[Contamination Characteristics and Ecological Risk Assessment of Pharmaceuticals and Personal Care Products in Drains Flowing into the Yellow River of Ningxia].

Pharmaceuticals and personal care products (PPCPs) are a group of emerging contaminants causing detrimental effects on aquatic living organisms even at low doses. To investigate the contamination characteristics and ecological risks of PPCPs in drains flowing into the Yellow River of Ningxia, 21 PPCPs were detected and analyzed using solid phase extraction and ultra-high performance liquid chromatography-mass spectrometry in this study. All 21 targeted compounds were detected in the drains, with total concentrations ranging from 47.52 to 1 700.96 ng·L-1. Ciprofloxacin, acetaminophen, benzophenone-3, and diethyltoluamide were the more commonly detected compounds, with detection frequencies exceeding 80%. The five highest-concentration PPCPs were acetaminophen, diethyltoluamide, caffeine, benzophenone-3, and levofloxacin, with the maximum concentrations of 597.21, 563.23, 559.00, 477.28, and 473.07 ng·L-1, respectively. Spatial analysis showed that the pollution levels of PPCPs in the drains of the four cities were different, with average concentrations of ∑PPCPs in the order of Yinchuan>Shizuishan>Wuzhong>Zhongwei. The total concentration of PPCPs before flowing into the Yellow River ranged from 124.82 to 1 046.61 ng·L-1. Source analysis showed that livestock and poultry breeding wastewater was the primary source for sulfadiazine and oxytetracycline, whereas medical wastewater was the primary source for levofloxacin and ciprofloxacin. The primary sources of triclocarban and triclosan were domestic sewage and industrial wastewater, whereas the primary source of caffeine and diethyltoluamide was domestic sewage. The pollution of diciofenac, cimetidine, triclocarban, and triclosan in the drains was positively correlated with the regional population and economic development level. The ecological risk assessment indicated that levofloxacin, diclofenac, gemfibrozil, benzophenone-3, and triclocarban posed high risks to aquatic organisms in drains flowing into the Yellow River. It is worthwhile to consider the mixture risk of the PPCPs that exhibited high risk at most sampling sites.

Uncovering the Power of HSCCC: Separation of Diastereomeric and Regioisomeric Styrylpyrones from the Stem Bark of Goniothalamus leiocarpus.

The plant Goniothalamus leiocarpus of the Annonaceae family is used as an alternative medicine in tropical regions. Applying high-speed counter current chromatography (HSCCC), eight new bioactive styrylpyrone isomers, including 6R,7S,8R,2'S-goniolactone B (1), 6S,7S,8S,2'S-goniolactone B (2), 6R,7R,8R,2'S-goniolactone B (3), 6R,7S,8S,2'S-goniolactone C (4), 6R,7S,8R,2'S-goniolactone C (5), 6S,7R,8S,2'S-goniolactone C (6), and two positional isomers, 6R,7R,8R,2'S-goniolactone G (7) and 6S,7R,8R,2'S-goniolactone G (8), were isolated from a chloroform fraction (2.1 g) of G. leiocarpus, which had a prominent spot by TLC analysis. The structures of the new compounds were elucidated by MS, NMR, IR, and UV spectra, and their absolute configurations were determined by Mosher's method, ECD, and X-ray diffraction analysis. The isolates are characteristic components found in plants of the genus Goniothalamus and consist of two structural moieties: a styrylpyrone and a dihydroflavone unit. The isolation of the eight new compounds demonstrates the effectiveness of HSCCC in separating the isomers of natural styrylpyrone. In a bioactivity assessment, compounds 1 and 6 exhibited cytotoxic effects against the human colon carcinoma cell lines LS513 and SW620 with IC50 values ranging from 1.6 to 3.9 µM. Compounds 1, 2, 7, and 8 showed significant synergistic activity against antibiotic-resistant Staphylococcus aureus strains.

Annonaceous Acetogenins Synergistically Inhibit Hepatocellular Carcinoma with Sorafenib.

