Effects of trigonelline, diosgenin, and Cistanche deserticola polysaccharide on the culture of female germline stem cells in vitro.

Female germline stem cells (FGSCs) are renewable sources of oocytes that play an indispensable role in re-establishing mammal fertility. Here, we have established FGSCs from neonatal mice, which exhibit characteristics of germline stem cells. We show that compared with monomeric trigonelline and diosgenin, macromolecular compounds Cistanche deserticola polysaccharides (CDPs) in Chinese herbal medicine can enhance the ability of FGSCs to differentiate into oocytes at appropriate concentrations while maintaining self-renewal in vitro. In contrast, trigonelline and diosgenin inhibited the expression of germ cell-specific genes while reducing cell proliferation activity. In summary, CDPs could induce the differentiation and self-renewal of FGSCs in vitro. The comparison of the effects of the active components of different types of Chinese medicine will provide a reference for the development of clinical drugs in the future, and help to elucidate the development process of FGSCs.

Nuciferine reduces inflammation induced by cerebral ischemia-reperfusion injury through the PI3K/Akt/NF-κB pathway.

BACKGROUND: Cerebral ischemia has the characteristics of high incidence, mortality, and disability, which seriously damages people's health. Cerebral ischemia-reperfusion injury is the key pathological injury of this disease. However, there is a lack of drugs that can reduce cerebral ischemia-reperfusion injury in clinical practice. At present, a few studies have provided some evidence that nuciferine can reduce cerebral ischemia-reperfusion injury, but its specific mechanism of action is still unclear, and further research is still needed. OBJECTIVE: In this study, PC12 cells and SD rats were used to construct OGD/R and MCAO/R models, respectively. Combined with bioinformatics methods and experimental verification methods, the purpose of this study was to conduct a systematic and comprehensive study on the effect and mechanism of nuciferine on reducing inflammation induced by cerebral ischemia-reperfusion injury. RESULTS: Nuciferine can improve the cell viability of PC12 cells induced by OGD/R, reduce apoptosis, and reduce the expression of inflammation-related proteins; it can also improve the cognitive and motor dysfunction of MCAO/R-induced rats by behavioral tests, reduce the area of cerebral infarction, reduce the release of inflammatory factors TNF-α and IL-6 in serum and the expression of inflammation-related proteins in brain tissue. CONCLUSION: Nuciferine can reduce the inflammatory level of cerebral ischemia-reperfusion injury in vivo and in vitro models by acting on the PI3K/Akt/NF-κB signaling pathway, and has the potential to be developed as a drug for the treatment of cerebral ischemia-reperfusion injury.

Reyanning Mixture on Asymptomatic or Mild SARS-CoV-2 Infection in Children and Adolescents: A Randomized Controlled Trial.

OBJECTIVE: To assess the effect and safety of Reyanning Mixture (RYN) in treating asymptomatic or mild severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in children and adolescents. METHODS: This is a prospective, open-label, randomized controlled trial. Patients aged 1-17 years and diagnosed with asymptomatic or mild coronavirus disease-2019 (COVID-19) were assigned to an intervention group (RYN plus standard care) and a control group (standard care) according to a randomization list. The primary outcomes were SARS-CoV-2 negative conversion time. Secondary outcomes included negative conversion rate on days 3 and 7, hospital length of stay, symptom relief rate, new-onset symptoms of asymptomatic infected patients, and progressive disease rate. The cycle threshold (Ct) values of ORF1ab or N genes were also tested. RESULTS: A total of 214 patients in the intervention group and 217 in the control group were analyzed. The SARS-CoV-2 negative conversion time was significantly shortened in the intervention group [5 days (interquartile range (IQR): 5-6) vs. 7 days (IQR: 6-7), P<0.01]. By days 3 and 7, the negative conversion rates were significantly higher in the intervention group (day 3: 32.7% vs. 21.2%, P=0.007; day 7: 75.2% vs. 60.8%, P=0.001). Ct values significantly increase on day 2 [ORF1ab gene: 35.62 (IQR: 29.17-45.00) vs. 34.22 (IQR: 28.41-39.41), P=0.03; N gene: 34.97 (IQR: 28.50-45.00) vs. 33.51 (IQR: 27.70-38.25), P=0.024] and day 3 [ORF1ab gene: 38.00 (IQR: 32.72-45.00) vs. 35.81 (IQR: 29.96-45.00), P=0.003; N gene: 37.16 (IQR: 32.01-45.00) vs. 35.26 (IQR: 29.09-45.00), P=0.01]. No significant difference was found in hospital length of stay between the two groups (P>0.05). Symptoms of cough were significantly improved (82.2% vs. 70.0%, P=0.02) and wheezing was significantly reduced (0.7% vs. 12.9%, P<0.01) in the intervention group compared with the control group. During the trial, no disease progression or serious adverse events were reported. CONCLUSION: Adding RYN to standard care may be a safe and effective treatment for children with asymptomatic and mild SARS-CoV-2 infection. (Registration No. ChiCTR2200060292).

