Identification of novel rheumatoid arthritis-associated MiRNA-204-5p from plasma exosomes

Wu Long-Fei,Zhang Qin,Mo Xing-Bo,Lin Jun,Wu Yang-Lin,Lu Xin,He Pei,Wu Jian,Guo Yu-Fan,Wang Ming-Jun,Ren Wen-Yan,Deng Hong-Wen,Lei Shu-Feng,Deng Fei-Yan 생화학분자생물학회 2022 Experimental and molecular medicine Vol.54 No.-

Rheumatoid arthritis (RA) is an autoimmune disease characterized by infiltration of immune cells in the synovium. However, the crosstalk of immune cells and synovial fibroblasts is still largely unknown. Here, global miRNA screening in plasma exosomes was carried out with a custom microarray (RA patients vs. healthy controls = 9:9). A total of 14 exosomal miRNAs were abnormally expressed in the RA patients. Then, downregulated expression of exosomal miR-204-5p was confirmed in both the replication (RA patients vs. healthy controls = 30:30) and validation groups (RA patients vs. healthy controls = 56:60). Similar to the findings obtained in humans, a decreased abundance of exosomal miR-204-5p was observed in mice with collagen-induced arthritis (CIA). Furthermore, Spearman correlation analysis indicated that plasma exosomal miR-204-5p expression was inversely correlated with disease parameters of RA patients, such as rheumatoid factor, erythrocyte sedimentation rate, and C-reactive protein. In vitro, our data showed that human T lymphocytes released exosomes containing large amounts of miR-204-5p, which can be transferred into synovial fibroblasts, inhibiting cell proliferation. Overexpression of miR-204-5p in synovial fibroblasts suppressed synovial fibroblast activation by targeting genes related to cell proliferation and invasion. In vivo assays found that administration of lentiviruses expressing miR-204-5p markedly alleviated the disease progression of the mice with CIA. Collectively, this study identified a novel RA-associated plasma exosomal miRNA-204-5p that mediates the communication between immune cells and synovial fibroblasts and can be used as a potential biomarker for RA diagnosis and treatment.

Ginsenoside Ro, an oleanolic saponin of Panax ginseng, exerts antiinflammatory effect by direct inhibiting toll like receptor 4 signaling pathway

Hong-Lin Xu,Guang-Hong Chen,Yu-Ting Wu,Ling-Peng Xie,Zhang-Bin Tan,Bin Liu,Hui-Jie Fan,Hong-Mei Chen,Gui-Qiong Huang,Min Liu,Ying-Chun Zhou 고려인삼학회 2022 Journal of Ginseng Research Vol.46 No.1

Background: Panax ginseng Meyer (P. ginseng), a herb distributed in Korea, China and Japan, exerts benefits on diverse inflammatory conditions. However, the underlying mechanism and active ingredients remains largely unclear. Herein, we aimed to explore the active ingredients of P. ginseng against inflammation and elucidate underlying mechanisms. Methods: Inflammation model was constructed by lipopolysaccharide (LPS) in C57BL/6 mice and RAW264.7 macrophages. Molecular docking, molecular dynamics, surface plasmon resonance imaging (SPRi) and immunofluorescence were utilized to predict active component. Results: P. ginseng significantly inhibited LPS-induced lung injury and the expression of proinflammatory factors, including TNF-a, IL-6 and IL-1b. Additionally, P. ginseng blocked fluorescence-labeled LPS (LPS488) binding to the membranes of RAW264.7 macrophages, the phosphorylation of nuclear factor-kB (NF-kB) and mitogen-activated protein kinases (MAPKs). Furthermore, molecular docking demonstrated that ginsenoside Ro (GRo) docked into the LPS binding site of toll like receptor 4 (TLR4)/myeloid differentiation factor 2 (MD2) complex. Molecular dynamic simulations showed that the MD2-GRo binding conformation was stable. SPRi demonstrated an excellent interaction between TLR4/MD2 complex and GRo (KD value of 1.16 × 10<SUP>-9</SUP> M). GRo significantly inhibited LPS488 binding to cell membranes. Further studies showed that GRo markedly suppressed LPS-triggered lung injury, the transcription and secretion levels of TNF-α, IL-6 and IL-1β. Moreover, the phosphorylation of NF-kB and MAPKs as well as the p65 subunit nuclear translocation were inhibited by GRo dose-dependently. Conclusion: Our results suggest that GRo exerts anti-inflammation actions by direct inhibition of TLR4 signaling pathway.

