Effects of cyanotoxins on nitrogen transformation in aquaculture systems with microplastics coexposure: Adsorption behavior, bacterial communities and functional genes.

Microcystin-LR (MC-LR) and microplastics (MPs) have attracted increasing attention as important new pollutants in freshwater fishery environments. However, there are few reports on the effects of long-term combined MC-LR and MPs pollution on nitrogen transformation and microbial communities in aquaculture ponds, and the resulting risks have yet to be determined. Therefore, in this study, traditional refractory MPs (polystyrene, PS), biodegradable MPs (polylactic acid, PLA) and MC-LR, which are common in freshwater fishery environments in China, were selected as pollutants to construct a microcosm that simulates freshwater aquaculture ponds. MC-LR coexposure to PS and PLA was tested to reveal the effects of these pollutants on nitrogen transformation and microbial communities in aquaculture ponds, as well as to elucidate the potential risks posed by traditional refractory MPs and biodegradable MPs to freshwater aquaculture ecosystems. The results revealed that the MPs had a relatively high adsorption rate for MC-LR and that PS presented a relatively high adsorption capacity, whereas PLA presented a relatively high desorption capacity. Single or combined MPs and MC-LR pollution disrupted the normal nitrogen cycle in the aquaculture system, causing an overall loss of nitrogen in the water, and denitrification and nitrogen fixation in the water were inhibited to a certain extent through the inhibition of nitrogen cycle-related functional genes, with the PS + MC-LR group having the greatest inhibitory effect. In addition, compared with single-pollutant exposure, combined exposure to MC-LR and MPs had a greater effect on the microbial community composition. Analysis of the integrated biomarker response (IBR) index revealed that the risk of combined exposure to MC-LR and PS was greater than that of single exposure, so this phenomenon merits further attention.

Breaking Latent Infection: How ORF37/38-Deletion Mutants Offer New Hope against EHV-1 Neuropathogenicity.

Equid alphaherpesvirus 1 (EHV-1) has been linked to the emergence of neurological disorders, with the horse racing industry experiencing significant impacts from outbreaks of equine herpesvirus myeloencephalopathy (EHM). Building robust immune memory before pathogen exposure enables rapid recognition and elimination, preventing infection. This is crucial for effectively managing EHV-1. Removing neuropathogenic factors and immune evasion genes to develop live attenuated vaccines appears to be a successful strategy for EHV-1 vaccines. We created mutant viruses without ORF38 and ORF37/38 and validated their neuropathogenicity and immunogenicity in hamsters. The ∆ORF38 strain caused brain tissue damage at high doses, whereas the ∆ORF37/38 strain did not. Dexamethasone was used to confirm latent herpesvirus infection and reactivation. Dexamethasone injection increased viral DNA load in the brains of hamsters infected with the parental and ∆ORF38 strains, but not in those infected with the ∆ORF37/38 strain. Immunizing hamsters intranasally with the ∆ORF37/38 strain as a live vaccine produced a stronger immune response compared to the ∆ORF38 strain at the same dose. The hamsters demonstrated effective protection against a lethal challenge with the parental strain. This suggests that the deletion of ORF37/38 may effectively inhibit latent viral infection, reduce the neuropathogenicity of EHV-1, and induce a protective immune response.

Alkaloids of Aconiti Lateralis Radix Praeparata inhibit growth of non-small cell lung cancer by regulating PI3K/Akt-mTOR signaling and glycolysis.

Aconiti Lateralis Radix Praeparata (Fuzi in Chinese) is widely used in the clinical treatment of tumors. This study aims to explore the active fractions and underlying mechanisms of Fuzi in the treatment of non-small cell lung cancer (NSCLC). Fuzi alkaloids (FZA) is prepared and found to inhibit the growth of NSCLC both in vitro and in vivo significantly. A total of 53 alkaloids are identified in FZA by UPLC-Q-TOF-MS. Proteomics experiment show that 238 differentially expressed proteins regulated by FZA are involved in amino acid anabolism, pyrimidine metabolism and PI3K/Akt-mTOR signaling pathway. Metabolomics analyses identify 32 significant differential metabolites which are mainly involved in amino acid metabolism, TCA cycle and other pathways. Multi-omics research combined with molecular biological assays suggest that FZA might regulate glycolysis through PI3K/Akt-mTOR pathway to treat NSCLC. The study lays a foundation for the anti-cancer investigation of Fuzi and provides a possible scientific basis for its clinical application.