Nutrient Metabolism Pathways Analysis and Key Candidate Genes Identification Corresponding to Cadmium Stress in Buckwheat through Multiomics Analysis.

Fagopylum tatarium (L.) Gaertn (buckwheat) can be used both as medicine and food and is also an important food crop in barren areas and has great economic value. Exploring the molecular mechanisms of the response to cadmium (Cd) stress can provide the theoretical reference for improving the buckwheat yield and quality. In this study, perennial tartary buckwheat DK19 was used as the experimental material, its key metabolic pathways in the response to Cd stress were identified and verified through transcriptomic and metabolomic data analysis. In this investigation, 1798 metabolites were identified through non-targeted metabolomic analysis containing 1091 up-regulated and 984down-regulated metabolites after treatment. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of differential metabolites was significantly enriched in galactose metabolism, glycerol metabolism, phenylpropane biosynthesis, glutathione metabolism, starch and sucrose metabolism. Linkage analysis detected 11 differentially expressed genes (DEGs) in the galactose metabolism pathway, 8 candidate DEGs in the lipid metabolism pathway, and 20 candidate DEGs in the glutathione metabolism pathway. The results of our study provided useful clues for genetically improving the resistance to cadmium by analyzing the molecular mechanism of cadmium tolerance in buckwheat.

Effect of acupuncture at Xinwu acupoint combined with loratadine and fluticasone propionate on symptom alleviation, nasal function, and serum histamine level in patients with allergic rhinitis.

OBJECTIVE: To determine the influence of acupuncture at the Xinwu acupoint combined with western medicine (loratadine and fluticasone propionate) on symptom alleviation, nasal mucociliary clearance velocity (MCV), and serum histamine level of patients with allergic rhinitis (AR). METHODS: A total of 122 patients with AR treated in Gansu province hospital of TCM and The Third People's Hospital of Gansu Province from April 2019 to April 2021 were retrospectively analyzed. Among them, 54 patients treated with loratadine and fluticasone propionate were assigned to the control group, and 68 patients treated with additional acupuncture at the Xinwu acupoint based on treatment of the control group were assigned to the observation group. The treatment efficacy of the two groups was compared, and the scores of main symptoms and nasal function were also compared before and after therapy. Additionally, the two groups were compared in the levels of histamine, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and immunoglobulin E (IgE) before and after therapy. RESULTS: After therapy, the observation group yielded a higher total effective rate than the control group (P=0.006) and had lower symptom scores than the control group (P<0.001). Additionally, the MCV of the two groups increased (P<0.001), and the nasal mucociliary transit time (MTT) and nasal resistance (NR) of both groups decreased (P<0.001) after therapy. The observation group showed a greatly better improvement of nasal function than the control group (P<0.001). Moreover, after therapy, the observation group showed lower histamine and IgE levels than the control group (P<0.01) and the observation group presented significantly lower levels than the control group, and had lower rhinoconjunctivitis Quality of Life Questionnaire (RQLQ) scores than the control group (P<0.001). The two groups were not different in the incidence of adverse reactions (P=0.886). CONCLUSION: Acupuncture at Xinwu acupoint combined with loratadine and fluticasone propionate can deliver a powerful efficacy on AR and alleviate the clinical symptoms, without increasing adverse reactions.

Efficacy of Lianhua Qingwen for children with SARS-CoV-2 Omicron infection: A propensity score-matched retrospective cohort study.

