Neurulation of the cynomolgus monkey embryo achieved from 3D blastocyst culture.

Neural tube (NT) defects arise from abnormal neurulation and result in the most common birth defects worldwide. Yet, mechanisms of primate neurulation remain largely unknown due to prohibitions on human embryo research and limitations of available model systems. Here, we establish a three-dimensional (3D) prolonged in vitro culture (pIVC) system supporting cynomolgus monkey embryo development from 7 to 25 days post-fertilization. Through single-cell multi-omics analyses, we demonstrate that pIVC embryos form three germ layers, including primordial germ cells, and establish proper DNA methylation and chromatin accessibility through advanced gastrulation stages. In addition, pIVC embryo immunofluorescence confirms neural crest formation, NT closure, and neural progenitor regionalization. Finally, we demonstrate that the transcriptional profiles and morphogenetics of pIVC embryos resemble key features of similarly staged in vivo cynomolgus and human embryos. This work therefore describes a system to study non-human primate embryogenesis through advanced gastrulation and early neurulation.

Primate gastrulation and early organogenesis at single-cell resolution.

Our understanding of human early development is severely hampered by limited access to embryonic tissues. Due to their close evolutionary relationship with humans, nonhuman primates are often used as surrogates to understand human development but currently suffer from a lack of in vivo datasets, especially from gastrulation to early organogenesis during which the major embryonic cell types are dynamically specified. To fill this gap, we collected six Carnegie stage 8-11 cynomolgus monkey (Macaca fascicularis) embryos and performed in-depth transcriptomic analyses of 56,636 single cells. Our analyses show transcriptomic features of major perigastrulation cell types, which help shed light on morphogenetic events including primitive streak development, somitogenesis, gut tube formation, neural tube patterning and neural crest differentiation in primates. In addition, comparative analyses with mouse embryos and human embryoids uncovered conserved and divergent features of perigastrulation development across species-for example, species-specific dependency on Hippo signalling during presomitic mesoderm differentiation-and provide an initial assessment of relevant stem cell models of human early organogenesis. This comprehensive single-cell transcriptome atlas not only fills the knowledge gap in the nonhuman primate research field but also serves as an invaluable resource for understanding human embryogenesis and developmental disorders.

2D Porphyrin-Based Covalent-Organic Framework/PEG Composites: A Rational Strategy for Photocatalytic Hydrogen Evolution.

Two-dimensional porphyrin-based covalent-organic frameworks (2D-por-COFs) have gained significant attention as attractive platforms for efficient solar light conversion into hydrogen production. Herein, it is found that introducing transition metal zinc and polyethylene glycol (PEG) into 2D-por-COFs can effectively improve the photocatalytic hydrogen evolution performance. The photocatalytic hydrogen evolution rate of ZnPor-COF is 2.82 times higher than that of H2Por-COF. Moreover, ZnPor-COF@PEG has the highest photocatalytic hydrogen evolution efficiency, which is 1.31 and 3.7 times that of pristine ZnPor-COF and H2Por-COF, respectively. The filling of PEG makes the layered structure of COFs more stable. PEG reduces the distortion and deformation of the carbon skeleton after the experiment of photocatalytic hydrogen evolution. The layered stacking and crystallization of 2D-por-COFs are also enhanced. Meanwhile, the presence of PEG also accelerates the transfer of excited electrons and enhances the photocatalytic hydrogen evolution activity. This strategy will provide valuable insights into the design of 2D-por-COFs as efficient solid photocatalysts for solar-driven hydrogen production.

Prognostic effects of previous cancer history on patients with major salivary gland cancer.