Characteristics of Different Obesity Metabolic Indexes and their Correlation with Insulin Resistance in Patients with Polycystic Ovary Syndrome.

This study is aimed to investigate the characteristics of different obesity metabolic indexes [body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), waist-to-height ratio (WHtR), cardiometabolic index (CMI), body roundness index (BRI), visceral adiposity index (VAI), and lipid aggregation products (LAP)] and their correlation with insulin resistance (IR) in patients with polycystic ovary syndrome (PCOS). The study was conducted on 140 subjects with PCOS and 133 control subjects aged 18-44 years. According to insulin resistance index (HOMA-IR) ≥ 2.69 and HOMA-IR < 2.69, PCOS group members were divided into insulin resistance group and non-insulin resistance group. Anthropometric and serological characteristics of the population with PCOS focused on calculating different obesity metabolic indexes and HOMA-IR. BMI, WC, WHR, WHtR, CMI, BRI, VAI, and LAP were significantly higher in PCOS patients than in the control group, and the differences were all statistically significant (P < 0.05); In the insulin resistance group of PCOS patients, BMI, WC, WHR, WHtR, CMI, BRI, VAI, and LAP were significantly higher than in the non-insulin resistance group (P < 0.05). In PCOS patients, BMI (r = 0.658, P < 0.001), WC (r = 0.0.662, P < 0.001), WHR (r = 0.377, P < 0.001), WHtR (r = 0.660, P < 0.001), CMI (r = 0.698, P < 0.001), BRI (r = 0.757, P < 0.001), VAI (r = 0.640, P < 0.001), and LAP (r = 0.767, P < 0.001) were positively correlated with IR. Obesity metabolic indexes associated with PCOS were elevated in the PCOS group compared to the control group, and in the PCOS insulin-resistant group compared to the non-insulin resistant group. Novel obesity metabolic indexes, especially CMI, BRI and LAP, might be more appropriate for evaluating the risk of concurrent IR in PCOS.

The Relationship Between Anti-Müllerian Hormone and Body Composition Components in Patients With Polycystic Ovary Syndrome of Reproductive Age.

To investigate and analyze the relationship between body composition components, including Body Mass Index (BMI), body fat percentage, waist-to-hip ratio, and visceral fat index, and Anti-Müllerian Hormone (AMH) levels in patients diagnosed with Polycystic Ovary Syndrome (PCOS), The study aims to provide a comprehensive understanding of how various aspects of body composition impact AMH levels. This study enrolled 167 women with PCOS of reproductive age. Serum AMH level and body composition were measured, and the correlation between body composition elements and AMH levels was analyzed. AMH level was negatively correlated with body weight, BMI, fat-free mass, body fat percentage, waist-hip ratio, and visceral fat level (P < 0.01). And negatively correlated with skeletal muscle ratio ( P = 0.003). AMH level remained significantly associated with BMI ( P = 0.028), body fat percentage ( P = 0.040), waist-hip ratio (P = 0.003), and visceral fat level ( P = 0.040) after age was included and a multiple linear regression model was established. After adjusting for age, BMI was still significantly associated with AMH (P = 0.029). At the same time, there was no obvious linear correlation between BSA and AMH. The results showed that AMH levels were significantly different among the three groups (9.53 ± 5.12 vs 6.98 ± 3.35 vs 6.38 ± 3.38, P < 0.001; ng/mL). The level of AMH in the non-central obesity group was higher than that in the central obesity group (9.68 ± 5.22vs7.09 ± 3.83, P < 0.001; ng/mL). In PCOS patients, those who are more obese have lower AMH levels, indicating poorer Ovarian Reserve. BMI may independently affect AMH levels, apart from age, BSA, and other factors. Ovarian function in centrally obese patients is poorer than in those with non-central obesity.

