Identification of the metabolites from co-cultures of marine Streptomyces sp. IMB18-531 and Cladosporium sp. IMB19-099 / 药学学报

A new siderophore chelate (1) and 8 known compounds were identified from the liquid co-cultures of the marine-derived Streptomyces sp. IMB18-531 and Cladosporium sp. IMB19-099 by a combination of chromatography methods, including C18 reversed-phase medium pressure chromatography, gel column chromatography and HPLC. Their structures were determined by spectroscopic analysis and chemical methods as aluminioxamine E (1), desferrioxamine E (2), ferrioxamine E (3), terragine E (4), capsimicin (5), cyclo(L-prolinyl-L-tyrosine) (6), anthranilic acid (7), (Z)-14-methylpentadec-9-enoic acid (8), and (Z)-hexadec-8-enoic acid (9). Compound 2 showed inhibitory activities against the expression of liver fibrosis related genes COL1A1, MMP2, and TIMP2. Compounds 5, 8, and 9 displayed antibacterial activities against methicillin-resistant Staphylococcus aureus, S. epidermidis and Bacillus subtilis, with MICs of 16-64 μg·mL-1. Compound 5 showed cytotoxicities against human pancreatic cancer MIA Paca-2 and human colon cancer HT-29 cell lines with IC50 of 2.9 and 6.3 μmol·L-1, respectively.

The Signaling Pathway of Growth Differentiation Factor 15 and Its Receptor GFRAL / 中国生物化学与分子生物学报

Growth differentiation factor 15 (GDF15), a member of the transforming growth factor β (TGF-β) superfamily, is a new class of dimeric polypeptides with very low homology with other TGF-β family members. GDF15 was originally found in activated macrophages where it was secreted into the body circulation in two different cellular pathways. Moreover, GDF15 as a stress protein is widely involved in many signal pathways such as phosphoinositide 3-kinase / protein kinase B, extracellular signal-regulated kinase, c-Jun N-terminal kinase and nuclear factor-κB, and so on, and thus involved in the regulation of various disease processes. In addition, GDF15 as a new type of stress molecule acts as a biomarker and plays a regulatory role in obesity, weight loss, cancer development, cardiovascular disease, inflammation and autoimmune diseases. Glial-derived neurotrophic factor receptor alpha-like (GFRAL) is the specific receptor of GDF15, and the molecular basis of its activity is to conduct signal transduction through GFRAL-dependent binding into multimers. The intervention of the GDF15-GFRAL signaling pathway has a great application potential in the development of weight-loss drugs and cancer prognosis recovery drugs. This review focuses on the recent progress of GDF15-GFRAL and its related signaling pathways, the molecular structure of GDF15 and GFRAL, and the mechanism of GDF15-GFRAL signaling pathway, which reveals the role and regulatory ability of GDF15 as a biomarker in the development of disease and provides new insights in potential and treatment strategies of regulating the GDF15-GFRAL signaling pathway in related diseases.

Effect of ATAD3A gene silencing on the proliferation, invasion and migration of A375 human melanoma cells and its mechanisms / 中华皮肤科杂志