Inhibitory Effects of Phenolic Alkaloids of Menispermum Dauricum on Gastric Cancer in Vivo

Zhang, Hong-Feng,Wu, Di,Du, Jian-Kuo,Zhang, Yan,Su, Yun-Ming Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.24

The present study was conducted to investigate effects and mechanisms of action of phenolic alkaloids of Menispermum dauricum (PAMD) on gastric cancer in vivo. In vitro, cell apoptosis of human gastric cancer cell line SGC-7901 was observed using fluorescence staining. In vivo, a mice model was constructed to observe tumor growth with different doses. Cell apoptosis was examined using flow cytometry and K-RAS protein expression using Western blotting. The mRNA expression of P53, BCL-2, BAX, CASPASE-3, K-RAS was examined by real-time PCR. PAMD significantly suppressed tumor growth in the xenograft model of gastric cancer in a dose-dependent manner (p<0.01). Functionally, PAMD promoted cell apoptosis of the SGC-7901 cells and significantly increased the rate of cell apoptosis of gastric tumor cells (p<0.05). Mechanically, PAMD inhibited the expression of oncogenic K-RAS both at the mRNA and protein levels. In addition, PAMD affected the mRNA expression of the cell apoptosis-related genes (P53, BCL-2, BAX, CASPASE-3). PAMD could suppress gastric tumor growth in vivo, possibly through inhibiting oncogenic K-RAS, and induce cell apoptosis possibly by targeting the cell apoptosis-related genes of P53, BCL-2, BAX, CASPASE-3.

The Lung Function Impairment in Non-Atopic Patients With Chronic Rhinosinusitis and Its Correlation Analysis

Linghao Zhang,Lu Zhang,Chun-Hong Zhang,Xiao-Bi Fang,Zhen-Xiao Huang,Qing -Yuan Shi,Li-Ping Wu,Peng Wu,Zhen-Zhen Wang,Zhi-Su Liao 대한이비인후과학회 2016 Clinical and Experimental Otorhinolaryngology Vol.9 No.4

Objectives. Chronic rhinosinusitis (CRS) is common disease in otorhinolaryngology and will lead to lower airway abnormality. However, the only lung function in CRS patients and associated factors have not been much studied. Methods. One hundred patients with CRS with nasal polyps (CRSwNP group), 40 patients with CRS without nasal polyps (CRSsNP group), and 100 patients without CRS were enrolled. The difference in lung function was compared. Meanwhile, CRSwNP and CRSsNP group were required to undergo a bronchial provocation or dilation test. Additionally, subjective and objective outcomes were measured by the visual analogue scale (VAS), 20-item Sino-Nasal Outcome Test (SNOT-20), Lund-Mackay score, Lund-Kennedy endoscopic score. The correlation and regression methods were used to analyze the relationship between their lung function and the above parameters. Results. The forced expiratory volume in 1 second (FEV1) and forced expiratory flow between 25% and 75% of forced vital capacity (FEF25-75) of CRSwNP group were significantly lower than other groups (P<0.05). On peak expiratory flow, there was no difference between three groups. In CRSwNP group, FEV1 was negatively correlated with peripheral blood eosinophil count (PBEC) and duration of disease (r=–0.348, P=0.013 and r=–0.344, P=0.014, respectively), FEF25-75 negatively with VAS, SNOT-20 (r=–0.490, P=0.028 and r=–0.478, P=0.033, respectively) in CRSsNP group. The incidence of positive bronchial provocation and dilation test was lower in CRSwNP group (10% and 0%, respectively), with both 0% in CRSsNP group. The multiple linear regression analysis indicated that change ratio of FEV1 before and after bronchial provocation or dilation test were correlated with PBEC in CRSwNP group (β=0.403, P=0.006). Conclusion. CRS leading to impaired maximum ventilation and small airway is associated with the existence of nasal polyp. Lung function impairments can be reflected by PBEC, duration, VAS, and SNOT-20. In CRSwNP patients, PBEC is independent predictor of FEV1 change ratio.