LanGui tea, an herbal medicine formula, protects against binge alcohol-induced acute liver injury by activating AMPK-NLRP3 signaling.

BACKGROUND: LanGui tea, a traditional Chinese medicine formulation comprising of Gynostemma pentaphyllum (Thunb.) Makino, Cinnamomum cassia (L.) J. Presl, and Ampelopsis grossedentata (Hand-Mazz) W.T. Wang, has yet to have its potential contributions to alcoholic liver disease (ALD) fully elucidated. Consequently, the objective of this research is to investigate the protective properties of LanGui tea against binge alcohol-induced ALD and the mechanisms underlying its effects. METHODS: An experimental model of acute alcohol-induced liver disease was performed to assess the protective effects of extract of LanGui tea (ELG) at both 50 and 100 mg.kg-1 dosages on male C57BL/6 mice. Various parameters, including hepatic histological changes, inflammation, lipids content, as well as liver enzymes and interleukin 1ß (IL-1ß) in the serum were measured. The pharmacological mechanisms of ELG, specifically its effects on adenosine monophosphate-(AMP)-activated protein kinase (AMPK) and NLR family pyrin domain containing 3 (NLRP3) signaling, were investigated through Western blotting, qRT-PCR, ELISA, immunohistochemistry, immunofluorescence analyses, and by blocking the AMPK activity. RESULTS: ELG demonstrated a mitigating effect on fatty liver, inflammation, and hepatic dysfunction within the mouse model. This effect was achieved by activating AMPK signaling and inhibitingNLRP3 signaling in the liver, causing a reduction in IL-1ß generation. In vitro studies further confirmed that ELG inhibited cell damage and IL-1ß production in ethanol-induced hepatocytes by enhancing AMPK-NLRP3 signaling. Conversely, the pharmacological inhibition of AMPK activity nearly abrogated such alteration. CONCLUSIONS: Thus, LanGui tea emerges as a promising herbal therapy for ALD management involving AMPK-NLRP3 signaling.

Modulation of cellular metabolism and alleviation of bacterial dysbiosis by Aconiti Lateralis Radix Praeparata in non-small cell lung cancer treatment.

BACKGROUND: Non-small cell lung cancer (NSCLC) is a highly prevalent and fatal form of lung cancer. In China, Aconiti Lateralis Radix Praeparata (Fuzi in Chinese), derived from the lateral root of Aconitum carmichaeli Debx. (Ranunculaceae, Aconitum), is extensively prescribed to treat cancer in traditional medicine and clinical practice. However, the precise mechanism by which Fuzi treats NSCLC remains unknown. PURPOSE: This article aims to assess the efficacy of Fuzi against NSCLC and elucidate its underlying mechanism. METHODS: Marker ingredients of Fuzi decoction were quantified using UPLC-TSQ-MS. The effectiveness of Fuzi on NSCLC was evaluated using a xenograft mouse model. Subsequently, a comprehensive approach involving network pharmacology, serum metabolomics, and 16S rDNA sequencing was employed to investigate the anti-NSCLC mechanism of Fuzi. RESULTS: Pharmacological evaluation revealed significant tumour growth inhibition by Fuzi, accompanied by minimal toxicity. Network pharmacology identified 29 active Fuzi compounds influencing HIF-1, PI3K/Akt signalling, and central carbon metabolism in NSCLC. Integrating untargeted serum metabolomics highlighted 30 differential metabolites enriched in aminoacyl-tRNA biosynthesis, alanine, aspartate, and glutamate metabolism, and the tricarboxylic acid (TCA) cycle. Targeted serum metabolomics confirmed elevated glucose content and reduced levels of pyruvate, lactate, citrate, α-ketoglutarate, succinate, fumarate, and malate following Fuzi administration. Furthermore, 16S rDNA sequencing assay showed that Fuzi ameliorated the dysbiosis after tumorigenesis, decreased the abundance of Proteobacteria, and increased that of Firmicutes and Bacteriodetes. PICRUSt analysis revealed that Fuzi modulated the pentose phosphate pathway of the gut microbiota. Spearman correlation showed that Proteobacteria and Escherichia_Shigella accelerated the TCA cycle, whereas Bacteroidota, Bacteroides, and Lachnospiraceae_NK4A136_group suppressed the TCA cycle. CONCLUSIONS: This study firstly introduces a novel NSCLC mechanism involving Fuzi, encompassing energy metabolism and intestinal flora. It clarifies the pivotal role of the gut microbiota in treating NSCLC and modulating the TCA cycle. Moreover, these findings offer valuable insights for clinical practices and future research of Fuzi against NSCLC.