OBJECTIVES: To explore the prognostic effects of previous cancer history on patients with major salivary gland cancer (SGC). SUBJECTS AND METHODS: SGC patients with (sec-SGC) and without (one-SGC) a previous cancer from the SEER database were identified. Cox proportional hazards regression (CoxPH) models were used to compare the prognosis between sec-SGC and one-SGC patients. Subgroup analyses for sec-SGC patients by gender, previous cancer types, previous cancer histology, and cancer diagnosis interval (CDI) were performed. Two CoxPH models were constructed to distinguish sec-SGC patients with different prognostic risks. RESULTS: 9098 SGC patients were enrolled. Overall, sec-SGC patients (adjusted HR [aHR] = 1.26, p < 0.001), especially those with a CDI ≤ 5 years (aHR = 1.47, p < 0.001), had worse overall survival (OS) than one-SGC patients. In subgroup analysis, only sec-SGC patients with a previous head and neck cancer who were female (aHR = 2.38, p = 0.005), with a CDI ≤ 5 years (aHR = 1.65, p = 0.007) or with a previous squamous cell carcinoma (aHR = 6.52, p < 0.001) had worse OS. Our models successfully differentiated all sec-SGC patients into high-, intermediate- and low-risk groups with different prognosis. CONCLUSIONS: Sec-SGC patients with different previous cancer types, gender, CDI and previous cancer histology had varied prognosis. The models we constructed could help differentiate the prognosis of sec-SGC patients with different risks.

Developing a Predictive Model for Minimal or Mild Endometriosis as a Clinical Screening Tool in Infertile Women: Uterosacral Tenderness as a Key Predictor.

STUDY OBJECTIVE: To develop a noninvasive predictive model based on patients with infertility for identifying minimal or mild endometriosis. DESIGN: A retrospective cohort study. SETTING: This study was conducted at a tertiary referral center. PATIENTS: A total of consecutive 1365 patients with infertility who underwent laparoscopy between January 2013 and August 2020 were divided into a training set (n = 910) for developing the predictive model and a validation set (n = 455) to confirm the model's prediction efficiency. The patients were randomly assigned in a 2:1 ratio. INTERVENTIONS: Sensitivities, specificities, area under the curve, the Hosmer-Lemeshow goodness of fit test, Net Reclassification Improvement index, and Integrated Discrimination Improvement index were evaluated in the training set to select the optimum model. In the validation set, the model's discriminations, calibrations, and clinical use were tested for validation. MEASUREMENTS AND MAIN RESULTS: In the training set, there were 587 patients with minimal or mild endometriosis and 323 patients without endometriosis. The combination of clinical parameters in the model was evaluated for both statistical and clinical significance. The best-performing model ultimately included body mass index, dysmenorrhea, dyspareunia, uterosacral tenderness, and serum cancer antigen 125 (CA-125). The nomogram based on this model demonstrated sensitivities of 87.7% and 93.3%, specificities of 68.6% and 66.4%, and area under the curve of 0.84 (95% confidence interval 0.81-0.87) and 0.85 (95% confidence interval 0.80-0.89) for the training and validation sets, respectively. Calibration curves and decision curve analyses also indicated that the model had good calibration and clinical value. Uterosacral tenderness emerged as the most valuable predictor. CONCLUSION: This study successfully developed a predictive model with high accuracy in identifying infertile women with minimal or mild endometriosis based on clinical characteristics, signs, and cost-effective blood tests. This model would assist clinicians in screening infertile women for minimal or mild endometriosis, thereby facilitating early diagnosis and treatment.

The disease recurrence perception scale for patients with inflammatory bowel disease: Instrument development and cross-sectional validation study.

Disease recurrence perception plays a key role in disease management and subsequent disease recurrence prevention. However, there are no specific tools for assessing disease recurrence perception in patients with inflammatory bowel disease (IBD) characterized by alternating remission and recurrence. To develop and validate an instrument for measuring disease recurrence perception of patients with IBD, the study was conducted in two steps: (1) instrument development and (2) psychometric tests. A total of 623 patients with IBD participated in the study. The common sense model of illness self-regulation (CSM) was used as a framework for instrument development. The administered version contained 48 items intended to be relevant to at least one of the six dimensions of the model. Based on preliminary analyzes, 12 items were deleted leaving 36 items for more detailed psychometric and factor analyzes. The Cronbach's alpha coefficient of the total 36-item instrument was 0.915. The content validity indexes at item and scale levels were satisfactory. The test-retest reliability of the total instrument was 0.870. Exploratory principal components analysis (n = 278) was used to identify six components congruent with intended CSM constructs that accounted for 62.6% of total item variance. Confirmatory factor analysis (n = 345) found acceptable fit for the six factor measurement model (χ2/df = 1.999, GFI = 0.846, NFI = 0.855, IFI = 0.922, TLI = 0.910, CFI = 0.921, RMSEA = 0.054). Overall, the DRPSIBD demonstrated satisfactory reliability and validity to warrant further development as a measure of disease recurrence perception of patients with IBD.

