GLK transcription factors accompany ELONGATED HYPOCOTYL5 to orchestrate light-induced seedling development in Arabidopsis.

Light-induced de-etiolation is an important aspect of seedling photomorphogenesis. GOLDEN2 LIKE (GLK) transcriptional regulators are involved in chloroplast development, but to what extent they participate in photomorphogenesis is not clear. Here, we show that ELONGATED HYPOCOTYL5 (HY5) binds to GLK promoters to activate their expression, and also interacts with GLK proteins in Arabidopsis (Arabidopsis thaliana). The chlorophyll content in the de-etiolating Arabidopsis seedlings of the hy5 glk2 double mutants was lower than that in the hy5 single mutant. GLKs inhibited hypocotyl elongation, and the phenotype could superimpose on the hy5 phenotype. Correspondingly, GLK2 regulated the expression of photosynthesis and cell elongation genes partially independent of HY5. Before exposure to light, DE-ETIOLATED 1 (DET1) affected accumulation of GLK proteins. The enhanced etioplast development and photosystem gene expression observed in the det1 mutant were attenuated in the det1 glk2 double mutant. Our study reveals that GLKs act downstream of HY5, or additive to HY5, and are likely quantitatively adjusted by DET1, to orchestrate multiple developmental traits during the light-induced skotomorphogenesis-to-photomorphogenesis transition in Arabidopsis.

Targeted chemical profiling for p-HAP glycosides by using molecular networking and comparative analysis of their contents between Artemisia japonica and Artemisia capillaris.

A total of 65 phenolic acid compounds were annotated or identified by UHPLC-MS/MS method, among them, 17 p-HAP (p-hydroxyacetophenone) glycosides were firstly targeted profiled based on molecular networking. Their characteristic product ions of MS/MS spectra were found and examined on the guideline of targeted isolation. As a result, a new p-HAP glycoside was thus obtained and determined as 2'-O-caffeoyl-p-HAP-4-O-ß-D-glucopyranoside (33) based on 1D and 2D NMR data. Besides, multicomponents quantitative analysis indicated the distinct regional variability in chemicals distribution of A. japonica, and meanwhile, the contents of p-HAP glycosides from A. japonica were higher than those in A. capillaris as a whole, which further suggested the potential medicinal value of A. japonica.

Utilization of complementary and alternative medicine by perimenopausal women in China: a cross-sectional study.

OBJECTIVE: To investigate the utilization of complementary and alternative medicine (CAM) by perimenopausal women and explore factors associated with their preference. METHODS: A cross-sectional study was conducted involving 558 perimenopausal women in Wuhan, China. Online questionnaires were available from December 2021 to April 2022. Instruments used were the International Complementary and Alternative Medicine Questionnaire and the Greene Climacteric Scale. The association between scores and participants' sociodemographic characteristics and perimenopausal symptoms was analyzed using multiple linear regression. RESULTS: The overall utilization of CAM by perimenopausal women was 50.4% (281/558). Perimenopausal symptoms affecting the frequency of CAM usage included anxiety (adjusted ß = 0.18), somatic (adjusted ß = 0.07), and vasomotor symptoms (adjusted ß = 0.76), P < 0.05. The most commonly used CAM were massage (37.7%), herbal therapies (19.9%), and dietary supplements (18.2%). Walking (58.6%) was the most popular self-practice therapy. Perimenopausal women consulted Western (38.4%) and Chinese medicine physicians (34.8%) and dentists (36.0%) most frequently. CONCLUSIONS: Perimenopausal women experience discomforts during this age-related stage, and their use of CAM to manage symptoms is common. The usage and preferences of these management strategies are worthy of further exploration. There is a need to understand cost-effective and appropriate management interventions to improve the quality of life of perimenopausal women.

Breastfeeding Practices of Women Returning to Full-Time Employment in China: Prevalence and Determinants.