Objective:To evaluate the effect of ATPase family AAA-domain containing protein 3A (ATAD3A) gene silencing on the proliferation, invasion and migration of A375 human melanoma cells.Methods:From August to December in 2019, melanoma and paracancerous tissues were collected from 3 patients with pathologically diagnosed melanoma in People′s Hospital of Chongqing Yubei District, and Western blot analysis was performed to measure the protein expression of ATAD3A in the above tissues. Cultured A375 human melanoma cells were divided into 2 groups to be infected with a lentiviral vector carrying shATAD3A (shATAD3A group) and an empty vector (shCtrl group) respectively, and real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis were performed to verify the interference efficiency. Cell counting kit-8 (CCK8) assay and colony formation assay were performed to compare cell proliferative ability and colony-formation ability respectively between the 2 groups, and Transwell invasion assay and wound healing assay to compare invasive and migratory abilities respectively between the above 2 groups. Western blot analysis was performed to determine the expression of cell self-renewal-related proteins (NANOG, SRY-related high-mobility-group box protein SOX2, octamer-binding protein 4[OCT4]) and invasion- and migration-related proteins (matrix metalloproteinase 2[MMP2], vimentin, zinc-finger transcription factor SLUG) in the 2 groups. Two-independent-sample t test was used to compare the experimental indices between the 2 groups. Results:Western blot analysis showed that ATAD3A was significantly highly expressed in the 3 melanoma tissues compared with the paracancerous tissues ( t = 10.825, P < 0.001) . qRT-PCR and Western blot analysis showed that the mRNA and protein expression of ATAD3A in A375 cells was significantly lower in the shATAD3A group (0.230 ± 0.073, 0.279 ± 0.267, respectively) than in the shCtrl group (1.000 ± 0.244, 0.867 ± 0.115, respectively; t = 9.461, 8.595, respectively; P < 0.001 or = 0.002) , indicating that the ATAD3A gene-silenced A375 cell line was successfully constructed. Colony formation assay revealed that the colony-formation rate was significantly lower in the shATAD3A group than in the shCtrl group (22.667% ± 2.510% vs. 43.667% ± 5.030%, t = 6.464, P = 0.003) , and CCK-8 assay showed that the cellular proliferative activity significantly decreased from day 2 to day 4 in the shATAD3A group compared with the shCtrl group. Wound healing assay showed significantly slower wound healing and decreased wound healing rate from the 12 th hour (32.920% ± 4.642% vs. 49.302% ± 1.448%, t = 5.835, P = 0.004) to the 24 th hour in the shATAD3A group compared with the shCtrl group, and Transwell invasion assay revealed significantly decreased number of invasive cells in the lower Transwell chambers in the shATAD3A group compared with the shCtrl group (68.330 ± 13.050 vs. 234.330 ± 19.139, t = 12.411, P < 0.001) . Western blot analysis showed that the protein expression of NANOG, SOX2, OCT4, MMP2, vimentin, SLUG was significantly lower in the shATAD3A group than in the shCtrl group ( P < 0.05 or 0.001) . Conclusion:ATAD3A is highly expressed in melanoma tissues, and ATAD3A gene silencing can inhibit the proliferation, invasion and migration abilities of melanoma A375 cells.

Study on effects of ferulic acid combined with adipose derived mesenchymal stem cells on apoptosis of hepatic stellate cells by regulating TGF-β/Smad signaling transduction pathway / 重庆医学

Objective To investigate the effects of ferulic acid (FA) and adipose-derived mesenchymal stem cells(ADMSCs)on rat hepatic stellate cells (HSCs) by regulating TGF-β/smad signal transduction pathway.Methods HSCs were divided into 4 groups:blank control group,FA control group,ADMSCs control group and FA+ADMSCs group.The apoptosis rate of HSCs in each group was detected by flow cytometry.The expression levels of TGF-β1,Smad2,Smad3 and Smad7 mRNA in HSCs were detected by qRT-PCR.The TGF-β1 and Smad7 protein levels,and phosphorylated-Smad2/3 (p-Smad2/3)expression were detected by Western blot.Results Compared with the other 3 groups,the apoptosis rate of HSCs in the FA+ADMSCs group was significantly increased(P＜0.05),while the expression levels of TGF-β1,Smad2 and Smad3 mRNA were significantly decreased,and the Smad7 mRNA expression was increased;moreover,the expression levels of TGF-β1 protein and p-Smad2/3 were significantly decreased,while the Smad7 protein expression was significantly increased(P＜0.05).Conclusion FA can enhance the effect of ADMSCs for down-regulating the TGF-β1 expression in HSCs,and then leads to the decrease of downstream p-Smad2/3 activity and Smad7 expression increase,thus participates in promoting cellular apoptosis.

