Identifying biomarkers of ginseng medicines with different natures on heart failure.

ETHNOPHARMACOLOGICAL RELEVANCE: The nature of Chinese medicine is a unique index to measure its efficacy. Generally, treating the hot syndrome with cold nature medicine and vice versa. Ginseng medicines, a renowned Chinese medicine known for its qi tonifying action, encompasses various herbal materials such as ginseng, red ginseng, and black ginseng (GS, RG, and BG, respectively), ginseng leaves (GL), and American ginseng (AG), which exhibited different natures, thought contained similar ginsenosides. This traditional effect of GS and RG "reinvigorate the pulse for relieving qi depletion". It is closely linked to anti-heart failure (HF), HF is a clinical manifestation of deficiency of "heart-qi". However, the elucidation of the mechanism underlying the anti-HF effects of ginseng medicines with different natures remains a significant challenge. AIM OF THE STUDY: To elucidate pharmacological mechanisms underlying the effect of ginseng medicines on HF, and to identify biomarkers associated with their various natures. Furthermore, it provides the basis for the different applications of ginseng medicines with various natures. MATERIALS AND METHODS: This study established a rat model of HF induced by isoproterenol (ISO) combined with a specific diet. Four representative hot/cold herbs were selected as compared references for the medicine natures. The divergent effects of these herbs on the HF model were investigated by analyzing RNA-seq data to identify genes expressed differentially. Additionally, pathways associated with medicine natures were obtained using KEGG. Furthermore, UPLC-QqQ-MS/MS, as well as ELISA, were used to measure indexes associated with the nervous system, energy metabolisms, and endocrinology systems, such as BNP, CK, IL-1, T3, T4, cAMP, cGMP, AD, adrenal hormones (DOC, CORT, and COR), progestogens (pregnenolone, P, 17-OH-PR, and 17-OH-P), androgens (DHEA, A4, and T), and estrogens hormones (E2). RESULTS: All ginseng medicines demonstrated varying levels of efficacy in alleviating HF and GS exhibited a significant protective effect on HF. The ginseng medicines with qi tonifying primarily achieve their effect by enhancing the levels of adrenal hormones (DOC, CORT, and COR), T4, elevation of cAMP/cGMP, and activation of AchE. Warm nature qi tonifying ginseng medicines increased the levels of 17-OH-PR and P while decreasing 17-OH-P and the ratio of E2/T. On the other hand, cold nature qi tonifying ginseng medicines decreased the levels of A4 and T while increasing the ratio of E2/T. CONCLUSION: Overall, the effects of warm nature ginseng medicines are stronger on HF compared to cold nature ginseng medicines. Our research firstly reported that the E2/T ratio, progestogens (17-OH-PR, 17-OH-P, and P), and androgens (A4 and T) have been identified as significant biomarkers for discerning the mechanism differences of ginseng medicines with differences natures in treatment of HF.

The Impact of Doctor-Nurse Complementary Model Combined with Disease Tracking on Stroke Patients.

