Anti-osteoporotic effects of Yi Mai Jian on bone metabolism of ovariectomized rats.

Yi Mai Jian herbal formula (YMJ) is formulated with Eucommiae Folium, Astragali Radix, Ligustri Lucidi Fructus, and Elaeagnus Fructus to improve bone function in traditional Chinese medicine. The anti-osteoporotic effects of YMJ in bone metabolism were evaluated in ovariectomized (OVX) rats. The skeletal structure of the femur and vertebrae was analyzed after treating OVX rats with YMJ for 114 days. The results showed that YMJ significantly increased the bone mineral density (BMD) and trabecular number (Tb. N) of the femur and 5th lumbar vertebrae and reduced trabecular separation (Tb. Sp). Moreover, trabecular bone volume/total tissue volume (BV/TV), bone stiffness, and maximum femur load were significantly increased. The serum concentrations of NTX1 and PYD were significantly decreased. According to these results, YMJ could ameliorate osteoporosis in ovariectomized rats. Eucommiae Folium and Elaeagnus Fructus inhibited osteoclast differentiation, Ligustri Lucidi Fructus inhibited calcium reabsorption, Astragali Radix stimulated osteoblast proliferation, and Astragali Radix and Eucommiae Folium stimulated mineralization. Therefore, the combination of the four herbs into one formula, YMJ, could alleviate bone remodeling caused by low estrogen levels. We suggest that YMJ could be a healthy food candidate for preventing post-menopausal osteoporosis.

The chromatographic fingerprinting study and genotoxicity evaluation of aqueous extract from Cyclocarya paliurus leaves.

Cyclocarya paliurus leaf is a medicinal and edible homologous plant, which possess various bioactive components with significant health benefits. However, the quality and safety of the aqueous extract from Cyclocarya paliurus leaves (CPLAE) vary greatly due to the raw materials and preparation technology. At present, chromatographic fingerprinting has been widely used for qualitative and quantitative analysis of traditional Chinese medicine (TCM). In this study, a method combining high performance liquid chromatography (HPLC) fingerprint with quantitative analysis was established and successfully applied to the characterization and quality evaluation of the CPLAE. In addition, the genetic safety of the CPLAE was evaluated by genotoxicity tests, including Ames test, chromosomal aberration test of Chinese hamster lung (CHL) cell in vitro, and bone marrow micronucleus test in mice. The results showed that 10 batches of CPLAE samples were analyzed by high performance liquid chromatography coupled with mass spectrometry (HPLC-MS), and the similarity of chromatographic fingerprint of each batch was above 0.961, indicating good similarity. At the same time, the 6 compounds with high absorption strength in the chromatogram were quantitatively analyzed. The results showed that all 6 compounds had good regression (R2=1.000) in the test range, and the recoveries ranged from 96.25% to 102.46%. The results of the 3 genotoxicity tests showed that the highest dose of CPLAE had no genotoxicity. In conclusion, the newly established chromatographic fingerprint and multi-component quantitative analysis method is stable and accurate, and can be used for the identification and quality evaluation of the CPLAE. Moreover, the CPLAE has the characteristics of safety and high quality as functional materials in food.

A study on the effectiveness of sodium selenite in treating cadmium and perfluoro octane sulfonic (PFOS) poisoned zebrafish (Danio rerio).

Perfluoro octane sulfonate (PFOS) and cadmium (Cd) are toxic elements in the environment. As a micronutrient trace element, selenium (Se) can mitigate the adverse effects induced by PFOS and Cd. However, few studies have examined the correlation between Se, PFOS and Cd in fish. The present study focused on the antagonistic effects of Se on PFOS+Cd-induced accumulation in the liver of zebrafish. The fish was exposed to PFOS (0.08mg/L), Cd (1mg/L), PFOS+ Cd (0.08 mg/L PFOS+1 mg/L Cd), L-Se (0.07mg/L Sodium selenite +0.08mg/L PFOS+1mg/L Cd), M-Se (0.35mg/L Sodium selenite + 0.08mg/L PFOS+ 1 mg/L Cd), H-Se (1.75 mg/L Sodium selenite + 0.08 mg/L PFOS+ 1mg/L Cd) for 14d. The addition of selenium to fish exposed to PFOS and Cd has been found to have significant positive effects. Specifically, selenium treatments can alleviate the adverse effects of PFOS and Cd on fish growth, with a 23.10% improvement observed with the addition of T6 compared to T4. In addition, selenium can alleviate the negative effects of PFOS and Cd on antioxidant enzymes in zebrafish liver, thus reducing the liver toxicity caused by PFOS and Cd. Overall, the supplementation of selenium can reduce the health risks to fish and mitigate the injuries caused by PFOS and Cd in zebrafish.