Enhancing the Electrochemiluminescence of Porphyrin via Crystalline Networks of Metal-Organic Frameworks for Sensitive Detection of Cardiac Troponin I.

Porphyrins easily aggregate due to unfavorable π-π accumulation, causing luminescent quenching in the aqueous phase and subsequently reducing luminescent efficiency. It is a feasible way to immobilize porphyrin molecules through metal-organic framework materials (MOFs). In this study, 5,10,15,20-tetrakis (4-carboxyphenyl) porphyrin (TCPP) was introduced into the metal-organic skeleton (PCN-224) as a ligand. The result showed that the electrochemiluminescence (ECL) and photoluminescence (PL) efficiency of the MOF skeleton was 8.2 and 6.5 times higher than TCPP, respectively. Impressively, the periodic distribution of porphyrin molecules in the MOF framework can overcome the bottleneck of porphyrin aggregation, resulting in the organic ligand TCPP participating in the electron transfer reaction. Herein, based on the PCN-224, a sandwich-type ECL immunosensor was constructed for the determination of cardiac troponin I (cTnI). It provided sensitive detection of cTnI in the range of 1 fg/mL to 10 ng/mL with a detection limit of 0.34 fg/mL. This work not only innovatively exploited a disaggregation ECL (DIECL) strategy via the crystalline framework of MOF to enhance the PL and ECL efficiency of porphyrin but also provided a promising ECL platform for the ultrasensitive monitoring of cTnI.

Transmit-Receive Sparse Synthesis of Linear Frequency Diverse Array in Range-Angle Space Using Genetic Algorithm.

Unlike conventional phased array (PA), frequency diversity array (FDA) can perform the beampattern synthesis not only in an angle dimension but also in a range dimension by introducing an additional frequency offset (FO) across the array aperture, thus greatly enhancing the beamforming flexibility of an array antenna. Nevertheless, an FDA with uniform inter-element spacing that consists of a huge number of elements is required when a high resolution is needed, which results in a high cost. To substantially reduce the cost while almost maintaining the antenna resolution, it is important to conduct a sparse synthesis of FDA. Under these circumstances, this paper investigated the transmit-receive beamforming of a sparse-fda in range and angle dimensions. In particular, the joint transmit-receive signal formula was first derived and analyzed to resolve the inherent time-varying characteristics of FDA based on a cost-effective signal processing diagram. In the sequel, the GA-based low sidelobe level (SLL) transmit-receive beamforming of the sparse-fda was proposed to generate a focused main lobe in a range-angle space, where the array element positions were incorporated into the optimization problem. Numerical results showed that 50% of the elements can be saved for the two linear FDAs with sinusoidally and logarithmically varying frequency offsets, respectively termed as sin-FO linear-FDA and log-FO linear-FDA, with only a less than 1 dB increment in SLL. The resultant SLLs are below -9.6 dB, and -12.9 dB for these two linear FDAs, respectively.

Developmental trajectories of health-promoting behaviours among pregnant women: A longitudinal study.

