Luteolin Alleviates Liver Fibrosis in Rat Hepatic Stellate Cell HSC-T6: A Proteomic Analysis.

Background: Traditional Chinese medicine (TCM) with single or compound materials is an effective cure for liver fibrosis. Hepatic stellate cells (HSCs) play a key role in liver fibrosis pathology and have become a novel drug target for this condition. Methods: CCK-8 assay was used to determine the cytotoxicity of four components, SYPA, HSYPA, Apigenin, and Luteolin, from Deduhonghua-7 powder on HSC-T6 cells. Transforming Growth Factor ß 1 (TGFß1)-induced fibrotic cell model and CCI4-induced fibrotic rat model were constructed, the expression of fibrosis-related genes, the pathological changes and serum biochemical markers were evaluated. Proteomic analysis was performed to determine the mechanism by which luteolin attenuated liver fibrosis, which were further confirmed by Western blot. Results: Luteolin attenuates liver fibrosis in HSC-T6 cells and luteolin decreases the liver fibrosis index level in vivo. A total of 5000 differentially expressed proteins (DEPs) were obtained using proteomic analysis. KEGG analysis found that DEPs were concentrated in various metabolic pathways, including DNA replication and repair and lysosomal signaling. GO analysis showed that molecular functions included the activity and binding of various enzymes, related cellular components included the extracellular space, lysosomal lumen, mitochondrial matrix, and nucleus, and biological processes included collagen organization and biosynthesis and the positive regulation of cell migration. Western blot results showed that CCR1, CD59, and NAGA were downregulated in TGFß1 treatment, while upregulated both in Lut2 and Lut10 treatment. Meanwhile, eight proteins, ITIH3, MKI67, KIF23, DNMT1, P4HA3, CCDC80, APOB, FBLN2, that were upregulated in TGFß1 treatment, while downregulated both in Lut2 and Lut10 treatment. Conclusion: Luteolin was shown to have a strong protective effect on liver fibrosis. CCR1, CD59, and NAGA may promote liver fibrosis while ITIH3, MKI67, KIF23, DNMT1, P4HA3, CCDC80, APOB, and FBLN2 may facilitate protection against fibrosis.

A MnAl double adjuvant nanovaccine to induce strong humoral and cellular immune responses.

At present, the most widely used aluminum adjuvants have poor ability to induce effective Th1 type immune responses. Existing evidence suggests that manganese is a potential metal adjuvant by activating cyclic guanosine phospho-adenosine synthase (cGAS)-interferon gene stimulator protein (STING) signaling pathway to enhance humoral and cellular immune response. Hence, the effective modulation of metal components is expected to be a new strategy to improve the efficiency of vaccine immunization. Here, we constructed a manganese and aluminum dual-adjuvant antigen co-delivery system (MnO2-Al-OVA) to enhance the immune responses of subunit vaccines. Namely, the aluminum hydroxide was first fused on the surface of the pre-prepared MnO2 nanoparticles, which were synthesized by a simple redox reaction with potassium permanganate (KMnO4) and oleic acid (OA). The engineered MnO2-Al-OVA could remarkably promote cellular internalization and maturation of dendritic cells. After subcutaneous vaccination, MnO2-Al-OVA rapidly migrated into the lymph nodes (LNs) and efficiently activate the cGAS-STING pathway, greatly induced humoral and cellular immune responses. Of note, our findings underscore the importance of coordination manganese adjuvants in vaccine design by promoting the activation of the cGAS-STING-IFN-I pathway. With a good safety profile and facile preparation process, this dual-adjuvant antigen co-delivery nanovaccine has great potential for clinical translation prospects.

Heat shock cognate 70 protein like-2 protein in camphor pollen is one of the major culprits of asthma.