Sorafenib was first approved as the standard treatment for advanced hepatocellular carcinoma (HCC). Despite providing an advantage in terms of patient survival, sorafenib has shown poor clinical efficacy and severe side effects after long-term treatment. Thus, combination treatment is a potential way to increase the effectiveness and reduce the dose-limiting toxicity of sorafenib. Extracts of the seeds of Annona montana have shown synergistic antitumor activity with sorafenib, and seven annonaceous acetogenins, including three new acetogenins, muricin P (2), muricin Q (3), and muricin R (4), were isolated from the extracts by bioguided fractionation and showed synergy with sorafenib. The structures of these compounds were determined using spectroscopic and chemical methods. Annonacin (1) and muricin P (2), which reduced intracellular ATP levels and promoted apoptosis, exhibited synergistic cytotoxicity with sorafenib in vitro. In vivo, annonacin (1) displayed synergistic antitumor activity by promoting tumor cell apoptosis. Moreover, the potential mechanism of annonacin (1) was predicted by transcriptomic analysis, which suggested that SLC33A1 is a potential target in HCC. Annonacin (1) might be a novel candidate for combination therapy with sorafenib against advanced HCC.

Natural Product BO-1 as an Inner Responsive Molecule Inhibits Antimicrobial-Resistant Staphylococcus aureus via Synergism.

Multidrug-resistant Staphylococcus aureus, a Gram-positive bacterium that causes several difficult-to-treat human infections, is a considerable threat to global healthcare. We hypothesize that there exist inner responsive molecules (IRMs) which can function synergistically with antibiotics to restore the sensitivity of resistant bacteria to existing antibiotics without inducing new antibiotic resistance. An investigation of the extracts of the Chinese medicinal herb Piper betle L. led to the isolation of six benzoate esters, BO-1-BO-6. Among these, BO-1 as a distinct IRM displayed considerable synergism by potentiating antibacterial activity against five antibiotic-resistant S. aureus strains. Mechanistic studies demonstrated that BO-1 acted as a suppressing drug resistance IRM via inhibiting efflux activity. A combination of BO-1 with ciprofloxacin significantly inhibited resistance to this antibiotic and reversed its resistance in the S. aureus strain. Furthermore, BO-1 effectively enhanced the activity of ciprofloxacin against the efflux fluoroquinolone-resistant S. aureus strain SA1199B that caused infection in two animal models and significantly decreased the inflammatory factors IL-6 and C-reactive protein of the infected mice, thereby showing the practice utility of this approach.

[Pollution Characteristics and Risk Assessment of Typical Endocrine Disrupting Chemicals in Drains Flowing into the Yellow River of Ningxia].

Solid-phase extraction and ultra-high performance liquid chromatography-mass spectrometry were used to detect and analyze the contamination of nine typical endocrine disrupting chemicals (EDCs) in drains flowing into the Yellow River of Ningxia. Sources of these EDCs were analyzed, and their risks were assessed. The results showed that EDCs were detected in drains at all 33 sampling sites, with total concentrations （ΣEDCs） of 82.28-1730.09 ng·L-1. Among phenolic compounds, bisphenol A (BPA) and nonylphenol (NP) were two that were more commonly detected EDCs, with the detection rates above 90%; estrone (E1) and estriol (E3) were more commonly detected estrogenic compounds, both with detection rates of 79%. On a spatial scale, the average concentrations of ΣEDCs in the drains in Shizuishan and Yinchuan were much higher than those in Wuzhong and Zhongwei. Concentrations of ΣEDCs at the sampling sites before flowing into Yellow River ranged from 82.28 to 979.82 ng·L-1. The source analysis showed that industrial wastewater and domestic sewage were two primary sources for BPA, whereas industrial wastewater was the primary source for OP. The primary sources of E1 and E3 were livestock and poultry breeding wastewater and domestic sewage, respectively. Risk assessment results showed that EDCs in drains flowing into the Yellow River posed low or moderate ecological risk but high risk for estrogenic activity at all sampling sites.

Synthesis and anti-SARS-CoV-2 evaluation of lipid prodrugs of ß-D-N4-hydroxycytidine (NHC) and a 3'-fluoro-substituted analogue of NHC.