BACKGROUND: Returning to work after childbirth is a common reason for women to stop breastfeeding. This study aimed to assess breastfeeding practices and breastfeeding support available to employed women in China, and factors affecting breastfeeding duration. METHODS: A cross-sectional survey of 1,243 breastfeeding women employed full-time was conducted. Participants completed a sociodemographic questionnaire and the Workplace Breastfeeding Support Scale (WBSS). Multiple linear regression analysis was used to explore the factors that are associated with breastfeeding. RESULTS: The mean exclusive breastfeeding duration and any breastfeeding duration of full-time employed women were 5.7 ± 0.5 months and 9.8 ± 1.5 months, respectively. The total WBSS score was 46.6 ± 5.3 (M ± SD). Generally, women perceived coworkers (M [SD] = 4.8 [1.0]) and supervisors (M [SD] = 5.7 [1.2]) to be supportive of breastfeeding. Lower scores on the WBSS were related to lack of technical and facility support, indicating no access to a refrigerator to store breast milk (M [SD] = 2.0 [1.5]) or to a breast pump (M [SD] = 1.7 [1.7]). Similarly, a private area for expressing breast milk (M [SD] = 1.4 [1.0]) was unavailable. Maternity leave, residential province, ethnicity, education level, average monthly household income, main reason for stopping exclusive breastfeeding, commute time greater than 1 hour, and the total WBSS score were also factors influencing breastfeeding duration of the full-time employed women. CONCLUSIONS: There were gaps in breastfeeding practices and workplace breastfeeding support of Chinese full-time employed women when compared with the World Health Organization recommendations. Occupational health providers should consider these findings when developing programs to support breastfeeding in the workplace.

PD-1/PD-L1 combined with LAG3 is associated with clinical activity of immune checkpoint inhibitors in metastatic primary pulmonary lymphoepithelioma-like carcinoma.

Primary pulmonary lymphoepithelioma-like carcinoma (PLELC) is an Epstein-Barr virus (EBV)-related, rare subtype of non-small-cell lung cancer (NSCLC). Immune checkpoint inhibitors (ICI) show durable responses in advanced NSCLC. However, their effects and predictive biomarkers in PLELC remain poorly understood. We retrospectively analyzed the data of 48 metastatic PLELC patients treated with ICI. Pretreated paraffin-embedded specimens (n = 19) were stained for PD-1, PD-L1, LAG3, TIM3, CD3, CD4, CD8, CD68, FOXP3, and cytokeratin (CK) by multiple immunohistochemistry (mIHC). Next-generation sequencing was performed for 33 PLELC samples. Among patients treated with ICI monotherapy (n = 30), the objective response rate (ORR), disease control rate (DCR), median progression-free survival (mPFS), and overall survival (mOS) were 13.3%, 80.0%, 7.7 months, and 24.9 months, respectively. Patients with PD-L1 ≥1% showed a longer PFS (8.4 vs. 2.1 months, p = 0.015) relative to those with PD-L1 <1%. Among patients treated with ICI combination therapy (n = 18), ORR, DCR, mPFS, and mOS were 27.8%, 100.0%, 10.1 months, and 19.7 months, respectively. Patients with PD-L1 ≥1% showed a significantly superior OS than those with PD-L1 <1% (NA versus 11.7 months, p = 0.001). Among the 19 mIHC patients, those with high PD-1/PD-L1 and LAG3 expression showed a longer PFS (19.0 vs. 3.9 months, p = 0.003). ICI also showed promising efficacy for treating metastatic PLELC. PD-L1 may be both predictive of ICI treatment efficacy and prognostic for survival in PLELC. PD-1/PD-L1 combined with LAG3 may serve as a predictor of ICI treatment effectiveness in PLELC. Larger and prospective trials are warranted to validate both ICI activity and predictive biomarkers in PLELC. This study was partly presented as a poster at the IASLC 20th World Conference on Lung Cancer 2019, 7-10 September 2019, Barcelona, Spain.

Phylogenomic conflict analyses in the apple genus Malus s.l. reveal widespread hybridization and allopolyploidy driving diversification, with insights into the complex biogeographic history in the Northern Hemisphere.