AIMS AND OBJECTIVES: The aim of this study is to estimate differences in health-promoting behaviour levels throughout the pregnancy trimesters, to identify distinct patterns of health-promoting behaviour trajectories during pregnancy and to evaluate predictors of these trajectory group memberships. BACKGROUND: Promoting healthy lifestyle behaviours during pregnancy could decrease devastating outcomes for the mother and foetus. However, there is currently limited insight into the dynamics of health-promoting behaviours during pregnancy. DESIGN: An observational longitudinal study. METHODS: 158 pregnant women were recruited from June 2020 to June 2021 in Qingdao, China. The Health-Promoting Lifestyle Profile was used to assess health-promoting behaviours. Latent growth model was performed to compare health-promoting behaviours at different time points. Group-based trajectory model was applied to identify health-promoting behaviour trajectories. Multinomial logistic regression was adopted to determine the predictors of trajectory group memberships. We used the STROBE checklist to report this study. RESULTS: The entire sample of pregnant women experienced a significant increase in health-promoting behaviours during pregnancy. Three trajectories were identified including a 'low-increase behaviour trajectory (20.1% of sample)', a 'moderate-increase behaviour trajectory (58.0% of sample)' and a 'stable then increased behaviour trajectory (21.9% of sample)'. Low maternal sense of coherence, lack of pre-pregnancy exercise habit, artificial insemination and low monthly family income were significantly associated with the low-increase behaviour trajectory. High self-efficacy and pre-pregnancy exercise habit were significantly associated with the stable then increased behaviour trajectory. CONCLUSIONS: Pregnant women exhibit different health-promoting behaviours throughout the pregnancy trimesters. Meanwhile, three trajectories were identified among pregnant women. Thus, more attentions should be paid on early identification and targeted intervention in a future study. RELEVANCE OF CLINICAL PRACTICE: Healthcare providers should pay closer and earlier attention to identify women in the low-increase trajectory subgroup at the outset of pregnancy. Similarly, increased efforts should be made to improve maternal self-efficacy and develop good pre-pregnancy exercise habit in future study.

Effects of e-health interventions on health outcomes in patients with rheumatoid arthritis: a systematic review and meta-analysis.

AIMS AND OBJECTIVES: To evaluate the effects of e-health interventions on disease activity, self-efficacy, pain and quality of life among patients with rheumatoid arthritis (RA). BACKGROUND: Prior systematic reviews have only reported the quality and features of e-health interventions in patients with RA. However, the effect of e-health interventions in patients with RA is unclear. DESIGN: Systematic review and meta-analysis. METHODS: This review was conducted following the PRISMA guideline. We searched 5 databases, including PubMed, EMBASE, CINAHL, Scopus and the Cochrane library. The risk of bias was assessed using the Cochrane risk of bias tool. The quality of the evidence was assessed via the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) criteria. Using a random-effects model adopted the standardised mean difference (SMD) with a 95% confidence interval (CI). A chi-squared test and an I2 test were used to assess heterogeneity. Subgroup analyses were conducted based on different controls. RESULTS: A total of 9 randomised control trials were included in this study. Compared with the control group, disease activity of the e-health group significantly decreased (SMD with 95% CI: -.17 [-.30, -.04], p = .01, I2  = 1%). Meanwhile, trials with usual care control had a larger effect on disease activity (SMD with 95% CI: -.21 [-.40, -.02], p = .03, I2  = 38%). The effect of e-health interventions on self-efficacy was controversial; pain and quality of life were negative in the currently included studies. The quality of evidence was rated as low for disease activity and very low for pain, self-efficacy and quality of life. CONCLUSIONS: The effect of e-health interventions on disease activity was statistically significant. More well-designed randomised controlled trials are still needed to verify the effects in the future. RELEVANCE TO CLINICAL PRACTICE: This study shows the potential value of e-health in improving health outcomes in patients with RA.

Aromaticity-Enhanced pH-Responsive Electrochemiluminescence of Cyclopentadienols.