BACKGROUND: Lianhua Qingwen Granules or Capsules (LHQW) has accumulated much research evidence in the fight against the coronavirus disease 2019 (COVID-19) epidemic. However, there are still few data on its efficacy and safety in children with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. PURPOSE: To evaluate the efficacy and safety of LHQW in children with SARS-CoV-2 Omicron infection. METHODS: We conducted a single-center, propensity-score matched retrospective cohort study of children with SARS-CoV-2 Omicron infection in Shanghai New International Expo Center mobile cabin hospital between April 1st and June 1st, 2022. Eligible patients received either LHQW granules/capsules plus supportive care (LHQW group) or supportive care alone (control group). The primary outcome was the negative conversion time of nucleic acid. Secondary outcomes included the negative conversion rate of nucleic acid, the length of hospital stay, clinical disease progression, and cycle threshold [Ct] values for SARS-CoV-2 open reading frame [ORF1ab] or nucleocapsid [N] genes. RESULTS: Overall, 2808 patients were enrolled, and 346 patients in each group were included in the analysis. Among the propensity-score matched groups, LHQW treatment was associated with an accelerated negative conversion time of nucleic acid (median: 5 d vs. 6 d, Hazard ratio: 1.25, 95% CI: 1.08 - 1.46, Log-rank p < 0.001), a higher negative conversion rate of nucleic acid (Day 2 - 6: 2.9% vs. 0.6%, p = 0.036; 29.8% vs. 5.5%, p < 0.001; 42.5% vs. 24.3%, p < 0.001; 51.4% vs. 31.5%, p < 0.001; 63.3% vs. 55.2%, p = 0.030), shorter hospital stay (median: 10 d vs. 11 d, Hazard ratio: 1.50, 95% CI: 1.29 - 1.74, Log-rank p < 0.001), and lower rates of asymptomatic infection progressing to mild (37.9% vs. 46.5%, p = 0.021). CONCLUSION: Our study suggested that LHQW treatment was associated with faster clinical recovery in children with SARS-CoV-2 Omicron infection.

Efficacy and safety of Reyanning mixture in patients infected with SARS-CoV-2 Omicron variant: A prospective, open-label, randomized controlled trial.

BACKGROUND: A wave of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant rapidly resulted in a steep increase in the infected population and an overloaded healthcare system. Effective medications for Omicron are currently limited. The previous observational study supports the efficacy and safety of Reyanning (RYN) mixture in the treatment of coronavirus disease 2019 (COVID-19). PURPOSE: To evaluate the efficacy of RYN in asymptomatic and mildly infected patients with SARS-CoV-2 infection. STUDY DESIGN AND METHODS: This study was a prospective, open-label, randomized controlled trial. We consecutively recruited 2830 patients from Shanghai New International Expo Center mobile cabin hospital and randomized them in a 1:1 ratio to receive RYN plus standard care or receive standard care alone. The primary outcomes were the negative conversion of nucleic acid. Secondary outcomes included the hospital duration, new-onset symptoms, proportion of disease progression, and the viral load measured by the cycle threshold (Ct) value. RESULTS: A total of 1393 patients in the intervention group and 1407 patients in the control group completed the study. The negative conversion time of nucleic acid was significantly shortened in the intervention group (median: 6 d vs. 7 d, Hazard ratio: 0.768, 95CI %: 0.713-0.828, p < 0.0001). The negative conversion rate of nucleic acid was significantly higher in the intervention group (Day 3: 32.4% vs. 18.3%; Day7: 65.3% vs. 55.2%, p < 0.001). The hospitalization duration was significantly shortened in the intervention group (median: 8 d vs. 9 d, Hazard ratio: 0.759, 95% CI: 0.704-0.818, p < 0.0001). The proportion of new-onset fever (2.4% vs. 4.1%, p = 0.012), coughing (12.2% vs. 14.8%, p = 0.046), and expectoration (6.0% vs. 8.0%, p = 0.032) in the intervention group was significantly lower. RYN treatment increased Ct values and reduced the viral load. No disease progression and serious adverse events were reported during the study. CONCLUSION: RYN is a safe and effective treatment that can accelerate virus clearance and promote disease recovery in asymptomatic and mild Omicron infections.

Pharmacological mechanism and therapeutic efficacy of Icariside II in the treatment of acute ischemic stroke: a systematic review and network pharmacological analysis.

