Erzhiwan Ameliorates Restraint Stress- and Monobenzone-Induced Depigmentation in Mice by Inhibiting Macrophage Migration Inhibitory Factor and 8-Hydroxy-2-Deoxyguanosine.

Purpose: Vitiligo is an autoimmune disease that results in the loss of epidermal melanocytes. The treatments for patients with vitiligo remain lacking. Erzhiwan (EZW), a traditional Chinese Medicine composed of Ligustri Lucidi Fructus and Ecliptae Herba, was used to ameliorate depigmentation since ancient China. This study aims to investigate the effect of EZW on vitiligo-related depigmentation. Methods: A vitiligo-related depigmentation mouse model was induced by monobenzone and restraint stress. The experimental depigmentation mice were treated with EZW. Histological observation of skin was conducted. Cutaneous oxidative damage and inflammation were determined. A network pharmacology analysis was carried out. Results: EZW reduced depigmentation score (p<0.01), cutaneous inflammatory infiltration (p<0.01), and CD8α-positive expression (p<0.01), and increased cutaneous melanin content in experimental depigmentation mice. EZW reduced stress reaction in experimental depigmentation mice (p<0.01). EZW inhibited 8-hydroxy-2-deoxyguanosine (8-OHdG)-related DNA oxidative damage in the skin (p<0.05, p<0.01). In addition, EZW reduced cutaneous macrophage migration inhibitory factor (MIF)-CD74-NF-κB signaling (p<0.01). The network pharmacology analysis demonstrated that EZW regulated necroptosis, apoptosis, and FoxO signaling pathways in vitiligo. An in vitro experiment showed that the main ingredient of EZW, specnuezhenide, protected against monobenzone and MIF-induced cell death in HaCaT cells (p<0.01). Conclusion: EZW ameliorates restraint stress- and monobenzone-induced depigmentation via the inhibition of MIF and 8-OHdG signaling. The findings provide a data basis of an utilization of EZW in vitiligo.

Exploring the effects and mechanism of peony pollen in treating benign prostatic hyperplasia.

Paeonia suffruticosa is widely cultivated globally due to its medicinal and ornamental value. Peony pollen (PP) is commonly used in Chinese folk medicine to make tea to treat benign prostatic hyperplasia (BPH), but its molecular mechanism against BPH is yet to be comprehended. The objective of this research was to experimentally verify the effect of PP in the treatment of BPH and to preliminarily reveal its mechanism of action on BPH using network pharmacology methods. The results revealed that PP could decrease prostate volume and prostate index, serum testosterone (T), dihydrotestosterone (DHT), and estradiol (E2) levels. Moreover, it could improve prostate tissue structure in BPH model animals as well. Additionally, database searches and disease target matching revealed 81 compounds in PP. Of these, 3, 7, 8, 2'-tetrahydroxyflavone, Chrysin, Wogonin, Limocitrin, and Sexangularetin were the top five compounds associated with the therapeutic effects of BPH. Furthermore, 177 therapeutic targets for BPH were retrieved from databases of Swiss Target, DisGeNET, Drugbank, Genecards, OMIM, TTD, and Uniprot. In contrast, core targets AKT1, EGFR, IL6, TNF, and VEGFA were obtained by PPI network diagram. Molecular docking also showed that the main efficacy components and potential core targets in PP had good binding capacity. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomics (KEGG) analysis established that the effect of PP in BPH therapy was mainly through regulating the expression levels of protein kinase B on phosphatidylinositol 3-kinase and phosphatidylinositol 3-kinase-protein kinase B pathways. Additionally, Western blot experiments also exhibited a significant elevation in the activated PI3K and AKT proteins in the model (Mod) group relative to the control (Con) group, and the expression of these activated proteins was significantly reduced after PP administration. In summary, this research provides a scientific basis for employing PP to treat BPH, preliminarily reveals its mechanism of action and potential targets, and lays the foundation for further research and development.

Purification, Identification, Chelation Mechanism, and Calcium Absorption Activity of a Novel Calcium-Binding Peptide from Peanut (Arachis hypogaea) Protein Hydrolysate.