Due to significantly tackling the problems of aggregation-caused quenching and water insolubility, aggregation-induced emission electrochemiluminescence (AIE-ECL) has emerged as a research highlight in aqueous detection and sensing. Herein, we reported a series of cyclopentadienols featuring excellent AIE-ECL properties on the basis of an enhanced aromaticity strategy. In detail, substituents profoundly determined ECL emission by affecting the characteristic absorption peak intensity ratio in UV-vis spectra and lowest unoccupied molecular orbital (LUMO)-highest occupied molecular orbital (HOMO) energies. It was found that 1,2,3,4,5-pentafluorophenyl cyclopentadienol (PFCD) containing an electron-withdrawing fluorine substituent, the maximum R/B band ratio, and a smaller LUMO-HOMO band gap demonstrated the best ECL performance. Meanwhile, such an AIE-ECL system displayed a wide response range toward pH (4-12) with a good linear relationship. Our research not only enriched polycyclic aromatic hydrocarbon-based AIE-ECL systems but also established an efficient pH sensor in the aqueous phase.

Comparative transcriptome analysis reveals different defence responses during the early stage of wounding stress in Chi-Nan germplasm and ordinary Aquilaria sinensis.

BACKGROUND: Agarwood is a valuable Chinese medicinal herb and spice that is produced from wounded Aquilaria spp., is widely used in Southeast Asia and is highly traded on the market. The lack of highly responsive Aquilaria lines has seriously restricted agarwood yield and the development of its industry. In this article, a comparative transcriptome analysis was carried out between ordinary A. sinensis and Chi-Nan germplasm, which is a kind of A. sinensis tree with high agarwood-producing capacity in response to wounding stress, to elucidate the molecular mechanism underlying wounding stress in different A. sinensis germplasm resources and to help identify and breed high agarwood-producing strains. RESULTS: A total of 2427 and 1153 differentially expressed genes (DEGs) were detected in wounded ordinary A. sinensis and Chi-Nan germplasm compared with the control groups, respectively. KEGG enrichment analysis revealed that genes participating in starch metabolism, secondary metabolism and plant hormone signal transduction might play major roles in the early regulation of wound stress. 86 DEGs related to oxygen metabolism, JA pathway and sesquiterpene biosynthesis were identified. The majority of the expression of these genes was differentially induced between two germplasm resources under wounding stress. 13 candidate genes related to defence and sesquiterpene biosynthesis were obtained by WGCNA. Furthermore, the expression pattern of genes were verified by qRT-PCR. The candidate genes expression levels were higher in Chi-Nan germplasm than that in ordinary A. sinensis during early stage of wounding stress, which may play important roles in regulating high agarwood-producing capacity in Chi-Nan germplasm. CONCLUSIONS: Compared with A. sinensis, Chi-Nan germplasm invoked different biological processes in response to wounding stress. The genes related to defence signals and sesquiterepene biosynthesis pathway were induced to expression differentially between two germplasm resources. A total of 13 candidate genes were identified, which may correlate with high agarwood-producting capacity in Chi-Nan germplasm during the early stage of wounding stress. These genes will contribute to the development of functional molecular markers and the rapid breeding highly of responsive Aquilaria lines.

Genome-wide investigation and expression analysis of the AP2/ERF family for selection of agarwood-related genes in Aquilaria sinensis (Lour.) Gilg.

Aquilaria sinensis is an important non-timber tree species for producing high-value agarwood, which is widely used as a traditional medicine and incense. Agarwood is the product of Aquilaria trees in response to injury and fungal infection. The APETALA2/ethylene responsive factor (AP2/ERF) transcription factors (TFs) play important roles in plant stress responses and metabolite biosynthesis. In this study, 119 AsAP2/ERF genes were identified from the A. sinensis genome and divided into ERF, AP2, RAV, and Soloist subfamilies. Their conserved motif, gene structure, chromosomal localization, and subcellular localization were characterized. A stress/defense-related ERF-associated amphiphilic repression (EAR) motif and an EDLL motif were identified. Moreover, 11 genes that were highly expressed in the agarwood layer in response to whole-tree agarwood induction technique (Agar-Wit) treatment were chosen, and their expression levels in response to methyl jasmonate (MeJA), salicylic acid (SA), or salt treatment were further analyzed using the quantitative real time PCR (qRT-PCR). Among the 11 genes, eight belonged to subgroup B-3. All 11 genes were significantly upregulated under salt treatment, while eight genes were significantly induced by both MeJA and SA. In addition, the gene clusters containing these upregulated genes on chromosomes were observed. The results obtained from this research not only provide useful information for understanding the functions of AP2/ERF genes in A. sinensis but also identify candidate genes and gene clusters to dissect their regulatory roles in agarwood formation for future research.

