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1.
iScience ; 27(1): 108690, 2024 Jan 19.
Article in English | MEDLINE | ID: mdl-38235340

ABSTRACT

Prenatal stress has been extensively documented as a contributing factor to adverse cardiac development and function in fetuses and infants. The release of glucocorticoids (GCs), identified as a significant stressor, may be a potential factor inducing cardiac hypertrophy. However, the underlying mechanism remains largely unknown. Herein, we discovered that corticosterone (CORT) overload induced cardiac hypertrophy in embryonic chicks and fetal mice in vivo, as well as enlarged cardiomyocytes in vitro. The impaired mitochondria dynamics were observed in CORT-exposed cardiomyocytes, accompanied by dysfunction in oxidative phosphorylation and ATP production. This phenomenon was found to be linked to decreased mitochondrial fusion protein mitofusin 2 (MFN2). Subsequently, we found that CORT facilitated the ubiquitin-proteasome-system-dependent degradation of MFN2 with an enhanced binding of appoptosin to MFN2, serving as the underlying cause. Collectively, our findings provide a comprehensive understanding of the mechanisms by which exposure to stress hormones induces cardiac hypertrophy in fetuses.

2.
Environ Pollut ; 342: 123049, 2024 Feb 01.
Article in English | MEDLINE | ID: mdl-38042470

ABSTRACT

Smoldering can achieve effective disposal of sewage sludge (SS) with high moisture content at low energy input, providing social and economic benefits. However, smoldering is accompanied by the emission of high concentrations of CO/NOx, and thus, it requires sufficient attention. This study comprehensively investigates the effects of SS characteristics and experimental parameters on CO/NOx emissions and smoldering characteristics. Results showed that when the moisture content of SS increases from 35% to 50%, CO concentration increases while NOx formation is simultaneously inhibited. After airflow rate exceeds 5 cm/s, the concentrations of CO and NOx begin to decrease. When SS concentration is increased to 20%, the emission concentration of gas pollutants is directly increased. However, high temperatures inhibit the formation of NOx. When the particle size range is 180-270 µm, the formation of CO/NOx is promoted. Finally, a back propagation (BP) neural network model is constructed with SS characteristics and experimental parameters as input conditions, and CO/NOx emission concentration, smoldering velocity, and smoldering temperature as output parameters. The BP neural network model can effectively predict the emission concentration of CO/NOx and smoldering characteristics, providing support for intelligent control scenarios related to SS smoldering, it will help to further explore the great potential of smoldering treatment.


Subject(s)
Air Pollutants , Environmental Pollutants , Sewage , Air Pollutants/analysis , Temperature
3.
Environ Res ; 238(Pt 2): 117271, 2023 12 01.
Article in English | MEDLINE | ID: mdl-37793589

ABSTRACT

Co-pyrolysis of sewage sludge (SS) and tobacco stalk (TS) was conducted, where TS are rich in inorganic elements. The effects of the mixing ratio and pyrolysis temperature on pyrolysis products were investigated through the tube furnace system. The components of gas and tar were determined. In addition, the N/S ratio, pore distribution characteristics, surface functional group characteristics, aromatisation characteristics and inorganic element content of char were studied. Results show that the addition of TS to SS has a significant synergistic effect. Particularly, when the ratio of SS to TS is 1:3, the pyrolysis gas yield is the largest. The inorganic elements with a catalytic effect can promote the conversion of char and tar into pyrolysis gas. However, with the increase in pyrolysis temperature, the synergistic effect will gradually weaken. The increase in pyrolysis temperature will promote the development of mesopores in char but will reduce the order degree of the char structure. The addition of TS in SS will further increase the number of mesopores in char, and the obtained char carbon structure is more ordered. When the pyrolysis temperature is 600 °C, the tar yield is the highest, and the higher the mixing ratio of SS, the better the formation of polyaromatic compounds and aliphatic hydrocarbons.