Mechanical behavior and microstructure of porcine brain tissues under pulsed electric fields.

Pulsed electric fields are extensively utilized in clinical treatments, such as subthalamic deep brain stimulation, where electric field loading is in direct contact with brain tissue. However, the alterations in brain tissue's mechanical properties and microstructure due to changes in electric field parameters have not received adequate attention. In this study, the mechanical properties and microstructure of the brain tissue under pulsed electric fields were focused on. Herein, a custom indentation device was equipped with a module for electric field loading. Parameters such as pulse amplitude and frequency were adjusted. The results demonstrated that following an indentation process lasting 5 s and reaching a depth of 1000 µm, and a relaxation process of 175 s, the average shear modulus of brain tissue was reduced, and viscosity decreased. At the same amplitude, high-frequency pulsed electric fields had a smaller effect on brain tissue than low-frequency ones. Furthermore, pulsed electric fields induced cell polarization and reduced the proteoglycan concentration in brain tissue. As pulse frequency increased, cell polarization diminished, and proteoglycan concentration decreased significantly. High-frequency pulsed electric fields applied to brain tissue were found to reduce impedance fluctuation amplitude. This study revealed the effect of pulsed electric fields on the mechanical properties and microstructure of ex vivo brain tissue, providing essential information to promote the advancement of brain tissue electrotherapy in clinical settings.

Spectral Characteristics and Functional Responses of Phospholipid Bilayers in the Terahertz Band.

Understanding the vibrational information encoded within the terahertz (THz) spectrum of biomolecules is critical for guiding the exploration of its functional responses to specific THz radiation wavelengths. This study investigated several important phospholipid components of biological membranes-distearoyl phosphatidylethanolamine (DSPE), dipalmitoyl phosphatidylcholine (DPPC), sphingosine phosphorylcholine (SPH), and lecithin bilayer-using THz time-domain spectroscopy. We observed similar spectral patterns for DPPC, SPH, and the lecithin bilayer, all of which contain the choline group as the hydrophilic head. Notably, the spectrum of DSPE, which has an ethanolamine head group, was different. Interestingly, density functional theory calculations confirmed that the absorption peak common to DSPE and DPPC at approximately 3.0 THz originated from a collective vibration of their similar hydrophobic tails. Accordingly, the cell membrane fluidity of RAW264.7 macrophages with irradiation at 3.1 THz was significantly enhanced, leading to improved phagocytosis. Our results highlight the importance of the spectral characteristics of the phospholipid bilayers when studying their functional responses in the THz band and suggest that irradiation at 3.1 THz is a potential non-invasive strategy to increase the fluidity of phospholipid bilayers for biomedical applications such as immune activation or drug administration.

Artemisinin and Ambrosia trifida extract aggravate the effects of short freeze-thaw stress in winter rye (Secale cereale) seedlings.

The freeze-thaw and allelopathy from alien giant ragweed (Ambrosia trifida L.) and artemisinin have led to a serious stress to plants, influencing the agricultural quality and crop yield in north-east China. Yet, little is known how allelopathy affect plants under the freeze-thaw process. In this study, the characteristics in winter rye (Secale cereale L.) seedlings were investigated by laboratory simulation. The results showed that during the freezing process, application of artemisinin and A. trifida extract significantly increased the soluble protein content and accelerated lipid peroxidation, while they significantly inhibited antioxidant enzymes, photosynthesis and respiration (P <0.05). During the thawing process, the freezing pressure decreased, and activities of antioxidant enzymes were significantly improved to mitigate artemisinin and A. trifida extract induced stress (P <0.05). In addition, the sensitivity of the investigated metabolic processes in winter rye seedlings were highest to artemisinin and A. trifida extract in the freezing process. This study suggested that the stress response induced by artemisinin and A. trifida extract on winter rye seedlings in the freezing process was greater than that in the thawing process.

Modeling nutrient flows from land to rivers and seas - A review and synthesis.