Association between pregnancy intention and smoking or alcohol consumption in the preconception and pregnancy periods: A systematic review and meta-analysis.

AIMS: To investigate the association between pregnancy intention and smoking or alcohol consumption in preconception and pregnancy periods. BACKGROUND: Suboptimal lifestyle such as smoking and alcohol consumption can lead to devastating outcomes on the maternal and foetus. Pregnancy intention exerts a significant effect on promoting healthy lifestyle behaviours. However, no reliable evidences confirmed pregnancy intention was associated with smoking and alcohol consumption before and during pregnancy. DESIGN: Systematic review and meta-analysis. METHODS: We performed a comprehensive search from databases including PubMed, Cochrane, Web of Science, IEEE Xplore, MEDLINE, ProQuest and Scopus from the inception of these databases up to November, 2020. All eligible studies exploring the association between pregnancy intention and smoking or alcohol consumption were included. The fixed- or random effect pooled measure was used to estimate the odds ratio (OR) or risk ratio (RR) and 95% CI. In addition, the PRISMA checklist was used in this meta-analysis. RESULTS: A total of 23 studies were included in this systematic review and meta-analysis. During pregnancy, the findings suggested that women with unplanned pregnancy were 68% more likely to consume cigarettes (OR = 1.68, 95% CI = 1.44-1.95) and 44% more likely to consume alcohol (OR = 1.44, 95% CI = 1.15-1.81) than those women with planned pregnancy. Meanwhile, during preconception, women with unplanned pregnancy were 30% more likely to consume cigarettes (OR = 1.30, 95% CI = 1.10-1.53) and 20% more likely to consume alcohol (OR = 1.20, 95% CI = 1.01-1.42) than those women with planned pregnancy. CONCLUSION: The findings suggested that women with unplanned pregnancy were more likely to follow unhealthy behaviours such as smoking and alcohol consumption before and during pregnancy. Health professionals should consider the women's desire for pregnancy to decrease preconception and pregnancy smoking or alcohol consumption in future studies. RELEVANCE OF CLINICAL PRACTICE: Pregnancy intention is the key determinant of smoking and alcohol consumption during preconception and pregnancy periods. Offering effective contraception in primary healthcare setting could prevent unplanned pregnancy. Meanwhile, popularising minimal alcohol consumption and comprehensive smoke-free legislation would be beneficial to improve reproductive outcomes.

Transcriptomics and Metabolomics Analyses Reveal Defensive Responses and Flavonoid Biosynthesis of Dracaena cochinchinensis (Lour.) S. C. Chen under Wound Stress in Natural Conditions.

Dracaena cochinchinensis has special defensive reactions against wound stress. Under wound stress, D. cochinchinensis generates a resin that is an important medicine known as dragon's blood. However, the molecular mechanism underlying the defensive reactions is unclear. Metabolomics and transcriptomics analyses were performed on stems of D. cochinchinensis at different timepoints from the short term to the long term after wounding. According to the 378 identified compounds, wound-induced secondary metabolic processes exhibited three-phase characteristics: short term (0-5 days), middle term (10 days-3 months), and long term (6-17 months). The wound-induced transcriptome profile exhibited characteristics of four stages: within 24 h, 1-5 days, 10-30 days, and long term. The metabolic regulation in response to wound stress mainly involved the TCA cycle, glycolysis, starch and sucrose metabolism, phenylalanine biosynthesis, and flavonoid biosynthesis, along with some signal transduction pathways, which were all well connected. Flavonoid biosynthesis and modification were the main reactions against wound stress, mainly comprising 109 flavonoid metabolites and 93 wound-induced genes. A group of 21 genes encoding CHS, CHI, DFR, PPO, OMT, LAR, GST, and MYBs were closely related to loureirin B and loureirin C. Wound-induced responses at the metabolome and transcriptome level exhibited phase characteristics. Complex responses containing primary metabolism and flavonoid biosynthesis are involved in the defense mechanism against wound stress in natural conditions, and flavonoid biosynthesis and modification are the main strategies of D. cochinchinensis in the long-term responses to wound stress.