ß-D-N4-hydroxycytidine (NHC, EIDD-1931) is a nucleoside analogue that exhibits broad spectrum antiviral activity against a variety of RNA viruses. Herein, we report the synthesis of a series of lipid prodrugs of NHC and a novel 3'-fluoro modified NHC analogue, and evaluation of their antiviral activity against five variants of SARS-CoV-2. All lipid prodrugs showed potent antiviral activity against the tested SARS-CoV-2 variants with EC50 values in the range of 0.31-3.51 µM, which were comparable to those of NHC or higher than those of remdesivir and molnupiravir. An increase in the cytostatic activity of the lipid prodrugs was found, but prodrug 2d proved equally selective as molnupinavir. The 3'-F analogue of NHC (6) only displayed minor antiviral activity against the SARS-CoV-2 Omicron variant (EC50 = 29.91 µM), while no activity was found for other variants at the highest concentration tested. The promising antiviral data of the lipid prodrugs of NHC suggest that they deserve further investigation as new anti-SARS-CoV-2 drugs.

The insecticidal effect of the botanical insecticide chlorogenic acid on Mythimna separata (Walker) is related to changes in MsCYP450 gene expression.

The oriental armyworm Mythimna separata (Walker) (Lepidoptera: Noctuidae) can feed on the leaves of many crops, resulting in vast areas of damage and severe losses. Therefore, this insect has become a significant agricultural pest in north Asia. In this study, we fed 3rd instar larvae with artificial diets containing different concentrations of chlorogenic acid and found a significant lethal effect and the mortality increased with increasing chlorogenic acid concentration. Next, we measured the sublethal effect of chlorogenic acid at LC20 on the growth and development of M. separata larvae. The durations of the 4th and 5th instar were longer than those of the control group (prolonged by 0.8 and 0.6 days, respectively), and the 6th instar was shorter (by 1.1 days). The total survival rate, pupation rate, eclosion rate, sex ratio, and oviposition amount in the LC20 chlorogenic acid-treated group were significantly lower than those in the control group. Furthermore, transcriptome analysis of 3rd instar larvae fed various concentrations of chlorogenic acid revealed that several MsCYP450 genes were significantly up-regulated, and this finding was further validated by qRT-PCR. In addition, various concentrations of chlorogenic acid and different treatment times significantly affected the enzyme activity of CYP450 in 3rd instar larvae. Importantly, dietary ingestion of dsMsCYP450 significantly reduced the mRNA level of MsCYP450 genes and increased mortality in the presence of chlorogenic acid. Our results revealed that MsCYP6B6, MsCYP321A7, and MsCYP6B7-like play an essential role in the detoxification of chlorogenic acid by M. separata. This study provides evidence of control effect by botanical insecticide chlorogenic acid on M. separata, and potential detoxification mechanism mediated by P450 of botanical insecticide in arthropods.

[Contamination Characteristics and Ecological Risk Assessment of Pharmaceuticals and Personal Care Products (PPCPs) in the Third Drain of Ningxia].

To investigate the contamination characteristics and potential ecological risks of pharmaceuticals and personal care products (PPCPs) in the Third Drain of Ningxia, 14 PPCPs were detected and analyzed using solid-phase extraction coupled with ultra-high performance liquid chromatography-tandem mass spectrometry. The results showed that these 14 PPCPs were detected in the Third Drain and its confluent streams, with total concentrations of 117.74-1947.64 ng·L-1 and 63.94-4509.39 ng·L-1, respectively. Detection ratios of gemfibrozil (GEM), caffeine (CAF), avobenzone (BM-DBM), benzophenone-3 (BP-3), 3-(4-methylbenzylidene)-camphor (4-MBC), and diethyltoluamide (DEET) were 100% in the drain. The highest-concentration pharmaceutical was GEM (7.78-721.84 ng·L-1), followed by CAF (41.74-246.86 ng·L-1), and the highest-concentration personal care product was DEET (3.17-219.91 ng·L-1), followed by BP-3 (56.92-150.14 ng·L-1). Concentrations of PPCPs at different sampling points exhibited spatial differences. The total PPCPs concentration increased dramatically and reached a maximum value after flowing through Pingluo County, then showed a decreasing trend downstream. Correlation analysis showed that 4-MBC was significantly positively correlated with COD (P<0.01). IBU, XMTD, TCC, and TCS were significantly correlated with NH4+-N (P<0.05). DIC, BF, CBZ, and DEET were significantly correlated with TN (P<0.05). The results indicated that concentrations of PPCPs were closely related to water quality indexes. Risk assessment showed that DIC, IBU, GEM, CBZ, CAF, and BP-3 had high risks, whereas BM-DBM, TCC, and TCS had moderate risks.

