Recent progress on the application of compound formulas of traditional Chinese medicine in clinical trials and basic research in vivo for chronic liver disease.

ETHNOPHARMACOLOGICAL RELEVANCE: Chronic liver diseases mainly include chronic viral liver disease, metabolic liver disease, cholestatic liver disease (CLD), autoimmune liver disease, and liver fibrosis or cirrhosis. Notably, the compound formulas of traditional Chinese medicine (TCM) is effective for chronic liver diseases in clinical trials and basic research in vivo, which provide evidence of chronic liver disease treatment with integrated TCM and traditional Western medicine. AIM OF THE REVIEW: This paper aims to provide a comprehensive review of the compound formulas of TCM for treating different chronic liver diseases to elucidate the composition, main curative effects, and mechanisms of these formulas and research methods. MATERIALS AND METHODS: Different keywords related to chronic liver diseases and keywords related to the compound formulas of TCM were used to search the literature. PubMed, Scopus, Web of Science, and CNKI were searched to screen out original articles about the compound formulas of TCM related to the treatment of chronic liver diseases, mainly including clinical trials and basic in vivo research related to Chinese patent drugs, classic prescriptions, proven prescriptions, and hospital preparations. We excluded review articles, meta-analysis articles, in vitro experiments, articles about TCM monomers, articles about single-medicine extracts, and articles with incomplete or uncertain description of prescription composition. Plant names were checked with MPNS (http://mpns.kew.org). RESULTS: In this review, the clinical efficacy and mechanism of compound formulas of TCM were summarized for the treatment of chronic viral hepatitis, nonalcoholic fatty liver disease, CLD, and liver fibrosis or cirrhosis developed from these diseases and other chronic liver diseases. For each clinical trial and basic research in vivo, this review provides a detailed record of the specific composition of the compound formulas of TCM, type of clinical research, modeling method of animal experiments, grouping methods, medication administration, main efficacy, and mechanisms. CONCLUSION: The general development process of chronic liver disease can be summarized as chronic hepatitis, liver fibrosis or cirrhosis, and hepatocellular carcinoma. The compound formulas of TCM have some applications in these stages of chronic liver diseases. Owing to the continuous progress of medical technology, the benefits of the compound formulas of TCM in the treatment of chronic liver diseases are constantly changing and developing.

Obesity in children and adolescents and the risk of ovarian cancer: A systematic review and dose‒response meta-analysis.

OBJECTIVE: The relationship between obesity in children and adolescents and the risk of ovarian cancer remains controversial. The aim of this meta-analysis was to explore the exact shape of this relationship. METHODS: We conducted dose‒response meta-analyses of cohort and case‒control studies, including published studies derived from searches in the PubMed, Embase, Web of Science and Cochrane Library databases until October 2022. Pooled effect size estimates are expressed as relative risks (RRs) or odds ratios (ORs) with 95% confidence intervals (CIs) and were evaluated by fixed-effect models. A nonlinear dose‒response meta-analysis was performed by using a restricted cubic spline model. RESULTS: After screening 4215 publications, 10 studies were included in the present meta-analysis. Overall analyses revealed statistically significant associations of obesity in children and adolescents with ovarian cancer (adjusted RR = 1.19, 95% CI: 1.11 to 1.28, P < 0.001). Moreover, the association was consistently significant in most subgroup analyses, for example, using geographic stratification, the results remained stable both in the Americas(RR = 1.11; 95% CI: 1.01 to 1.21; P = 0.022) and Europe (RR = 1.46; 95% CI: 1.21 to 1.77; P<0.001). For the dose‒response analyses, the risk of ovarian cancer increased with the degree of obesity, and the trend increased rapidly when body mass index (BMI) was over 25.95 kg/m2. CONCLUSION: Our findings indicate that obesity in children and adolescents is a risk factor for ovarian cancer, and the risk increases with increasing BMI.

Malate-mediated CqMADS68 enhances aluminum tolerance in quinoa seedlings through interaction with CqSTOP6, CqALMT6 and CqWRKY88.

Aluminum (Al) stress in acidic soils has severe negative effects on crop productivity. In this study, the alleviating effect and related mechanism of malate on Al stress in quinoa (Chenopodium quinoa) seedlings were investigated. The findings indicated that malate alleviated the growth inhibition of quinoa seedlings under Al stress, maintained the enzymatic and nonenzymatic antioxidant systems, and aided resistance to the damage caused by excessive reactive oxygen species (ROS). Under Al stress, malate significantly increased the contents of chlorophyll and carotenoids in quinoa shoots by 103.8% and 240.7%, and significantly increased the ratios of glutathione (GSH)/oxidized glutathione (GSSG), and ascorbate (AsA)/dehydroascorbate (DHA) in roots by 59.9% and 699.2%, respectively. However, malate significantly decreased the superoxide radical (O2•-), hydrogen peroxide (H2O2), malondialdehyde (MDA) and Al contents in quinoa roots under Al stress by 32.7%, 60.9%, 63.1% and 49%, respectively. Moreover, the CqMADS family and the Al stress-responsive gene families (CqSTOP, CqALMT, and CqWRKY) were identified from the quinoa genome. Comprehensive expression profiling identified CqMADS68 as being involved in malate-mediated Al resistance. Transient overexpression of CqMADS68 increased Al tolerance in quinoa seedlings. More importantly, we found that CqMADS68 regulated the expression of CqSTOP6, CqALMT6 and CqWRKY88 and further demonstrated the interaction of CqMADS68 with CqSTOP6, CqALMT6 and CqWRKY88 by bimolecular fluorescence complementation (BIFC) experiments. Moreover, transient overexpression and physiological and biochemical analyses demonstrated that CqSTOP6, CqALMT6 and CqWRKY88 could also improve Al tolerance by maintaining the antioxidant capacity of quinoa seedlings. Taken together, these findings reveal that CqMADS68, CqSTOP6, CqALMT6 and CqWRKY88 may be important contributors to the Al tolerance regulatory network in quinoa, providing new insights into Al stress resistance.