BACKGROUND AND OBJECTIVE: Epimedii has long been used as a traditional medicine in Asia for the treatment of various common diseases, including Alzheimer's disease, cancer, erectile dysfunction, and stroke. Studies have reported the ameliorative effects of Icariside II (ICS II), a major metabolite of Epimedii, on acute ischemic stroke (AIS) in animal models. Based on network pharmacology, molecular docking, and molecular dynamics (MD) simulations, we conducted a systematic review to evaluate the effects and neuroprotective mechanisms of ICS II on AIS. METHODS: First, we have searched 6 databases using studies with ICS II treatment on AIS animal models to explore the efficacy of ICS II on AIS in preclinical studies. The literature retrieval time ended on March 8, 2022 (Systematic Review Registration ID: CRD42022306291). There were no restrictions on the language of the search strategy. Systematic review follows the Patient, Intervention, Comparison and Outcome (PICO) methodology and framework. SYCLE's RoB tool was used to evaluate the the risk of bias. In network pharmacology, AIS-related genes were identified and the target-pathway network was constructed. Then, these targets were used in the enrichments of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways and gene ontology (GO). Molecular docking and MD simulation were finally employed between ICS II and the potential target genes. RESULTS: Twelve publications were included describing outcomes of 1993 animals. The literature details, animal strains, induction models, doses administered, duration of administration, and outcome measures were extracted from the 12 included studies. ICS II has a good protective effect against AIS. Most of the studies in this systematic review had the appropriate methodological quality, but some did not clearly state the controlling for bias of potential study. Network pharmacology identified 246 targets with SRC, CTNNB1, HSP90AA1, MAPK1, and RELA as the core target proteins. Besides, 215 potential pathways of ICS II were identified, such as PI3K-Akt, MAPK, and cGMP-PKG signaling pathway. GO enrichment analysis showed that ICS II was significantly enriched in subsequent regulation such as MAPK cascade. Molecular docking and MD simulations showed that ICS II can closely bind with important targets. CONCLUSIONS: ICS II is a promising drug in the treatment of AIS. However, this systematic review reveals key knowledge gaps (i.e., the protective role of ICS II in women) that ICS II must address before it can be used for the treatment of human AIS. Our study shows that ICS II plays a protective role in AIS through multi-target and multi-pathway characteristics, providing ideas for the development of drugs for the treatment of AIS.

The phytochemical, corynoline, diminishes Aurora kinase B activity to induce mitotic defect and polyploidy.

Plants are a rich source for bioactive compounds. However, plant extracts can harbor a mixture of bioactive molecules that promote divergent phenotypes and potentially have confounding effects in bioassays. Even with further purification and identification, target deconvolution can be challenging. Corynoline and acetylcorynoline, are phytochemicals that were previously isolated through a screen for compounds able to induce mitotic arrest and polyploidy in oncogene expressing retinal pigment epithelial (RPE) cells. Here, we shed light on the mechanism by which these phytochemicals can attack human cancer cells. Mitotic arrest was coincident to the induction of centrosome amplification and declustering, causing multi-polar spindle formation. Corynoline was demonstrated to have true centrosome declustering activity in a model where A549 cells were chemically induced to have more than a regular complement of centrosomes. Corynoline could inhibit the centrosome clustering required for pseudo-bipolar spindle formation in these cells. The activity of AURKB, but not AURKA or polo-like kinase 4, was diminished by corynoline. It only partially inhibited AURKB, so it may be a partial antagonist or corynoline may work upstream on an unknown regulator of AURKB activity or localization. Nonetheless, corynoline and acetylcorynoline inhibited the viability of a variety of human cancer derived cell lines. These phytochemicals could serve as prototypes for a next-generation analog with improved potency, selectivity or in vivo bioavailability. Such an analog could be useful as a non-toxic component of combination therapies where inhibiting the chromosomal passenger protein complex is desired.

Uncertainty and sensitivity analysis of spatial distributed roughness to a hydrodynamic water quality model: a case study on Lake Taihu, China.

