CwJAZ4/9 negatively regulates jasmonate-mediated biosynthesis of terpenoids through interacting with CwMYC2 and confers salt tolerance in Curcuma wenyujin.

Plant JASMONATE ZIM-DOMAIN (JAZ) genes play crucial roles in regulating the biosynthesis of specialized metabolites and stressful responses. However, understanding of JAZs controlling these biological processes lags due to numerous JAZ copies. Here, we found that two leaf-specific CwJAZ4/9 genes from Curcuma wenyujin are strongly induced by methyl-jasmonate (MeJA) and negatively correlated with terpenoid biosynthesis. Yeast two-hybrid, luciferase complementation imaging and in vitro pull-down assays confirmed that CwJAZ4/9 proteins interact with CwMYC2 to form the CwJAZ4/9-CwMYC2 regulatory cascade. Furthermore, transgenic hairy roots showed that CwJAZ4/9 acts as repressors of MeJA-induced terpenoid biosynthesis by inhibiting the terpenoid pathway and jasmonate response, thus reducing terpenoid accumulation. In addition, we revealed that CwJAZ4/9 decreases salt sensitivity and sustains the growth of hairy roots under salt stress by suppressing the salt-mediated jasmonate responses. Transcriptome analysis for MeJA-mediated transgenic hairy root lines further confirmed that CwJAZ4/9 negatively regulates the terpenoid pathway genes and massively alters the expression of genes related to salt stress signaling and responses, and crosstalks of multiple phytohormones. Altogether, our results establish a genetic framework to understand how CwJAZ4/9 inhibits terpenoid biosynthesis and confers salt tolerance, which provides a potential strategy for producing high-value pharmaceutical terpenoids and improving resistant C. wenyujin varieties by a genetic approach.

Combinational study with network pharmacology, molecular docking and preliminary experiments on exploring common mechanisms underlying the effects of weijing decoction on various pulmonary diseases.

Objective: 'Homotherapy for heteropathy' is a theory by which different diseases with similar pathogenesis can be treated with one Chinese formula. We aimed to explore the key components and core targets of Weijing decoction (WJD) in treating various lung diseases, namely, pneumonia, chronic obstructive pulmonary disease (COPD), acute lung injury (ALI), pulmonary fibrosis, pulmonary tuberculosis and non-small cell lung cancer (NSCLC), via network pharmacology, molecular docking and some experiments. Significance: This is the first study on the mechanism of WJD in treating various lung diseases by 'homotherapy for heteropathy'. This study is helpful for the transformation of TCM formula and development of new drugs. Methods: Active components and therapeutic targets of WJD were obtained via TCMSP and UniProt databases. Targets of the six pulmonary diseases were harvested from the GeneCards TTD, DisGeNet, UniProt and OMIM databases. Drug-disease intersection targets, corresponding Venn diagrams, herb-component-target networks and protein-protein interaction networks were established. Furthermore, GO biological function and KEGG enrichment analysis were completed. Moreover, the binding activity between main compounds and core targets was measured through molecular docking. Finally, the xenograft NSCLC mouse model was established. Immune responses were evaluated by flow cytometry and mRNA expression levels of critical targets were measured by real-time PCR. Results: JUN, CASP3 and PTGS2 were the most critical targets in six pulmonary diseases. The active compounds beta-sitosterol, tricin and stigmasterol stably bound to many active sites on target proteins. WJD had extensive pharmacological regulation, involving pathways related to cancer, inflammation, infection, hypoxia, immunity and so on. Conclusions: Effects of WJD against various lung diseases involve lots of compounds, targets and pathways. These findings will facilitate further research as well as clinical application of WJD.

Facile preparation of a novel iminodisuccinate modified chitin and its excellent properties as a silver bioadsorbent and antibacterial agent.