AIMS: In recent decades, Cinnamomum camphora have gradually become the main street trees in Shanghai. This study aims to investigate the allergenicity of camphor pollen. MAIN METHODS: A total of 194 serum samples from patients with respiratory allergy were collected and analyzed. Through protein profile identification and bioinformatics analysis, we hypothesized that heat shock cognate protein 2-like protein (HSC70L2) is the major potential allergenic protein in camphor pollen. Recombinant HSC70L2 (rHSC70L2) was expressed and purified, and a mouse model of camphor pollen allergy was established by subcutaneous injection of total camphor pollen protein extract (CPPE) and rHSC70L2. KEY FINDINGS: Specific IgE was found in the serum of 5 patients in response to camphor pollen and three positive bands were identified by Western blotting. Enzyme-linked immunosorbent assay (ELISA), Immune dot blot and Western blot experiments confirmed that CPPE and rHSC70L2 can cause allergies in mice. Moreover, rHSC70L2 induces polarization of peripheral blood CD4+ T cells to Th2 cells in patients with respiratory allergies and mice with camphor pollen allergy. Finally, we predicted the T cell epitope of the HSC70L2 protein, and through the mouse spleen T cell stimulation experiment, we found that the 295EGIDFYSTITRARFE309 peptide induced T cells differentiation to Th2 and macrophages differentiation to the alternatively activated (M2) state. Moreover, 295EGIDFYSTITRARFE309 peptide increased the serum IgE levels in mice. SIGNIFICANCE: The identification of HSC70L2 protein can provide novel diagnostic and therapeutic targets for allergies caused by camphor pollen.

Triple functional mild photothermal improves gene editing of PD-L1 for enhanced antitumor immunity.

Traditional photothermal therapy ablates tumor cells by a high temperature (> 50 °C). Although it has shown good anti-tumor effect in animal models, the potential damages to healthy tissues and the unnecessary inflammatory reactions caused by the high temperature have hindered the clinical transitions of traditional photothermal therapy. In this study, we used polydopamine (PDA) as a mild photothermal material and control the maximum temperature below 45 °C, which not only avoided the side effects caused by a high temperature, but also ablated a fraction of tumor cells and produced tumor antigens. Meanwhile, the near-infrared (NIR) light also served as a "switch" to trigger the release of CRISPR/Cas9 RNP from Fe3O4 nanoparticles (Fe3O4 NPs) after their accumulation to tumor sites via magnetic targeting. The triple functional mild photothermal therapy achieved significant PD-L1 gene knockout efficiency in the tumor-bearing mice, reversed the condition of immunosuppression in the tumor microenvironment, led to a higher level of anti-tumor immune responses and effectively inhibited the growth of melanoma. We anticipate that this triple functional mild photothermal therapy would provide a potential new approach for the treatment of malignant tumors.

Structure elucidation and NMR assignments of a new sesquiterpene of volatile oil from Artemisia frigida Willd.

A new sesquiterpene, artefrigin (5), together with four known sesquiterpenes were isolated from the volatile oil of Artemisia frigida Willd. The structure of five was elucidated by spectroscopic methods, including UV, IR, HR-ESI-MS and extensive 1D and 2D NMR techniques.

Two new compounds from supercritical fluid extract of Artemisia integrifolia L.

Two new compounds, namely integrin A (1) and integrin B (2), were isolated from the supercritical fluid extract (SFE) of Artemisia integrifolia L. Their structures were elucidated by spectroscopic methods, including UV, IR, HR-ESI-MS and extensive 1D and 2D NMR techniques.

Static magnetic fields accelerate osteogenesis by regulating FLRT/BMP pathway.

OBJECTIVES: Static magnetic fields (SMF) have been proved to enhance osteogenic differentiation in mesenchymal stem cells (MSCs). However, the effect of SMF on mandibular condylar chondrocytes (MCCs) are less investigated, which contributes to the vertical formation of mandible. The purpose of the present study was to identify whether SMF accelerate the osteogenesis on mature condylar cartilage and explore the potential regulatory mechanism. METHODS: In this study, we presented a 280 mT SMF stimulation set-up to investigate the genomic effects of SMF exposure on MCCs differentiation and osteoblast-related factor secretion in vitro. Induced by Oricell™ for osteogenesis, MCCs from primary SD Rat were stimulated with or without SMF for cell culture. Cell proliferation was determined by CCK-8. The enhanced osteogenetic capacity of the SMF stimulated MCCs was identified by Alizarin red staining (ARS). Additionally, the effects of SMF on the expression of transmembrane protein marker (FLRT3), terminal differentiation markers (BMP2), and transcription factors (Smad1/5/8) were quantified by Western blot and immunofluorescence analysis. RESULTS: Compared with the control group, SMF decreased the proliferation of MCCs (p < 0.05) after 14 days osteogenesis-specific induction. The mineral synthesis of MCCs was upregulated by SMF (p < 0.0001). The expression of BMP2, Smad1/5/8 showed decrease trends while the protein level of FLRT3 acted in contrary manner (p < 0.05). CONCLUSIONS: Our findings emphasized the ability of osteogenesis positively respond to SMF stimulation by exhibiting enhanced differentiation via FLRT/BMP signaling.

Effective treatment of residue of acrylic acid production using a fluid-bed/fixed-bed system with low energy consumption.