Water quality modeling facilitates management of nutrient flows from land to rivers and seas, in addition to environmental pollution management in watersheds. In the present paper, we review advances made in the development of seven water quality models and highlight their respective strengths and weaknesses. Afterward, we propose their future development directions, with distinct characteristics for different scenarios. We also discuss the practical problems that such models address in the same region, China, and summarize their different characteristics based on their performance. We focus on the temporal and geographical scales of the models, sources of pollution considered, and the main problems that can be addressed. Summary of such characteristics could facilitate the selection of appropriate models for resolving practical challenges on nutrient pollution in the corresponding scenarios globally by stakeholders. We also make recommendations for model enhancement to expand their capabilities.

The collective vibrational modes of dihydropyridine in nifedipine studied by terahertz spectroscopy.

Cardiovascular pharmaceuticals have drawn huge attention in drug development. Nifedipine (NFD) is an important member of calcium channel blockers (CCB) with the structural characteristic of dihydropyridine (DHP), but the binding mechanism to its target remains an open question. Even though several analytical techniques have been used for structural characterizations, the information of collective vibrational behavior is still lacking. In this work, we use terahertz (THz) spectroscopy to investigate the spectral fingerprints of NFD, and quantitatively evaluate the temperature-induced frequency shifts. Combined with quantum chemical calculations, each THz fingerprint is attributed to specific collective vibrational modes. The collective vibrations of DHP are mainly distributed below 2.5 THz, which provides complementary information to understand the behavior of rigid DHP ring. The rotation of methyl group and the wagging of nitrophenyl group are widely distributed in the range of 1.0-4.0 THz, which is helpful for the conformational recognition between NFD and target molecule. THz spectroscopy is demonstrated to be suitable for characterizing the collective vibrational modes of DHP and elucidating the drug-target binding behavior from the perspective of noncovalent interactions. It has the potential to become a non-invasive technology for conformational analysis and pharmaceutical development.

Qu-Yu-Jie-Du Decoction Ameliorates Dextran Sulfate Sodium-Induced Colitis in Mice by Modulation of Neutrophils and Macrophage Infiltration.

Background: Inflammatory bowel disease (IBD) is becoming a global disease. A percentage of IBD patients will not react to therapy or will lose their response. Qu-Yu-Jie-Du Decoction (QYJD) is a traditional Chinese medicine formula commonly used for intestinal diseases. It has been reported that QYJD has an anti-inflammatory effect, but the mechanism is not fully understood. In this study, we mainly evaluated the anti-inflammatory effect of QYJD and explored the possible mechanisms. Methods: Twenty-four BALB/c mice were randomly divided into 4 groups according to their body weight, namely, the control group, the dextran sulfate sodium (DSS) group, the DSS + QYJD group, and the QYJD group. Mice were given 3% DSS drinking water freely, and at the same time, mice were given normal saline or QYJD (4.44 mg/g/d), respectively. Mental state, faeces, and weight were recorded every day. On the 10th day, the mice were sacrificed and collected for investigation. The length of the mice colon was measured. Histological analysis was used to detect the morphological changes in the colon. Immunohistochemistry was used to measure the infiltration of macrophages (F4/80, CD163) and neutrophils (Ly6G). Colorimetry was used to detect the myeloperoxidase (MPO) activity of colon tissues. ELISA was utilized to detect associated inflammatory cytokines and chemokines in colon tissues. Results: QYJD alleviated the weight loss and colitis symptoms of mice caused by DSS. QYJD fought against the shortening of the intestine caused by DSS; that is, it improved the decline of intestinal compliance in mice and had a protective effect on colon tissues. The mechanisms were related to downregulating macrophages and neutrophils in colon tissues of infiltration. Besides, QYJD simultaneously reduced the activity of myeloperoxidase activity (MPO) and the contents of IL-1ß, IL-6, TNF-α, TGF-ß, CCL2, and CXCL2 in colon tissues. Conclusions: QYJD can ameliorate DSS-induced colitis in mice and the mechanism is connected with a reduction in neutrophil and macrophage infiltration.

Effects of Different Types of Electric Fields on Mechanical Properties and Microstructure of Ex Vivo Porcine Brain Tissues.