Roughness is an important parameter in hydrodynamic and water quality modelling; it has direct effects on bottom shear stress which relied on sediment and vegetation. The varied roughness caused by spatial heterogeneity of sediment and vegetation may lead to uncertain simulation results. To investigate the effect of roughness uncertainty on the performance of hydrodynamic water quality models, a typical large shallow lake in China (Lake Taihu) was divided into eight areas for illustrating the effect of spatial variation of roughness on hydrodynamics and water quality. Total nitrogen (TN) was selected as the variable to calculate the uncertainty interval, and sensitive positions greatly affected by roughness as well as the appropriate range of roughness were explored by means of regional sensitive analysis (RSA). The results showed that roughness had the most significant effect on the bottom velocity. The uncertainty for water quality caused by roughness presented a striking spatial difference; the uncertainty interval for TN could be up to 1.3 mg/L. The posterior distribution of roughness was given to further narrowed the range of roughness, and the updated roughness range manifested that roughness value should be set higher in the area with thick sediment and abundant vegetation. It is of utmost importance to consider the comprehensive effects of sediment and vegetation in the determination of roughness. For certain lake areas with great water quality simulation error, the error could be effectively reduced by setting spatial distributed roughness. The optimization scheme was provided for the reasonable determination of roughness, so that the dynamic characteristic at the sediment-water interface could be represented synthetically. In this paper, the uncertainty and sensitivity of roughness in hydrodynamic water quality model are analyzed to provide reference for parameter setting of large shallow water lake model. For large scale lakes, parameters need to be modified according to the actual condition due to the spatial difference of friction coefficient at the bottom.

Applications of Molecular Imprinting Technology in the Study of Traditional Chinese Medicine.

Traditional Chinese medicine (TCM) is one of the most internationally competitive industries. In the context of TCM modernization and internationalization, TCM-related research studies have entered a fast track of development. At the same time, research of TCM is also faced with challenges, such as matrix complexity, component diversity and low level of active components. As an interdisciplinary technology, molecular imprinting technology (MIT) has gained popularity in TCM study, owing to the produced molecularly imprinted polymers (MIPs) possessing the unique features of structure predictability, recognition specificity and application universality, as well as physical robustness, thermal stability, low cost and easy preparation. Herein, we comprehensively review the recent advances of MIT for TCM studies since 2017, focusing on two main aspects including extraction/separation and purification and detection of active components, and identification analysis of hazardous components. The fundamentals of MIT are briefly outlined and emerging preparation techniques for MIPs applied in TCM are highlighted, such as surface imprinting, nanoimprinting and multitemplate and multifunctional monomer imprinting. Then, applications of MIPs in common active components research including flavonoids, alkaloids, terpenoids, glycosides and polyphenols, etc. are respectively summarized, followed by screening and enantioseparation. Related identification detection of hazardous components from TCM itself, illegal addition, or pollution residues (e.g., heavy metals, pesticides) are discussed. Moreover, the applications of MIT in new formulation of TCM, chiral drug resolution and detection of growing environment are summarized. Finally, we propose some issues still to be solved and future research directions to be expected of MIT for TCM studies.

Effects and Mechanism of Zishen Jiangtang Pill on Diabetic Osteoporosis Rats Based on Proteomic Analysis.

OBJECTIVE: To explore the effect and mechanism of ZJP on DOP rats by proteomic analysis. MATERIALS AND METHODS: After the establishment of diabetes model by Streptozocin (STZ, 60 mg/kg), 40 Wistar rats were equally divided into normal group, model group (diabetic rats), high-dose group (3.0 g/kg/d ZJP), and low-dose group (1.5 g/kg/d ZJP) and received treatment for 3 months. Histological changes in bone and pancreas tissues were observed by hematoxylin and eosin staining, electron microscopy, and immunofluorescence. Proteomic and bioinformatic analyses were performed to identify the differentially expressed proteins. The fingerprint and active ingredients of ZJP were identified via high-performance liquid chromatography (HPLC). RESULTS: Compared with the model group, ZJP could rescue the weight, fasting blood glucose, and fasting insulin of rats in both high-dose and low-dose group. ZJP could also improve the microstructures of pancreatic islet cells, bone mass, and trabecular and marrow cavities in DOP rats. Bioinformatic analysis suggested that ZJP might influence DOP via multiple pathways, mainly including ribosomes, vitamin digestion and absorption, and fat digestion and absorption. The primary active ingredients, including notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, icariin, and ginsenoside Rb1, were detected. CONCLUSION: ZJP could significantly improve the histomorphology and ultrastructure of bone and islets tissues and might serve as an effective alternative medicine for the treatment of DOP.

Nicotinamide riboside attenuates age-associated metabolic and functional changes in hematopoietic stem cells.