Subject(s)
Nicotiana , Sewage , Pyrolysis , Temperature , Carbon
4.
Chinese Journal of School Health ; (12): 1058-1062, 2023.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-985361

ABSTRACT

Objective@#Based on Newell s Model of Constraints, the present study aims to explore the effects of sports environment design on children s fundamental movement skills, and to provide certain theoretical and practical guidance for promoting children s movement development and carrying out sports activities.@*Methods@#From March to June 2022, using the method of cluster sampling, 153 typical developmental children from a private kindergarten in Xi an were selected, and were randomly assigned to the regular activity group, sports division A group, and sports division B group for a period of 12 weeks (twice a week, 1 hour each time) experimental intervention. Among them, the regular activity group engaged in regular physical activities according to the kindergarten plan; the sports division A group participated in daily physical activities after the sports division; and the sports division B group engaged in structured and autonomous physical activities after the sports division. Motorische Basiskompetenzen in Kindergarten (MOBAK-KG) scale was used to assess the level of fundamental movement skills. Chi square test, one way analysis of variance and repeated measures analysis of variance were used to test the effects of sports environment design on children s fundamental movement skills.@*Results@#After the experiment, children s manipulative skills,mobility skills, and fundamental movement skills improved significantly ( P <0.01); group and test time had a significant interaction on children s fundamental movement skills [ F (2,150) =113.07, P <0.01, η 2=0.60],the posttest score of fundamental movement skills of children in group B of sports division (12.08±1.82) was significantly higher than that of group A of sports division (10.71±2.56), regular activity group (8.57±4.16).@*Conclusion@#The sports environment design under the constraint model perspective can effectively promote the development of children s fundamental movement skills. Kindergartens, families and communities should coordinate to promote the development of children s fundamental movement skills.

5.
J Magn Reson ; 294: 122-127, 2018 09.
Article in English | MEDLINE | ID: mdl-30036812

ABSTRACT

The flip-back pulse combined with cross-polarization magic-angle spinning (CPMAS-FB) has been widely used to reduce the optimal repetition delay, resulting in enhancing sensitivity per unit time. Despite its common use in samples with long 1H T1 relaxation time, the experimental setup of the repetition delay in CPMAS-FB is not obvious. In this article, a simple model is used to derive the optimal repetition delay and expected sensitivity gain through the 1H T1 relaxation time and signal decay during 1H spinlock.

6.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-668039

ABSTRACT

Objective:To study the effects of insulin and berberine on osseointegration of titanium implant in type Ⅱ diabetic rats.Methods:10 out of 53 SD rats were randomly selected and selected into H group(healthy group)(n =10).The remaining rats were treated with high glucose diet plus low dose of streptozotocin(STZ) and 40 rats were induced into the models of type Ⅱ diabeties.The 40 dibetic rats were randomly divided into group B(berberine),group Ⅰ(insulin),group IB(insulin + berberine) and group N(without treatment).2 pure titanium implants were implanted into the bilateral femoral metaphysis of each rat.Mter 7,32,38,66 and 101 d,the body weight(BW) and fasting blood glucose(GLU) were measured.After 10 week,the rats were sacrifice,femoral metaphysis samples were subjected to Micro-CT scan,VG staining and implant pull-out test.Hard tissue slices were observed;bone volume fraction(BV/TV),Liang Houdu (Tb.Th),trabecular bone trabecular number(Tb.N),Liang Jianxi (Tb.Sp),trabecular bone trabecular bone volume (TBV),bone contact ratio(BIC) and the maximum pull-out strength (MPF) were measured.Results:There was no significant difference of BW and GLU between IB group and H group(P > 0.05),but the BW and GLU of IB group was better than those of Ⅰ group and B group,Ⅰ group and B group were better than those of group N (P < 0.05).BV/TV,Tb.Th,Tb.N,Tb.Sp,TBV,BIC and MPF of IB group were not statistically different from those of H group(P >0.05);but the indexes of IB group were higher than those of Ⅰ group and B group,those of Ⅰ group and B group were higher than those of N group (P < 0.05).Conclusion:The combination of insulin and berberine can effectively reduce blood glucose and can promote osseointegration of titanium implants in type Ⅱ diabetic rats.