Radiomics for predicting survival in patients with locally advanced rectal cancer: a systematic review and meta-analysis.

Background: Radiomics has recently received considerable research attention for providing potential prognostic biomarkers for locally advanced rectal cancer (LARC). We aimed to comprehensively evaluate the methodological quality and prognostic prediction value of radiomic studies for predicting survival outcomes in patients with LARC. Methods: The Cochrane, Embase, Medline, and Web of Science databases were searched. The radiomics quality score (RQS), Transparent Reporting of a Multivariable Prediction Model for Individual Prognosis or Diagnosis (TRIPOD) checklist, the Image Biomarkers Standardization Initiative (IBSI) guideline, and the Prediction Model Risk of Bias Assessment Tool were used to assess the quality of the selected studies. A further meta-analysis of hazard ratio (HR) regarding disease-free survival (DFS) and overall survival (OS) was performed. Results: Among the 358 studies reported, 15 studies were selected for our review. The mean RQS score was 7.73±4.61 (21.5% of the ideal score of 36). The overall TRIPOD adherence rate was 64.4% (251/390). Most of the included studies (60%) were assessed as having a high risk of bias (ROB) overall. The pooled estimates of the HRs were 3.14 [95% confidence interval (CI): 2.12-4.64, P<0.01] for DFS and 3.36 (95% CI: 1.74-6.49, P<0.01) for OS. Conclusions: Radiomics has potential to noninvasively predict outcome in patients with LARC. However, the overall methodological quality of radiomics studies was low, and the adherence to the TRIPOD statement was moderate. Future radiomics research should put a greater focus on enhancing the methodological quality and considering the influence of higher-order features on reproducibility in radiomics.

Deoxyschizandrin ameliorates obesity and non-alcoholic fatty liver disease: Involvement of dual Farnesyl X receptor/G protein-coupled bile acid receptor 1 activation and leptin sensitization.

Natural dual farnesyl X receptor (FXR)/G protein-coupled bile acid receptor 1 (TGR5) activators have received little attention in the management of metabolic diseases. Deoxyschizandrin (DS), a natural lignan, occurs in S. chinensis fruit and has potent hepatoprotective effects, whereas its protective roles and mechanisms against obesity and non-alcoholic fatty liver disease (NAFLD) are largely elusive. Here, we identified DS as a dual FXR/TGR5 agonist using luciferase reporter and cyclic adenosine monophosphate (cAMP) assays. DS was orally or intracerebroventricularly administrated to high-fat diet-induced obesity (DIO) mice, and methionine and choline-deficient L-amino acid diet (MCD diet)-induced non-alcoholic steatohepatitis to evaluate its protective effects. Exogenous leptin treatment was employed to investigate the sensitization effect of DS on leptin. The molecular mechanism of DS was explored by Western blot, quantitative real-time PCR analysis, and ELISA. The results showed that DS activated FXR/TGR5 signaling and effectively reduced NAFLD in DIO and MCD diet-fed mice. DS countered obesity in DIO mice by promoting anorexia and energy expenditure and reversing leptin resistance, involving both peripheral and central TGR5 activation and leptin sensitization. Our findings indicate that DS may be a novel therapeutic approach for alleviating obesity and NAFLD through regulating FXR and TGR5 activities and leptin signaling.

An AS-qPCR-based method for the detection of Alzheimer's disease-related SNPs.