In order to effectively deal with large amounts of complex organic pollutants in the harmful distillation residues with low energy consumption, a novel two-stage fluid-bed/fixed-bed system was designed to catalyze oxidation of acrylic acid production residue. The effects of fluid-bed temperature, gaseous hourly space velocity (GHSV), and oxygen excess rate on the purification of acrylic acid production residue in the two-stage fluid-bed/fixed-bed system were studied to prove the feasibility of the method. The chemical oxygen demand (COD) of the discharged liquid was <100 mg/L, and the volatile organic compounds (VOCs) of the discharged gas amounted to <10 mg/m3 with a fluid-bed temperature of 380°C, emulsified residue's GHSV of 0.28 L/(kgcat ·hr), and O2 excessive rate of more than 4.32. The result of techno-economics indicates the feasibility of the long-term operation of process. Results further illustrate the advantages of the proposed two-stage fluid-bed/fixed-bed system, which can treat acrylic acid production residue with high efficiency (COD < 100 mg/L, VOCs < 10 mg/m3 ) and low energy consumption (~24,856 kw·hr/ton) in the chemical industry. PRACTITIONER POINTS: A novel two-stage fluid-bed/fixed-bed system was developed for acrylic acid production residue treatment. No extra energy was required at low temperature in the two-stage fluid-bed/fixed-bed system. Purification of residue could be finished at low temperature by the catalytic pyrolysis and catalytic oxidation process. The two-stage system did not produce toxic gases and particulate matters.

Structure elucidation and antimicrobial activities of five compounds from Artemisia integrifolia L.

A new compound, integracid (1), together with four known compounds were isolated from the dichloromethane (CH2Cl2) extract from Artemisia integrifolia L. The structures of compounds (1-5) were elucidated by spectroscopic methods, including ultraviolet, infrared (IR), high resolution-electrospray ionization-mass spectrometry (HR-ESI-MS) and extensive one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) techniques, and by comparison with data reported in the references. Antibacterial activities of the compounds were evaluated against various bacteria.

Antihyperlipidemic Effect, Identification and Isolation of the Lipophilic Components from Artemisia integrifolia.

Artemisia integrifolia L (Compositae) is a medicinal and edible plant. To investigate its antihyperlipidemic effect, a crude lipophilic extract and the composing compounds were isolated and fractioned from the petroleum ether extract of aerial parts of A. integrifolia using column chromatography on silica gel. The anti-hyperlipidemia effect was studied in a rat model of acute hyperlipidemia, which was induced by triton WR-1339. A new compound, integrinol (4), together with nine known compounds, namely chamazulene (1), acetylenes (E)-2 (2), acetylenes (E)-3 (3), eugenol (5), palmitic acid (6), oleic acid (7), linoleic acid (8), linolenic acid (9) and 12,13-epoxylinolenic acid were isolated from the crude lipophilic extract of A. integrifolia. The LD50 value of the crude extract was more than 4g/kg. In Triton WR-1339-induced acute hyperlipidemia model, the crude lipophilic extract (200 mg/kg) significantly reduced total cholesterol (TC) by 70% (p ≤ 0.01) and triglycerides (TGs) by 94% (p ≤ 0.001). The fractioned compounds, such as chamazulene (1), acetylene-2 (2), and linolenic acid (9), used at 4 mg/kg dose, also significantly decreased the concentrations of TC (32%, 33% and 64%, respectively) and TGs (48%, 33% and 93%, respectively). These compounds (i.e., chamazulene, acetylenes (E)-2, and linolenic acid) were considered to be responsible for the bioactive antihyperlipidemic effect. In conclusion, the crude lipid extract of Artemisia integrifolia L could be used as a potential treatment to avert hyperlipidemia. Further studies to confirm these results in other models of hyperlipidemia (e.g., diet-induced obesity) are warranted.

Species and quantity of airborne pollens in Shanghai as monitored by gravitational and volumetric methods.