Objective: The incidence of stroke worldwide is increasing year by year. With the enhancement of public health awareness, people's demand for the quality of stroke rehabilitation is getting higher and higher, so better quality care measures are needed in the treatment of stroke. Based on this, this paper explores the impact of a new type of nursing care measure, the complementary health care model combined with condition tracking, on stroke patients. Methods: 238 stroke patients were randomly divided into a conventional group (n=119) and a combined group (n=119). 238 stroke patients were randomly divided into conventional group (n=119) and combined group (n=119). The conventional group received routine care, in which doctors and nursing carried out their own work without cooperation after the patients were admitted to the hospital; the combined group received a complementary health care model and condition tracking, in which doctors and nurses jointly checked the rooms, discussed cases, jointly formulated treatments and nursing care plans, and jointly formulated the patients' discharge and rehabilitation plans after the patients were admitted to the hospital. Before the intervention, at the time of discharge, and 6 months after discharge, the neurological function of the patients in both groups was assessed using the National Institutes of Health Stroke Scale (NIHSS) and the Fugl-Meyer (FMA) scale, the cognitive function of the patients in both groups was assessed using the Montreal Cognitive Assessment (MoCA) scale and the Measured Mental State Examination (MMSE), and the cognitive function of the patients in both groups was assessed using the General Self-Efficacy Scale (GSS) and the Montreal Cognitive Assessment (MCA) scale. General Self-Efficacy Scale (GSES) to assess self-efficacy, Exercise Adherence Questionnaire (EAQ) to assess adherence to functional exercise and Specific Quality of Life Scale (SSQoL-12) to assess the quality of life of patients in both groups, and the self-developed satisfaction with nursing care to assess patients' satisfaction with the care model. Results: Before the intervention, there was no difference in the National Institutes of Health Stroke Scale (NIHSS), the Fugl-Meyer Assessment (FMA), the Montreal Cognitive Assessment (MoCA), the Mental State Examination (MMSE), the General Self-Efficacy Scale (GSES), the Exercise Adherence Questionnaire (EAQ) and the Stroke-Specific Quality of Life Scale-12 (SSQoL-12) scores between the two groups (P > .05). At discharge and six months later, NIHSS scores continued to decrease in both groups, with the joint group being lower than the conventional group (P < .05); scores for all other items continued to increase, with the joint being higher than the conventional group (P < .05). Satisfaction with care was higher in the combined group than in the conventional group (P < .05). Conclusion: The complementary healthcare model combined with condition tracking can effectively promote the prognosis of rehabilitation of stroke patients, and has a positive effect in promoting the recovery of neurological and cognitive functions, strengthening self-efficacy, and improving the quality of life, which can be promoted in the clinic.

Pillararene-Based Stimuli-Responsive Supramolecular Delivery Systems for Cancer Therapy.

Cancer poses a significant challenge to global public health, seriously threatening human health and life. Although various therapeutic strategies, such as chemotherapy (CT), radiotherapy, phototherapy, and starvation therapy, are applied to cancer treatment, their limited therapeutic effect, severe side effects, and unsatisfactory drug release behavior need to be carefully considered. Thus, there is an urgent need to develop efficient drug delivery strategies for improving cancer treatment efficacy and realizing on-demand drug delivery. Notably, pillararenes, as an emerging class of supramolecular macrocycles, possess unique properties of highly tunable structures, superior host-guest chemistry, facile modification, and good biocompatibility, which are widely used in cancer therapy to achieve controllable drug release and reduce the toxic side effects on normal tissues under various internal/external stimuli conditions. This review summarizes the recent advance of stimuli-responsive supramolecular delivery systems (SDSs) based on pillararenes for tumor therapy from the perspectives of different assembly methods and hybrid materials, including molecular-scale SDSs, supramolecular nano self-assembly delivery systems, and nanohybrid SDSs. Moreover, the prospects and critical challenges of stimuli-responsive SDSs based on pillararenes for cancer therapy are also discussed.

Effects of repeated low-level red-light therapy on macular retinal thickness and microvascular system in children with myopia.

OBJECTIVE: The objective of the study was to use optical coherence tomography angiography (OCTA) to analyze the effects of repeated low-level red-light (LLLT) therapy on macular retinal thickness and the microvascular system in children with myopia to evaluate the safety of this therapy. METHODS: This prospective study included 40 school-age children with myopia (80 eyes), aged 7-14 years, who received therapy using a LLLT instrument. At baseline and therapy for 1 month, 3 months, 6 months, all children underwent comprehensive ophthalmological examinations, including slit-lamp examination, uncorrected visual acuity, best-corrected visual acuity, spherical equivalent degree, axial length, and OCTA. The vessel densities of the superficial retinal capillary plexus, macular inner retinal thickness, and full-layer retinal thickness were measured. RESULTS: The macular inner retinal thickness increased at 1 month and remained unchanged thereafter, It differed significantly in nine areas at 1, 3, and 6 months compared to the thicknesses before therapy (P < 0.05); however, we observed no significant differences between the different time points (P > 0.05). The macular full-layer retinal thickness increased at 1 month and remained unchanged thereafter; the changes showed significant differences at 1 month and 3 months compared to before therapy, for the inner nasal region (P < 0.05). The other eight areas showed significant differences at 1, 3, and 6 months compared with before therapy (P < 0.05); however, no significant difference was observed between the different time points after therapy (P > 0.05). The vessel density of the superficial retinal capillary plexus did not differ significantly among the four groups (P > 0.05). CONCLUSIONS: LLLT therapy was safe. The school-aged children exhibited macular thickening after LLLT therapy, which had no significant effect on macular microcirculation.