Expression level, clinical significance and target gene functions of circu-lating miR-103b in patients with pre-diabetes mellitus / 中国病理生理杂志

AIM:To investigate whether serum microRNA(miR)-103b plays a critical role in the pathogene-sis of type 2 diabetes mellitus(T2DM)and pre-diabetic syndrome.METHODS:Bioinformatic analysis was used for iden-tification of miR-103b and its targets,and the results were assessed by real-time PCR and receiver-operating characteristic (ROC)curve analysis in 48 patients with pre-diabetes mellitus(pre-DM),47 patients with noncomplicated diabetes melli-tus(NCDM),and 50 healthy individuals.RESULTS:miR-103b was significantly down-regulated in serum from the pa-tients with pre-DM and NCDM compared with healthy individuals.The ROC curve analysis found that the area under the curve(AUC)of miR-103b was 0.887(95% CI 0.809~0.944).The bioinformatic analysis has demonstrated that miR-103b has a high degree of site conservation among different mammalian species,such as Homo sapiens,Mus musculus,Rat-tus norvegicus,Pongo pygmaeus,Sus scrofa,etc.Fifty-three potential targets of miR-103b were predicted, most of which were involved in MAPK,Wnt,insulin and Ras signaling pathways,and enriched in various biological processes(such as phosphoprotein,DNA regulation transcription,cell growth and proliferation,apoptosis, cell cycle, etc), molecular func-tions(such as protein binding)and cell component(such as filamentous actin).CONCLUSION:Serum miR-103b can be used as an objective complement to traditional diagnosis of pre-diabetes,indicating important implications regarding the distinguish of the undiagnosed cases between diabetes and pre-diabetes by circulating miRNA.

Inter-regional, base-like and informatized support of field medical station during training / 医疗卫生装备

Objective To explore the inter-regional,base-like and informatized support of the field medical station during rotational training.Methods The field medical station information system developed by the hospital was introduced,which had the working mode involving in a set of system and two kinds of terminals.The problems of the information system were analyzed during iner-regional,base-like rotational training.Results The information system had its functions realized,and stills had to be improved in casualty information input flow,precision materials management and allocation standard of operating terminal.Conclusion The field medical station information system contributes to enhancing its service efficiency and informatization.

Advances on studies of miR-21 in vascular endothelial function and angiogenesis / 中国病理生理杂志

MicroRNAs ( miRNAs) are a class of non-coding , endogenous , single-stranded small RNA mole-cules composed of 19~25 nucleotides .miRNAs are widely involved in the process of human life activities .Recent studies have shown that part of miRNAs regulate the vascular endothelial function and angiogenesis .High expression of miRNA-21 is found to play important roles in the cell proliferation , cell apoptosis , cell growth and death of vascular endothelial cells . This review will focus on the recent progress related to miRNAs in vascular endothelial function and angiogenesis , providing a new insight in cardiovascular disease prevention , clinical diagnosis , prognosis and target therapeutics .

Inter-regional, base-like and informatized support of field medical station during training / 医疗卫生装备

Objective To explore the inter-regional,base-like and informatized support of the field medical station during rotational training.Methods The field medical station information system developed by the hospital was introduced,which had the working mode involving in a set of system and two kinds of terminals.The problems of the information system were analyzed during iner-regional,base-like rotational training.Results The information system had its functions realized,and stills had to be improved in casualty information input flow,precision materials management and allocation standard of operating terminal.Conclusion The field medical station information system contributes to enhancing its service efficiency and informatization.

Effects of miR-30 c on viability and migratory ability of HUVECs by tar-geting PAI-1 / 中国病理生理杂志

AIM: To investigate the effect of microRNA (miR)-30c on the viability and migratory ability of human umbilical vein endothelial cells (HUVECs) by targeting plasminogen activator inhibitor-1 (PAI-1).METHODS:The HUVECs were transfected with miR-30c mimic and inhibitor or negative control (NC), and then the expression levels of miR-30c, PAI-1 mRNA and protein were detected by RT-qPCR and Western blot.The viability and migratory ability of HUVECs were measured by CCK-8 assay and wound healing test .After bioinformatic analysis, the assessment of miR-30c binding to PAI-1 3’-UTR was carried out using a luciferase reporter gene assay .RESULTS:miR-30c directly down-regu-lated PAI-1 levels by binding to the 3’ UTR seed sequence of PAI-1 mRNA.Furthermore, transfection of a miR-30c mimic down-regulated the expression of PAI-1 at mRNA and protein levels, leading to enhanced migratory ability and viability of the HUVECs.However, transfection of a miR-30c inhibitor up-regulated the expression of PAI-1 at mRNA and protein le-vels, leading to decreased migratory ability and viability .CONCLUSION:Regulation of miR-30c level changes the migra-tory ability and viability of HUVECs by affecting the PAI-1 expression, indicating the involvement of miR-30c in modulating endothelial function .