Alzheimer's disease (AD) is one of the most serious neurodegenerative diseases in the world and has a strong genetic predisposition. At present, there is still no effective method for the early diagnosis and prevention of AD. Accumulating evidence shows the association of several loci with AD risk, such as apolipoprotein E (APOE) and translocase of outer mitochondrial membrane 40 (TOMM40). However, for routine disease diagnosis in clinics, genotype detection methods based on gene sequencing technology are time-consuming and excessively costly. Thus, in this study, we developed a high-sensitivity, low-cost, and convenient single nucleotide polymorphism (SNP) detection assay method based on allele-specific quantitative polymerase chain reaction (AS-qPCR) technology, which can be used to determine the SNP genotype in APOE and TOMM40. A total of 40 patients were recruited from the outpatient department of the memory clinic of Dongzhimen Hospital, Beijing University of Chinese Medicine. The SNP detection assay method includes three steps. First, positive plasmids with different genotypes (TT/CC/TC) in APOE rs429358, rs7412, and TOMM40 rs11556505 were prepared. Second, 3'-T/3'-C primers were designed to amplify these positive plasmids for each SNP site. Finally, we calculated the log10 of the copy number ratio for each positive plasmid, and the genotype interpretation interval was established. Based on this method, we investigated whether the SNPs in 40 patients could be accurately calculated using AS-qPCR technology. The accuracy of SNP detection was verified by PCR-Pooling sequencing. The results showed that SNP genotypes assessed by AS-qPCR technology corresponded perfectly to the results obtained by conventional DNA sequencing. We have developed a genotype detection method for AD based on AS-qPCR, which can be performed easily, rapidly, accurately, and at low cost. The method will contribute to the early diagnosis of patients with late-onset Alzheimer's and the detection of large clinical samples in the future.

Aconiti Lateralis Radix Praeparata as Potential Anticancer Herb: Bioactive Compounds and Molecular Mechanisms.

Aconiti Lateralis Radix Praeparata (Fuzi in Chinese) is a traditional herbal medicine widely used in China and other Asian countries. In clinical practice, it is often used to treat heart failure, rheumatoid arthritis, and different kinds of pains. Fuzi extract and its active ingredients exert considerable anticancer, anti-inflammatory, and analgesic effects. The main chemical substances of Fuzi include alkaloids, polysaccharides, flavonoids, fatty acids, and sterols. Among of them, alkaloids and polysaccharides are responsible for the anticancer efficacy. Most bioactive alkaloids in Fuzi possess C19 diterpenoid mother nucleus and these natural products show great potential for cancer therapy. Moreover, polysaccharides exert extraordinary tumor-suppressive functions. This review comprehensively summarized the active ingredients, antineoplastic effects, and molecular mechanisms of Fuzi by searching PubMed, Web of Science, ScienceDirect, and CNKI. The anticancer effects are largely attributed to inducing apoptosis and autophagy, inhibiting proliferation, migration and invasion, regulating body immunity, affecting energy metabolism, as well as reversing multidrug resistance. Meanwhile, several signaling pathways and biological processes are mainly involved, such as NF-κB, EMT, HIF-1, p38 MAPK, PI3K/AKT/mTOR, and TCA cycle. Collectively, alkaloids and polysaccharides in Fuzi might serve as attractive therapeutic candidates for the development of anticancer drugs. This review would lay a foundation and provide a basis for further basic research and clinical application of Fuzi.

Quality Evaluation of Decoction Pieces of Gardeniae Fructus Based on Qualitative Analysis of the HPLC Fingerprint and Triple-Q-TOF-MS/MS Combined with Quantitative Analysis of 12 Representative Components.