Phylogenomic evidence from an increasing number of studies has demonstrated that different data sets and analytical approaches often reconstruct strongly supported but conflicting relationships. In this study, 785 single-copy nuclear genes and 75 complete plastomes were used to infer the phylogenetic relationships and estimate the historical biogeography of the apple genus Malus sensu lato, an economically important lineage disjunctly distributed in the Northern Hemisphere and involved in known and suspected hybridization and allopolyploidy events. The nuclear phylogeny recovered the monophyly of Malus s.l. (including Docynia); however, the genus was supported to be biphyletic in the plastid phylogeny. An ancient chloroplast capture event in the Eocene in western North America best explains the cytonuclear discordance. Our conflict analysis demonstrated that ILS, hybridization, and allopolyploidy could explain the widespread nuclear gene tree discordance. One deep hybridization event (Malus doumeri) and one recent event (Malus coronaria) were detected in Malus s.l. Furthermore, our historical biogeographic analysis integrating living and fossil data supported a widespread East Asian-western North American origin of Malus s.l. in the Eocene, followed by several extinction and dispersal events in the Northern Hemisphere. We also propose a general workflow for assessing phylogenomic discordance and biogeographic analysis using deep genome skimming data sets.

Gunsight sutures significantly reduce surgical-site infection after ileostomy reversal compared with linear sutures.

BACKGROUND: Surgical-site infection (SSI) was one of the most common post-operative morbidities of ileostomy reversal. Although several skin-closure procedures had been developed to reduce the rate of SSI, the optimal procedure remains unclear. In this study, we compared the effect of two surgical techniques for wound closure following ileostomy reversal: gunsight suture (GS) and linear suture (LS). METHODS: A total of 233 patients who underwent loop ileostomy at the Sixth Affiliated Hospital of Sun Yat-sen University between January 2015 and December 2017 were enrolled into our study. These patients were divided into two groups: the LS group and the GS group. We compared the clinical characteristics between the two groups and analyzed the data using IBM SPSS to identify risk factors for SSI. RESULTS: Both groups successfully underwent surgery. The rate of SSI was significantly lower in the GS group (n = 2, 0.02%) than in the LS group (n = 16, 12.00%, P = 0.007). The length of hospital stay after the operation in the GS group was significantly shorter than that in the LS group (8.1 ± 3.2 vs 10.8 ± 5.4 days, P < 0.001). Multivariate analysis showed that GS was an independent protective risk factor for SSI (odds ratio = 0.212, P = 0.048). CONCLUSIONS: Compared with the LS technique, the GS technique can significantly decrease the rate of SSI and shorten the length of hospital stay after surgery. The GS technique may be recommended for wound closure following ileostomy reversal.

Experience predicts the duration of exclusive breastfeeding: The serial mediating roles of attitude and self-efficacy.

BACKGROUND AND AIMS: The relationship between prior breastfeeding experience and the duration of exclusive breastfeeding is of significant interest, but few studies have explored the mechanisms underlying this relationship. The purpose of this study is to address two hypotheses: (a) that attitude and self-efficacy mediate the relationship between previous breastfeeding experience and the duration of exclusive breastfeeding; and (b) that attitude and self-efficacy have serial mediation functions in this relationship. METHODS: The data collection process was divided into two stages. The original stage included 394 women hospitalized after delivery who completed socio-demographic questionnaires (including previous breastfeeding experience), the Iowa Infant Feeding Attitude Scale (IIFAS), and the Breastfeeding Self-efficacy Short-Form Scale (BSES-SF). Follow-up data about duration of exclusive breastfeeding were obtained at six months postpartum. RESULTS: Mediation analysis indicated that previous breastfeeding experience directly affected the duration of exclusive breastfeeding and that the rate of the total indirect effect was 22.878%. Self-efficacy mediated previous breastfeeding experience and duration of exclusive breastfeeding, whereas attitude and self-efficacy played the series mediational role between previous breastfeeding experience and duration of exclusive breastfeeding. Cluster analysis supported these results. CONCLUSIONS: The duration of exclusive breastfeeding can be promoted by improving breastfeeding attitude and self-efficacy in women without breastfeeding experience.

Correction to: The RNA binding protein RBMS3 inhibits the metastasis of breast cancer by regulating Twist1 expression.

In the original publication of this article [1], the molecular weight of RBMS3 was incorrectly noted as 38 KDa within Fig 1A, Fig 2A and Fig 2B. The figures have been updated to list the correct molecular weight of RBMS3 as 41 KDa.

Comparative transcriptomic analysis reveals the regulatory effects of inorganic nitrogen on the biosynthesis of Monascus pigments and citrinin.

