ATLL with BSA and the effect of metal ions on the interaction of drug-protein in vivo.

A New Strategy to Probe and Compare the Binding Modes of Two Perfluorocarboxylic Acids with Human Serum Albumin Based on Spectroscopic and Molecular Docking Methods.

Perfluorocarboxylic acids (PFCAs) have been widespread used for over half century as surfactants in commercial and industrial products because of their hydrophilic and hydrophobic peculiarity. Perfluoroundecanoic acid (PFUnA) and perfluorotridecanoic acid (PFTriA) are two representatives of long-chain PFCAs, and they were detected more frequently in human body recent years, however, the two PFCAs were found to express endocrine disruption effects, developmental toxicity and teratogenicity. In this study, we established a new strategy to probe the binding modes of PFUnA (PFTriA) with the most abundant protein human serum albumin (HSA) based on spectroscopic and molecular docking methods. Results showed that both PFUnA and PFTriA can quench the intrinsic fluorescence of HSA with one binding site by means of dynamic and static quenching procedure with a strong affinity and the order is PFUnA>PFTriA. On the basis of thermodynamic results, we knew that the main driving force of the interaction between PFUnA and HSA was electrostatic force (ΔH=-26.32 kJ·mol-1, ΔS=21.76 J·mol-1·K-1), while van der Waals interaction and halogen-bond played major roles in complexation process of PFTriA-HSA (ΔH=-39.69 kJ·mol-1, ΔS=-25.66 J·mol-1·K-1). The binding distance (r<8 nm) indicated that the non-radioactive energy transfer came into being from HSA to PFUnA (PFTriA). The binding process of PFUnA (PFTriA) with HSA caused conformational and some micro-environmental changes of HSA, but also led to a loss of helical stability through three-dimensional fluorescence and circular dichroism spectra (CD). Furthermore, site markers competitive experiments and molecular docking revealed that PFUnA and PFTriA had a high affinity into hydrophobic pocket of subdomain IIA in HSA through polar force, hydrophobic interaction and halogen-bond and so on, and the fluorophore Trp residues was located in the binding position which proved further the quenching of PFUnA and PFTriA on HSA fluorescence. The accurate and full basic data in the work are beneficial to clarify the binding mechanism of long-chain perfluorocarboxylic acids with serum protein in vivo, and provide essential theoretical clues for their toxicity assessment and toxicologic research.