7.
Eur J Pharm Sci ; 85: 47-52, 2016 Mar 31.
Article in English | MEDLINE | ID: mdl-26836368

ABSTRACT

Cocrystallization of an active pharmaceutical ingredient (API) with a cocrystal former (co-former) is widely used to tailor the physicochemical properties of parent APIs. For proton-pump inhibitors (PPIs), the isolation of cocrystals has not been widely investigated. Here, a 1:1 cocrystal of a PPI molecule, dexlansoprazole (DLS), was obtained by solvent crystallization with isonicotinamide (INM). The product was characterized by X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), solid-state and liquid NMR, as well as Fourier transform infrared spectroscopy (FTIR) techniques. A two-point R2(2)(9) hetero-synthon was proposed to exist in the cocrystal, where intermolecular hydrogen bonding occurs between NH, SO groups of DLS and amide of INM. The dissolution profiles of DLS and DLS-INM in water were also collected, and the results demonstrate the cocrystal exhibits superior apparent maximum solubility relative to the pure drug.


Subject(s)
Dexlansoprazole/chemistry , Niacinamide/chemistry , Solubility , Calorimetry, Differential Scanning/methods , Crystallization/methods , Hydrogen Bonding , Magnetic Resonance Spectroscopy/methods , Powder Diffraction/methods , Proton Pump Inhibitors/chemistry , Solvents/chemistry , Spectroscopy, Fourier Transform Infrared/methods , X-Ray Diffraction/methods
8.
J Cereb Blood Flow Metab ; 35(1): 86-94, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25335802

ABSTRACT

Human apolipoprotein E (APOE) exists in three isoforms ɛ2, ɛ3, and ɛ4, of which APOE4 is the main genetic risk factor of Alzheimer's disease (AD). As cerebrovascular defects are associated with AD, we tested whether APOE genotype has an impact on the integrity and function of the blood-brain barrier (BBB) in human APOE-targeted replacement mice. Using the quantitative in situ brain perfusion technique, we first found lower (13.0% and 17.0%) brain transport coefficient (Clup) of [(3)H]-diazepam in APOE4 mice at 4 and 12 months, compared with APOE2 and APOE3 mice, reflecting a decrease in cerebral vascularization. Accordingly, results from immunohistofluorescence experiments revealed a structurally reduced cerebral vascularization (26% and 38%) and thinner basement membranes (30% and 35%) in 12-month-old APOE4 mice compared with APOE2 and APOE3 mice, suggesting vascular atrophy. In addition, APOE4 mice displayed a 29% reduction in [(3)H]-d-glucose transport through the BBB compared with APOE2 mice without significant changes in the expression of its transporter GLUT1 in brain capillaries. However, an increase of 41.3% of receptor for advanced glycation end products (RAGE) was found in brain capillaries of 12-month-old APOE4 mice. In conclusion, profound divergences were observed between APOE genotypes at the cerebrovascular interface, suggesting that APOE4-induced BBB anomalies may contribute to AD development.


Subject(s)
Apolipoprotein E4/genetics , Blood-Brain Barrier/physiology , Cerebrovascular Circulation/genetics , Alzheimer Disease/genetics , Alzheimer Disease/physiopathology , Animals , Apolipoprotein E2/genetics , Apolipoprotein E3/genetics , Blood-Brain Barrier/metabolism , Brain/blood supply , Brain/metabolism , Brain/physiopathology , Diazepam/pharmacokinetics , Female , Humans , Male , Mice , Mice, Inbred C57BL , Perfusion , Protein Isoforms
9.
Bioorg Med Chem ; 22(8): 2442-6, 2014 Apr 15.
Article in English | MEDLINE | ID: mdl-24679673

ABSTRACT

A new inhibitor of TNF-α production (IC50=0.89 µM) named vialinin C (1) was isolated from dry fruiting bodies of an edible Chinese mushroom, Thelephora vialis. The structure of 1 was determined by high-resolution MS, NMR spectroscopic analysis, and confirmed by synthesis. Synthesis of ganbajunin B (5) obtained from the same origin was also described.