A novel calcium-binding peptide was purified from peanut protein hydrolysate using gel filtration chromatography and identified using HPLC-MS/MS. Its amino acid sequence was determined as Phe-Pro-Pro-Asp-Val-Ala (FPPDVA, named as FA6) with the calcium-binding capacity of 15.67 ± 0.39 mg/g. Then, the calcium chelating characteristics of FPPDVA were investigated using ultraviolet-visible absorption spectroscopy, fluorescence spectroscopy, Fourier transform infrared spectroscopy, particle size, and zeta potential. The results showed that FPPDVA interacted with calcium ions, the chelation of calcium ions induced FPPDVA to fold and form a denser structure, the calcium-binding sites may mainly involve oxygen atoms from the carboxyl residues of Asp and Ala, and Phe possessed contact energy and carbonyl residues of Val. Microstructure analysis showed that FPPDVA-calcium chelate exhibited a regularly ordered and tightly aggregated sheets or block structures. Additionally, FPPDVA-calcium chelate had good gastrointestinal digestive stability and thermal stability. The results of everted rat intestinal sac and Caco-2 cell monolayer experiments showed that FPPDVA-calcium chelate could promote calcium absorption and transport through the Cav1.3 and TRPV6 calcium channels. These data suggest that FPPDVA-calcium chelate possesses the potential to be developed and applied as calcium supplement.

Study on the mechanism of Danshen-Guizhi drug pair in the treatment of ovarian cancer based on network pharmacology and in vitro experiment.

Our study aims to explore the active components and mechanisms of the Danshen-Guizhi drug pair in treating ovarian cancer by network pharmacology and in vitro experiment. The "component-target-pathway" diagram of the Danshen-Guizhi drug pair was established by network pharmacology, and the effective active components, important targets as well as potential mechanisms of the Danshen-Guizhi drug pair were analyzed. The predicted results were verified by molecular docking and in vitro experiments. The main active components of the Danshen-Guizhi drug pair in the treatment of ovarian cancer are salviolone, luteolin, ß-sitosterol and tanshinone IIA. The main core target is PTGS2. The pathways involved mainly include the cancer pathway, PI3K-Akt signaling pathway, and IL-17 signaling pathway. The molecular docking results showed that salviolone and tanshinone IIA had good binding ability to the target. The expression of PTGS2 mRNA and PGE2 in ovarian cells were significantly inhibited by salviolone. The mechanism of the Danshen-Guizhi drug pair in the treatment of ovarian cancer may be regulating cell proliferation, apoptosis and tumor immunity. This provides a theoretical basis for the clinical development and application of the Danshen-Guizhi drug pair.

Peptides from walnut (Juglans mandshurica Maxim.) protect hepatic HepG2 cells from high glucose-induced insulin resistance and oxidative stress.

Oxidative stress is an important factor in the pathogenesis of insulin resistance (IR) in type 2 diabetes mellitus (T2DM). Bioactive peptides from nuts have been shown to alleviate oxidative stress and IR. However, the specific mechanisms underlying their activity remain unclear. This study investigated the protective effects of three novel peptides derived from Juglans mandshurica Maxim., LVRL, LRYL, and VLLALVLLR, against high glucose-induced IR and oxidative stress in HepG2 cells. The possible mechanisms underlying these effects were also elucidated. The walnut-derived peptides improved glucose consumption, glucose uptake, and glucose transporter type 4 (GLUT4) translocation, and elevated the phosphorylation of insulin receptor substrate 1 (IRS-1), phosphatidylinositol 3-kinase (PI3K), and protein kinase B (Akt). They also increased the activities of glutathione peroxidase (GSH-Px), catalase (CAT), and superoxide dismutase (SOD), the nuclear transport of nuclear factor E2-related factor 2 (Nrf2), and the protein expression of heme oxygenase-1 (HO-1). Furthermore, the walnut-derived peptides reduced high glucose-induced ROS overproduction and the phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. These results suggested that walnut-derived peptides protect HepG2 cells from high glucose-induced IR and oxidative stress by activating IRS-1/PI3 K/Akt and Nrf2/HO-1 signaling pathways.

[Effect of application of herbal medicine paste to "Tianshu" (ST25)on intestinal mobility and expression of vasoactive intestinal peptide and substance P in colonic myenteric plexus in rats with functional constipation].