Chitosan entrapping of sodium alginate / Lycium barbarum polysaccharide gels for the encapsulation, protection and delivery of Lactiplantibacillus plantarum with enhanced viability.

To preserve the viability of probiotics during digestion and storage, encapsulation techniques are necessary to withstand the challenges posed by adverse environments. A core-shell structure has been developed to provide protection for probiotics. By utilizing sodium alginate (SA) / Lycium barbarum polysaccharide (LBP) as the core material and chitosan (CS) as the shell, the probiotic load reached 9.676 log CFU/mL. This formulation not only facilitated continuous release in the gastrointestinal tract but also enhanced thermal stability and storage stability. The results obtained from Fourier transform infrared spectroscopy and thermogravimetric analysis confirmed that the addition of LBP and CS affected the microstructure of the gel by enhancing the hydrogen bond force, so as to achieve controlled release. Following the digestion of the gel within the gastrointestinal tract, the released amount was determined to be 9.657 log CFU/mL. The moisture content and storage stability tests confirmed that the encapsulated Lactiplantibacillus plantarum maintained good activity for an extended period at 4 °C, with an encapsulated count of 8.469 log CFU/mL on the 28th day. In conclusion, the newly developed core-shell gel in this study exhibits excellent probiotic protection and delivery capabilities.

Colquhounia root tablet in the treatment of idiopathic membranous nephropathy with subnephrotic proteinuria.

PURPOSE: Idiopathic membranous nephropathy (IMN) is the most frequent global cause of nephrotic syndrome in non-diabetic people. In clinical practice, An effective and mild treatment for IMN patients with subnephrotic proteinuria has been adopted. Colquhounia root tablet (CRT) is a traditional Chinese medicine that is widely used in China to treat glomerulopathies. In this study, the effectiveness and safety of CRT in the treatment of IMN with subnephrotic proteinuria have been determined by reviewing the clinical records of 44 patients with IMN. METHODS: Retrospective analysis of IMN patients with subnephrotic proteinuria treated with CRT in combination with ACEI/ARB or ACEI/ARB alone. The remission rate (complete or partial remission) was the main outcome observed, and proteinuria, estimated glomerular filtration rate (eGFR), serum albumin levels, and adverse effects were the secondary outcomes. RESULTS: This clinical trial included 44 patients, and the overall remission rates at months 6, 9, and 12 after treatment were 68.2% versus 27.3% (p = 0.016), 72.7% versus 36.4% (p = 0.015), and 77.3% versus 36.4% (p = 0.006) in the treatment and control groups, respectively. The application of CRT treatment was an independent predictor of proteinuria remission (p = 0.024). In addition, in patients who were positive for phospholipase A2 receptor (PLA2R) antibodies, the overall remission rate was higher in the treatment group than in the control group after 9 months of treatment (75% versus 23.08%, p = 0.017). CONCLUSION: This retrospective study illustrates that, based on supportive therapy, CRT could be effective in the treatment of IMN with subnephrotic proteinuria with a good safety profile at the same time.

Integrated metabolomics and transcriptomic analysis reveals metabolic changes of flavor compounds of Camellia assamica host plant after parasitized by Viscumarticulatum.