In this study, quality evaluation (QE) of 40 batches of decoction pieces of Gardeniae Fructus (GF) produced by different manufacturers of herbal pieces was performed by qualitative analysis of the HPLC fingerprint and ultra-fast liquid chromatography (UFLC)-triple-Q-TOF-MS/MS combined with quantitative analysis of multiple components, which we established previously for QE of traditional medicine. First, HPLC fingerprints of 40 samples were determined, and the common peaks in the reference fingerprint were assigned. Second, the components of the common peaks in the HPLC fingerprints were identified by UFLC-triple-Q-TOF-MS/MS. Finally, the contents of the components confirmed by reference substances were measured. The results showed that there were 28 common peaks in the HPLC fingerprints of 40 samples. The components of these 28 common peaks were identified as 13 iridoids, 4 crocins, 7 monocyclic monoterpenoids, 3 organic acids, and 1 flavonoid. Of these, a total of 12 components, including 7 iridoids of geniposide, shanzhiside, geniposidic acid, deacetyl asperulosidic acid methyl ester, gardenoside, scandoside methyl ester, and genipin gentiobioside, 2 crocins such as crocin I and crocin II, 1 monocyclic monoterpenoid of jasminoside B, 1 organic acid of chlorogenic acid, and 1 flavonoid of rutin, were unambiguously identified by comparison with reference substances. There were certain differences in the contents of these 12 components among 40 samples. The geniposide content ranged from 37.917 to 72.216 mg/g, and the total content of the 7 iridoids ranged from 59.931 to 94.314 mg/g.

Atractylon, a novel dopamine 2 receptor agonist, ameliorates Parkinsonian like motor dysfunctions in MPTP-induced mice.

BACKGROUND: Motor symptoms of Parkinson's disease (PD) are characterized by bradykinesia, resting tremor, rigidity, slow movement, impaired gait and postural instability, resulting from progressive loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc). Atractylon is a natural furan compound in Atractylodes rhizomes, exhibiting anticancer, anti-inflammation, antiviral and gastroprotective activities, and so on. However, it is still unknown whether atractylon is beneficial to motor dysfunctions of PD. METHODS: GPCR-targeted piggyBac-TANGO compound screening system, cAMP assay, and immunostaining of p-CREB and BDNF were used to identify dopamine 2 receptor (DRD2) activation. The effects of atractylon on motor deficits and gait disturbances, as well as tyrosine hydroxylase (TH) in the SNpc were investigated in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice. RESULTS: Atractylon treatment increased the eGFP expression in dose-dependent manner in piggyBac-TANGO assay, decreased cAMP production, and enhanced the levels of p-CREB and BDNF in DRD2 highly expresseding SY-SY5Y cells. In MPTP-induced mice, atractylon improved the slow movement, diminished voluntary locomotion, and abnormal gait parameters, such as duration, cadence, average speed, step cycle, stride length, and so on. Moreover, atractylon rescued the TH positive cells in SNpc and TH positive nerve fibers in striatum. CONCLUSIONS: Atractylon could effectively activate DRD2, attenuate motor deficits and gait disorders, and protect dopaminergic neurons in MPTP-induced PD mice. Our findings stretch out the therapeutic potential of atractylon for motor symptoms of PD.

Berbamine reduces body weight via suppression of small GTPase Rab8a activity and activation of paraventricular hypothalamic neurons in obese mice.

Small GTPase Rab8a is involved in fat-specific protein 27 (Fsp27) mediated lipid droplet accumulation in adipocytes. By screening inhibitors of Rab8a GTPase from a natural compound library, berbamine (BBM), a marketing drug for treatment of leukopenia in China, was identified to inhibit the activity of Rab8a GTPase and block the differentiation of 3T3-L1 adipocytes. Animal study showed that BBM could reduce body weight, improved glucose and lipid metabolic homeostasis in high-fat diet-induced obesity (DIO) C57BL/6 mice and db/db mice. Additional, BBM increased energy expenditure and inhibited food intake in mice but not in lean mice. Moreover, intracerebroventricular injection (i.c.v.) of BBM inhibited feeding behavior and increased c-Fos expression in paraventricular nucleus of the hypothalamus (PVH) of mice. Our data suggest that BBM may improve obesity through the inhibition of Rab8a GTPase activity and the activation of anorexigenic energy-sensing neuron in PVH.

Nomilin and Its Analogues in Citrus Fruits: A Review of Its Health Promotion Effects and Potential Application in Medicine.