[Contamination Characteristics and Ecological Risk Assessment of Antibiotics in the Third Drain of Ningxia].

Solid-phase extraction and ultra-high performance liquid chromatography-electrostatic field orbit hydrazine mass spectrometry were used to detect and analyze the contamination of nine antibiotics including sulfonamides, tetracyclines, macrolides, quinolones, and lincomycin in the third drain of Ningxia. Correlations between the antibiotic concentration and water quality indexes were explored, and the ecological risks were evaluated. The results showed that seven antibiotics were detected in the third drain and its confluent stream, with the total concentrations of 14.91-153.48 ng·L-1 and ND-39.37 ng·L-1, respectively. The detection ratios of clarithromycin, levofloxacin, ciprofloxacin, and lincomycin were 100% in the third drain. The highest-concentration antibiotic was levofloxacin (0.84-94.12 ng·L-1), followed by lincomycin (11.15-48.13 ng·L-1). Based on the spatial distribution analysis, after flowing through Pingluo County, total antibiotic concentrations in the third drain increased significantly and showed an increasing trend. The maximum concentration appeared in the Huinong section and showed a decreasing trend downstream. The total concentration of antibiotics before flowing into the Yellow River was 20.26 ng·L-1. Correlation analysis showed that levofloxacin was significantly positively correlated with NH4+-N and TN (P<0.01), erythromycin was significantly correlated with TN, and clarithromycin was significantly correlated with both NH4+-N and TN (P<0.05), which indicated that water quality index was closely related to antibiotic concentration. The results of the ecological risk assessment showed that levofloxacin and clarithromycin in the third drain posed certain ecological risks.

Reverse construction of dominant/secondary facets in Bi24O31Br10 photocatalysts for boosting electronic transfer.

In this paper, it is found that the preferential growth of secondary {117} facets of Bi24O31Br10 into dominant facets would lead to higher photocatalytic activity, although the original main {213} facet has a stronger molecular oxygen adsorption ability, which illustrates that the charge separation efficiency induced by dominant/secondary facet control plays a more important role than that of O2 adsorptive performance in improving activity.

Luminescent cluster-organic frameworks constructed from predesigned supertetrahedral {Ln4Zn6} secondary building units.

3d-4f heterometallic supertetrahedral clusters with the formula of Ln4Zn6(µ6-O)L4(CH3COO)6(NO3)4(CH3OH)4(H2O)2 (1-Ln, Ln = Eu, Gd, Tb, H3L = 2-(hydroxymethyl)-2-(pyridin-4-yl)-1,3-propanediol) have been successfully introduced as stable secondary building units (SBUs) to construct new cluster-organic frameworks with tunable emission, demonstrating a promising strategy for developing new optical materials.

Stimulus-specific hypothalamic encoding of a persistent defensive state.

Persistent neural activity in cortical, hippocampal, and motor networks has been described as mediating working memory for transiently encountered stimuli1,2. Internal emotional states, such as fear, also persist following exposure to an inciting stimulus3, but it is unclear whether slow neural dynamics are involved in this process. Neurons in the dorsomedial and central subdivisions of the ventromedial hypothalamus (VMHdm/c) that express the nuclear receptor protein NR5A1 (also known as SF1) are necessary for defensive responses to predators in mice4-7. Optogenetic activation of these neurons, referred to here as VMHdmSF1 neurons, elicits defensive behaviours that outlast stimulation5,8, which suggests the induction of a persistent internal state of fear or anxiety. Here we show that in response to naturalistic threatening stimuli, VMHdmSF1 neurons in mice exhibit activity that lasts for many tens of seconds. This persistent activity was correlated with, and required for, persistent defensive behaviour in an open-field assay, and depended on neurotransmitter release from VMHdmSF1 neurons. Stimulation and calcium imaging in acute slices showed that there is local excitatory connectivity between VMHdmSF1 neurons. Microendoscopic calcium imaging of VMHdmSF1 neurons revealed that persistent activity at the population level reflects heterogeneous dynamics among individual cells. Unexpectedly, distinct but overlapping VMHdmSF1 subpopulations were persistently activated by different modalities of threatening stimulus. Computational modelling suggests that neither recurrent excitation nor slow-acting neuromodulators alone can account for persistent activity that maintains stimulus identity. Our results show that stimulus-specific slow neural dynamics in the hypothalamus, on a time scale orders of magnitude longer than that of working memory in the cortex9,10, contribute to a persistent emotional state.