BACKGROUND: The prevalence of allergic diseases has markedly increased in the last decades. It is therefore important to assess the distribution of airborne pollen, the most important aeroallergen, for allergic disease prevention and control. OBJECTIVE: To identify the species and quantity of airborne pollens, and observe their distribution characteristics in Shanghai, using gravitational (Durham Sampler) and volumetric (Rotorod Sampler 40) methods simultaneously. In addition, the correlation between both methods was analyzed to provide effective preventive measures for pollen-sensitized individuals. METHOD: Pollen counts were monitored in the same area from November 1, 2009 to October 31, 2010 by samplers set at the same height and site. Pollen concentrations as well as any association between the two methods were determined. RESULTS: Two pollen concentration peaks in Shanghai were observed from March to May (spring) and September to October (autumn). In spring, tree pollen was the main species, with a predominance of Broussonetia. In autumn, grass pollen predominated, with mostly Humulus. Thirty-two species were identified by both gravitational and volumetric methods. Five and seven additional species were identified exclusively by the gravitational and volumetric methods, respectively. Pollen counts obtained from both devices were significantly correlated (P<0.05). CONCLUSIONS: Two methods were used simultaneously for the first time to monitor pollen counts in central urban Shanghai, showing two annual peaks. Broussonetia and Humulus were the predominant spring and autumn pollens, respectively. Pollen counts obtained by both methods were clearly correlated. Regional airborne pollen monitoring offers preventive measures for sensitized individuals and provides useful clinical information.

[Insulinotropic action of hippocampal cholinergic neurostimulating peptide mediated by activated type 3 muscarinic receptor in INS-1 cells].

OBJECTIVE: [corrected] To characterize the insulinotropic action of hippocampal cholinergic neurostimulating peptide (HCNP) and analyze the role of type 3 muscarinic receptor (M(3)R) pathway in the action of HCNP. METHODS: INS-1 cells were incubated in routine RPMI 1640 medium (control group), RPMI 1640 supplemented with 50 pg/ml synthetic HCNP (HCNP group), or HCNP-containing medium with the addition of PMA 18 h prior to insulin release assay. The insulin levels in the medium was measured using radioimmunoassay following stimulation with different concentrations of glucose. Real-time quantitative PCR was used for detecting the gene expression of HCNP-pp, choline acetyltransferase (ChAT) and M(3)R in HCNP group and control group. RESULTS: After stimulation with different concentrations of glucose (5.6 and 16.7 mmol/L), HCNP group showed significantly higher insulin levels than the control and HCNP+ PMA groups. Compared with those in the control group, the mRNA levels of HCNP-pp, ChAT, and M(3)R were all lowered in HCNP group. CONCLUSION: HCNP can promote insulin release in INS-1 cells by increasing ChAT activity and activating M(3)R, and this effect is inhibited by PMA.

[Clinical observation on acupuncture for treatment of abdominal postoperative pain].

OBJECTIVE: To compare the therapeutic effects between acupuncture based on syndrome differentiation and analgesic on abdominal postoperative pain. METHODS: One hundred cases of abdominal postoperative pain were randomly divided into two groups, 50 cases in each one. In acupuncture group, the treatment was applied according to meridian differentiation and point selection on the affected meridian. Ashi points near to the incision as the main points and those closely connected with Zangfu functions were selected, such as Yanglingquan (GB 34), Taichong (LR 3) and Zusanli (ST 36), etc. In medication group, muscular injection of Bucinnazine was administered. The severity of pain was evaluated with Visual Analogue Scale (VAS) before and after treatment. RESULTS: The remarkably effective rate in acupuncture group was 60.0% (30/50), which was markedly better than that 28.0% (14/50) in medication group (P < 0.01). VAS scores in 30 min and 4 h after treatment as well as 24 h after operation in two groups were all reduced remarkably as compared with those before treatment (all P < 0.01), indicating the satisfactory analgesia in treatment. VAS scores in acupuncture group were lower apparently than those in medication group in 30 min and 4 h after treatment (P < 0.01,P < 0.05). CONCLUSION: Acupuncture has quick analgesia in treatment of abdominal postoperative pain, which is superior to muscular injection of Bucinnazine because of its advantages of long-term and significant efficacy.

Myosin-X is required for cranial neural crest cell migration in Xenopus laevis.

Myosin-X (MyoX) belongs to a large family of unconventional, nonmuscle, actin-dependent motor proteins. We show that MyoX is predominantly expressed in cranial neural crest (CNC) cells in embryos of Xenopus laevis and is required for head and jaw cartilage development. Knockdown of MyoX expression using antisense morpholino oligonucleotides resulted in retarded migration of CNC cells into the pharyngeal arches, leading to subsequent hypoplasia of cartilage and inhibited outgrowth of the CNC-derived trigeminal nerve. In vitro migration assays on fibronectin using explanted CNC cells showed significant inhibition of filopodia formation, cell attachment, spreading and migration, accompanied by disruption of the actin cytoskeleton. These data support the conclusion that MyoX has an essential function in CNC migration in the vertebrate embryo.