With age, hematopoietic stem cells (HSC) undergo changes in function, including reduced regenerative potential and loss of quiescence, which is accompanied by a significant expansion of the stem cell pool that can lead to haematological disorders. Elevated metabolic activity has been implicated in driving the HSC ageing phenotype. Here we show that nicotinamide riboside (NR), a form of vitamin B3, restores youthful metabolic capacity by modifying mitochondrial function in multiple ways including reduced expression of nuclear encoded metabolic pathway genes, damping of mitochondrial stress and a decrease in mitochondrial mass and network-size. Metabolic restoration is dependent on continuous NR supplementation and accompanied by a shift of the aged transcriptome towards the young HSC state, more youthful bone marrow cellular composition and an improved regenerative capacity in a transplant setting. Consequently, NR administration could support healthy ageing by re-establishing a more youthful hematopoietic system.

Effectiveness and safety of traditional Chinese medical therapy for cancer-related fatigue: a systematic review and Meta-analysis of randomized controlled trials.

OBJECTIVE: To assess the effectiveness and safety of traditional Chinese medical therapy for cancer-related fatigue. METHODS: We systematically searched eight electronic databases up to June 2017 for randomized clinical trials of traditional Chinese medical therapy for cancer-related fatigue. Two authors independently extracted data and assessed the risk bias of the included trials using the Cochrane Handbook. Data were analyzed by RevMan 5.2 software. RESULTS: A total of 23 trials involving 1832 participants identified with cancer-related fatigue were included. Twenty trials reported a beneficial effect of traditional Chinese medical therapy on cancer-related fatigue. On pooling the data from Chinese herbal medicine therapy and acupuncture or moxibustion therapy, respectively, significant differences were found between experimental groups and control groups. Fatigue improvement rates showed significant differences between traditional Chinese medical therapy and control groups [odds ratio (OR), 7.62; 95% confidence interval (CI), 3.75-15.49; P < 0.000 01; and OR, 3.78; 95% CI, 2.29-6.23; P < 0.000 01). Fatigue change scores also showed significant differences between the two groups (mean difference, -0.91; 95% CI, -0.16 to -0.65; P < 0.000 01). Eleven trials demonstrated that traditional Chinese medical therapy improved the quality of life of cancer patients. No severe adverse effects occurred in traditional Chinese medical therapy groups. CONCLUSION: Meta-analysis showed that Chinese medical therapy seems to be effective and safe in the treatment of cancer-related fatigue.

Anti-neuraminidase activity of chemical constituents of Balanophora involucrata.

Balanophora involucrata J. D. Hooker has been known to possess potential anti-inflammatory and antibacterial activities; however, its antiviral activity has not been evaluated so far. In order to find new neuraminidase inhibitors (NAIs), the neuraminidase (NA) inhibition activity of different B. involucrata extracts was evaluated. In this study, an in vitro NA inhibition assay was performed to identify which extract of B. involucrata exhibits (maximal) inhibitory activity against NA. Ultra high performance liquid chromatography/quadrupole time-of-flight-tandem mass spectroscopy (MS/MS) and molecular docking techniques were used to identify the specific compounds responsible for the anti-influenza activity of the extract, and to explore the potential natural NAIs. The ethyl acetate extract of B. involucrata exhibited significant inhibitory activity against NA with 50% inhibitory concentration (IC50 ) value of 159.5 µg/mL. Twenty compounds were identified according to the MS/MS spectra; among them two compounds (quercitrin and phloridzin) showed obvious inhibitory activity against NA, with IC50 of 311.76 and 347.32 µmol/L, respectively. This study suggested that B. involucrata can be a potential natural source of NAIs and may be useful in the fight against ferocious influenza viruses.

Combined Blockade of Smad3 and JNK Pathways Ameliorates Progressive Fibrosis in Folic Acid Nephropathy.