All-inorganic open frameworks based on gigantic four-shell Ln@W8@Ln8@(SiW12)6 clusters.

A series of POM-based all-inorganic open frameworks, H9[Ln9W8(µ4-O)12(µ2-O)24(H2O)24](SiW12O40)3·60H2O (1-Ln, Ln = La, Pr and Nd), have been fabricated and structurally characterized for the first time. The unique architectures are constructed from nanoscale four-shell Ln@W8@Ln8@(SiW12)6 clusters. Additionally, the porous nature of the frameworks has also been investigated.

[Mechanisms and Efficiencies of Removal of PPCPs by Pilot River Water Bypass Treatment Process].

The pharmaceuticals and personal care products (PPCPs) released into urban rivers are triggering certain ecological risks. The current study investigated the removal efficiencies of 30 frequently detected PPCPs by two river water bypass treatment processes (CS-BAF-UF-Ozone and CS-MBR-Ozone), and investigated the removal mechanism and eco-toxicological risk variation of target compounds via section-removal investigation and risk quotient model, respectively. Results indicated that both processes could efficiently remove the target PPCPs; the removal rates of tetracyclines and caffeine were>90% in the biological sections, while sulfonamides, fluoroquinolones, and other pharmaceuticals could only be efficiently removed when the COD of the influent and the water temperature were comparatively higher; the ozone process had particular effect on removing these compounds. The cumulative removal rate of all PPCPs during the whole process was higher than 92.5%. The total risk quotient (RQtot) of target PPCPs could be efficiently reduced by the bypass treatment processes; the RQtot decreased from 12.6 in the influent river water to 0.2 in the ozone effluent, with a removal rate of 98.4%.

Gene Correction Reverses Ciliopathy and Photoreceptor Loss in iPSC-Derived Retinal Organoids from Retinitis Pigmentosa Patients.

Retinitis pigmentosa (RP) is an irreversible, inherited retinopathy in which early-onset nyctalopia is observed. Despite the genetic heterogeneity of RP, RPGR mutations are the most common causes of this disease. Here, we generated induced pluripotent stem cells (iPSCs) from three RP patients with different frameshift mutations in the RPGR gene, which were then differentiated into retinal pigment epithelium (RPE) cells and well-structured retinal organoids possessing electrophysiological properties. We observed significant defects in photoreceptor in terms of morphology, localization, transcriptional profiling, and electrophysiological activity. Furthermore, shorted cilium was found in patient iPSCs, RPE cells, and three-dimensional retinal organoids. CRISPR-Cas9-mediated correction of RPGR mutation rescued photoreceptor structure and electrophysiological property, reversed the observed ciliopathy, and restored gene expression to a level in accordance with that in the control using transcriptome-based analysis. This study recapitulated the pathogenesis of RPGR using patient-specific organoids and achieved targeted gene therapy of RPGR mutations in a dish as proof-of-concept evidence.

[UPLC-Q-TOF/MS for rapid analysis of chemical constituents in Sanhuang tablets].

The qualitative analysis method of ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) was established for the chemical constituents in Sanhuang tablets. Waters ACQUITY BEH C18 (2.1 mm×100 mm, 1.7 µm) column was used with 0.1% formic acid solution (A)-0.1% formic acid acetonitrile (B) as the mobile phase for gradient elution. The flow rate was 0.2 mL•min⁻¹; the sample volume was 1 µL and the column temperature was 30 ℃. The high-resolution quadrupole time-flight mass spectrometry was used as detector with electrospray ion source in both positive and negative models, and the dry gas temperature was 325 ℃. Based on the analysis of mass spectrometry and literature reports, 38 compounds were confirmed, including 1 alkaloid, 1 dianthrone compound, 6 tannins, 7 anthraquinone glycosides, 6 anthraquinones and 17 flavonoids. Liquid chromatography-mass spectrometry method is simple, reliable and rapid to identify the chemical compositions of Sanhuang tablets, and it is helpful to reveal its chemical constituents and pharmacodynamic substances.