The Relationship Between Plant-Based Diet and Risk of Digestive System Cancers: A Meta-Analysis Based on 3,059,009 Subjects.

Background and Objectives: Diets containing red or processed meat are associated with a growing risk of digestive system cancers. Whether a plant-based diet is protective against cancer needs a high level of statistical evidence. Methods: We performed a meta-analysis of five English databases, including PubMed, Medline, Embase, Web of Science databases, and Scopus, on October 24, 2021 to identify published papers. Cohort studies or case-control studies that reported a relationship between plant-based diets and cancers of the digestive system were included. Summary effect-size estimates are expressed as Risk ratios (RRs) or Odds ratios (ORs) with 95% confidence intervals and were evaluated using random-effect models. The inconsistency index (I2) and τ2 (Tau2) index were used to quantify the magnitude of heterogeneity derived from the random-effects Mantel-Haenszel model. Results: The same results were found in cohort (adjusted RR = 0.82, 95% CI: 0.78-0.86, P < 0.001, I2 = 46.4%, Tau2 = 0.017) and case-control (adjusted OR = 0.70, 95% CI: 0.64-0.77, P < 0.001, I2 = 83.8%, Tau2 = 0.160) studies. The overall analysis concluded that plant-based diets played a protective role in the risk of digestive system neoplasms. Subgroup analyses demonstrated that the plant-based diets reduced the risk of cancers, especially pancreatic (adjusted RR = 0.71, 95% CI: 0.59-0.86, P < 0.001, I2 = 55.1%, Tau2 = 0.028), colorectal (adjusted RR = 0.76, 95% CI: 0.69-0.83, P < 0.001, I2 = 53.4%, Tau2 = 0.023), rectal (adjusted RR = 0.84, 95% CI: 0.78-0.91, P < 0.001, I2 = 1.6%, Tau2 = 0.005) and colon (adjusted RR = 0.88, 95% CI: 0.82-0.95, P < 0.001, I2 = 0.0%, Tau2 = 0.000) cancers, in cohort studies. The correlation between vegan and other plant-based diets was compared using Z-tests, and the results showed no difference. Conclusions: Plant-based diets were protective against cancers of the digestive system, with no significant differences between different types of cancer. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42022322276, Identifier: CRD42022322276.

Mechanisms of calcium sulfate in alleviating cadmium toxicity and accumulation in pak choi seedlings.

Gypsum (calcium sulfate dihydrate, CaSO4 ·2H2O) is commonly applied to improve soil quality and nutrient supply. Previous studies also suggested it is a cost-effective soil amendment in alleviating cadmium (Cd) toxicity and accumulation in plants. The aim of this study was to investigate how this is achieved. We used pak choi as our research material because it is a popular vegetable in Asia, and as a leafy vegetable, it accumulates higher Cd level than other types of vegetable. Under Cd stress, application of CaSO4 promoted pak choi seedling growth, decreased the oxidative stress in roots, reduced Cd accumulation, and enhanced the photosynthesis in shoots. We revealed the inhibition of Cd2+ absorption by CaSO4 is largely due to the competition between Ca2+ and Cd2+ for ion channels or transporter. Moreover, under Cd stress, CaSO4 facilitated the sulphate assimilation, increased the biosynthesis of phytochelatins, and activated the expression of transporters for vacuolar sequestration. Together, CaSO4 could benefit plant growth and enhance Cd tolerance by suppressing Cd root uptake and lowering the Cd content in cytoplasm.

Plasma membrane-localized protein BcHIPP16 promotes the uptake of copper and cadmium in planta.