Nomilin is one of the major limonoids, which are plant secondary metabolites also known as tetranortriterpenoids. Nomilin is found mostly in common edible citrus fruits including lemons, limes, oranges, grapefruits, mandarins, along with traditional Chinese medicines derived from citrus fruits, such as tangerine seed, tangerine peel, fructus aurantii immaturus, etc. A number of studies have demonstrated that nomilin and its analogues exhibit a variety of biological and pharmacological activities. These include anti-cancer, immune-modulatory, anti-inflammatory, anti-obesity, anti-viral, anti-osteoclastogenic, anti-oxidant, and neuro-protective effects. Thus, nomilin and its analogues have emerged as a potential therapy for human diseases. The purpose of this review is to chronicle the evolution of nomilin research from examining its history, structure, occurrence, to its pharmacological and disease-preventing properties as well as its potential utilization in medicine and food science.

Screening and identification of Euphorbiae pekinensis Rupr. anti-angiogenic multi-components with UPLC-QTOF-MS in zebrafish.

Euphorbia pekinensis Rupr. (EP) (Euphorbiaceae), as Traditional Chinese Medicine (TCM), exhibits therapeutic effects on tumors in clinical practice. Anti-angiogenesis may be an underlying molecular mechanism of EP's actions. However, the anti-angiogenic active ingredients of EP remain unclear. The screening and analysis of anti-angiogenic agents were essential for the sufficient utilization and development of EP. Thus, we established a UPLC-QTOF-MS method based on a transgenic zebrafish model to screen anti-angiogenesis activity components in EP. UPLC-QTOF-MS was used to characterize compounds from EP and in vivo compounds in Tg (flk1: mCherry) zebrafish larvae treated with EP. Based on the identification results, five components were selected, and their anti-angiogenesis activity were investigated via assessment of intersegmental blood vessels during the development of the transgenic zebrafish. Three of these components (3,3'-O-dimethoxy ellagic acid, quercetin, and ingenol) are active components of EP with anti-angiogenic effects. Among them, 3, 3'-O-dimethoxy ellagic acid and ingenol were first demonstrated with anti-angiogenesis effects. UPLC-PDA analysis was performed on EP water extracts to determine anti-angiogenesis active ingredients quantitatively. In the concentration range of 100-200 µg/mL, EP and the active ingredient compositions, mixed according to the content of EP, had equivalent anti-angiogenesis activities. These experimental results indicate that the UPLC-QTOF-MS method, combined with a transgenic zebrafish model, is rapid, sensitive and reliable. The combination in TCM offers the potential to achieve certain effect levels with lower concentrations of the individual compound.

Characterization of the Therapeutic Effects of Novel Chimeric Antigen Receptor T Cells Targeting CD38 on Multiple Myeloma.

Multiple myeloma (MM) is a tumor type characterized by the unregulated proliferation of clonal plasma cells in the bone marrow. Immunotherapy based on chimeric antigen receptor T cell (CAR-T) therapy has achieved exciting success in the treatment of hematological malignant tumors. CD38 is highly and evenly expressed in MM and is an attractive target for MM treatment. Here, we successfully constructed two novel second-generation CAR-T cells targeting CD38 by retroviral vector transduction. CD38 CAR-T cells could be activated effectively after stimulation with CD38-positive tumor cells and secrete cytokines such as IFN-γ and TNF-α to promote tumor cell apoptosis in in vitro experiments. Real-time fluorescence monitoring experiments, luciferase detection experiments and flow cytometry experiments revealed the efficient and specific killing abilities of CD38 CAR-T cells against CD38-positive tumor cells. The proliferation ability of CD38 CAR-T cells in vitro was higher than that of untransduced T cells. Further antitumor experiments in vivo showed that CD38 CAR-T cells could be quickly activated to secrete IFN-γ and eliminate tumors. Thus, novel CD38-targeted second-generation CAR-T cells have efficient and specific antitumor activity and may become a novel therapy for the clinical treatment of MM.

High methane emissions from thermokarst lakes on the Tibetan Plateau are largely attributed to ebullition fluxes.