Tea is one of the most popular beverages, it has many health benefits and flavor properties due to the presence of numerous secondary metabolites. Camellia assamica is also a main source of tea, which is mainly planted in the regions of southwest China. In this study, a non-targeted and targeted metabolomics analysis and sensory evaluation on tea leaves with and without mistletoe (Viscum articulatum) was carried out using liquid chromatography-mass spectrometry. RNA-seq-based transcriptomic analysis was conducted in parallel on the same samples, subsequently gene expression and metabolic differentiation were also investigated. Tea leaves with mistletoe presented much lower contents of (-)-catechin, (-)-epicatechin, (-)-gallocatechin gallate and (-)-epicatechin gallate, but significantly higher levels of free amino acids including Arg, Asp, GABA and Gln than that without mistletoe. Transcriptomic analysis also confirmed the main differentially expressed genes (DEGs) containing phenylpropanoid and flavonoid biosynthesis were down-regulated, but genes of amino acid biosynthesis were up-regulated. qRT-PCR analysis further revealed that the relative expression of CsCHS, CsC4H, CsANS, CsLAR, and CsF3H was hindered, while CsglyA and CsilvE expression was increased.

CsRVE1 promotes seasonal greening of albino Camellia sinensis cv. Huangkui by activating chlorophyll biosynthesis.

Seasonal greening is a crucial survival strategy for albino tea cultivars, during which dysfunctional chloroplasts recover and chlorophyll biosynthesis increases in albino leaves. However, the regulatory mechanisms of seasonal greening in albino tea plants remain unclear. Here, we report that CsRVE1, a nuclear-located Myb-like transcription factor, can positively modulate the seasonal greening of albino Camellia sinensis cv. Huangkui leaves by activating the expression of genes involved in light harvesting and chlorophyll biosynthesis. The transcriptional expression of CsRVE1 increased during seasonal greening and was tightly correlated with increases in the expression of genes involved in light harvesting (CsLhcb) and chlorophyll biosynthesis (CsCHLH, CsHEMA1 and CsCAO). In vivo and in vitro molecular analyses showed that CsRVE1 can directly bind to the promoters of CsLhcb, CsCHLH and CsPORA, eventually leading to chlorophyll accumulation in tea leaves. Furthermore, transient suppression of CsRVE1 in tea leaves led to a decrease in target gene expression. In contrast, the overexpression of CsRVE1 in Arabidopsis led to chlorophyll increases and the activation of AtLhcb, AtPORA, AtCHLH, etc. These results identify CsRVE1 as an important promoter of seasonal greening that functions by regulating genes involved in chlorophyll biosynthesis in albino tea plants and shed new light on the regulatory mechanisms of leaf phenotypes in plants.

Role of Endophytic Bacteria in the Remobilization of Leaf Nitrogen Mediated by CsEGGT in Tea Plants (Camellia sinensis L.).

As an important economic plant, tea (Camellia sinensis) has a good economic value and significant health effects. Theanine is an important nitrogen reservoir, and its synthesis and degradation are considered important for nitrogen storage and remobilization in tea plants. Our previous research indicated that the endophyte CsE7 participates in the synthesis of theanine in tea plants. Here, the tracking test confirmed that CsE7 tended to be exposed to mild light and preferentially colonized mature tea leaves. CsE7 also participated in glutamine, theanine, and glutamic acid circulatory metabolism (Gln-Thea-Glu) and contributed to nitrogen remobilization, mediated by the γ-glutamyl-transpeptidase (CsEGGT) with hydrolase preference. The reisolation and inoculation of endophytes further verified their role in accelerating the remobilization of nitrogen, especially in the reuse of theanine and glutamine. This is the first report about the photoregulated endophytic colonization and the positive effect of endophytes on tea plants mediated and characterized by promoting leaf nitrogen remobilization.

γ-Glutamyl-transpeptidase CsGGT2 functions as light-activated theanine hydrolase in tea plant (Camellia sinensis L.).