A novel iminodisuccinate modified chitin (ICH) was prepared using crab shells via a one-step facile procedure. The ICH with grafting degree of 1.46 and deacetylation degree of 47.68 % possessed maximum adsorption capacity of 2572.41 mg/g for silver ions (Ag(I)).The ICH also exhibited good selectivity and reusability. The adsorption followed better with the Freundlich isotherm model, while fitted well with both the Pseudo-first-order and Pseudo-second-order kinetics models. The characteristical results showed that the excellent Ag(I) adsorption capability of ICH should be attributed to both looser porous microstructure as well as additional functional groups-grafting molecular. Moreover, the Ag-loaded ICH (ICH-Ag) showed remarkable antibacterial properties against six typical pathogenic bacteria strains (Escherichia coli, Pseudomonas aeruginosa, Enterobacter aerogenes, Salmonella typhimurium, Staphylococcus aureus, and Listeria monocytogenes), with the corresponding 90 % minimal inhibitory concentrations ranged 0.426-0.685 mg/mL. Further study on the silver release, microcell morphology, and metagenomic analysis suggested that many Ag nanoparticles were formed after the Ag(I) adsorption, and the antibacterial mechanisms of the ICH-Ag involved both cell membranes destruction and intracellular metabolism disturbing. This research presented a coupling solution of crab shell wastes treatment with chitin-based bioadsorbents preparation, metal removal and recovery, as well as antibacterial agent production.

Network Pharmacology-Based Exploration on the Intervention of Qinghao Biejia Decoction on the Inflammation-Carcinoma Transformation Process of Chronic Liver Disease via MAPK and PI3k/AKT Pathway.

Chronic liver disease(CLD) is a slow-developing and long-term disease that can cause serious damage to the liver. Thus far, it has been associated with viral hepatitis, non-alcoholic fatty liver disease(NAFLD), alcoholic liver disease(ALD), hepatic fibrosis(HF), liver cirrhosis (LC), and liver cancer. Qinghao Biejia Decoction (QBD) is a classic ancient Chinese herbal prescription with strong immune-enhancing, anti-inflammatory, and anti-tumor effects. In this study, we used a network pharmacology approach to investigate the molecular mechanisms of QBD in the inflammation-carcinoma transformation process of chronic liver disease. Two key drug targets, MAPK1 and PIK3CA, were screened using network pharmacology and molecular docking techniques, revealing dihydroartemisinin, artesunate, 12-O-Nicotinoylisolineolone, caffeic acid, and diincarvilone A as active ingredients involved in QBD mechanisms. The main signaling pathways involved were the PI3K-AKT signaling pathway and MAPK signaling pathway. In summary, our results indicated that QBD affects the inflammatory transformation of chronic liver disease through MAPK1 and PIK3CA and signaling pathways MAPK and PI3K/AKT. These data provide research direction for investigating the mechanisms underlying the inflammation-carcinoma transformation process in QBD for chronic liver disease.

Qualitative and quantitative analysis of the major components in Qinghao Biejia decoction by UPLC-Orbitrap Fusion-MS/MS and UPLC-QQQ-MS/MS and evaluation of their antibacterial activities.

OBJECTIVE: In the present study, the chemical components of Qinghao Biejia decoction (QBD) were qualitatively and quantitatively analyzed using UPLC-Orbitrap Fusion-MS/MS and UPLC-QQQ-MS/MS techniques, followed by identification of each component's origin and evaluation of the antibacterial activity of QBD and its components. METHODS: High-resolution mass spectrometry was used to obtain information on the precise molecular weight, retention time, and fragmentation ion peaks of the compounds used to identify the components of QBD and establish a method for their quantification. In vitro assays including determination of the minimal inhibitory concentration and growth curves were used to assess the antibacterial activity of QBD and its components. RESULTS: A total of 39 components, including fatty acids, phenolic acids, amino acids, flavonoids, coumarins, terpenoids, and alkaloids, were identified by UPLC-Orbitrap Fusion-MS/MS. A high-performance analytical method was also established to quantify 12 components of QBD. The content of mangiferin was relatively high (estimated to be 814 µg/g). The results of the antibacterial assays indicated that mangiferin exhibits antibacterial effects against two strains causing respiratory tract infections. CONCLUSIONS: The present study suggests that mangiferin may serve as a natural compound which shows high antibacterial activity. The results can aid the discovery and analysis of the active antimicrobial components present in QBD and further provide a reference for quality assessment of multi-component herbal prescriptions.