Electrotherapy plays a crucial role in regulating neuronal activity. Nevertheless, the relevant therapeutic mechanisms are still unclear; thus, the effects of electric fields on brain tissue's mechanical properties and microstructure need to be explored. In this study, focusing on the changes in mechanical properties and microstructure of ex vivo porcine brain tissues under different types of electric fields, directional and alternating electric fields (frequencies of 5, 20, 50, and 80 Hz, respectively) integrate with a custom-designed indentation device. The experimental results showed that for the ex vivo brain tissue, the directional electric field (DEF) can reduce the elastic properties of brain tissue. Simultaneously, the DEF can increase the cell spacing and reduce the proteoglycan content. The transmission electron microscope (TEM) analysis observed that the DEF can reduce the integrity of the plasma membrane, the endoplasmic reticulum's stress response, and the myelin lamella's separation. The alternating electric field (AEF) can accelerate the stress relaxation process of brain tissue and change the time-dependent mechanical properties of brain tissue. Meanwhile, with the increase in frequency, the cell spacing decreased, and the proteoglycan content gradually approached the control group without electric fields. TEM analysis observed that with the increase in frequency, the integrity of the plasma membrane increases, and the separation of the myelin lamella gradually disappears. Understanding the changes in the mechanical properties and microstructure of brain tissue under AEF and DEF enables a preliminary exploration of the therapeutic mechanism of electrotherapy. Simultaneously, the essential data was provided to support the development of embedded electrodes. In addition, the ex vivo experiments build a solid foundation for future in vivo experiments.

Clinical treatment of lumbar disc herniation.

OBJECTIVE: To investigate the clinical effect of radiofrequency ozone and injection of anti-inflammatory analgesic solution into the internal orifice of nerve root combined with traditional Chinese medicine hook operation in the treatment of lumbar disc herniation. METHODS: Patients with lumbar disc herniation who were admitted to our hospital on December 20, 2017 and June 19, 2019 were selected as the main research objects, and the included patients were divided into control group, basic group and comprehensive group by random number table method. Control group was treated with radiofrequency ozone therapy, basic group was treated with injection of anti-inflammatory analgesic solution into the internal orifice of nerve root in addition to the control group, comprehensive group was treated with traditional Chinese medicine hook operation in addition to the basic group. The clinical treatment effects were observed. RESULTS: A total of 153 patients were included in this study, including 40 in the control group, 40 in the basic group, and 73 in the comprehensive group. The results showed that the NRS scores of control group were 3±0.98, 2±0.93 and 2±0.85 at 1 month, 3 months and 1 year after treatment, respectively. NRS scores in the basic group were 3±0.18, 2±0.33, and 2±0.15, respectively. NRS scores in the comprehensive group were 2±0.78, 1±0.54, and 1±0.77, respectively. Compared with the control group, there were significant differences in basic group and comprehensive group at each time point (P < 0. 05). At the same time, compared with the basic group, the NRS score of the comprehensive group was statistically different (P < 0.05). CONCLUSION: Radiofrequency ozone and injection of anti-inflammatory analgesic solution into the internal orifice of nerve root combined with hook operation can obtain good short-term and medium-term effects in the treatment of lumbar disc herniation. It is a safe and effective minimally invasive treatment method. KEY WORDS: Internal orifice of nerve root, Lumbar disc herniation, Ozone.

Effects of ultrasonic on structure, chain conformation and morphology of pectin extracted from Premna microphylla Turcz.

In this study, ultrasonic effects on structure, chain conformation and morphology of pectin extracted from Premna microphylla Turcz (PEP) and its probable mechanism were investigated. In the process of ultrasonic treatments, the chains of PEP were fractured rapidly within the initial 10 min and then the degradation rate gradually slowed down. The primary structure of PEP nearly remained unchanged after ultrasonic degradation. The rigid semi-flexible chains of PEP were converted into flexible chains, flexible coils, even compact coils. Sonication at low intensity for short time made PEP molecular chains curly collapse and tighten up. Long duration sonication at high intensity generated excessive small rigidness segments that mutually aggregated because of hydrogen bonds and inhibited the self-coiling of PEP chains. Atomic force microscopy (AFM) analysis supported the conformation transition of PEP chains. The results provided a fundamental basis for orientation design and process control of PEP structure.

Impact of repeated freeze-thaw cycles environment on the allelopathic effect to Secale cereale L. seedlings.