Theanine is an important secondary metabolite endowing tea with umami taste and health effects. It is essential to explore the metabolic pathway and regulatory mechanism of theanine to improve tea quality. Here, we demonstrated that the expression patterns of CsGGT2 (γ-glutamyl-transpeptidase), participated in theanine synthesis in vitro in our previous research, are significantly different in the aboveground and underground tissues of tea plants and regulated by light. Light up-regulated the expression of CsHY5, directly binding to the promoter of CsGGT2 and acting as an activator of CsGGT2, with a negative correlation with theanine accumulation. The enzyme activity assays and transient expression in Nicotiana benthamiana showed that CsGGT2, acting as bifunctional protein, synthesize and degrade theanine in vitro and in planta. The results of enzyme kinetics, Surface plasmon resonance (SPR) assays and targeted gene-silencing assays showed that CsGGT2 had a higher substrate affinity of theanine than that of ethylamine, and performed a higher theanine degradation catalytic efficiency. Therefore, light mediates the degradation of theanine in different tissues by regulating the expression of the theanine hydrolase CsGGT2 in tea plants, and these results provide new insights into the degradation of theanine mediated by light in tea plants.

UV-B regulates seasonal greening of albino leaves by modulating CsHY5-inhibiting chlorophyll biosynthesis in Camellia sinensis cv. Huangkui.

Seasonal greening is crucial for albino plants but the underlying regulatory mechanism is unclear, especially concerning light regulation as one of the most important environmental factors for light-sensitive albino tea plants. Here, we report that the UV-B signal regulates the seasonal greening process of albino leaves by modulating CsHY5-inhibiting chlorophyll biosynthesis in Camellia sinensis cv. Huangkui. Reduction of solar UV-B in plantation promoted the seasonal greening of albino 'HK' leaves by inhibiting CsHY5 transcription and activating genes involved in light-harvesting CsLhlb and the chlorophyll biosynthetic pathway (CsCHLH, CsHEMA1, and CsPORA), leading to enrichment of chlorophyll accumulation and recovery of dysfunctional chloroplasts. In contrast, indoor supplementary UV-B exposure reduced chlorophylls by activating CsHY5 but inhibiting chlorophyll biosynthetic genes. In vivo and in vitro molecular analyses showed that CsHY5 can directly bind to the promoters of CsLhlb, CsCHLH, CsHEMA1, and CsPORA. These results indicate that CsHY5 acts as a repressor for the seasonal greening of the albino tea plants in response to the UV-B signal. This is the first study that investigates the regulatory role of the CsHY5-mediated UV-B signal in regulating the seasonal greening of the albino tea plant, which improves our understanding of light regulation in leaf phenotypes of higher plants.

The Effect of Brief Mindfulness Meditation on Suicidal Ideation, Stress and Sleep Quality.

OBJECTIVES: Suicide is the fourth leading cause of death for individuals aged 15-29 years, and early intervention on suicidal ideation and risk factors should be priortized. Brief mindfulness meditation (BMM) is convenient and cost-effective in improving physical and mental well-being, but less is known about its efficacy for suicidal ideation, stress and sleep quality. We investigated the effects of BMM on suicidal ideation, stress, and sleep quality for individuals with suicide risk. METHODS: Sixty-four college students with high suicidal ideation (aged 18-30 years) were randomly allocated to either a BMM (n = 32) or control group (n = 32). The BMM was based on Anapanasati and core mindfulness concepts. Sixty participants completed all scheduled sessions including pretest, one month of intervention or waiting, and posttest. Suicidal ideation was measured with the Beck Scale for Suicidal Ideation. Stress was evaluated using the Perceived Stress Scale and salivary cortisol levels. Sleep was measured using the Pittsburgh Sleep Quality Index and actigraphy accompanied with 7-day sleep diaries. RESULTS: Post-intervention, the BMM group showed significant decrease in suicidal ideation with a large effect size; the decrease showed a medium effect size in the control group. The BMM group, but not the control group, showed significant decrease in morning salivary cortisol and sleep latency, and improved sleep efficiency. CONCLUSIONS: BMM could help reduce suicidal ideation, stress, and sleep disturbance for individuals with high suicidal ideation and it may implicate effective suicide prevention strategy.

Duration and Determinants of Delayed Diagnosis with Tuberculosis in Shenzhen, China: A Cross-Sectional Study.