Subject(s)
Benzofurans/chemical synthesis , Parabens/chemical synthesis , Terphenyl Compounds/metabolism , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Agaricales/chemistry , Agaricales/metabolism , Benzofurans/chemistry , Magnetic Resonance Spectroscopy , Molecular Conformation , Parabens/chemistry , Terphenyl Compounds/chemistry , Terphenyl Compounds/isolation & purification , Tumor Necrosis Factor-alpha/metabolism
10.
J Magn Reson ; 239: 75-80, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24424008

ABSTRACT

A novel method to realize rapid repetition of (1)H NMR experiments at ultra-fast MAS frequencies is demonstrated. The ultra-fast MAS at 110kHz slows the (1)H-(1)H spin diffusion, leading to variations of (1)H T1 relaxation times from atom to atom within a molecule. The different relaxation behavior is averaged by applying (1)H-(1)H recoupling during relaxation delay even at ultra-fast MAS, reducing the optimal relaxation delay to maximize the signal to noise ratio. The way to determine optimal relaxation delay for arbitrary relaxation curve is shown. The reduction of optimal relaxation delay by radio-frequency driven recoupling (RFDR) was demonstrated on powder samples of glycine and ethenzamide with one and multi-dimensional NMR measurements.


Subject(s)
Amino Acids/chemistry , Nuclear Magnetic Resonance, Biomolecular/methods , Algorithms , Electromagnetic Fields , Glycine/analysis , Histidine/analysis , Signal-To-Noise Ratio
11.
PLoS One ; 8(12): e80931, 2013.
Article in English | MEDLINE | ID: mdl-24349023

ABSTRACT

Tumor necrosis factor alpha (TNF-α), a central mediator of the inflammatory response, is released from basophilic cells and other cells in response to a variety of proinflammatory stimuli. Vialinin A is a potent inhibitor of TNF-α production and is released from RBL-2H3 cells. Ubiquitin-specific peptidase 5 (USP5), a deubiquitinating enzyme, was identified as a target molecule of vialinin A and its enzymatic activity was inhibited by vialinin A. Here we report production of TNF-α is decreased in USP5 siRNA-knockdown RBL-2H3 cells, compared with control cells. The finding of the present study strongly suggests that USP5 is one of the essential molecules for the production of TNF-α in RBL-2H3.


Subject(s)
Terphenyl Compounds/metabolism , Tumor Necrosis Factor-alpha/metabolism , Animals , Blotting, Western , Cell Line , Endopeptidases/metabolism , Interleukin-4/metabolism , RNA, Small Interfering , Rats , Reverse Transcriptase Polymerase Chain Reaction
12.
BMC Gastroenterol ; 13: 140, 2013 Sep 23.
Article in English | MEDLINE | ID: mdl-24053454

ABSTRACT

BACKGROUND: Non-alcoholic steatohepatitis (NASH) is a condition that occurs during the progression of non-alcoholic fatty liver disease. Effective therapy for NASH is still lacking. In this study, we investigated the effects of Ursodeoxycholic acid (UDCA) in the treatment of NASH. METHODS: Western and Chinese databases were searched by independent investigators using appropriate MESH headings to identify randomized, controlled Western and Chinese clinical trials, published between January 1990 and October 2012, testing the effects of UDCA in patients with NASH. Patient characteristics and trial endpoints were analyzed, with quality assessment according to widely acknowledged criteria. P < 0.05 was defined as statistically significant in all trials. RESULTS: Twelve qualified randomized clinical trials, including six from China and involving 1160 subjects, were selected. Seven of these trials assessed the effects of UDCA Monotherapy, with the other five testing combinations of UDCA with vitamin E, polyene phosphatidylcholine, silymarin, glycyrrhizin and tiopronin. The duration of therapy ranged from 3 to 24 months, with two studies using high doses of UDCA (23-35 mg/kg/d). The average quality point was 2.69, and was significantly lower in articles from China than in those from Western countries (2.2 ± 0.4 vs. 3.8 ± 1.1, respectively, p < 0.05). UDCA Monotherapy significantly improved liver function in five studies and improved steatosis and fibrosis in two studies. All five studies assessing UDCA combination therapy showed significant improvements liver function, while two studies also improved steatosis and inflammation. One study of high-dose UDCA showed significant improvements in ALT, γGT and liver fibrosis, whereas the other study showed no significant change in ALT and liver pathology. CONCLUSIONS: UDCA therapy is effective in NASH, especially when combined with other drugs. However, the low quality of these studies and the heterogeneity of their results precluded further meta-analysis. Additional carefully designed clinical trials are needed, especially in China.