Immune checkpoints and immunotherapy in non-small cell lung cancer: Novel study progression, challenges and solutions.

Lung cancer is the most common type of cancer with the highest mortality rate worldwide. Non-small cell lung cancer (NSCLC) accounts for ~85% of the total number of lung cancer cases. In the past two decades, immunotherapy has become a more promising treatment method than traditional treatments (surgery, radiotherapy and chemotherapy). Immunotherapy has been shown to improve the survival rate of patients and to have a superior effect when controlling lung cancer than traditional therapy. However, only a small number of patients can benefit from immunotherapy, and not all patients who qualify experience long-term benefits. In the clinic, the objective response rate of programmed cell death protein 1 treatment without the prior screening of patients is only 15-20%. Immunotherapy is associated with both opportunities and challenges for patients with NSCLC. The current challenges of immunotherapy include the lack of accurate biomarkers, inevitable resistance and insufficient understanding of immune checkpoints. In previous years, several methods for overcoming the challenges posed by immunotherapy have been proposed, but combination therapy is the most suitable choice. A large number of studies have shown that the combination of drugs can significantly improve their efficacy, compared with monotherapy, and that some therapeutic combinations have been approved by the Food and Drug Administration for the treatment of NSCLC. Traditional Chinese medicine (TCM) is a traditional medical practice in China that can play an important role in immunotherapy. Most agents used in TCM originate from plants, and have the advantages of low toxicity and multiple targets. In addition, TCM includes a unique class of drugs that can improve autoimmunity. Therefore, TCM may be a promising treatment method for all types of cancer.

Metabolomics and integrated network pharmacology analysis reveal Tricin as the active anti-cancer component of Weijing decoction by suppression of PRKCA and sphingolipid signaling.

Currently, conventional methods of treating non-small cell lung cancer (NSCLC) have many disadvantages. An alternative effective therapy with minimal adverse reactions is urgently needed. Weijing decoction (WJD), which is a classic ancient Chinese herbal prescription, has been used successfully to treat pulmonary system diseases containing lung cancer in the clinic. However, the key active component and target of Weijing decoction are still unexplored. Therefore, for the first time, our study aims to investigate the pharmacological treatment mechanism of Weijing decoction in treating NSCLC via an integrated model of network pharmacology, metabolomics and biological methods. Network pharmacology results conjectured that Tricin is a main bioactive component in this formula which targets PRKCA to suppress cancer cell growth. Metabolomics analysis demonstrated that sphingosine-1-phosphate, which is regulated by sphingosine kinase 1 and sphingosine kinase 2, is a differential metabolite in plasma between the WJD-treated group and the control group, participating in the sphingolipid signaling. In vitro experiments demonstrated that Tricin had vital effects on the proliferation, pro-apoptosis, migration and colony formation of Lewis lung carcinoma cells. Through a series of validation assays, Tricin inhibited the tumor growth mainly by suppressing PRKCA/SPHK/S1P signaling and antiapoptotic signaling. On the other hand, Weijing formula could inhibit the tumor growth and prolong the survival time. A high dosage of Tricin was much more potent in animal experiments. In conclusion, we confirmed that Weijing formula and its primary active compound Tricin are promising alternative treatments for NSCLC patients.

Acute effects of CH3NH3PbI3 perovskite on Scenedesmus obliquus and Daphnia magana in aquatic environment.

CH3NH3PbI3 is one of the most widely studied and most promising photoelectric conversion materials for large-scale application. However, once it is discharged into the aquatic environment, it will release a variety of lethal substances to the aquatic organisms. Herein, two typical aquatic pollution indicators, Scenedesmus obliquus (a typical phytoplankton) and Daphnia magna (a typical zooplankton), were used to assess the acute effects of CH3NH3PbI3 perovskite on aquatic organisms. The results showed that, when the initial CH3NH3PbI3 perovskite level (CPL) was 40 mg L-1 or higher, the growth of S. obliquus would be remarkably inhibited with significant decreases of chlorophyll content and protein content. And when the CPL was over 5 mg L-1, the survival of D. magna would be notably threatened. Specifically, the 72 h EC-50 of CH3NH3PbI3 perovskite to S. obliquus was calculated as 37.21 mg L-1, and the 24 h LC-50 of this perovskite to D. magna adults and neonates were calculated as 37.53 mg L-1 and 18.55 mg L-1, respectively. Moreover, remarkably solution pH declination and large amounts of lead bio-accumulation was observed in the both acute experiments, which could be the main reasons causing the above acute effects. Considering the strong acute effects of these CH3NH3PbI3 perovskite materials and their attractive application prospect, more attentions should be paid on their harmness to the environment.

