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OBJECTIVE@#To investigate whether Blimp1 plays an anti-apoptosis role in myeloma by interfering with ATF4/CHOP cell apoptosis pathway induced by endoplasmic reticulum stress, and to explore the anti-myeloma mechanism of aspirin.@*METHODS@#The bone marrow fluid of 40 newly diagnosed multiple myeloma patients without treatment and 30 control people with relatively normal bone marrow was collected. Flow cytometry was used to separated the normal and abnormal plasma cells, LV-Blimp1-RNAi (40051-2) recombinant lentivirus down-regulates the expression of Blimp-1 in U266 cell line and detected the changes of the expression of ATF4 and CHOP gene. U266 cells were stimulated by aspirin at different concentrations (0, 0.5, 2.5, 5.0 mmol/L) in vitro. Then the effect of aspirin on proliferation of U266 cells was measured by CCK-8 assay, the mRNA expression levels of Blimp1, ATF4 and CHOP in four groups were detected by real-time PCR.@*RESULTS@#The expression level of Blimp1 in phenotype abnormal plasma cells was significantly increased as compared with normal cells, while the expression of ATF4 and CHOP in phenotype abnormal plasma cells was significantly decreased as compared with normal cells (P＜0.05). In the case of MOI=100, the transfection efficiency of U266 cells was beyond 80% as detected by fluorescence microscopy. Compared with blank conrol and negatine control groups, Blimp1 mRNA expression level in positive group was significantly reduced while ATF4 and CHOP expression significantly increased. CCK-8 showed that the proliferation activity of U266 cells could be inhibited by aspirin, which showed a time-and dose-dependent manner; at the same time, the expression level of Blimp1 in U266 cells were decreased with the increasing of aspirin concentration, while the expression level of ATF4 and CHOP was increased with the increasing of aspirin concentration.@*CONCLUSIONS@#Blimp1 may display the anti-apoptosis of myeloma cells through interfering with ATF4/CHOP signaling pathway; low dose of aspirin may play anti-myeloma effect by inhibiting the expression of Blimp1 in myeloma cells.
Subject(s)
Humans , Activating Transcription Factor 4 , Apoptosis , Cell Line, Tumor , Cell Proliferation , Multiple Myeloma , Positive Regulatory Domain I-Binding Factor 1 , Signal TransductionABSTRACT
OBJECTIVE@#To explore the expression of Blimp1, ATF4 and CHOP in bone marrow mononuclear cells from patients with multiple myeloma as well as the effect of aspirin on their expression.@*METHODS@#Sixty untreated patients with multiple myeloma and 30 patients with relatively normal bone marrow were selected. Mononuclear cells from the bone marrow fluid were separated using Ficoll separation solution. CD138 plasma cells were sorted by immunomagnetic beads method. RT-PCR was used to detect the expression levels of Blimp1, ATF4 and CHOP mRNA in U266 cells cultured in vitro. The cells were divided into blank control group, negative control group (no-loaded virus transfection) and positive experimental group [LV-Blimp1-RNAi (40051-2) transfection] by lentivirus transfection. RT-PCR was used to detect the expression of Blimp1, ATF4 and CHOP mRNA in cells of different groups. U266 cells were stimulated in vitro with different concentrations of aspirin solution (0, 0.5 mmol/L, 2.5 mmol/L, 5.0 mmol/L) for 24, 48 h and 72 h, respectively. The ability of cell proliferation in different groups was measured by CCK-8. U266 cells were stimulated with different concentrations of aspirin for 48 hours. And the mRNA expression of Blimp1, ATF4 and CHOP was detected by RT-PCR.@*RESULTS@#Compared with plasma cells in normal group, the expression of Blimp1 mRNA in CD138 plasma cells of MM patients significantly increased (8.040±1.878), and the mRNA expression levels of ATF4 and CHOP significantly decreased (0.735±0.089; 0.837±0.062) (P＜0.05). U266 cells were cultured in vitro. Compared with the blank control group and the negative control group, the mRNA expression level of Blimp in the positive experimental group was significantly down-regulated after infection with LV-Blimp1-RNAi (40051-2) lentiviral expression vector (0.637±0.021). ATF4 and CHOP mRNA expression levels were significantly increased (1.452 ± 0.027; 1.721 ± 0.038) (P＜0.05). The proliferation of U266 cells decreased after stimulation with aspirin. In the range of (0.5-5) mmol/L, aspirin could significantly inhibit the proliferation of U266 cells. The inhibition effect of aspirin was increased along with prolongation of time and increase of concentrations. After aspirin stimulation of different concentrations for 48 hours, the expression level of Blimp1 in U266 cells decreased with increasing of drug concentration, while the expression levels of ATF4 and CHOP increased with increasing of drug concentration.@*CONCLUSION@#Inhibition of Blimp1 expression in multiple myeloma cells can promote the expression of ATF4 and CHOP. Aspirin can inhibit the proliferation activity of myeloma cells by down-regulating Blimp1 expression in myeloma cells and up-regulating ATF4 and CHOP expression, therefore plays an anti-tumor rote.