Subject(s)
Cholagogues and Choleretics/therapeutic use , Fatty Liver/drug therapy , Ursodeoxycholic Acid/therapeutic use , Humans , Non-alcoholic Fatty Liver Disease , Treatment Outcome
13.
World J Gastroenterol ; 19(28): 4576-81, 2013 Jul 28.
Article in English | MEDLINE | ID: mdl-23901235

ABSTRACT

AIM: To investigate the association between Helicobacter pylori (H. pylori) infection and the prevalence of Crohn's disease (CD). METHODS: Subjects were selected from patients admitted the gastrointestinal (GI) department at The First Affiliated Hospital School of Medicine (Zhejiang University) for abdominal pain, hematochezia, diarrhea and other GI symptoms between January 2008 and September 2012. CD was diagnosed by endoscopy and biopsy. H. pylori infection was detected by a (14)C-urea breath test and culturing of the biopsy sample. Demographic, anthropometric and serologic data were collected for each patient. H. pylori infection rate was compared between CD and control groups, followed by a subgroup analysis based on extent and severity of CD. Student's t, Mann-Whiney U, and χ(2) tests were used to analyze the data. RESULTS: A total of 447 patients were analyzed, including 229 in the CD group and 248 in the control group. There were no significant differences in age, sex, and rates of hypertension or diabetes. However, the CD group showed significantly higher rates of smoking history (34.9% vs 18.1%), alcohol intake (17.4% vs 8.1%), white blood cell count (9.7 ± 2.9 × 10(9)/L vs 4.3 ± 0.9 × 10(9)/L), and C-reactive protein (36.3 ± 20.8 mg/L vs 5.5 ± 2.3 mg/L) but lower body mass index (24.5 ± 2.0 kg/m(2) vs 26.0 ± 2.2 kg/m(2)) than the control group. The H. pylori infection rate in the CD group was 27.1%, significantly lower than that of 47.9% in the control group. Furthermore, the H. pylori infection rates in patients with colonic, small intestine, ileocolonic and extensive CD were 31.1%, 28.9%, 26.8% and 25.9% respectively, all of which were significantly lower than in the control group. Finally, the H. pylori infection rates in patients with remission, moderate and severe CD were 34.3%, 30.7% and 22.0% respectively, which were also significantly lower than in the control group. CONCLUSION: Lower H. pylori infection in CD patients suggests a correlation between bacterial infection and CD, suggesting caution when considering H. pylori eradication in CD patients.


Subject(s)
Crohn Disease/epidemiology , Helicobacter Infections/epidemiology , Helicobacter pylori/isolation & purification , Adult , Biopsy , Breath Tests , Chi-Square Distribution , China , Crohn Disease/diagnosis , Crohn Disease/therapy , Endoscopy, Gastrointestinal , Female , Helicobacter Infections/diagnosis , Helicobacter Infections/microbiology , Helicobacter Infections/therapy , Humans , Male , Middle Aged , Prevalence , Prognosis , Remission Induction , Retrospective Studies , Risk Factors , Serologic Tests , Severity of Illness Index
14.
Chin Med J (Engl) ; 126(16): 3151-5, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23981628