Background: Early diagnosis and timely treatment of tuberculosis are critical for disease control and management. However, diagnostic delay remains severe around the world. We aim to evaluate the duration and factors associated with diagnostic delay of tuberculosis in Shenzhen, China. Methods: We conducted a face-to-face interview to collect the whole care-seeking process of patients diagnosed with active TB in Shenzhen, China, from April 1 to September 30, 2021. The duration from symptom onset to confirmed diagnosis was recorded. The risk factors of diagnostic delay were identified by binary stepwise logistic regression analysis. Results: Among 288 confirmed TB cases, 170 (59.0%) were delayed diagnosis. The median diagnostic delay was 39.5 days. Median patient delay was 23 days and health system delay was 7 days. Income ≤315USD/month (OR = 2.97 [95% CI: 1.15-7.69]), cough (OR = 3.00 [95% CI: 1.16-7.76]), weight loss (OR = 15.59 [95% CI: 1.85-131.56]), use of traditional Chinese Medicine (OR = 5.03 [95% CI: 1.04-24.31]) and over-the-counter cough syrup (OR = 2.73 [95% CI: 1.10-6.76]) were significant risk factors for patient delay. Fever (OR = 0.13[95% CI: 0.04-0.48]) and hemoptysis (OR = 0.06 [95% CI0.01-0.30]) were protective factors for patient delay. Cough (OR = 2.85 [95% CI: 1.49-5.49]) and availability of chest X-ray (OR = 0.21[CI: 0.11-0.39]) were factors associated with health system delay. Conclusion: Delayed diagnosis of tuberculosis remains an unresolved problem. Patients with low income, self-treatment with over-the-counter medicine and accepting TCM suffered from a higher risk of patient delay. It is important to give more help to the vulnerable people and strengthen tuberculosis knowledge among primary health providers. Keeping all health providers alert to TB symptoms can facilitate earlier TB diagnosis and better disease control.

Comparative Study on Medicinal Natures (qi) of Black Ginseng, Red Ginseng, and Ginseng Leaves Based on Typical Deficiency-Heat Syndrome Rat Model.

To elucidate the medicinal nature of black ginseng (BG) by comparison of the effects of four Chinese herbs with different medicinal natures on the deficiency-heat syndrome rat model which was established by intragastric administration of traditional Chinese drugs with hot nature, the appearance indexes, biochemical indexes, and pathological sections of thyroid and stomach were examined. In addition, the seven short-chain fatty acids (SCFAs) in rat feces were also determined by headspace gas chromatography-mass spectrometry to reveal the action mechanism of the drugs with different natures. Results indicated that all the 4 drugs could exhibit similar actions in regulating the biochemical indexes of triiodothyronine (T3), thyroxine (T4), thyrotropin-releasing hormone (TRH), thyroid-stimulating hormone (TSH), and corticosterone (CORT) representing the hypothalamus-pituitary-thyroid (HPT) and hypothalamus-pituitary-adrenal (HPA) axes of the animal. However, cold-natured cortex phellodendri (HB) and ginseng leaves (GLs) showed stronger downregulation of the AChE activity of the nervous system. Red ginseng (RG) and BG tested exhibited stronger upregulation of the liver Na+-K+-ATPase activity. Principal component analysis (PCA) showed that GLs are similar to those of HB which belongs to the cold-nature drug, whereas BG showed closer to RG which attributes to a warm-nature drug. Thus, BG could be ascribed to a warm-nature drug. Further research disclosed that RG and BG mainly regulated the acetic acid and GL and HB primarily modulated the isovaleric acid and hexanoic acid in rat feces, which could be the features of drugs with warm or cold nature on the regulation of SCFAs in rats. It is for the first time that the medicinal nature of BG and its effect on the SCFAs were examined.

Preparation and related properties of melanin iron supplement.