Allelopathy, as environmental stress, plays a prominent role in stress ecotoxicity, and global warming directly increases freeze-thaw cycles (FTCs) frequency in the winter. Yet, the effect between FTCs environment and allelopathy stress is rarely known, and the interaction of allelopathy stresses lacks consideration. Here, we addressed interactions between artemisinin stress (AS) and A. trifida extract stress (AES) under Non-FTCs and FTCs environments. The results found that AS and AES had an antagonistic relation under Non-FTCs environment, while a strong synergism and cooperation under FTCs environment affect the growth and physiology in S. cereale seedlings. Besides, AS and AES under FTCs environment had more inhibition on the growth of roots and shoots, chlorophylls, photosynthetic parameters, and relative water content; while more promotion on malondialdehyde, soluble sugar, and soluble protein. Moreover, the antioxidant enzyme activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX) were increased by AS and AES, showing a good resistance of S. cereale seedlings to allelopathy stress, but FTCs environment significantly weakened this resistance. Thus, the allelopathic effect of AS and AES on S. cereale seedlings was significantly emphasized by FTCs environment.

Cropland degradation and nutrient overload on Hainan Island: A review and synthesis.

As the only "tropical base of agricultural production" in China, Hainan lsland is vigorously developing high-value agriculture and is becoming the province with the highest proportion of cash crops. However, this intensive farming with large nutrient inputs has caused cropland degradation, nitrogen (N) and phosphorus (P) overloads and water pollution, which have been reversed to initiate the construction of free trade ports. Here, we systematically review the status, driving factors, and environmental impacts of cropland degradation and nutrient overload with quantified evaluations and compared with other global tropics. Over the last 30 years, the soil pH in Hainan decreased by 0.3 units, and the soil organic carbon (SOC) decreased by 20%. This soil degradation has consequently aggravated nutrient losses, caused low use efficiency, and has required farmers add additional large nutrient to maintain harvests. P overuse is more serious than N overuse in Hainan due to the misuse of high P content compound fertilizers. The current N and P usage densities were 4% and 66% higher than the national average per crop season, i.e., 301 kg N ha-1 and 98 kg P ha-1, respectively, and the application rates were even higher for vegetables, i.e., 43% and 115% higher than the national average for vegetables. Consequently, water quality degradation occurred. The nutrient contents of several estuaries have exceeded the Class III standards. Potential improvement strategies are proposed: (i) Organic materials must be recycled to curb the declines in SOC and pH, and more benefits would be obtained by together use of biochar. (ii) Nutrient quotas must be implemented to balance nutrient budgets and reduce excessive surpluses and losses. (iii) The service functions of ecological protection zones for water and soil conservation must be strengthened. These strategies also apply to other global tropics that face similar challenges of soil and ecological degradation.

Study on phenolic acids of Lonicerae japonicae Flos based on ultrahigh performance liquid chromatography-tandem mass spectrometry combined with multivariate statistical analysis.

Lonicerae japonicae Flos, a traditional Chinese medicine, has the function of evacuating heat and detoxifying. To promote the optimization of Lonicerae japonicae Flos germplasms and improve the quality of medicinal materials, 55 batches of five Lonicerae japonicae Flos germplasms with the same origin were collected during different periods, a UHPLC-TOF-MS method was established, and 22 kinds of phenolic acids were found and qualitatively analysed. Seventeen phenolic acids were selected for quantitative analysis by UHPLC-QqQ-MS/MS, and the quantitative results were analysed by principal component analysis, orthogonal partial least squares-discriminant analysis, and partial least squares discriminant analysis. The contents of phenolic acids in periods S1-S6 were found to be significantly different. There were also significant differences in the accumulation of phenolic acids in Lonicerae japonicae Flos during different growth periods. Ferulic acid, 5-O-caffeoylquinic acid, and caffeic acid were determined to be important components to distinguish the different growth periods of Lonicerae japonicae Flos. There were significant differences in the phenolic acid content of different germplasms of Lonicerae japonicae Flos, and the total amount of 17 phenolic acids and total acids (chlorogenic acid, 3,5-di-O-caffeoylquinic acid, and 4,5-di-O-caffeoylquinic acid) in "Hua Jin No. 6" was highest, so the quality of "Hua Jin No. 6" was better than that of the four other germplasms. In addition, chlorogenic acid methyl ester and caffeic acid were the smallest markers in combination to distinguish the five germplasms of Lonicerae japonicae Flos.

The effects of electric fields on the mechanical properties and microstructure of ex vivo porcine brain tissues.