Recyclable magnetic chitosan microspheres with good ability of removing cationic dyes from aqueous solutions.

Sr3.8Fe25.7O70.4-chitosan magnetic microparticles (Sr3.8Fe25.7O70.4-CMNs) with a core-shell structure were synthesized, characterized and applied for the removal of two model cationic dyes. The results showed that these magnetic microparticles possess fast adsorption rate and high adsorption efficiency for both crystal violet (CV) and basic red 9 (BR9) at a temperature ranging 30 °C to 40 °C and suitable pH range (pH ≥ 7). The maximum removal efficiency for CV and BR9 attained to 94.5% and 97.5% in 30 min, which was significantly faster and higher than that of chitosan (<50% in 60 min) (P<0.01). And its maximum adsorption capacity for CV and BR9 reached 29.46 mg/g and 32.16 mg/g, respectively. The adsorption process of Sr3.8Fe25.7O70.4-CMNs follows the Langmuir isotherm with a high correlation coefficient (R2 > 0.97) and the pseudo-second-order model. Additionally, the synthesized Sr3.8Fe25.7O70.4-CMNs were easy to regeneration and reuse, and the removal rate remained above 90% after 5 recycle times. This study would provide a new more environmental friendly material and method for the treatment of wastewater containing toxic dyes.

[Effect of Electroacupuncture Intervention on Rehabilitation of Upper Limb Motor Function in Patients with Ischemic Stroke].

OBJECTIVE: To observe the effect of electro-elongated needle intervention on rehabilitation of upper limb motor function in patients with ischemic stroke. METHODS: A total of 100 ischemic stroke patients were equally and randomly divided into manual acupuncture group and electro-elongated needle group (n=50 cases/group). For patients of the manual acupuncture group, filiform needles were respectively inserted into Jianyu (LI 15), Jiquan (HT 1), Shousanli (LI 10), Neiguan (PC 6) and Hegu (LI 4) on the affected side, manipulated with lifting-thrusting reinforcing or reducing method for 40 min, once daily for two weeks except the weekends, and for those of the electro-elongated needle group, elongated needles were repeatedly penetrated from LI 15 to Binao (LI 14), and from LI 10 to Waiguan (TE 5) on the affected side, followed by electrical stimulation for 40 min, once daily for two weeks except weekends. The therapeutic effect of manual acupuncture and electro-elongated needle for upper extremity rehabilitation was assessed according to the modified upper-extremity Fugl-Meyer Assessment (FMA) and Wolf Motor Function Test (WMFT) separately. The surface electromyogram (sEMG) of the deltoid muscle was recorded by using a Megawin Surface Electrogram System and the root mean square (RMS) of the amplitude of sEMG was used to evaluate the functional state of the deltoid muscle. RESULTS: After the treatment, the score of FMA and WMFT in both manual acupuncture and electro-elongated needle groups were significantly increased compared with pre-treatment in the same one group (P<0.01), suggesting an improvement of the motor function of the upper limbs, but RMS ratios of the amplitude of sEMG were evidently decreased in both groups (P<0.05, P<0.01), suggesting a relief of the abnormal muscular tension. The therapeutic effects of the electro-elongated needle were obviously superior to those of manual acupuncture in up-regulating FMA and WMFT scores and down-regulating RMS ratio (P<0.01). CONCLUSION: Electro-elongated needle therapy can effectively improve the motor function of upper limb in ischemic stroke patients and has a better effect in comparison with simple manual acupuncture.