OBJECTIVE: To observe the effect of acupoint application at bilateral "Tianshu" (ST25) on intestinal mobility and immunoactivity of vasoactive intestinal peptide (VIP) and substance P (SP) in colonic myenteric plexus of rats with functional constipation (FC), so as to analyze its mechanisms underlying improving FC. METHODS: Forty male SD rats were randomly divided into 4 groups, namely normal control, model, acupoint application and medication, with 10 rats in each group. The FC model was established by gavage of Loperamide Hydrochloride suspension fluid (0.5 mg/mL, 3 mg·kg-1·d-1) for 7 days. Herbal medicine paste (composed of Rheum Officinale, Sodium Sulfate, Mangnolia Officinalis, etc.) was applied to bilateral ST25 for 6 h, once daily for 4 weeks. Rats of the medication group were treated by gavage of Mosapride suspension fluid (0.15 mg/mL, 1.58 mg·kg-1·d-1) for 4 weeks. After the treatment, the rats were deprived of water for 12 hours, and then treated by gavage of 2 mL of activated carbon suspension, followed by recording the first black defecation time and the number of fecal particles and water content of feces within 6 h so as to assess the intestinal mobility. The immunoactivity and average surface density of VIP and SP positive granules in the colonic myenteric plexus were detected by immunohistochemistry. RESULTS: Compared with the normal control group, the first black defecation time was significantly prolonged, and the number and water content of fecal particles within 6 h, and the expression and the average surface density of VIP and SP were significantly reduced in the model group (P<0.01). After the treatment and compared with the model group, the first black defecation time was shortened, and the fecal water content and fecal particle number within 6 h, as well as the expression and the average surface density of VIP and SP were considerably increased in both acupoint application and medication groups (P<0.01). There were no significant differences between the acupoint application and medication groups in all the indexes mentioned above after the interventions (P>0.05). CONCLUSION: Acupoint application may improve the intestinal motility in FC rats, which may be asso-ciated with its effects in up-regulating the immunoactivity of VIP and SP in colonic myenteric plexus of the large intestine.

Kinetic and equilibrium of U(VI) biosorption onto the resistant bacterium Bacillus amyloliquefaciens.

This study evaluated U(VI) biosorption properties by the resistant bacterium, Bacillus amyloliquefaciens, which was isolated from the soils with residual radionuclides. The effect of biosorption factors (uptake time, pH, ionic concentration, biosorbent dosage and temperature) on U(VI) removal was determined by batch experiments. The uptake processes were characterized by using SEM, FTIR, and XPS. The experimental data of U(VI) biosorption were fitted by the pseudo-second-order. The maximum uptake capacity was 179.5 mg/g at pH 6.0 by Langmuir model. The thermodynamic results: ΔGо, ΔHо and ΔSо for uptake processes were calculated as -6.359 kJ/mol, 14.20 kJ/mol and 67.19 J/mol/K, respectively. The results showed that the biosorption of Bacillus amyloliquefaciens will be an ideal method to remove radionuclides.

Arid1a regulates response to anti-angiogenic therapy in advanced hepatocellular carcinoma.

BACKGROUND & AIMS: AT-rich interaction domain 1a (Arid1a), a component of the chromatin remodeling complex, has emerged as a tumor suppressor gene. It is frequently mutated in hepatocellular carcinoma (HCC). However, it remains unknown how Arid1a suppresses HCC development and whether Arid1a deficiency could be exploited for therapy, we aimed to explore these questions. METHODS: The expression of Arid1a in human and mouse HCCs was determined by immunohistochemical (IHC) staining. Gene expression was determined by quantitative PCR, ELISA or western blotting. Arid1a knockdown HCC cell lines were established by lentiviral-based shRNA. Tumor angiogenesis was quantified based on vessel density. The regulation of angiopoietin (Ang2) expression by Arid1a was identified by chromatin immunoprecipitation (ChIP) assay. The tumor promoting function of Arid1a loss was studied with a xenograft model in nude mice and diethylnitrosamine (DEN)-induced HCC in Arid1a conditional knockout mice. The therapeutic values of Ang2 antibody and sorafenib treatment were evaluated both in vitro and in vivo. RESULTS: We demonstrate that Arid1a deficiency, occurring in advanced human HCCs, is associated with increased vessel density. Mechanistically, loss of Arid1a causes aberrant histone H3K27ac deposition at the angiopoietin-2 (Ang2) enhancer and promoter, which eventually leads to ectopic expression of Ang2 and promotes HCC development. Ang2 blockade in Arid1a-deficient HCCs significantly reduces vessel density and tumor progression. Importantly, sorafenib treatment, which suppresses H3K27 acetylation and Ang2 expression, profoundly halts the progression of Arid1a-deficient HCCs. CONCLUSIONS: Arid1a-deficiency activates Ang2-dependent angiogenesis and promotes HCC progression. Loss of Arid1a in HCCs confers sensitivity to Ang2 blockade and sorafenib treatment. LAY SUMMARY: AT-rich interaction domain 1a (Arid1a), is a tumor suppressor gene. Arid1a-deficiency promotes Ang2-dependent angiogenesis leading to hepatocellular carcinoma progression. Arid1a-deficiency also sensitizes tumors to Ang2 blockade by sorafenib treatment.