Chemical constituents from branch of Fraxinus sieboldiana / 中国中药杂志

Using a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, macroporous adsorbent resin, and reversed-phase HPLC, 115 compounds including diterpenes, sesquiterpenes, treterpenes, coumarins, lignans, fatty acid derivatives, and simple aromatic derivatives were isolated from an ethanol extract of branch of Fraxinus sieboldiana (Oleaceaue), and their structures of the compounds were elucidated by spectroscopic methods including 1 D, 2D NMR and MS techniques. Among them, 41 compounds were new. In previous reports, we have been described the isolation, structure elucidation, and bioactivities of the 41 new compounds and 22 known orii including 8 coumarins, 4 phenolic and 12 phenylethanoidal glycosides. As a consequence, we herein reported the isolation and structure elucidation of the remaining 50 known compounds including 8- hydroxy-12-oxoabieta-9(11),13-dien-20-oic 8, 20-lactone(1), 6beta-hydroxyfcrruginol(2),(+)-pisiferic acid(3), (+)-pisiferal(4),(+)-7-dehydroabiet6none(5), 1-oxomiltirone(6), subdigitatone(7), linarionoside B(8), (9S)-linarionoside B(9), (3R,9R)-3-hydroxy-7,8-dihydro-beta-ionol 9-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside(10), ursolic acid(11), betulinic acid(12), euscaphic acid(13), (+)-syringaresinol(14), (+)-fraxiresinol(15), (+)-1-hydroxysyringaresinol(16), pinoresinol(17), medioresinol(18), 8-acetoxypinoresinol(19), epipinoresinol(20), (-)-olivil(21), (+)-cyclo-olivil(22), 3,3'-dimethoxy-4,4',9-trihydroxy-7,9'-epoxylignan-7'-one(23),(+)-1-hydroxypinoresinol 4'-O-beta-D-glucopyranoside (24), (+)-1-hydroxypinoresinol 4"-O-beta-D-glucopyranoside(25),(+)-syringaresinol O-beta-D-glucopyranoside (26), liriodendrin (27), ehletianol D(28), icariside E5(29) (-)-(7R, 8R)-threo-1-C-syringylglycerol(30),(-)-(7R, 8S)-erythro-guaiacylglycerol (31),(-)-(7R, 8R)-threo-guaiacylglycerol(32), 3-(4-beta-D-glucopyranosyloxy-3-methoxy)-phenyl-2E-propenol(33),2,3-dihydroxy-l-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone(34), 2,3-dihydroxy-1-(4-hydroxy-3-methoxyphenyl)-1-propanone (35), 3-hydroxy-l-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone(36), omega-hydroxypropioguaiacone(37), sinapyladehyde(38), trans-p-hydroxycinnamaldehyde(39), syringic acid(40), vanilic acid(41), vanillin(42), 4-hydroxy-benzaldehyde (43), (24R)-24-ethyl-5alpha-cholestane-3beta,5,6beta-triol(44), beta-sitosterol(45), daucosterol(46), 2,6-dimethoxy-I,4-benzoquinone(47), 2,6-dimethoxy-pyran-4-one(48), 1-(beta-D-ribofuranosyl)uracil(49), and mannitol(50). Compouds 1-7,12,18,28-37,44 and 48 were obtained from the genus Fraxinus for the first time.