Acute kidney injury leading to chronic kidney disease through tubulointerstitial fibrosis is a major challenge in nephropathy. Several signaling pathways promote interstitial fibrosis; however, effective suppression of fibrosis may require blockade of more than one pathway. This study investigated whether blockade of Smad3 and c-Jun N-terminal kinase (JNK) signaling gives added suppression of interstitial fibrosis in folic acid nephropathy. A single high dose of folic acid (FA) causes acute tubular damage in C57BL/6J mice followed by interstitial fibrosis and chronic renal impairment. Co-activations of Smad3 and JNK signaling occur in both tubular epithelial cells and myofibroblasts in areas of tubulointerstitial damage and fibrosis in both murine FA-induced nephropathy and human IgA nephropathy. Groups of mice were treated with a Smad3 inhibitor (SIS3), a JNK inhibitor (SP600125), or a combination from day 6 after FA administration until being killed on day 28. Each drug efficiently inhibited its specific target (Smad3 phosphorylation or c-Jun phosphorylation) without affecting the other pathway. Given alone, each drug partially reduced renal fibrosis, whereas the combination therapy gave an additive and profound protection from renal fibrosis and improved renal function. Inhibition of Smad3 and/or JNK signaling activities prevented down-regulation of PGC-1α in tubular epithelial cells and up-regulation of PGC-1α in myofibroblasts during FA-induced renal fibrosis and inflammation. The expression of PGC-1α was upregulated in Smad3 -/- NRK52E cells while downregulated in Smad3 -/- NRK49F cells, suggesting that Smad3 signaling may regulate expression of PGC-1α in renal tubular epithelial cells and fibroblasts in distinct fashion. In vivo and cell culture studies also indicate that Smad3 and JNK signaling cooperate to cause mitochondrial dysfunction and cell damage in tubular epithelial cells via direct actions on the transcription of PGC-1α. These pathways also act cooperatively to promote renal fibroblast proliferation in tempo-spatial fashion. In conclusion, we have identified a potential combination therapy for progressive renal fibrosis which operates, in part, through modifying mitochondrial function.

Jatrorrhizine inhibits mammary carcinoma cells by targeting TNIK mediated Wnt/ß-catenin signalling and epithelial-mesenchymal transition (EMT).

BACKGROUND: Traf2 and Nck interacting serine protein kinase (TNIK) is a tumour target protein which its high expression is closely related to the occurrence and development of mammary carcinoma cells. Molecular docking revealed that jatrorrhizine, a protoberberine alkaloid, exhibits good binding affinity and interaction with TNIK. However, the underlying mechanisms of jatrorrhizine targeting TNIK inhibits the proliferation and metastasis of breast cancer cells remain unclear. METHODS: To figure out the mechanisms in vitro and in vivo, the CRISPR/Cas9 technology was used to knockout TNIK gene and detected qualitatively by immunofluorescence and immunoblotting assay. The MTT cell viability assay for cytotoxicity test, the apoptosis were detected by flow cytometry, the migration and invasion were evaluated by colony formation, wound healing assay and cell invasion assay, respectively. Anticancer effects were further corroborated by 4T1/Luc homograft tumour model. RESULTS: The results showed that targeted knockout of TNIK that attenuated Wnt/ß-catenin signalling and epithelial-mesenchymal transition (EMT) expression, the effects were potentiated by the addition of jatrorrhizine. Moreover, jatrorrhizine distinctly inhibited the proliferation of MDA-MB-231, MCF-7 and 4T1 cells with IC50 values of 11.08 ± 1.19 µM, 17.11 ± 4.54 µM and 22.14 ± 2.87 µM, induced mitochondrial dysfunction and early apoptosis involving mitochondrial apoptotic pathway. These results were further corroborated by the 4T1 tumour-bearing mice, which showed that jatrorrhizine significantly suppressed the proliferation and metastasis of mammary carcinoma cells without obvious toxicity. CONCLUSION: These findings provide an overall perspective that jatrorrhizine potentially restrains TNIK regulating Wnt/ß-catenin signalling and EMT expression for mammary cancer targeted therapy.

Leigongteng (Radix et Rhizoma Tripterygii) via compatibility with Jinqiancao (Herba Lysimachiae): its toxicity-reduced efficacy in H22-bearing mice.