1,3-Benzodioxole-based fibrate derivatives as potential hypolipidemic and hepatoprotective agents.

A series of target compounds 1,3-benzodioxole-based fibrate derivatives were designed and synthesized. All the target compounds were preliminarily evaluated by hyperlipidemia mice induced by Triton WR-1339, in which compound 12 displayed a greater anti-hyperlipidemia activity than other compounds as well as positive drug fenofibrate (FF). 12 showed a significant reduction of plasma lipids, such as triglycerides (TG), total cholesterol (TC) and low-density lipoprotein cholesterin (LDL-C), in high fat diet (HFD) induced hyperlipidemic mice. In addition, hepatic transaminases (AST and ALT) were ameliorated after administration of 12, in particular the AST, and the histopathological examination showed that 12 improved the hepatic lipid accumulation. The expression of PPAR-α involved in lipids metabolism was up-regulated in the liver tissues of 12-treated group. Other significant activity such as antioxidant, and anti-inflammation was confirmed and reinforced the effects of 12 as a potential hypolipidemia and hepatoprotective agent.

Optimization of UC-MSCs cold-chain storage by minimizing temperature fluctuations using an automatic cryopreservation system.

The effective long-term cryopreservation of human mesenchymal stem cells is an essential prerequisite step and represents a critical approach for their sustained supply in basic research, regenerative medicine, and tissue engineering applications. Off-the-shelf availability of human umbilical cord-derived mesenchymal stromal cells (UC-MSCs) for regenerative medicine application requires the development of nontoxic, safe, and efficient protocols for cryopreservation. In the long-term low-temperature storage process of cells, traditional manual storage has a great impact on cell activity, recovery, and function due to repeated exposure of cells to room temperature. To minimize the effect of fluctuation in ambient temperature on stored cells, we designed an automatic cryopreservation system that handles cells under controlled temperatures. In this work, UC-MSCs were utilized to investigate and compare the influence of manual and automatic cryopreservation approaches. To simulate the manual process, the UC-MSCs were transferred back and forth repeatedly (up to 400 times) between the liquid nitrogen (LN2) tank (-150 °C) and room temperature by a robotic arm. Similarly, the UC-MSCs from the same batch were collected and transferred repeatedly between two storage units by the automatic cryopreservation system, where the cells were maintained below-150 °C throughout the cold chain process. Viability, percent recovery, adherence capability, cell proliferation, and multilineage differentiation ability were assessed for UC-MSCs. Compared to the manual approach, UC-MSCs handled by the automatic system demonstrated higher viability, percent recovery, and cell proliferation, as well as improved adherence to culture plate with greater potential in multilineage differentiation after 400 temperature cycles. The described entire cold chain system may provide a powerful tool to develop safe, reliable and efficient protocols for manufacturing and banking of UC-MSCs, improving their off-the-shelf availability for regenerative medicine applications.

Simultaneous determination of multiple active 2-(2-phenylethyl)chromone analogues in agarwood by HPLC, QAMS, and UPLC-MS.