Enhancing Functional Expression of β-glucosidase in Pichia pastoris by Co-expressing Protein Disulfide Isomerase

Jian-hong Zhang,Dan Wu,Jian Chen,Jing Wu 한국생물공학회 2011 Biotechnology and Bioprocess Engineering Vol.16 No.6

The expression of heterologous proteins may exert severe stress on the host cells at different levels. Protein folding and disulfide bond formation were identified as rate-limited steps in recombinant protein secretion in yeast cells. For the production of β-glucosidase in Pichia pastoris, final β-glucosidase activity reached 1,749 U/mL after fermentation optimization in a 3 L bioreactor, while the specific activity decreased from 620to 467 U/mg, indicating a potential protein misfolding. To solve this problem, protein disulfide isomerase, a chaperone protein which may effectively regulate disulfide bond formation and protein folding, was co-expressed with β-glucosidase. In the co-expression system, a β-glucosidase production level of 2,553 U/mL was achieved and the specific activity of the enzyme reached 721 U/mg, which is 1.54 fold that of the control.

Genetic Analysis of Seed Number per Pod in Brassica napus Using Augmented North Carolina (NC) 2

Li Wu Zhang,Ping Wu Liu,Deng Feng Hong,Guang Sheng Yang 한국유전학회 2008 Genes & Genomics Vol.30 No.3

Seed number per pod is one of the three important components of yield in rapeseed. An experiment with the augmented NCⅡ design was conducted for investigating the heterosis, combining ability and genetic model of this trait. Results revealed that this trait was fitted with additive-dominance genetic model by using the variance analysis of ( Wr + Vr) and (Wr - Vr). Higher general predictability ratio (0.693) supported that both additive and non-additive genes governed this trait. The moderate estimates of narrow-sense heritability (0.588) further confirmed this. The gene distribution estimates indicated the occurrence of asymmetry. At least one major group of genes controlled the inheritance of this trait. Per se performance of parents is a reflection of their "gca" (general combining ability) effects in most F1s. Among the genotypes Y106 ranked as the top general combiner followed by HZ398 and HZ396. High estimates of mid-parent heterosis and potential ratio were observed in HZ398/Y106, indicating that it would be worthwhile to utilize these diverse genotypes as parents for the development of improved hybrids or segregants.

Corrigendum to "Ginsenoside Ro, an oleanolic saponin of Panax ginseng, exerts anti-inflammatory effect by direct inhibiting toll like receptor 4 signaling pathway" [J Ginseng Res 46 (2022) 156-166]

Xu, Hong-Lin,Chen, Guang-Hong,Wu, Yu-Ting,Xie, Ling-Peng,Tan, Zhang-Bin,Liu, Bin,Fan, Hui-Jie,Chen, Hong-Mei,Huang, Gui-Qiong,Liu, Min,Zhou, Ying-Chun The Korean Society of Ginseng 2022 Journal of Ginseng Research Vol.46 No.2

miR-140-3p Knockdown Suppresses Cell Proliferation and Fibrogenesis in Hepatic Stellate Cells via PTEN-Mediated AKT/mTOR Signaling

Shi-Min Wu,Tian-Hong Li,Hao Yun,Hong-Wu Ai,Ke-Hui Zhang 연세대학교의과대학 2019 Yonsei medical journal Vol.60 No.6

Purpose: Liver fibrosis is a major cause of morbidity and mortality and the outcome of various chronic liver diseases. Activationof hepatic stellate cells (HSCs) is the key event in liver fibrosis. Studies have confirmed that miR-140-3p plays a potential regulatoryeffect on HSC activation. However, whether miR-140-3p mediates the liver fibrosis remains unknown. Materials and Methods: Expression of miR-140-3p was detected by real-time quantitative PCR (qPCR). Cell proliferation wasmeasured by MTT, while cell apoptosis rate was determined via flow cytometry. Western blot assay was used to detect the expressionof cleaved PARP. The fibrogenic effect was evaluated by expression of α-smooth muscle actin and desmin. Functional experimentswere performed in transforming growth factor β1 (TGF-β1)-induced HSC-T6 cells with transfection of anti-miR-140-3pand/or siPTEN. Target binding between miR-140-3p and PTEN was predicted by the TargetScan database and identified usingluciferase reporter assay and RNA immunoprecipitation. Results: TGF-β1 induced the activation of HSC-T6 cells, and miR-140-3p expression varied according to HSC-T6 cell activationstatus. Knockdown of miR-140-3p reduced cell proliferation and the expressions of α-SMA and desmin, as well as increasedapoptosis, in TGF-β1-induced HSC-T6 cells, which could be blocked by PTEN silencing. Additionally, inactivation of the AKT/mTOR signaling pathway stimulated by miR-140-3p knockdown was abolished when silencing PTEN expression. PTEN was negativelyregulated by miR-140-3p via direct binding in HSC-T6 cells. Conclusion: miR-140-3p is an important mediator in HSC-T6 cell activation, and miR-140-3p knockdown suppresses cell proliferationand fibrogenesis in TGF-β1-induced HSC-T6 cells, indicating that miR-140-3p may be a potential novel molecular targetfor liver fibrosis.