ABSTRACT

OBJECTIVE: To review the current advances on the role of uncoupling protein (UCP) in the pathogenesis and progress of nonalcoholic fatty liver disease (NAFLD). DATA SOURCES: A comprehensive search of the PubMed literature without restriction on the publication date was carried out using keywords such as UCP and NAFLD. STUDY SELECTION: Articles containing information related to NAFLD and UCP were selected and carefully analyzed. RESULTS: The typical concepts, up-to-date findings, and existing controversies of UCP2 in NAFLD were summarized. Besides, the effect of a novel subtype of UCP (hepatocellular down regulated mitochondrial carrier protein, HDMCP) in NAFLD was also analyzed. Finally, the concept that any mitochondrial inner membrane carrier protein may have, more or less, the uncoupling ability was reinforced. CONCLUSIONS: Considering the importance of NAFLD in clinics and UCP in energy metabolism, we believe that this review may raise research enthusiasm on the effect of UCP in NAFLD and provide a novel mechanism and therapeutic target for NAFLD.


Subject(s)
Fatty Liver/etiology , Ion Channels/physiology , Mitochondrial Proteins/physiology , Animals , Fatty Acids, Nonesterified/metabolism , Fatty Liver/metabolism , Humans , Mitochondrial Proteins/analysis , Mitochondrial Proteins/chemistry , Non-alcoholic Fatty Liver Disease , Uncoupling Protein 2
15.
Bioorg Med Chem Lett ; 23(15): 4328-31, 2013 Aug 01.
Article in English | MEDLINE | ID: mdl-23791076

ABSTRACT

Vialinin A, a small compound isolated from the Chinese mushroom Thelephora vialis, exhibits more effective anti-inflammatory activity than the widely used immunosuppressive drug tacrolimus (FK506). Here, we show that ubiquitin-specific peptidase 5/isopeptidase T (USP5/IsoT) is a target molecule of vialinin A, identified by using a beads-probe method. Vialinin A inhibited the peptidase activity of USP5/IsoT and also inhibited the enzymatic activities of USP4 among deubiquitinating enzymes tested. Although USPs are a member of thiol protease family, vialinin A exhibited no inhibitions for other thiol proteases, such as calpain and cathepsin.


Subject(s)
Anti-Inflammatory Agents/chemistry , Endopeptidases/chemistry , Protease Inhibitors/chemistry , Terphenyl Compounds/chemistry , Animals , Anti-Inflammatory Agents/metabolism , Cell Line , Endopeptidases/genetics , Endopeptidases/metabolism , Protease Inhibitors/metabolism , Protein Binding , Rats , Recombinant Proteins/biosynthesis , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Terphenyl Compounds/metabolism
16.
Zhongguo Zhong Xi Yi Jie He Za Zhi ; 33(12): 1658-64, 2013 Dec.
Article in Chinese | MEDLINE | ID: mdl-24517065

ABSTRACT

OBJECTIVE: To study the effect of Erigeron Breviscapus (EB) at different concentrations and different intervention time points on the mRNA and protein expression of OPG/RANKL/RANK in MG63 osteoblast-like cells and RAW264. 7 pre-osteoclast cells cultured in vitro, thus exploring roles EB played in bone rebuilding and its mechanisms. METHODS: MG63 osteoblast-like cells and RAW264.7 pre-osteoclast cells were cultured in vitro. The 3rd passage cells were divided into the control group and different experimental groups. Total RNA and protein were respectively isolated from cells treated with different concentrations of EB (0, 0.001, 0.01, 0.1, and 1.0 mg/mL) for 48 h. Meanwhile, the protein was extracted from 0 and 1 mg/mL EB groups at 12, 24, and 48 h respectively. Expression of OPG mRNA and RANKL mRNA in MG63 osteoblast-like cells, and expression of RANK mRNA in RAW264.7 pre-osteoclast cells were detected by semi-quantitative RT-PCR. Expression of OPG protein and RANKL protein in MG63 osteoblast-like cells, and expression of RANK protein in RAW264. 7 pre-osteoclast cells were detected by Western blot. RESULTS: Along with increased EB concentration, expression of OPG mRNA and protein in MG63 osteoblast-like cells was gradually lowered (P < 0.05) after 48-h intervention of EB, the expression of RANKL mRNA and protein in MG63 osteoblast-like gradually increased (P < 0.05); the expression of RANK mRNA in RAW264.7 pre-osteoclast cells increased (P < 0.05). But the expression of RANK mRNA was slightly lower in the 0.1 mg/mL EB group than in the 0.01 mg/mL EB group, and the expression of RANK protein in RAW264.7 pre-osteoclast cells gradually increased (P < 0.05). After treatment with 1 mg/mL EB for 12, 24, 48 h, the expression of OPG protein in MG63 osteoblast-like cells gradually decreased as time went by (P < 0.05), and the expression of RANKL protein in MG63 osteoblast-like and RANK protein in RAW264.7 pre-osteoclast cells gradually increased (P < 0.05). The expression of RANKL protein in RAW264.7 pre-osteoclast cells increased as time went by (P < 0.05). CONCLUSION: EB could inhibit the expression of OPG in osteoblasts in a dose- and time-dependent manner, promote the expression of RANKL in osteoblasts and the secretion of RANK in pre-osteoclast, indicating EB might play roles in promoting bone resorption.