Identification of tetracenomycin X from a marine-derived Saccharothrix sp. guided by genes sequence analysis / 药学学报

The crude extracts of the fermentation broth from a marine sediment-derived actinomycete strain, Saccharothrix sp. 10-10, showed significant antibacterial activities against drug-resistant pathogens. A genome-mining PCR-based experiment targeting the genes encoding key enzymes involved in the biosynthesis of secondary metabolites indicated that the strain 10-10 showed the potential to produce tetracenomycin-like compounds. Further chemical investigation of the cultures of this strain led to the identification of two antibiotics, including a tetracenomycin (Tcm) analogs, Tcm X (1), and a tomaymycin derivative, oxotomaymycin (2). Their structures were identified by spectroscopic data analysis, including UV, 1D-NMR, 2D-NMR and MS spectra. Tcm X (1) showed moderate antibacterial activities against a number of drug-resistant pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE) pathogens, with the MIC values in the range of 32-64 microg x mL(-1). In addition, 1 also displayed significant cytotoxic activities against human cancer cell lines, including HL60 (leukemia), HepG2 (liver), and MCF-7 (breast) with the IC 50 values of 5.1, 9.7 and 18.0 micromol x L(-1), respectively. Guided by the PCR-based gene sequence analysis, Tcm X (1) and oxotomaymycin (2) were identified from the genus of Saccharothrix and their 13C NMR data were correctly assigned on the basis of 2D NMR spectroscopic data analysis for the first time.

Halogenated natural products from the marine-derived actinobacteria and their halogenation mechanism / 药学学报

In the last decade, along with the development of taxonomy research in marine-derived actinobacteria, more and more halogenated natural products were discovered from marine actinobacteria. Most of them showed good biological activity and unique structure compared to those from land. The special halogenation mechanism in some compounds' biosynthesis has drawn great attention. So in this review, we focus on the halogenated natural products from marine actinobacteria and their halogenation mechanisms.

Apply research of MRI combined with 1H-MRS in meningiomas / 国际肿瘤学杂志

Objective To evaluate the roles of magnetic resonance imaging and proton magnetic resonance spectroscopy(1H-MRS) in the diagnosis of meningiomas. Methods 98 patients with meningiomas underwent conventional pre-contrast MR and contrast MR. Among them, 28 cases had two dimensional single voxel or multi voxel 1 H-MRS simultaneously both in the lesion's region and the contralateral side. Results On precontrast MR images of 98 cases, T1 WI showed 58.1% (61/105) isointensities, 31.4% (33/105) faintly low intensities and 10. 5% (11/105) mixed intensities; T2WI showed 40. 0% (42/105) isointensities, 41.0%(43/105) hyperintensities, 10.5% (11/105) faintly low intensities and 8.5% (9/105) mixed intensities. After administration of Gd-DTPA, the solid part of the tumors exhibited various enhancement in all the 98 cases.28 cases of MRS exhibited specific different spectral peaks, including increased of choline-containing compounds(Cho), absent or decreased of acetylaspartate(NAA), and the unchanged of creatine(Cr). The value of NAA, Cr, Cho, NAA/Cr, Cho/Cr, NAA/Cho in the tumor center of meningioma were 0. 09 ± 0.06,0.31 ± 0. 22, 0.46 ± 0. 16, 0.33 ± 0. 42, 1.50 ± 0. 68, 0. 15 ± 0.08, compared with the contralateral normal region, Cr has no significant difference (P ＞ 0. 05), NAA, Cho, NAA/Cr, Cho/Cr, NAA/Cho had significantly differences(P ＜ 0.05). Conclusion Conventional pre-contrast MR and contrast MR is the most important dignostic means for meningiomas, 1H-MRS combined with MRI can improve the diagnostic accuracy of meningiomas.