As a popular tool for regulating the physiological conditions of the brain and treating brain diseases, electrotherapy has become increasingly mature in clinical applications. However, the mechanical properties and microstructure of the brain that change with weak electric fields are often overlooked. Thus, the mechanical behaviors of the brain tissue, which play a critical role in modulating the brain form and brain function, need to be taken into account. Herein, the direct current electric fields were combined with a customized indentation device and simultaneously focused on the changes in the mechanical properties and microstructure of ex vivo porcine brain tissues under electric fields. The experimental results showed that the electric fields reduced the shear modulus and viscosity and increased the relaxation rate of ex vivo porcine brain tissues. Moreover, electric fields polarized the cell bodies and reduced proteoglycan content in the cortex. The TEM observation confirmed that the electric fields deepened the degree of endoplasmic reticulum expansion and decreased the structural integrity of the cell membrane and myelin sheath. This study confirmed the effect of electric fields on ex vivo brain tissues; concurrently, it created comparable space in microscopic structure/compositions and mechanical parameters for future deeper brain experiments under stress-electric field coupling.

Spatiotemporal characteristics of agricultural nitrogen and phosphorus emissions to water and its source identification: A case in Bamen Bay，China.

The spatiotemporal characteristics and sources identification of agricultural nitrogen (N) and phosphorus (P) emissions to the gulf are rarely reported in tropical regions of China, mainly due to the lack of local reliable data and quantitative tools for spatiotemporal changes. In this study, we constructed a high-resolution NUFER (NUtrient Flow in food chains, Environment and Resources use) model based on geology, meteorology, land use data, statistical data, and field investigation to quantify the spatiotemporal characteristics and sources of N and P emissions. Bamen Bay (BMB), a bay with a mangrove national wetland Park in the Hainan Island, China, was chosen as a case study. The results showed that agricultural N emission to water in 2018 increased fivefold compared to 1990. Leaching was the main method of agricultural N emission and was mainly distributed in farms in the west and north. The contribution of N emission from crop system to water increased 20.3% in 28 years. Poultry and fruits have contributed the most to N output, and the trend is continuing. P emission to water increased sevenfold compared 1990. The contribution of P emission from animal system to water increased from 86.8% in 1990 to 90.1% in 2018 due to low removal rate of livestock manure. P emission was mainly via direct discharge of manure, mainly distributed in livestock breeding sites near the bay. Poultry has consistently contributed the most to P output in 28 years, accounting for 49.1% in 2018. Fertilizers and fodder were the largest sources of N and P. The average N and P loss rates of BMB were 5.32 t km2 yr-1 and 0.26 t km2 yr-1. The future agricultural transformation is essential, and it is necessary to reduce the application of N fertilizer and increase the removal rate of livestock manure. These results can provide reference for other typical agricultural pollution bays in exploring the spatiotemporal characteristics of N and P emissions to water and the identification of agricultural sources.

He-Chan Pian inhibits the metastasis of non-small cell lung cancer via the miR-205-5p-mediated regulation of the GREM1/Rap1 signaling pathway.

BACKGROUND: He-Chan Pian (HCP), a traditional Chinese medicinal formula, shows promising efficacy for the treatment of lung cancer. PURPOSE: Gremlin (GREM1) plays an important role in gastrointestinal tumor metastasis; however, little is known about its role in lung cancer. We determined the mechanism underlying the protective effect of HCP against metastasis in a mouse model of non-small cell lung cancer (NSCLC) and demonstrated the role of GREM1. METHODS: Ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) was used to analyze the herbal components and metabolites from the serum of HCP-treated mice. The tumor, liver, and kidney were examined histologically, and the antitumor effects and toxicity of HCP were evaluated. Levels of epithelial-mesenchymal transition (EMT)-associated transcription factors were measured using western blotting in tumors from five groups (i.e., model, HCP [L], HCP [M], HCP [H], and positive control [cisplatin, DDP]). Differentially expressed proteins and genes were identified using protein chip and sequencing analyzes, respectively. Short hairpin RNAs and overexpression plasmids were introduced into cells to evaluate the effects of GREM1. To evaluate proliferation, migration, and invasion, the expression levels of proteins involved in the Rap1 pathway and EMT were measured in vitro. Xenograft tumors with overexpression-GREM1 (OE-GREM1) in A549 cells were examined for cell proliferation. A dual-luciferase assay was performed to verify the direct interaction of GREM1 with miR-205-5p in lung cancer. RESULTS: Thirty-six ingredients and bioactive constituents detected in the serum of HCP-treated mice were identified as the key compounds involved in the inhibition of tumor growth. Animal experiments revealed that HCP significantly decreased tumor volumes and had no adverse effects on the liver or kidney or side effects. GREM1 upregulation was closely related to tumor metastasis and was regulated by miR-205-5p, as confirmed using a dual-luciferase reporter assay. OE-GREM1 promoted A549 cell migration and invasion, promoted EMT, and increased the expression of Rap1 pathway intermediaries, whereas shGREM1 had the opposite effects. Furthermore, the effects of OE-GREM1 on proliferation in the A549 xenograft mouse model were attenuated, although HCP has an inhibitory effect on tumors. CONCLUSION: Our results suggest that HCP contributes to the inhibition of NSCLC metastasis via the Gremlin/Rap1 signaling pathway regulated by miR-205-5p.