Role of lncRNAs as Novel Biomarkers and Therapeutic Targets in Ovarian Cancer.

Ovarian cancer (OC) is the leading cause of death among all gynecological malignancies in the world and its underlying mechanism is still unclear. Compared with research on microRNAs, research on long non-coding RNAs (lncRNAs) is still in its infancy. Studies in recent years have demonstrated that lncRNAs exhibit multiple biological functions in various stages of OC development. In this review, we conclude that lncRNAs are closely involved in the pathogenesis of OC. The expression of lncRNAs indicates the early diagnosis, prognosis, and response to chemotherapy of OC. An attractive approach to treatment of OC is lncRNA small interfering RNA or acting as a plasmid targeting the expression of toxic genes, which is a novel step toward a major breakthrough in the treatment of human OC. E2-regulated lncRNA and its polymorphism, methylation, are also involved in OC. Further research efforts are needed before fully identifying, characterizing, and elucidating the actual functions of lncRNAs in OC at the molecular level and putting them into clinical practice.

The lncRNA H19 mediates breast cancer cell plasticity during EMT and MET plasticity by differentially sponging miR-200b/c and let-7b.

Metastasis is a multistep process by which tumor cells disseminate from their primary site and form secondary tumors at a distant site. The pathophysiological course of metastasis is mediated by the dynamic plasticity of cancer cells, which enables them to shift between epithelial and mesenchymal phenotypes through a transcriptionally regulated program termed epithelial-to-mesenchymal transition (EMT) and its reverse process, mesenchymal-to-epithelial transition (MET). Using a mouse model of spontaneous metastatic breast cancer, we investigated the molecular mediators of metastatic competence within a heterogeneous primary tumor and how these cells then manipulated their epithelial-mesenchymal plasticity during the metastatic process. We isolated cells from the primary mammary tumor, the circulation, and metastatic lesions in the lung in TA2 mice and found that the long noncoding RNA (lncRNA) H19 mediated EMT and MET by differentially acting as a sponge for the microRNAs miR-200b/c and let-7b. We found that this ability enabled H19 to modulate the expression of the microRNA targets Git2 and Cyth3, respectively, which encode regulators of the RAS superfamily member adenosine 5'-diphosphate (ADP) ribosylation factor (ARF), a guanosine triphosphatase (GTPase) that promotes cell migration associated with EMT and disseminating tumor cells. Decreasing the abundance of H19 or manipulating that of members in its axis prevented metastasis from grafts in syngeneic mice. Abundance of H19, GIT2, and CYTH3 in patient samples further suggests that H19 might be exploited as a biomarker for metastatic cells within breast tumors and perhaps as a therapeutic target to prevent metastasis.

Functional response properties of VIP-expressing inhibitory neurons in mouse visual and auditory cortex.

Despite accounting for about 20% of all the layer 2/3 inhibitory interneurons, the vasoactive intestinal polypeptide (VIP) expressing neurons remain the least thoroughly studied of the major inhibitory subtypes. In recent studies, VIP neurons have been shown to be activated by a variety of cortico-cortical and neuromodulatory inputs, but their basic sensory response properties remain poorly characterized. We set out to explore the functional properties of layer 2/3 VIP neurons in the primary visual (V1) and primary auditory cortex (A1), using two-photon imaging guided patch recordings. We found that in the V1, VIP neurons were generally broadly tuned, with their sensory response properties resembling those of parvalbumin (PV) expressing neurons. With the exception of response latency, they did not exhibit a significant difference from PV neurons across any of the properties tested, including overlap index, response modulation, orientation selectivity, and direction selectivity. In the A1, on the other hand, VIP neurons had a strong tendency to be intensity selective, which is a property associated with a subset of putative pyramidal cells and virtually absent in PV neurons. VIP neurons had a best intensity that was significantly lower than that of PV and putative pyramidal neurons. Finally, sensory evoked spike responses of VIP neurons were delayed relative to pyramidal and PV neurons in both the V1 and A1. Combined, these results demonstrate that the sensory response properties of VIP neurons do not fit a simple model of being either PV-like broadly tuned or pyramidal-like narrowly tuned. Instead, the selectivity pattern varies with sensory area and can even be, as in the case of low sound intensity responsiveness, distinct from both PV and pyramidal neurons.