Subject(s)
Drugs, Chinese Herbal/pharmacology , Erigeron , Osteoblasts/metabolism , Osteoclasts/metabolism , Osteoprotegerin/metabolism , Animals , Cell Differentiation , Cell Line , Humans , Mice , Osteoblasts/drug effects , Osteoclasts/drug effects , RANK Ligand/metabolism , RNA, Messenger/genetics , Receptor Activator of Nuclear Factor-kappa B/metabolism
17.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-440029

ABSTRACT

Objective To prepare an experimental encephalomyocarditis virus ( EMCV) vaccine and study its immunogenicity .Methods EMCVs were cultured in Vero cells and viral titer was detected by micro cytopathy method .A series of procedures including concentration , filtration and ultracentrifugation were conducted to purify EMCVs .Formaldehyde and β-propiolactone were used for viral inactivation and their efficacy was evaluated .The median lethal dose ( LD50 ) of EMCV was measured by using Reed-Muench method.The serum antibody titers in BALB/c mice vaccinated with different immunization strategies were analyzed through microneutralization assay .Then the immunized mice were challenged with 100LD50 , 50LD50 and 25LD50 of EMCV by intraperitoneal injection .Results The viruses could be inactivated by β-propiolactone at a volume ratio of 1 ∶4000 for 12 hours at 4℃.The LD50 of EMCV was 17 CCID50/ml.The immunized mice produced neutralizing antibodies and displayed resistance to EMCV infection.Conclusion The experimental EMCV vaccine could induce protective antibodies in mice .

18.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-436457

ABSTRACT

Objective To explore and develop methods for encephalomyocarditis virus (EMCV) identification.Methods According to the genetic sequence VR-129B of EMCV recorded in the GenBank,five gene fragments were selected to design primer sequence pairs.RNA was extracted to run RT-PCR,and then the products of amplification were identified by agarose gel electrophoresis.The results of DNA sequences were compared with the sequences in GenBank of the same EMCV strains.Antiserum was prepared based on the EMCV cultured in RK cells for establishing indirect immunofluorescence assay (IIFA) and neutralization test method,and verification for precision and specificity of the two methods were carried out after it.Antiserum that was prepared with GST-VP1 and GST-VP2 expressed in E.coli was reacted with the purified EMCV in Western blot test.Results By sequencing and comparing,the similarity of DNA fragments between the obtained and the GenBank recorded was reached 98% to 100%.The antiserum of No.20100901 batch that was chosen as the first antibody at a dilution of 1 ∶ 160 to develop IIFA brought about a better specificity.The neutralization titers of 20100901 batch antiserum was 1 ∶ 30 211 measured by fixing virus and diluting serum method,which showed good specificity and precision.The results of the Western blot test showed two clear bands above and under 33×103 respectively,which matched the theoretical value.Conclusion The RT-PCR,indirect immunofluorescence,neutralization test and Western blot method for EMCV strains identification were established initially.