Integrative Analysis of MALT1 as a Potential Therapeutic Target for Prostate Cancer and its Immunological Role in Pan-Cancer.

Background: Mucosa-associated lymphoma antigen 1 (MALT1) is an oncogene in subsets of diffuse large B cell lymphoma (DLBCL) and mucosa-associated lymphoid tissue type (MALT) lymphoma. However, the role of MALT1 across cancers, especially in prostate cancer is still poorly understood. Methods: Here, we used several public datasets to evaluate MALT1 expression. Then, PCa cell lines and nude mice were used to investigate the cellular functions in vitro and in vivo. Microarray data were downloaded from The Cancer Genome Atlas and MALT1 was subjected to gene set enrichment analysis (GSEA) and Gene Ontology (GO) analysis to identify the biological functions and relevant pathways. Additionally, the correlations between MALT1 expression and mismatch repair (MMR) gene mutation, immune checkpoint gene expression, tumor mutational burden (TMB), and microsatellite instability (MSI) were investigated by Pearson correlation analysis. Moreover, the correlation between MALT1 expression and tumor immune infiltration was analyzed by the Tumor Immune Evaluation Resource (TIMER) database. Results: MALT1 overexpression was significantly correlated with MMR gene mutation levels and crucially promoted proliferation and colony genesis while reducing PCa cell apoptosis levels in vivo and in vitro. MALT1 expression showed strong correlations with immune checkpoint genes, TMB, and MSI in most cancers. The GO analysis indicated that MALT1-coexpressed genes were involved in heterotypic cell-cell adhesion, actin filament-based movement regulation, and action potential regulation. GSEA revealed that MALT1 expression was associated with several signaling pathways, including the NF-κB signaling, Wnt/ß-catenin and TGF-ß signaling pathways, in PCa. Additionally, MALT1 expression was significantly correlated with the infiltration of immune cells, including B cells, CD8+ T cells, dendritic cells and macrophages, and negatively correlated with CD4+ cell infiltration in PCa. Conclusion: MALT1 expression is higher in pancancer samples than in normal tissues. MALT1 promoted proliferation and colony genesis while reducing PCa cell apoptosis levels, and MALT1 suppression could inhibit xenograft tumor establishment in nude mice. Furthermore, MALT1 expression is closely related to the occurrence and development of multiple tumors in multiple ways. Therefore, MALT1 may be an emerging therapeutic target for a variety of cancers especially PCa.

Identification of Chemical Components of Qi-Fu-Yin and Its Prototype Components and Metabolites in Rat Plasma and Cerebrospinal Fluid via UPLC-Q-TOF-MS.

Qi-Fu-Yin, a traditional Chinese medicine formula, has been used to treat Alzheimer's disease (AD, a neurodegenerative disorder) in clinical setting. In this study, the chemical components of Qi-Fu-Yin and its prototype components and metabolites in rat plasma and cerebrospinal fluid, after oral administration, were preliminarily characterized via ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS). A total of 180 compounds, including saponins, flavonoids, organic acids, sucrose esters, oligosaccharide esters, phthalides, phenylethanoid glycosides, alkaloids, xanthones, terpene lactones, ionones, and iridoid glycoside, were tentatively characterized. For the first time, 51 prototypical components and 26 metabolites, including saponins, phthalides, flavonoids, sucrose esters, organic acids, alkaloids, ionones, terpene lactones, iridoid glycoside, and their derivatives, have been tentatively identified in the plasma. Furthermore, 10 prototypical components (including butylidenephthalide, butylphthalide, 20(S)-ginsenoside Rh1, 20(R)-ginsenoside Rh1, and zingibroside R 1) and 6 metabolites were preliminarily characterized in cerebrospinal fluid. These results were beneficial to the discovery of the active components of Qi-Fu-Yin anti-AD.