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Objective To determine the protective role of keratinocyte growth factor 2 (KGF-2) on dextran sodium sulfate (DSS)-induced murine colitis and investigate the mechanism of its effect on intestinal mucosal barrier.Methods A total of 36 male C57BL/c mice were divided into 4 groups:normal control group,DSS model group,model + KGF-2 (5 mg/kg) group and model + KGF-2 (10 mg/kg)group.Groups except the control were added 3.5 ％ DSS in drinking water.Disease activity index (DAD,weight change,colon length loss and histological score were detected to evaluate the protective effect of KGF-2 on colitis.Serum fluorescein isothiocyanate-dextran (FITC-D) permeability were assayed by multiscan spectrum.In order to explore the protective role of KGF-2 on murine intestinal mucosal barrier,ZO-1 and occludin protein concentration were detected by immunohistochemical staining and Western blot.Meanwhile,cytokines including TNF-α,IL-6,IL-10,TGF-1β,IFN-γ and IL-1β in colonictissue were detected by ELISA.Results Compared with DSS-induced colitis model group,10 mg/kg KGF-2 significantly reduced DAI (P =0.021 1),weight loss (P =0.017 6),colon length loss (4.956 ±0.2583 vs.6.289±0.215 7,P =0.001 1),histologicalscore (12.17±1.222 vs.7.000±0.6325,P =0.001 1),and FITC-D permeability (168.5 ± 27.01 vs.14.62 ± 1.812,P =0.004 7) and reversed the downregulation of tight junction (TJ) proteins (ZO-1:0.158 6 ± 0.010 51 vs.0.387 9 ± 0.028 64,P＜0.000 1;occluding:0.300 5 ± 0.026 56 vs.0.445 0 ± 0.056 62,P =0.043 4).Significant decrease of TNF-α (68.93 ± 3.379 vs.40.41 ± 1.576,P＜0.000 1) and increase of IL-6 (3 755 ± 309.8 vs.5 640 ± 418.0,P =0.004 7) and IL-10 (304.0 ± 21.47 vs.521.2 ± 49.40,P =0.002 4) levels were noted in the 10 mg/kg KGF-2 treated mice.The effect of the KGF-2 was dose-dependent.Conclusions KGF-2 could ameliorate DSS-induced colitis and it may be associated with the decrease of the damage of mucosal barrier structure and funcntion by preventing ZO-1 and occludin from downregulating.The protective effect of KGF-2 on intestinal barrier function may also be exerted by inhibition of TNF-α and stimulation of IL-6 and IL-10 secretion.
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In the present study, two new trinor-guaiane sesquiterpenes, named clavuridins B (1), and A (2), along with three known sesquiterpenes (3-5), were isolated from the Xisha soft coral Clavularia viridis. Their structures and absolute configurations were determined on the basis of spectroscopic analysis, X-ray diffraction analysis with Cu Kα radiation and by comparison with related model compounds. Compounds 1 and 3-5 were evaluated for their cytotoxic activity.