In this study, BM-Fe (black sesame melanin-iron complex) was prepared and characterized. The results showed that the carboxyl hydroxyl group of BSM (black sesame melanin) participated in the chelation of iron ions. EDS (energy dispersive spectroscopy) and XPS (X-ray photoelectron spectroscopy) results confirmed the presence of iron ions in BM-Fe. The results of DLS (dynamic light scattering) showed that the average particle sizes of BSM and BM-Fe were 844.9 nm and 294.3 nm, respectively, indicating that the structure of BM-Fe with a smaller particle size was formed after the binding of iron ions with the active group on BSM. Finally, the in vitro iron dissolution, iron ion identification, in vitro iron ion reduction, antioxidant activity, cytotoxicity and moisture resistance properties of BM-Fe and FST (ferrous sulfate tablets, a commonly used iron supplement) were comprehensively compared. The results showed that BSM combined with iron instead of physically mixing, and BM-Fe was easily reduced in the gastrointestinal environment. BM-Fe had good bioavailability and retained the excellent characteristics (such as oxidation resistance and biocompatibility) of BSM, and had the potential to be applied in the treatment of iron-deficiency-related diseases. In summary, BM-Fe prepared in this study not only retained the excellent characteristics of BSM but also had a good effect on iron supplementation, high bioavailability and low side effects. Comprehensive analysis showed that the performance of BM-Fe prepared in this study was similar to or even better than that of the control (FST). Thus, BM-Fe is expected to become a new comprehensive multi-functional iron supplement and has a broad developmental prospect.

Characterization of CsTSI in the Biosynthesis of Theanine in Tea Plants (Camellia sinensis).

Theanine is a unique major amino acid in tea plants responsible for umami taste and mental health benefits of tea. However, theanine biosynthesis and physiological role in tea plants are not fully understood. Here, we demonstrate that tea plant theanine synthetase is encoded by a glutamine synthetase gene CsTSI. The expression pattern of CsTSI is closely correlated with theanine and glutamine levels in various tissues. CsTSI transcripts were accumulated in root tip epidermal cells, pericycle and procambial cells, where CsTSI presents as a cytosolic protein. Ectopic expression of the gene in Arabidopsis led to greater glutamine and theanine production than controls when fed with ethylamine (EA). RNAi knockdown or overexpression of CsTSI in tea plant hairy roots reduced or enhanced theanine and glutamine contents, respectively, compared with controls. The CsTSI recombinant enzymes used glutamate as an acceptor and ammonium or EA as a donor to synthesize glutamine and theanine, respectively. CsTSI expression in tea roots responded to nitrogen supply and deprivation and was correlated with theanine contents. This study provides fresh insights into the molecular basis for the biosynthesis of theanine, which may facilitate the breeding of high-theanine tea plants for improving the nutritional benefit of tea.

A 3,000-year-old, basal S. enterica lineage from Bronze Age Xinjiang suggests spread along the Proto-Silk Road.

Salmonella enterica (S. enterica) has infected humans for a long time, but its evolutionary history and geographic spread across Eurasia is still poorly understood. Here, we screened for pathogen DNA in 14 ancient individuals from the Bronze Age Quanergou cemetery (XBQ), Xinjiang, China. In 6 individuals we detected S. enterica. We reconstructed S. enterica genomes from those individuals, which form a previously undetected phylogenetic branch basal to Paratyphi C, Typhisuis and Choleraesuis-the so-called Para C lineage. Based on pseudogene frequency, our analysis suggests that the ancient S. enterica strains were not host adapted. One genome, however, harbors the Salmonella pathogenicity island 7 (SPI-7), which is thought to be involved in (para)typhoid disease in humans. This offers first evidence that SPI-7 was acquired prior to the emergence of human-adapted Paratyphi C around 1,000 years ago. Altogether, our results show that Salmonella enterica infected humans in Eastern Eurasia at least 3,000 years ago, and provide the first ancient DNA evidence for the spread of a pathogen along the Proto-Silk Road.

Ambient Ultraviolet B Signal Modulates Tea Flavor Characteristics via Shifting a Metabolic Flux in Flavonoid Biosynthesis.