OBJECTIVE: To observe toxicity-reduced effects of Leigongteng (Radix et Rhizoma Tripterygii) (LGT) via compatibility with Jinqiancao (Herba Lysimachiae) (JQC) in H22-bearing mice and investigate the possible underlying mechanism, and further explore whether JQC can enhance LGT-evoked anti-tumor effect. METHODS: H22-bearing mice were orally administered with LGT alone and its compatibility with JQC, and tumors, serum, livers and kidneys were collected to evaluate the toxicity-reduced efficacy and the possible mechanism. RESULTS: LGT evoked significantly elevated biochemical indicators including serum alanine / aspartate transaminase (ALT/AST), creatinine (Cr) and urea nitrogen (BUN) as well as pathological damage in mice, which were all obviously reversed by JQC via compatibility at the ratios from 4/1 to 1/4. Further analysis indicated that pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-α), and malondialdehyde (MDA) levels significantly decreased, while anti-inflammatory cytokine interleukin (IL)-10, and glutathione (GSH), GSH-s transferase (GST), GSH peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) levels all increased in livers and kidneys of mice. Besides, after compatibility with JQC at the ratios of 4/1, 2/1, 1/1, 1/2 and 1/4, LGT-decreased tumor weight was further decreased by 48.4%, 57.3%, 54.0%, 49.3% and 52.9%, respectively (all P < 0.01). CONCLUSION: JQC could reduce LGT-induced hepatotoxicity and nephrotoxicity, and enhance the antitumor efficacy via compatibility with JQC, and the toxicity-reduced mechanism could involve inhibiting hepatic and kidney oxidative stress and inflammation.

An Integrated LC-MS-Based Strategy for the Quality Assessment and Discrimination of Three Panax Species.

: The quality assessment and discrimination of Panax herbs are very challenging to perform due to the complexity and variability of their chemical compositions. An integrated strategy was established using UHPLC-Q-Exactive/HRMS and HPLC-ESI-MS/MS to achieve an accurate, rapid, and comprehensive qualitative and quantitative analysis of Panax japonicas (PJ), Panax japonicus var. major (PM), and Panax zingiberensis (PZ). Additionally, discrimination among the three species was explored with partial least squares⁻discriminant analysis (PLS-DA) and orthogonal partial least squares⁻discriminant analysis (OPLS-DA) score plots. A total of 101 compounds were plausibly or unambiguously identified, including 82 from PJ, 78 from PM, and 67 from PZ. Among them, 16 representative ginsenosides were further quantified in three herbs. A clear discrimination between the three species was observed through a multivariate statistical analysis on the quantitative data. Nine compounds that allowed for discrimination between PJ, PM, and PZ were discovered. Notably, ginsenoside Rf (G-Rf), ginsenoside F3 (G-F3), and chikusetsu saponin IV (CS-IV) were the three most important differential compounds. The research indicated that the integrated LC-MS-based strategy can be applied for the quality assessment and discrimination of the three Panax herbs.

Detoxication mechanisms of Radix Tripterygium wilfordii via compatibility with Herba Lysimachia christinae in S180-bearing mice by involving Nrf2.

The combined administration between Radix Tripterygium wilfordii Hook F (LGT) and Herba Lysimachia christinae Hance (JQC) belongs to mutual detoxication compatibility of seven emotions in traditional Chinese medicine (TCM) theory. However, until now, the compatibility detoxication mechanisms remain unknown. The present study was undertaken to observe detoxication mechanisms of LGT through compatibility with JQC in tumor-bearing mice by involving NF-E2-related factor 2 (Nrf2)-mediated antioxidant defenses. In addition, influence of compatibility on antitumor activity was also investigated here. Our results demonstrated that compatibility with JQC administration significantly reversed LGT-elevated serum alanine/aspartate transaminase (ALT/AST) levels and alleviated hepatocytes' swelling or degeneration damage, and at the ratio 2/1 (LGT/JQC) produced the strongest detoxication effect. Besides, compatibility with JQC administration reversed not only LGT-elevated hepatic malondialdehyde (MDA) and tumor necrosis factor-α (TNF-α) but also the LGT lowered GSH, glutathione-s transferase (GST), glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT), and interleukin (IL)-10 levels. Furthermore, compatibility with JQC administration significantly up-regulated protein expression of Nrf2 and mRNA expression of it regulated downstream antioxidant genes such as heme oxygenase-1 (HO-1), NAD(P)H: quinone oxidoreductase-1 (NQO1), and glutamate cysteine ligase catalytic subunit (GCLC). In addition, compatibility with JQC further decreased LGT-decreased tumor weight and at the ratio 2/1 (LGT/JQC) also exerted the strongest synergistic effect. Collectively, through compatibility with JQC exerted detoxication effect on LGT-induced hepatotoxicity and the mechanisms could be at least partly attributed to up-regulation of Nrf2 and its downstream signals, thereby enhancing antioxidant defenses, and inhibiting lipid peroxidation, oxidative stress, and inflammation. Additionally, at the ratio 2/1 (LGT/JQC) exerted the strongest effects on both detoxication and synergism.