Subject(s)
Animals , Anthozoa , Chemistry , Biological Products , Chemistry , Pharmacology , Magnetic Resonance Spectroscopy , Molecular Structure , Sesquiterpenes, Guaiane , Chemistry , PharmacologyABSTRACT
AIM:To observe the therapeutic effect of stachydrine hydrochloride on experimental acute cerebral infarction in rats and to explore the underlying mechanisms .METHODS:SD rats ( n=75) were randomly divided into 5 groups:sham group, cerebral infarction model group , and stachydrine hydrochloride (10 mg/kg, 20 mg/kg and 40 mg/kg) treatment groups .After the establishment of cerebral infarction model , the rats were given stachydrine hydrochloride at dose of 10 mg/kg, 20 mg/kg or 40 mg/kg by gavage daily for 14 d.The impairment of neurological function in each group was scored .The cerebral infarction volume and brain water content were measured .Moreover , the protein levels of β-cate-nin, cyclin D1, glycogen synthase kinase 3β(GSK-3β) and p-GSK-3βin the brain tissues were detected by Western blot . RESULTS:Compared with cerebral infarction group , the score of neurological function impairment , cerebral infarction volume and brain water content were significantly decreased in stachydrine hydrochloride treatment groups .In addition , the protein levels of β-catenin, cyclin D1 and p-GSK-3βwere markedly increased after stachydrine hydrochloride treatment . CONCLUSION:Stachydrine hydrochloride protects against experimental acute cerebral infarction through activation of Wnt/β-catenin signaling pathway .
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AIM:To observe the therapeutic effect of stachydrine hydrochloride on experimental acute cerebral infarction in rats and to explore the underlying mechanisms .METHODS:SD rats ( n=75) were randomly divided into 5 groups:sham group, cerebral infarction model group , and stachydrine hydrochloride (10 mg/kg, 20 mg/kg and 40 mg/kg) treatment groups .After the establishment of cerebral infarction model , the rats were given stachydrine hydrochloride at dose of 10 mg/kg, 20 mg/kg or 40 mg/kg by gavage daily for 14 d.The impairment of neurological function in each group was scored .The cerebral infarction volume and brain water content were measured .Moreover , the protein levels of β-cate-nin, cyclin D1, glycogen synthase kinase 3β(GSK-3β) and p-GSK-3βin the brain tissues were detected by Western blot . RESULTS:Compared with cerebral infarction group , the score of neurological function impairment , cerebral infarction volume and brain water content were significantly decreased in stachydrine hydrochloride treatment groups .In addition , the protein levels of β-catenin, cyclin D1 and p-GSK-3βwere markedly increased after stachydrine hydrochloride treatment . CONCLUSION:Stachydrine hydrochloride protects against experimental acute cerebral infarction through activation of Wnt/β-catenin signaling pathway .
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Objective:To analyze the distribution characteristics of IL-13 gene rs20541C/T site polymorphism in Guangxi population and compare the distribution differences among different populations.Methods: IL-13 genotypes were examined by using SNaPshot technique and DNA sequencing in 275 Guangxi people and analyzed the distribution frequencies of allele and genotype in this site.The result compared with the allele and genotype of other populations(Europeans,Beijingers,Japanses and Africans).Results:The polymorphism of rs20541C/T of IL-13 gene in Guangxi population existed.The genotype frequencies of CC,CT and TT were 40.0%,46.2% and 13.8% respectively.The frequencies of C and T allele were found to be 63.1% and 36.9%.The polymorphism had no significant difference between male and female(P>0.05).Compared rs20541C/T of IL-13 gene with those of HapMap-CEU, HapMap-YRI and Tianjin people,the distribution frequency of genotypes was significantly different(P<0.05).In this site,there were significant differences of allele frequency when compared with the other five populations(P<0.05).Conclusion: There are different degrees of diversity of rs20541C/T polymorphism of IL-13 gene among different races and regions.