Role of integrin-linked kinase in renal tubular epithelial-mesenchymal transition and the regulatory effect of urokinase on its expression in mice with obstructive nephropathy / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of integrin-linked kinase (ILK) on renal tubular epithelial-mesenchymal transition and the regulatory effect of urokinase on LIK expression in mice with obstructive nephropathy.</p><p><b>METHODS</b>Normal male mice were randomly divided into sham-operated group (n=20), unilateral ureteral obstruction (UUO) group (n=28), and UUO with urokinase treatment group (uPA, n=28), and UUO was induced surgically in the latter two groups. The mice were sacrificed on days l, 3, 7 and 14 after the surgery, and renal interstitial fibrosis (RIF) was graded according to the result of Masson staining. The expression of ILK in the renal tissues of the rats was examined by immunofluorescence staining and Western blotting, and the expression of E-cadherin was detected by immunohistochemistry. RT-PCR was used to examine the mRNA expressions of ILK, E-cadherin and alpha-smooth muscle actin (alpha-SMA).</p><p><b>RESULTS</b>The expressions of ILK mRNA and protein were significantly increased in UUO group, but significantly decreased by treatment with uPA (P<0.05). The expression of alpha-SMA mRNA level was significantly increased, while E-cadherin decreased in mice with UUO on day 3 after the surgery. Treatment with uPA significantly inhibited such effects (P<0.05).</p><p><b>CONCLUSION</b>ILK plays an important role in renal interstitial fibrosis by mediating epithelial-mesenchymal transition. Urokinase attenuates renal tubulointerstitial fibrosis in mice with UUO possibly by inhibiting ILK expression and preventing tubular epithelial-mesenchymal transition.</p>

Effects of urokinase on renal interstitial fibrosis and transforming growth factor-beta1 in the kidney of rats with chronic cyclosporine A nephropathy / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of urokinase on renal interstitial fibrosis and transforming growth factor-beta1 (TGF-beta1) in the kidney of rats with chronic cyclosporine A nephropathy.</p><p><b>METHODS</b>Male Sprague-Dawley rats on low-salt diet were randomly divided into control (VH), CsA-treated (CsA), CsA+2000 U/kg.day uPA (CsA+U2) and CsA+6000 U.kg.3 days (CsA+U6) groups. The rats were given CsA intragastrically for 4 weeks to prepare CsA-induced chronic nephropathy model. Masson staining was used to examine fibrin deposition. Western blotting and reversal transcription polymerase chain reaction were employed to evaluate urokinase-type plasminogen activator (uPA) and TGF-beta1 protein and gene expressions, respectively.</p><p><b>RESULTS</b>CsA can increase fibrin deposition and the expression of TGF-beta1 in the renal tissue, which were significantly reduced after uPA treatment (P<0.05).</p><p><b>CONCLUSION</b>Continuous low-dose uPA treatment can reduce renal interstitial fibrosis in rats possibly in association with its inhibitory effect on TGF-beta1 expression.</p>

Effect of urokinase on renal interstitial inflammation in rats with chronic cyclosporine A nephropathy and its possible mechanism / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the protective effect of urokinase on renal interstitial inflammation and fibrosis in rats with chronic cyclosporine A (CsA)-induced nephropathy.</p><p><b>METHODS</b>Male SD rats were fed on low salt diet (0.05% sodium) for 7 days and randomized into 4 groups for treatment with CsA, CsA+continuous low-dose uPA (U2), intermittent CsA+ high-dose uPA (U6) or vehicle (control group). In the former 3 groups, the rats were subjected to daily intragastric administration of CsA (25 mg/kg) for 4 weeks to establish CsA-induced chronic nephropathy model, and those in U2 and U6 groups were given uPA at 2000 U/kg daily or at 6000 U/kg every 3 days, respectively. Four weeks after the treatment, the renal function and 24-h proteinuria were assessed, and Masson staining was used for examining fibrin deposition. Semi-quantitative immunohistochemical staining was employed for evaluation of ED-1-positive cells, urokinase-type plasminogen activator (uPA) and transforming growth factor-beta1 (TGF-beta 1).</p><p><b>RESULTS</b>Four weeks after the treatment, the CsA-treated rats showed significantly elevated serum creatinine (Scr), blood urea nitrogen (BUN) and increased urine proteins. Continuous administration of low-dose uPA resulted in significantly reduced Scr, BUN and 24-h urine protein excretion, while intermittent high-dose uPA treatment did not produce such changes. CsA increased fibrin deposition, total number of macrophages in renal interstitium and TGF-beta1 expression in the renal tissue, which were significantly reduced in U2 group (P<0.05) but not in U6 group (P>0.05).</p><p><b>CONCLUSION</b>Continuous administration of low-dose uPA may reduce interstitial fibrin deposition and alleviate renal interstitial inflammation in rats with chronic CsA nephropathy, possibly by reducing the number of macrophages and TGF-beta1 expression in the renal tissue.</p>