Monascus spp. and its secondary metabolites have been widely applied in foods and medicines for thousands of years in eastern Asia. Nitrogen sources are essential nutrients for the growth and metabolism of Monascus spp. Our previous study found that inorganic nitrogen sources (especially NH4Cl and NH4NO3) promoted the biosynthesis of Monascus pigments (MPs) and inhibited the production of citrinin. The objective of the present study was to investigate the regulatory mechanism of inorganic nitrogen on the biosynthesis of MPs and citrinin by the comparative transcriptional approach (RNA sequencing combined with RT-qPCR). Results indicated that the submerged fermentation of M. purpureus M3103 with NH4Cl or NH4NO3 as the sole nitrogen source can significantly increase the yields of MPs (especially for Monascus orange and red pigments) and decrease citrinin production, compared with the organic nitrogen source (peptone group). Comparative transcriptomic profiling by RNA sequencing found that the numbers of differentially expressed genes (DEGs) between different experimental groups-M group (peptone group) vs. ML group (NH4Cl group), and M group (peptone group) vs. MX group (NH4NO3 group), were 722 and 1287, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that genes involved in carbon and nitrogen metabolism, biosynthesis of amino acids were up-regulated by NH4Cl and NH4NO3, which would produce more biosynthetic precursors for MPs. Whereas, the inorganic nitrogen source (both of NH4Cl and NH4NO3) down-regulated the expression levels of genes involved in tyrosine metabolism. In addition, NR analysis indicated that the essential genes and transcription factors involved in the biosynthesis pathway of citrinin were down-regulated by NH4Cl and NH4NO3. These results indicated that NH4Cl or NH4NO3 as a nitrogen source for M. purpureus M3103 can significantly promote the precursor synthesis of Monascus pigments, but reduce the transcription of polyketide synthase for citrinin, and therefore significantly increase Monascus pigments production and decrease citrinin formation. These findings will facilitate a comprehensive understanding of the regulatory mechanisms of inorganic nitrogen in the biosynthesis of secondary metabolites in M. purpureus, and would benefit the application of M. purpureus in the production of MPs.

[Coridius Decoction increases penile erection hardness, IIEF-5 scores and testosterone level in patients with erectile dysfunction].

OBJECTIVE: To explore the effect of Coridius Decoction on penile erection hardness, IIEF-5 scores and the testosterone level in ED patients. METHODS: We selected 120 ED patients diagnosed and treated in our hospital between July 2018 and January 2020 and, using the random number table, divided them into a control (n = 55) and an observation (n = 65), the former treated with oral sildenafil and the latter with warm Coridius Decoction in addition, both for 8 weeks and followed up for 6 months. We compared the TCM syndrome scores, clinical effects, penile erection hardness, IIEF-5 scores, testosterone (T) level and adverse reactions between the two groups of patients. RESULTS: The TCM syndrome score, compared with the baseline, was significantly decreased in the observation (9.81 ± 0.61 vs 17.63 ± 1.16, P < 0.05) and the control group (17.56 ± 1.23 vs 13.18 ± 0.75, P < 0.05) after treatment, even lower in the former than in the latter group (P < 0.01). The total therapeutic effectiveness rate was markedly higher in the observation group than in the control (92.31% ï¼»60/65ï¼½ vs 80.00% ï¼»44/55ï¼½, P < 0.05). After medication, the erection hardness score (EHS) was dramatically higher than the baseline in the observation (4.21 ± 0.55 vs 2.55 ± 0.73, P < 0.01) and the control group (3.14 ± 0.54 vs 2.61 ± 0.73, P < 0.01), and so were the IIEF-5 score (18.58 ± 5.26 vs 12.00 ± 4.68, P < 0.05 and 15.29 ± 4.70 vs 11.94 ± 5.54, P < 0.05) and the T level (ï¼»13.27 ± 4.21ï¼½ vs ï¼»9.43 ± 4.31ï¼½ nmol/L, P < 0.05 and ï¼»10.74 ± 4.15ï¼½ vs ï¼»9.01 ± 4.72ï¼½ nmol/L, P < 0.05), both even higher in the former than in the latter group (P < 0.01). There were no statistically significant differences in the incidence rates of adverse reactions between the observation and control groups (6.15% ï¼»4/65ï¼½ vs 3.64% ï¼»2/55ï¼½, P = 0.834). CONCLUSIONS: Coridius Decoction is safe and effective for the treatment of ED, which can significantly improve the clinical symptoms, increase penile erectile hardness and the T level, and repair the erectile function of the patient.