Fabrication of Photopolymer Hierarchical Micronanostructures by Coupling Electrospinning and Photolithography for SERS Substrates

Wen-Yi Zhang,Xin-Ze Xiao,Chao Lv,Jia Zhao,Gong Wang,Xuan Gu,Ran Zhang,Bin-Bin Xu,Dan-Dan Zhang,Ai-Wu Li,Yong-Lai Zhang,Hong-Bo Sun 한국고분자학회 2013 Macromolecular Research Vol.21 No.3

Reported here is the fabrication of photopolymer hierarchical micronanostructures through a combinative process of electrospinning and subsequent photolithography. Electrospun SU-8 (epoxy-based negative photoresist)nanofiber films have been patterned into gratings with periods of 100, 200, 300, and 400 μm, respectively. Deposition of a silver nanolayer on these interlaced nanofiber films would lead to the formation of various plasmonic nanostructures,and therefore, giving rise to abundant surface-enhanced Raman scattering (SERS) “hot spots”. In the detection of Rhodamine 6G (R6G), probing molecule, the resultant SERS substrates show both high sensitivity and good reproducibility. The SERS enhancement factor could reach as high as ~108, indicating high efficiency. The fabrication of patterned, highly efficient SERS substrates may hold a great promise for the integration of SERS substrates in various microdevices such as microfluidic chips.

Efficiently targeted therapy of glioblastoma xenograft via multifunctional biomimetic nanodrugs

Zhipeng Yao,Xiaochun Jiang,Hong Yao,Yafeng Wu,Fan Zhang,Cheng Wang,Chenxue Qi,Chenhui Zhao,Zeyu Wu,Min Qi,Jia Zhang,Xiaoxiang Cao,Zhichun Wang,Fei Wu,Chengyun Yao,Songqin Liu,Shizhang Ling,Hongping Xi 한국생체재료학회 2022 생체재료학회지 Vol.26 No.4

Background: Glioblastoma multiforme (GBM) is a fatal malignant primary brain tumor in adults. The therapeutic efficacy of chemotherapeutic drugs is limited due to the blood-brain barrier (BBB), poor drug targeting, and short biological half-lives. Multifunctional biomimetic nanodrugs have great potential to overcome these limitations of chemotherapeutic drugs. Methods: We synthesized and characterized a biomimetic nanodrug CMS/PEG-DOX-M. The CMS/PEG-DOX-M effectively and rapidly released DOX in U87 MG cells. Cell proliferation and apoptosis assays were examined by the MTT and TUNEL assays. The penetration of nanodrugs through the BBB and anti-tumor efficacy were investigated in the orthotopic glioblastoma xenograft models. Results: We showed that CMS/PEG-DOX-M inhibited cell proliferation of U87 MG cells and effectively induced cell apoptosis of U87 MG cells. Intracranial antitumor experiments showed that free DOX hardly penetrated the BBB, but CMS/PEG-DOX-M effectively reached the orthotopic ntracranial tumor through the BBB and significantly inhibited tumor growth. Immunofluorescence staining of orthotopic tumor tissue sections confirmed that nanodrugs promoted apoptosis of tumor cells. This study developed a multimodal nanodrug treatment system with the enhanced abilities of tumor-targeting, BBB penetration, and cancer-specific accumulation of chemotherapeutic drugs by combining chemotherapy and photothermal therapy. It can be used as a flexible and effective GBM treatment system and it may also be used for the treatment of other central nervous systems (CNS) tumors and extracranial tumors.