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<p><b>OBJECTIVE</b>To investigate the expression level of B lymphocyte-induced maturation protein-1(Blimp-1) mRNA in bone marrow mononuclear cells(BMMNC) of multiple myeloma(MM) patients and its clinical significance.</p><p><b>METHODS</b>Fluorescent quantitative real-time PCR(qRT-PCR) was used to measure Blimp-1 mRNA expression in BMMNC and flow cytometry(FCM) was performed to detect the number of malignant plasma cells in bone marrow of MM group (39 newly-diagnosed and untreated patients) and IDA group (5 IDA patients). The clinical data of all the patients' were collected, and the 39 patients in MM group were divided into 2 subgroups: in BD group 20 cases were treated with bortezomib-based regimen and in VOD group 19 patients were treated with VAD regimen. The age, sex, clinseal stage and type between the 2 subgroups were not statistically different. Blimp-1 mRNA expression level in BMMNC of MM patients was detected by qRT-PCR after 3 treatment cycles.</p><p><b>RESULTS</b>The expression levels of Blimp-1 mRNA in BMMNC of IDA patients and MM patients divided into 3 groups according to ISS were (0.00047±0.00027), ISS I(0.09543±0.02800), Ⅱ(0.13606±0.04162),Ⅲ (0.21202±0.03940), separately. There was statistical difference among the 4 groups(F=56.929,P<0.05) and there was significant difference between any 2 groups of these 4 groups(P<0.05). Significant positive correlation was found between Blimp-1 mRNA expression level and the number of malignant plasma cells, serum monoclonal proteins (M protein), β2-microglobulin(β2-MG), lactic dehydrogenase(LDH), C-reactive protein(CRP)(P<0.05). There was significant negative correlation between Blimp-1 mRNA and hemoglobin (Hb) level (P<0.05). After 3 cycles of chemotherapy, Blimp-1 mRNA level of patients with a >50% reduction of M protein was significantly lower than that of patients whose M protein did not decrease significantly(P<0.05). After 3 treatment cycles, Blimp-1 mRNA expression in BMMNC in BD group was significantly lower than that in VAD group [(0.02388±0.00871) vs (0.04823±0.00219), P<0.05].</p><p><b>CONCLUSION</b>The Blimp-1 mRNA expression level in BMMNC may reflect the tumor burden in MM patients, which related with ISS, and positively correlated with the malignant plasma cell number, M protein, β2-MG, LDH, CRP level, and negatively correlated with Hb. The change of Blimp-1 mRNA expression level in BMMNC relates with the extent of M protein reduction, suggesting it may be used as a marker for response to therapy. Bortezomib may have effect on malignant plasma cells by suppressing Blimp-1 mRNA expression.</p>
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<p><b>OBJECTIVE</b>To explore temn possible correlation between the computer occupational semen quality.</p><p><b>METHODS</b>We included in this study 224 sterile males (118 computermen and 106 non-computermen) treated in our clinic of male sterility and 125 normal fertile men as controls, and analyzed such parameters as semen liquefaction time and sperm density, vitality and motility according to the WHO standard.</p><p><b>RESULTS</b>Compared with the normal controls, there was a significant decrease in the semen volume and sperm density, vitality and motility (P < 0.05-0.01) and a marked increase in liquefaction time in the infertile computermen and non-computermen (P < 0.01). The semen volume and sperm vitality and motility were significantly lower in the infertile computermen than in the infertile non-eomputermen (P < 0.05). The three parameters were also significantly decreased in the 0-5, 6-10 and > 10 h/d computer use groups. Similarly, significantly lower sperm density, vitality and motility were observed in the > 10 hid group than in the 0-5 and 6-10 h/d groups (P < 0.05-0.01), but with no statistical difference between the latter two (P > 0.05).</p><p><b>CONCLUSION</b>Computer occupation is associated with abnormal semen quality, and long-term computer use ( > 10 h/d) may be one of the factors of male infertility.</p>
Subject(s)
Humans , Male , Computers , Infertility, Male , Diagnosis , Occupational Exposure , Occupations , Semen Analysis , Sperm Count , Sperm MotilityABSTRACT
Objective To investigate the bone metabolism of children with leukemia and its relationship to tumor necrosis factor-?(TNF-?)pre-and post-treatment.Methods Serum bone gla protein(BGP),the excretion rate of urianry deoxypyridinoline(DPD)and TNF-? were measured in 35 children with leucocythemia pre-and post-treatment and 30 normal children(control group).Results The results showed that BGP,the excretion rate of DPD and TNF-? in children with leucocythemia before treatment were significantly higher than those in control group.However,the excretion rate of DPD decreased after chemotherapy but still significantly higher than that of control group(P0.05).Conclusions In children with leukemia,there are bone absorption and bone dissolution before chemotherapy,while there is osteogenesis dysfunction and bone absorption after chemotherapy.TNF-? can be used as a parametre for evaluating treatment effect and monitoring relapse.