A Web-based Survey of Marital Quality and Job Satisfaction among Chinese Nurses.

PURPOSE: This study aimed to use a web-based survey to investigate the correlation between job satisfaction and marital quality and to identify the association of demographics with job satisfaction and marital quality. METHODS: Married nurses (N = 2,296) completed the questionnaires. Correlations and linear regression analyses were carried out. RESULTS: Both marital quality and job satisfaction were relatively low. Additionally, marital quality was positively correlated with job satisfaction. Age, marital status (in years) and average daily hours spent with spouse had positive impact on job satisfaction. Multiple linear regression analyses showed that age, monthly income, average daily hours spent with spouse and marital quality were positively associated with job satisfaction. CONCLUSION: Because of the shifts nurses working, there is little time for nurses to spend with their spouses and family. It is recommended that hospital leaders could provide more flexibility with nurses' shift choices so nurses can arrange their work-life balance better. Other considerations like reducing workload and reducing working hours should be promoted as options. Nurse managers could offer counseling services including strategies to cope with the balance between work and life. This effort could improve job satisfaction and reduce the rate of turnover of nurses.

KIAA1429 acts as an oncogenic factor in breast cancer by regulating CDK1 in an N6-methyladenosine-independent manner.

Most N6-methyladenosine (m6A) associated regulatory proteins (i.e., m6A writer, eraser, and reader proteins) are involved in the pathogenesis of various cancers, mostly in m6A-dependent manners. As a component in the m6A 'writers', KIAA1429 is reported to be an RNA-binding protein and involved in the m6A modification, mRNA splicing and processing. Till now, the functions of KIAA1429 in tumorigenesis and related mechanism have not been reported. In the present study, we found KIAA1429 was highly expressed in breast cancer tissues, but frequently down-regulated in non-cancerous breast tissues. The overall survival of breast cancer patients with high-expression KIAA1429 was significantly shorter than those with low-expression KIAA1429. Then, we demonstrated that KIAA1429 was associated with breast cancer proliferation and metastasis in vivo and in vitro. The potential targeting genes of KIAA1429 in breast cancer were identified by RNA immunoprecipitation sequencing. One of these genes is cyclin-dependent kinase 1 (CDK1), which plays an oncogenic role in cancers. Furthermore, we confirmed that KIAA1429 played its oncogenic role in breast cancer by regulating CDK1 by an m6A-independent manner. 5'-fluorouracil was found to be very effective in reducing the expression of KIAA1429 and CDK1 in breast cancer. These findings indicated that KIAA1429 could promote breast cancer progression and was correlated with pathogenesis. It may represent a promising therapeutic strategy on breast cancer, especially in combination with CDK1 treatment.

Microbial communities and volatile metabolites in different traditional fermentation starters used for Hong Qu glutinous rice wine.