19.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-231625

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of Erigeron Breviscapus (EB) at different concentrations and different intervention time points on the mRNA and protein expression of OPG/RANKL/RANK in MG63 osteoblast-like cells and RAW264. 7 pre-osteoclast cells cultured in vitro, thus exploring roles EB played in bone rebuilding and its mechanisms.</p><p><b>METHODS</b>MG63 osteoblast-like cells and RAW264.7 pre-osteoclast cells were cultured in vitro. The 3rd passage cells were divided into the control group and different experimental groups. Total RNA and protein were respectively isolated from cells treated with different concentrations of EB (0, 0.001, 0.01, 0.1, and 1.0 mg/mL) for 48 h. Meanwhile, the protein was extracted from 0 and 1 mg/mL EB groups at 12, 24, and 48 h respectively. Expression of OPG mRNA and RANKL mRNA in MG63 osteoblast-like cells, and expression of RANK mRNA in RAW264.7 pre-osteoclast cells were detected by semi-quantitative RT-PCR. Expression of OPG protein and RANKL protein in MG63 osteoblast-like cells, and expression of RANK protein in RAW264. 7 pre-osteoclast cells were detected by Western blot.</p><p><b>RESULTS</b>Along with increased EB concentration, expression of OPG mRNA and protein in MG63 osteoblast-like cells was gradually lowered (P < 0.05) after 48-h intervention of EB, the expression of RANKL mRNA and protein in MG63 osteoblast-like gradually increased (P < 0.05); the expression of RANK mRNA in RAW264.7 pre-osteoclast cells increased (P < 0.05). But the expression of RANK mRNA was slightly lower in the 0.1 mg/mL EB group than in the 0.01 mg/mL EB group, and the expression of RANK protein in RAW264.7 pre-osteoclast cells gradually increased (P < 0.05). After treatment with 1 mg/mL EB for 12, 24, 48 h, the expression of OPG protein in MG63 osteoblast-like cells gradually decreased as time went by (P < 0.05), and the expression of RANKL protein in MG63 osteoblast-like and RANK protein in RAW264.7 pre-osteoclast cells gradually increased (P < 0.05). The expression of RANKL protein in RAW264.7 pre-osteoclast cells increased as time went by (P < 0.05).</p><p><b>CONCLUSION</b>EB could inhibit the expression of OPG in osteoblasts in a dose- and time-dependent manner, promote the expression of RANKL in osteoblasts and the secretion of RANK in pre-osteoclast, indicating EB might play roles in promoting bone resorption.</p>


Subject(s)
Animals , Humans , Mice , Cell Differentiation , Cell Line , Drugs, Chinese Herbal , Pharmacology , Erigeron , Osteoblasts , Metabolism , Osteoclasts , Metabolism , Osteoprotegerin , Metabolism , RANK Ligand , Metabolism , RNA, Messenger , Genetics , Receptor Activator of Nuclear Factor-kappa B , Metabolism
20.
Cell Immunol ; 279(2): 140-4, 2012 Oct.
Article in English | MEDLINE | ID: mdl-23246504

ABSTRACT

Vialinin A is an extremely potent inhibitor of tumor necrosis factor (TNF)-α release from RBL-2H3 cells. The present study investigated in detail the inhibitory effects of vialinin A and its analog, 5',6'-dimethyl-1,1':4',1″-terphenyl-2',3',4,4″-tetraol (DMT), on TNF-α. Vialinin A and DMT inhibited the release of TNF-α from RBL-2H3 cells in a dose-dependent manner, but had no effect on ß-hexosaminidase activity. Also, vialinins had little effect on TNF-α mRNA levels. Intriguingly, vialinins inhibited TNF-α production at low concentrations, but not shown a dose-dependency. The potent inhibitory activities of vialinins against TNF-α production and release suggest promising new candidate pathways for anti-inflammatory agents.


Subject(s)
Cell Degranulation/drug effects , Terphenyl Compounds/pharmacology , Tumor Necrosis Factor-alpha/metabolism , beta-N-Acetylhexosaminidases/metabolism , Animals , Anti-Inflammatory Agents/pharmacology , Cell Line, Tumor , Female , Mice , Mice, Inbred BALB C , RNA, Messenger/biosynthesis , Rats , Tumor Necrosis Factor-alpha/genetics
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