Tea leaves contain an extraordinarily high level of flavonoids that contribute to tea health benefits and flavor characteristics, but the regulatory mechanism of ambient ultraviolet B (UV-B) on tea flavonoid enrichment remains unclear. Here, we report that ambient UV-B modulates tea quality by inducing a metabolic flux in flavonoid biosynthesis. UV-B absence decreased bitter- and astringent-tasting flavonol glycosides (kaempferol-7-O-glucoside, myricetin-3-O-glucoside, and quercetin-7-O-glucoside) but increased non-galloylated catechins. Conversely, supplementary UV-B increased flavonols and decreased catechins in tea leaves. These responses were achieved via CsHY5, which mediates the UV-B-induced MYB12 activation and binds to the promoters of flavonoid biosynthetic genes (CsFLS, CsLARa, and CsDFRa), leading to flavonoid changes. Transcriptomic data indicated that UV-B-induced tea flavonoid regulation is responsive to multiple biotic and abiotic environmental stresses. These findings improve our understanding of light-regulated tea astringency and bitterness underlying shading effects and seasonal light changes and provide novel insights into tea cultivation management and processing.

Strategies for Remodeling the Tumor Microenvironment Using Active Ingredients of Ginseng-A Promising Approach for Cancer Therapy.

The tumor microenvironment (TME) plays a key role in promoting the initiation and progression of tumors, leading to chemoradiotherapy resistance and immunotherapy failure. Targeting of the TME is a novel anti-tumor therapeutic approach and is currently a focus of anti-tumor research. Panax ginseng C. A. Meyer (ginseng), an ingredient of well-known traditional Asia medicines, exerts beneficial anti-tumor effects and can regulate the TME. Here, we present a systematic review that describes the current status of research efforts to elucidate the functions and mechanisms of ginseng active components (including ginsenosides and ginseng polysaccharides) for achieving TME regulation. Ginsenosides have variety effects on TME, such as Rg3, Rd and Rk3 can inhibit tumor angiogenesis; Rg3, Rh2 and M4 can regulate the function of immune cells; Rg3, Rd and Rg5 can restrain the stemness of cancer stem cells. Ginseng polysaccharides (such as red ginseng acidic polysaccharides and polysaccharides extracted from ginseng berry and ginseng leaves) can regulate TME mainly by stimulating immune cells. In addition, we propose a potential mechanistic link between ginseng-associated restoration of gut microbiota and the tumor immune microenvironment. Finally, we describe recent advances for improving ginseng efficacy, including the development of a nano-drug delivery system. Taken together, this review provides novel perspectives on potential applications for ginseng active ingredients as anti-cancer adjuvants that achieve anti-cancer effects by reshaping the tumor microenvironment.

Identification of a Novel c-Myc Inhibitor 7594-0037 by Structure-Based Virtual Screening and Investigation of Its Anti-Cancer Effect on Multiple Myeloma.

INTRODUCTION: Multiple myeloma (MM) is an extremely malignant and incurable hematological cancer. Increased expression of the c-Myc oncoprotein is closely associated with shorter overall survival of MM patients, implying that c-Myc is a potential therapeutic target. MAIN METHODS: We identified a potential c-Myc inhibitor 7594-0037 by structure-based virtual screening from the ChemDiv database. CCK8 assay and flow cytometry were used to detect MM cell viability, cell cycle and apoptosis. Q-PCR and Western blot were used to measure corresponding mRNA and protein expression levels. Protein stability assay measured the stability of c-Myc. RESULTS: Compound 7594-0037 exhibited stronger anti-proliferative activity against MM cells, and induced MM cell cycle G2 phase arrest and apoptosis. More importantly, compound 7594-0037 overcame myeloma resistance to bortezomib and exhibited a synergistic effect with bortezomib, resulting in increased MM cell death. The mechanism consists of compound 7594-0037 facilitating c-Myc protein degradation via decreasing the c-Myc S62 phosphorylation levels mediated by PIM1 kinase. Molecular dynamics simulation with the c-Myc/7594-0037 complex showed that compound 7594-0037 bound tightly to the N-terminus of c-Myc, and blocked the binding interaction of the two termini of c-Myc, which resulted in c-Myc entering into an unstable state. CONCLUSION: Overall, our study provides preliminary data for compound 7594-0037, which can be used as a novel c-Myc inhibitor and is a potential candidate therapeutic drug for multiple myeloma.