Cadmium (Cd) is one of the toxic heavy metals in soil, which not only suppresses crop production but also threatens human health. In this study, we aim to clarify the biological function of Cd-related gene BcHIPP16, so as to provide potential genetic solutions to decrease the Cd levels of pak choi. Tissue expression analysis showed that BcHIPP16 expressed in almost all the plant bodies. The transcriptional level of BcHIPP16 in roots was higher than that in shoots, which was significantly induced by copper (Cu) deficiency and Cd exposure conditions. Subcellular localization revealed that BcHIPP16 localized in plasma membrane. Expressing BcHIPP16 in yeast cells improved the sensitivity to Cu and Cd and improved their accumulation in yeast. Furthermore, the Cu and Cd content of Arabidopsis seedlings were increased and complemented, respectively when expressing BcHIPP16 in wild type (WT) and hip16 mutants. Non-invasive Micro-test Technology (NMT) was used to measure the real-time Cd2+ influx from the root surface of BcHIPP16 transgenic Arabidopsis lines, and the result demonstrated that BcHIPP16 promoted Cd2+ influx into Arabidopsis root cells. Taken together, our study showed that BcHIPP16 contributed to absorbing nutrient metal Cu and heavy metal Cd in planta.

Involvement of several putative transporters of different families in ß-cyclocitral-induced alleviation of cadmium toxicity in quinoa (Chenopodium quinoa) seedlings.

Cadmium (Cd) has a serious negative impact on crop growth and human food security. This study investigated the alleviating effect of ß-cyclocitral, a potential heavy metal barrier, on Cd stress in quinoa seedlings and the associated mechanisms. Our results showed that ß-cyclocitral alleviated Cd stress-induced growth inhibition in quinoa seedlings and promoted quinoa seedling root development under Cd stress. Moreover, it maintained the antioxidant system of quinoa seedlings, including the enzymatic, i.e., superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), and nonenzymatic, i.e., reduced glutathione (GSH) and ascorbic acid (ASA), antioxidants, which eliminate the damage from excessive reactive oxygen species (ROS). Our results showed that ß-cyclocitral could reduce the amount of Cd absorbed by roots. Furthermore, we systematically identified five transporter families from the quinoa genome, and the RT-qPCR results showed that ZIP, Nramp and YSL gene families were downregulated by ß-cyclocitral to reduce Cd uptake by roots. Thus, ß-cyclocitral promoted the growth, photosynthetic capacity and antioxidant capacity of the aboveground parts of quinoa seedlings. Taken together, these results suggested that the ß-cyclocitral-induced decrease in Cd uptake may be caused by the downregulation of several selected transporter genes.

Ferrous iron-induced increases in capitate glandular trichome density and upregulation of CbHO-1 contributes to increases in blinin content in Conyza blinii.

MAIN CONCLUSION: Ferrous iron can promote the development of glandular trichomes and increase the content of blinin, which depends on CbHO-1 expression. Conyza blinii (C. blinii) is a unique Chinese herbal medicine that grows in Sichuan Province, China. Because the habitat of C. blinii is an iron ore mining area with abundant iron content, this species can be used as one of the best materials to study the mechanism of plant tolerance to iron. In this study, C. blinii was treated with ferrous-EDTA solutions at different concentrations, and it was found that the tolerance value of C. blinii to iron was 200 µM. Under this concentration, the plant height, root length, biomass, and iron content of C. blinii increased to the maximum values, and the effect was dependent on the upregulated expression of CbHO-1. At the same time, under ferrous iron, the photosynthetic capacity and capitate glandular trichome density of C. blinii also significantly increased, providing precursors and sites for the synthesis of blinin, thus significantly increasing the content of blinin. These processes were also dependent on the high expression of CbHO-1. Correlation analysis showed that there were strong positive correlations between iron content, capitate glandular trichome density, CbHO-1 gene expression, and blinin content. This study explored the effects of ferrous iron on the physiology and biochemistry of C. blinii, greatly improving our understanding of the mechanism of iron tolerance in C. blinii.

The jasmonate-responsive transcription factor CbWRKY24 regulates terpenoid biosynthetic genes to promote saponin biosynthesis in Conyza blinii H. Lév.

Conyza blinii H. Lév., the most effective component is saponin, is a biennial medicinal material that needs to be overwintered. WRKY transcription factors family is a large protein superfamily that plays a predominant role in plant secondary metabolism, but their characteristics and functions have not been identified in C. blinii. The CbWRKY24 sequence was selectedfrom the transcriptome database of the C. blinii leaves constructed in our laboratory. Phylogenetic tree analysis revealed that it was associated with AaWRKY1 which can regulate artemisinin synthesis in Artemisia annua. Expression analysis in C. blinii revealed that CbWRKY24 was mainly induced by methyl jasmonate (MeJA) and cold treatments. Transcriptional activity assay showed that it had an independent biological activity. Overexpression of CbWRKY24 in transient transformed C. blinii resulted in improved totalsaponins content, which was attributed to upregulate the expression level of keys genes from mevalonate (MVA) pathway in transient transformed plants compared to wild type (WT) plants. Meanwhile, overexpression the CbWRKY24 in transient transformed tomato fruits showed that the transcript level of related genes in lycopene pathway decreased significantly when compared to WT tomatofruits. Additionally, the MeJA-response-element was found in the promoter regions of CbWRKY24 and the histochemical staining experiments showed that promoter had GUS activity in transiently transformed tobacco leaves. In summary, our results indicated that we may have found a transcription factor that can regulate the biosynthesis of terpenoids in C. blinii.