Green tea polyphenol treatment attenuates atherosclerosis in high-fat diet-fed apolipoprotein E-knockout mice via alleviating dyslipidemia and up-regulating autophagy.

BACKGROUND: Green tea polyphenol (GTP) is a polyphenol source from green tea that has drawn wide attention owing to epidemiological evidence of its beneficial effects in the prevention of cardiovascular disease; the underlying molecular mechanisms of these effects are not well understood. This study aimed to investigate the effects of GTP treatment on autophagy regulation in the vessel wall and lipid metabolism of HFD-fed male ApoE-knockout mice. METHODS: Adult male ApoE-knockout mice (n = 30) fed with a high-fat diet (HFD) were treated with either vehicle or GTP (3.2 or 6.4 g/L) administered via drinking water for 15 weeks, and C57BL/6J mice fed with standard chow diet (STD) were used as the control group. Metabolic parameters, expression of key mRNAs and proteins of hepatic lipid metabolism and autophagy in the vessel wall of mice were determined after the 15-week treatment. RESULTS: A HFD induced atherosclerosis formation and lipid metabolism disorders as well as reduced autophagy expression in the vessel wall of ApoE-knockout mice, but GTP treatment alleviated the lipid metabolism disorders, decreased the oxLDL levels in serum, and increased the mRNA and protein expressions of hepatic PPARα and autophagy markers (LC3, Beclin1 and p62) in the vessel wall of ApoE-knockout mice. CONCLUSIONS: Our findings suggest that GTP supplementation showed marked suppression of atherogenesis through improved lipid metabolism as well as through a direct impact on oxLDL and autophagy flux in the vessel wall.

Oral contraception following abortion: A systematic review and meta-analysis.

Oral contraceptives (OCs) following induced abortion offer a reliable method to avoid repeated abortion. However, limited data exist supporting the effective use of OCs postabortion. We conducted this systematic review and meta-analysis in the present study reported immediate administration of OCs or combined OCs postabortion may reduce vaginal bleeding time and amount, shorten the menstruation recovery period, increase endometrial thickness 2 to 3 weeks after abortion, and reduce the risk of complications and unintended pregnancies.A total of 8 major authorized Chinese and English databases were screened from January 1960 to November 2014. Randomized controlled trials in which patients had undergone medical or surgical abortions were included. Chinese studies that met the inclusion criteria were divided into 3 groups: administration of OC postmedical abortion (group I; n = 1712), administration of OC postsurgical abortion (group II; n = 8788), and administration of OC in combination with traditional Chinese medicine postsurgical abortion (group III; n = 19,707).In total, 119 of 6160 publications were included in this analysis. Significant difference was observed in group I for vaginal bleeding time (P = 0.0001), the amount of vaginal bleeding (P = 0.03), and menstruation recovery period (P < 0.00001) compared with the control groups. Group II demonstrated a significant difference in vaginal bleeding time (P < 0.00001), the amount of vaginal bleeding (P = 0.0002), menstruation recovery period (P < 0.00001), and endometrial thickness at 2 (P = 0.003) and 3 (P < 0.00001) weeks postabortion compared with the control group. Similarly, a significant difference was observed in group III for reducing vaginal bleeding time (P < 0.00001) and the amount of vaginal bleeding (P < 0.00001), shortening the menstruation recovery period (P < 0.00001), and increasing endometrial thickness 2 and 3 weeks after surgical abortion (P < 0.00001, all).Immediate administration of OCs postabortion may reduce vaginal bleeding time and amount, shorten the menstruation recovery period, increase endometrial thickness 2 to 3 weeks after abortion, and reduce the risk of complications and unintended pregnancies.