Ebullition has been shown to be an important pathway for methane (CH4) emissions from inland waters. However, the CH4 fluxes and their magnitudes in thermokarst lakes remain unclear due to limited research data, especially on the Tibetan Plateau (TP). The magnitude and regulation of two CH4 pathways, ebullition and diffusion, were investigated in 32 thermokarst lakes on the TP during the summer of 2020. CH4 emissions from thermokarst lakes on the TP showed significant spatiotemporal heterogeneity. Diffusion fluxes in lakes averaged 2.6 mmol m-2 d-1 (ranging from 0.003 to 48.4 mmol m-2 d-1), and ebullition fluxes in lakes averaged 6.6 mmol CH4 m-2 d-1 (ranging from 0.002 to 140.0 mmol m-2 d-1). Together, these ebullition fluxes contributed 66.1 ± 24.9% (ranging 5.4 to 100.0%) to the total (diffusion + ebullition) CH4 emissions, indicating the importance of ebullition as a major CH4 transport mechanism on the TP. In general, thermokarst lakes with higher CH4 diffusion fluxes and ebullition fluxes occurred in alpine meadows (2.5 ± 5.3 mmol m-2 d-1; 8.2 ± 20.6 mmol m-2 d-1), followed by alpine steppes (0.6 ± 5.3 mmol m-2 d-1; 0.7 ± 10.8 mmol m-2 d-1) and desert steppes (0.2 ± 0.2 mmol m-2 d-1; 0.6 ± 0.8 mmol m-2 d-1). The organic matter contents in water and sediment were found to be important factors influencing the seasonal variations in CH4 diffusion fluxes. However, the ebullition CH4 fluxes did not show a clear seasonal variation pattern. Our findings highlight the importance of considering the large spatiotemporal variations in ebullition CH4 fluxes to improve the accuracy of large-scale estimations of CH4 fluxes in thermokarst lakes on the TP. Greater insight into these aspects will increase the understanding of CH4 dynamics in thermokarst lakes on the TP, which is essential for forecasting and climate impact assessments and to better constrain feedback to climate warming.

Novel BCMA-OR-CD38 tandem-dual chimeric antigen receptor T cells robustly control multiple myeloma.

BCMA-targeting chimeric antigen receptor (CAR)-T cell therapy has shown remarkable clinical efficacy against multiple myeloma, yet antigen escape and tumor relapse still occur after the use of these therapies. Designing CAR-T therapies that targets multiple antigens simultaneously seems a feasible way to avoid antigen escape, and it has been extensively studied elsewhere. Here, we report novel BCMA-OR-CD38 Tan CAR T cells that can trigger robust cytotoxicity against target cells expressing either BCMA or CD38. We demonstrate that, in in vitro studies, these BCMA-OR-CD38 Tan CAR T cells exhibit similar CAR expression, superior cytotoxicity and antigen-stimulated T cell proliferation as compared to single-targeted CAR T cells or CD38-OR-BCMA Tan CAR T cells. Importantly, these BCMA-OR-CD38 Tan CAR-T cells can achieve complete tumor clearance in myeloma-bearing mice with no relapse observed through the course of these experiments. Finally, this BCMA-OR-CD38 Tan CAR was fully compatible with existing clinical grade T cell manufacturing procedures and can be implemented using current clinical protocols. Taken together, our results present an effective solution to the challenge of antigen escape in BCMA CAR T-cell therapies.

Co-Expression of miR155 or LSD1 shRNA Increases the Anti-Tumor Functions of CD19 CAR-T Cells.

Chimeric antigen receptor (CAR) T cells targeting CD19 antigen have produced remarkable clinical outcomes for cancer patients. However, identifying measures to enhance effector function remains one of the most challenging issues in CD19-targeted immunotherapy. Here, we report a novel approach in which a microRNA (miRNA) or short-hairpin RNA (shRNA) cassette was integrated into CAR-expressing retroviral vectors. Using this system, we generated anti-CD19 CAR-T cells co-expressing miR155 or LSD1 shRNA and found that anti-CD19 CAR-T cells with miR155 upregulation or LSD1 downregulation exhibited increased anti-tumor functions in vitro and in vivo. Transcriptional profiling analysis by RNA sequencing revealed the targets of miR155 and LSD1 in anti-CD19 CAR-T cells. Our experiments indicated that introduction of miRNA or shRNA expression into anti-CD19 CAR T-cells might be an effective strategy to improve the anti-tumor effects of CAR-T cell therapy.