Hong Qu glutinous rice wine (HQGRW), as one of the most typical representatives of Chinese rice wine, is generally brewed from glutinous rice by adding two traditional wine fermentation starters-Hong Qu (HQ) and Bai Qu (BQ). The objective of this study was to determine the microbial communities and volatile metabolites of different traditional fermentation starters for HQGRW, and elucidate the potential correlation between microbiota and volatile metabolites. Both heatmap and principal component analysis (PCA) revealed the significant variances in volatile profiles among different wine starters. Microbiological analysis based on high-throughput sequencing (HTS) technology demonstrated that both of bacterial and fungal communities varied significantly in different starters. HQ was dominated mainly by bacteria of Bacillus ginsengihumi (20.17%), Pantoea sp. (10.39%), Elizabethkingia sp. (5.52%), Streptococcus sp. (5.03%) Brevundimonas sp. (3.03%), Rickettsia prowazekii (2.94%), Thermus thermophilus (2.54%), Bacillus amyloliquefaciens (1.48%), Bacillus aryabhattai (1.42%); fungi of Monascus purpureus (39.7%), Aspergillus niger (27.35%), Xeromyces bisporus (8.39%), Aspergillus penicillioides (6.89%), Aspergillus flavus (2.33%) and Pichia farinose (0.79%). By contrast, BQ contained much higher proportions of bacteria of Lactococcus lactis (10.45%), Lactobacillus brevis (9.99%), Pediococcus pentosaceus (8.29%), Weissella paramesenteroides (6.69%), Lactobacillus fermentum (4.83%), Gluconobacter thailandicus (3.93%), Lactobacillus alimentarius (3.59%), fungi of Rhizopus arrhizus (31.47%), Saccharomycopsis fibuligera (27.86%), Aspergillus niger (20.81%), Issatchenkia orientalis (3.79%), Saccharomycopsis malanga (3.15%), Clavispora lusitaniae (2.29%), Candida tropicalis (1.47%), Saccharomyces cerevisiae (1.11%) and Rhizopus microsporus (0.57%). Furthermore, core functional microbiota that might contribute to volatile flavour development was explored through Spearman's correlation-based network analysis. Lactobacillus brevis, Lactobacillus alimentarius, Lactobacillus plantarum and Aspergillus niger were found to be strongly associated with acid compounds (FDR adjusted P < 0.01), while Pichia sp., Candida sp., Monascus purpureus, Lactobacillus brevis and Lactobacillus alimentarius were positively correlated with concentrations of aromatic esters associated with fruity and floral notes (FDR adjusted P < 0.01), implying that these microorganisms might make significant contributions to the flavour of rice wine. These findings demonstrated that the aromatic quality of HQGRW may be critically influenced by the microbiota in traditional fermentation starters. To conclude, this study would contribute to the development of novel defined starter cultures for improving the aromatic quality of HQGRW.

The RNA binding protein RBMS3 inhibits the metastasis of breast cancer by regulating Twist1 expression.

BACKGROUND: Metastasis remains the biggest obstacle for breast cancer treatment. Therefore, identification of specific biomarker of metastasis is very necessary. The RNA binding protein 3 (RBMS3) acts as a tumor suppressor in various cancers. Whereas, its role and underlying molecular mechanism in breast cancer is far from elucidated. METHODS: Quantitative real-time PCR and western blots were carried out to determine the expression of RBMS3 in breast cancer cells and tissues. Transwell and in vivo metastasis assay were conducted to investigate the effects of RBMS3 on migration, invasion and metastasis of breast cancer cells. Transcriptome sequencing was applied to screen out the differential gene expression affected by RBMS3. RNA immunoprecipitation assay combined with luciferase reporter assay were performed to explore the direct correlation between RBMS3 and Twist1 mRNA. RESULTS: RBMS3 was downregulated in breast cancer and ectopic expression of RBMS3 contributed to inhibition of cell migration, invasion in vitro and lung metastasis in vivo. Furthermore, RBMS3 negatively regulated Twsit1 expression via directly binding to 3'-UTR of Twist1 mRNA, and thereby decreased Twist1-induced expression of matrix metalloproteinase 2 (MMP2). Additionally, Twist1-induced cell migration, invasion and lung metastasis could be reversed by the upregulation of RBMS3. CONCLUSIONS: In summary, our study revealed a novel mechanism of the RBMS3/Twsit1/MMP2 axis in the regulation of invasion and metastasis of breast cancer, which may become a potential molecular marker for breast cancer treatment.

Rapamycin enhances the sensitivity of ER­positive breast cancer cells to tamoxifen by upregulating p73 expression.

A total of 70% of breast cancers express the estrogen receptor (ER)α; therefore, targeting the ER may be an effective endocrine therapy with which to inhibit breast cancer growth. Tamoxifen is the most common­used clinically used drug for the treatment of advanced or metastatic ER­positive (ER+) breast cancer. However, a substantial proportion of patients become resistant to endocrine therapies. To overcome this limitation, in this stud, we sought to maximize the benefits associated with tamoxifen therapy via drug combination strategies. We demonstrated that rapamycin, an FDA­approved mammalian target of rapamycin (mTOR) inhibitor, enhanced the effects of endocrine therapy with tamoxifen, and the concentration of tamoxifen required for ER+ breast cancer cell growth inhibition was substantially reduced. Moreover, treatment with rapamycin plus tamoxifen significantly inhibited tumor growth in vivo. In addition, this synergistic effect may be mediated by the induction of p73. We revealed a novel mechanism in which p73 increases ERα expression by directly binding to the promoter region of the ERα gene. Taken together, the findings of this study indicate that combination therapy with rapamycin and tamoxifen underlying p73­mediated ERα expression may provide new insight into the drug combination for the treatment of ER+ breast cancer.