[International research trends of pharmacognosy on molecular level --based on analysis of SCI literature].

The techniques and methods of molecular biology have been widely applied in pharmacognosy fields. International development trends of pharmacognosy studies on molecular level were analyzed by bibliometric methods using the SCIE database on Web of Science, the literature distribution, national distribution, agency distribution, periodicals distribution, and hot research topics were described using multivariate statistical analysis and multidimensional scaling analysis method,etc. The number of international pharmacognosy literature on molecular level is increasing year by year. USA, China and Japan have close cooperation, and focus on molecular identification and genetic diversity. Chinese scientists issued high-impact factor journals papers and high citations amount in the international forefront. The international pharmacognosy research on molecular level has developed rapidly. Chinese research has a significant influence.The molecular mechanism of the formation of Dao-di Herbs may become the next hotspot.

Qualitative and Quantitative Analysis of Lignan Constituents in Caulis Trachelospermi by HPLC-QTOF-MS and HPLC-UV.

A high-performance liquid chromatography coupled with quadrupole tandem time-of-flight mass (HPLC-QTOF-MS) and ultraviolet spectrometry (HPLC-UV) was established for simultaneous qualitative and quantitative analysis of the major chemical constituents in Caulis Trachelospermi, respectively. The analysis was performed on an Agilent Zorbax Eclipse Plus C18 column (4.6 mm×150 mm, 5 µm) using a binary gradient system of water and methanol, with ultraviolet absorption at 230 nm. Based on high-resolution ESI-MS/MS fragmentation behaviors of the reference standards, the characteristic cleavage patterns of lignano-9, 9'-lactones and lignano-8'-hydroxy-9, 9'-lactones were obtained. The results demonstrated that the characteristic fragmentation patterns are valuable for identifying and differentiating lignano-9,9'-lactones and lignano-8'-hydroxy-9,9'-lactones. As such, a total of 25 compounds in Caulis Trachelospermi were unambiguously or tentatively identified via comparisons with reference standards or literature. In addition, 14 dibenzylbutyrolatone lignans were simultaneously quantified in Caulis Trachelospermi by HPLC-UV method. The method is suitable for the qualitative and quantitative analyses of dibenzylbutyrolatone lignans in Caulis Trachelospermi.

Active components with inhibitory activities on IFN-γ/STAT1 and IL-6/STAT3 signaling pathways from Caulis Trachelospermi.

Initial investigation for new active herbal extract with inhibiting activity on JAK/STAT signaling pathway revealed that the extract of Caulis Trachelospermi, which was separated by 80% alcohol extraction and subsequent HP-20 macroporous resin column chromatography, was founded to strongly inhibit IFN-γ-induced STAT1-responsive luciferase activity (IFN-γ/STAT1) with IC50 value of 2.43 µg/mL as well as inhibiting IL-6-induced STAT3-responsive luciferase activity (IL-6/STAT3) with IC50 value of 1.38 µg/mL. Subsequent study on its active components led to the isolation and identification of two new dibenzylbutyrolactone lignans named 4-demethyltraxillaside (1) and nortrachelogenin 4-O-ß-D-glucopyranoside (2), together with six known compounds. The lignan compounds 1-4 together with other lignan compounds isolated in previous study were tested the activities on IFN-γ/STAT1 and IL-6/STAT3 pathways. The following result showed that the main components trachelogenin and arctigenin had corresponding activities on IFN-γ/STAT1 pathway with IC50 values of 3.14 µM and 9.46 µM as well as trachelogenin, arctigenin and matairesinol strongly inhibiting IL-6/STAT3 pathway with IC50 values of 3.63 µM, 6.47 µM and 2.92 µM, respectively.

[Application of associated symptom and sign-based acupoint selection in acupuncture-moxibustion treatment].

Based on traditional acupuncture-moxibustion treatment ideas, with differentiation of channels and collaterals as main part and feature, the important role of associated symptom and sign-based acupoint selection in acupuncture-moxibustion treatment is explained from angles of philosophy and medicine. Combined with clinical experience of acupuncture and moxibustion, the category of associated symptom and sign-based acupoint selection is explained in detail to make sure the accuracy of acupuncture-moxibustion differentiation. It could show uniqueness and advantage of theory and clinic in acupuncture-moxibustion and provide theoretical references in making acupuncture-moxibustion prescription to improve effectiveness of clinical treatment.