INTRODUCTION: Chromones are the major constituents of agarwood and are considered to be directly related to its quality. Agarotetrol, a chromone derivative, is a Chinese Pharmacopoeia content detection index. However, comprehensive high-performance liquid chromatography (HPLC), quantitative analysis of multiple components by a single marker (QAMS), and ultra-performance liquid chromatography mass spectrometry (UPLC-MS) analyses of this pharmacopeial plant material have never been performed. Moreover, reports regarding the separation and detection of multiple active 2-(2-phenylethyl)chromone analogues from this plant material are surprisingly scarce. OBJECTIVE: To establish a simple, reliable, and effective HPLC method utilising both diode array and MS detection for the simultaneous determination of multiple active chromone analogues in agarwood. METHODS: Four 2-(2-phenylethyl)chromones were isolated from methanol extracts of agarwood. After optimising the extraction, separation, and analytical conditions, validation of the developed analytical method indicated good linearity, satisfactory precision, and good recovery. On this basis, a method for the quantitative analysis of multiple components by a single marker was established. The four 2-(2-phenylethyl)chromones were identified by nuclear magnetic resonance spectroscopic analysis and UPLC coupled to electrospray ionisation quadrupole-time-of-flight MS. CONCLUSIONS: The behaviour of the chromones characterised by MS fragmentation indicated a loss of molecular CO and the formation of m/z 121 compounds by the cleavage of CH2 -CH2 bonds between the chromone and phenyl moieties. Three detection methods were successfully used in this study for agarwood detection, and this protocol may potentially be used as a tool for the quality control of agarwood.

Application of X-ray diffraction and energy dispersive spectroscopy in the isolation of sulfated polysaccharide from Porphyra haitanensis and its antioxidant capacity under in vitro digestion.

BACKGROUND: The separation and purification of Porphyra haitanensis polysaccharide (PHP), and the determination of changes in molecular weight (Mw) and antioxidant capacity after in vitro digestion, were undertaken. RESULTS: Analysis of two polysaccharide fractions (PHP0.5-1-UF and PHP1.0-1-UF) by various techniques showed that they were very pure sulfated polysaccharides without pigment or protein. PHP0.5-1-UF was filamentous or 'tape-like' sheets, whereas PHP1.0-1-UF had some filaments and large numbers of rounded aggregates. The Mw of PHP, PHP0.5-1-UF and PHP1.0-1-UF was 2.06 × 106 (±2.02%), 6.68 × 106 (±3.17%), and 1.14 × 106 (±3.44%) (g mol-1 ), respectively. After in vitro digestion, the Mw of PHP, PHP0.5-1-UF, and PHP1.0-1-UF decreased. Their antioxidant capacities were markedly higher than before digestion, especially PHP0.5-1-UF and its digestion products, which might be related to the reductions in Mw. CONCLUSION: These findings provide a greater understanding of the separation and purification of sulfated polysaccharides and the influence of digestion on biological activity. They also contribute to the practical application of sulfated polysaccharides in functional foods. © 2021 Society of Chemical Industry.

[Genetic analysis of a case with MEF2C deletion in association with 5q14.3 microdeletion syndrome].

OBJECTIVE: To explore the genetic basis for a child with febrile seizures. METHODS: Peripheral venous blood samples were taken from the child and his parents for the analysis of chromosomal karyotype and dynamic variant of the FMR1 gene. The family trio was also subjected to target capture and next generation sequencing (NGS) with a gene panel related to developmental retardation, mental retardation, language retardation, epilepsy and special facial features. RESULTS: The child was found to have a normal karyotype by conventional cytogenetic analysis (400 bands). No abnormal expansion was found with the CGG repeats of the FMR1 gene. NGS revealed that the child has carried a heterozygous c.864+1 delG variant of the MEF2C gene, which may lead to abnormal splicing and affect its protein function. The same variant was found in neither parent, suggesting that it has a de novo origin. Based on the American College of Medical Genetics and Genomics standards and guidelines, c.864+1delG variant of MEF2C gene was predicted to be pathogenic (PVS1+PS2+PM2). CONCLUSION: MEF2C, as the key gene for chromosome 5q14.3 deletion syndrome which was speculated as a cause for febrile seizures, has an autosomal dominant effect. The c.864+1delG variant of the MEF2C gene may account for the febrile seizures in this patient.