The regulation mechanisms of soluble starch and glycerol for production of azaphilone pigments in Monascus purpureus FAFU618 as revealed by comparative proteomic and transcriptional analyses.

Monascus spp. have been used for thousands of years as a traditional food additive in China. This mold can produce many different types of commercially valuable secondary metabolites of biological activity. Soluble starch and glycerol are the two principal carbon sources universally utilized by Monascus for the production of beneficial metabolites. In this study, the effects and regulation mechanisms of soluble starch and glycerol for M. purpureus FAFU618 on Monascus azaphilone pigments (MonAzPs) were investigated through ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS/MS), comparative proteomics and quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR). The production of intracellular and extracellular pigments was significantly different between the soluble starch group (SSG) and glycerol group (GCG). Additionally, the components of intracellular pigments revealed by UPLC-QTOF-MS/MS showed that Monascin and Ankaflavin increased significantly in the GCG, while Rubropunctatin and Monascorubrin increased in the SSG. Differentially expressed proteins of mycelia between SSG and GCG were analyzed by two-dimensional gel electrophoresis (2-DE) and MALDI-TOF/TOF MS. We identified 27 proteins with statistically altered expression, of which 18 proteins associated with the EMP (glycolytic pathway), translation, energy generation, proteolysis, etc. were up-regulated, and 9 proteins, including ribosomal proteins, heat shock proteins (HSPs) and others, were down-regulated in GCG. Meanwhile, the expression levels of MonAzP biosynthetic genes were also analyzed by RT-qPCR, and the results showed that mppA, mppC, mppR1 and mppR2 were down-regulated, whereas genes MpPKS5, MpFasA2, MpFasB2, mppB, mppD and mppE were up-regulated. Collectively, these findings illustrate that the regulation of MonAzPs is not only closely related to the expression levels of certain proteins in the polyketide synthesis pathway but also closely related to the concentration of primary metabolism-generated molecules that are used as substrates for polyketide synthesis. The present study provides insights into the regulation of different carbon sources on the metabolism of MonAzPs in M. purpureus FAFU618. These results may promote further development of functional foods or medicines from Monascus spp. fermented products.

Mining of a phospholipase D and its application in enzymatic preparation of phosphatidylserine.

Phosphatidylserine (PS) is useful as the additive in industries for memory improvement, mood enhancement and drug delivery. Conventionally, PS was extracted from soybeans, vegetable oils, egg yolk, and biomass; however, their low availability and high extraction cost were limiting factors. Phospholipase D (PLD) is a promising tool for enzymatic synthesis of PS due to its transphosphatidylation activity. In this contribution, a new and uncharacterized PLD was first obtained from GenBank database via genome mining strategy. The open reading frame consisted of 1614 bp and potentially encoded a protein of 538-amino-acid with a theoretical molecular mass of 60 kDa. The gene was successfully cloned and expressed in Escherichia coli. Its enzymatic properties were experimentally characterized. The temperature and pH optima of PLD were determined to be 60°C and 7.5, respectively. Its hydrolytic activity was improved by addition of Ca2+ at 5 mM as compared with the control. The enzyme displayed suitable transphosphatidylation activity and PS could be synthesized with L-serine and soybean lecithin as substrates under the catalysis of PLD. This PLD enzyme might be a potential candidate for industrial applications in PS production. To the best of our knowledge, this is the first report on genome mining of PLDs from GenBank database.

Three new cytotoxic oleanane triterpenoid saponins from Cylindrokelupha dalatensis.

Three new minor oleanane triterpenoid saponins, cylindrosides B (1), C (2), and D (3), were isolated from the seed of Cylindrokelupha dalatensis using chromatographic method. Their structures were established on the basis of the chemical and spectroscopic evidences. They displayed significant antitumor activity in vitro against HL60 cancer cell lines and IC50 values were 7.15 ± 0.63, 10.07 ± 0.97, and 4.74 ± 0.57 µM, respectively, by MTT method.