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<p><b>OBJECTIVE</b>To observe the clinical efficacy of drug toothpaste containing 2% Zanthoxylum Nitidum extract in reducing accumulation of dental plaque and occurrence of gingivitis in adults.</p><p><b>METHODS</b>Adopting double-blind, stratified and parallel design, the status of dental plaque and gingivitis in 200 healthy adults enrolled in the trial (100 in the treated group took the drug toothpaste to clean teeth, and 100 in the control group use common toothpaste instead) were assessed at before experiment, 3 months and 6 months after experiment.</p><p><b>RESULTS</b>At the end of the 3rd month and the 6th month, as compared with the control group, the L e-Silness Gingival Index (GI) in the treated group was decreased by 9.1% (P > 0.05) and 13.9 % (P< 0.5), in the same period, the Quigley-Hein(Turesky Modification) Plaque Index (PlI) decreased by 18.7% and 22.6% (P<0.05), respectively.</p><p><b>CONCLUSION</b>Toothpaste containing Zanthoxylum Nitidum extract can obviously decrease the incidence of dental plaque and enhance gingival health.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Dental Plaque , Drug Therapy , Double-Blind Method , Drugs, Chinese Herbal , Therapeutic Uses , Gingivitis , Drug Therapy , Phytotherapy , Toothpastes , Chemistry , Zanthoxylum , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate the oral health knowledge, attitudes and behavior of urban, rural and suburban adults in Deyang, PR China, and analyze the influence of several variables on the utilization of dental services.</p><p><b>METHODS</b>1010 adults with 35-44 years old from urban, rural and suburban areas were interviewed by Questionnaire for Adults, WHO Integrated Package for Basic Oral Care.</p><p><b>RESULTS</b>More than half of the surveyed adults claimed "having pain of mouth during the last 1 year". More urban adults (61.3%) used fluoridate toothpaste than suburban and rural adults (6.9%, 16.8%). 61.0% and 71.5% of rural and suburban adults had never seen a dentist, compared with 32.0% of urban adults. Logistic regression analysis showed that urban adults having pain in mouth during the last year were more likely to visit a dentist in the last 2 years.</p><p><b>CONCLUSION</b>Although the oral health attitude is good among the people in Deyang, the knowledge is poor. Dental service utilization was not enough and the situation was also worse for rural adults.</p>
Subject(s)
Adult , Humans , Male , Attitude to Health , China , Dental Care , Health Behavior , Health Knowledge, Attitudes, Practice , Oral Health , Rural Population , Surveys and Questionnaires , ToothpastesABSTRACT
<p><b>OBJECTIVE</b>To investigate the influence of a broad range of environmental conditions on initial rates of hydrogen peroxide produced by Streptococcus oralis (S. oralis).</p><p><b>METHODS</b>For each rate measurement, 1 ml aliquots of 10(12) cells/L mid-logarithmic phase S. oralis in TSBY were centrifuged and respectively washed by phosphate buffer containing 0.01-10 mmol/L glucose or sucrose, phosphate buffer with 5.0-7.5 pH or Bis-Tris buffer containing 0.01-100 mmol/L Ca(2+), 0.01-100 mmol/L F(-) or 0.01-100 mmol/L HFPO(3)(-). After S. oralis was cultured in respective buffer for 10, 20 and 30 min at 37 degrees C, the concentration of hydrogen peroxide in supernatant was assayed spectrophotometrically in 96-well micro-plate by ABTS-HRP at A(405).</p><p><b>RESULTS</b>Synthesis rate of hydrogen peroxide by S. oralis was 7.48 micromol/L per minute without carbohydrate, the synthesis rate of hydrogen peroxide by S. oralis increased with 0.01-10 mmol/L glucose and 0.01-10 mmol/L sucrose, but there was no statistically significant difference in synthesis rate among the carbohydrate groups. The rates of H2O2 synthesis were inhibited in the buffer at pH 5.0-6.0, compared with pH 7.0 (P < 0.05). Ca(2+) had little influence on the rate of H2O2 synthesis. IC(50) of H2O2 synthesis rates by S. oralis responded to FHPO(3)(-) and F(-) were 12.65 mmol/L and 1.90 mmol/L respectively.</p><p><b>CONCLUSIONS</b>Environmental conditions may influence the synthesis rate of H2O2 by S. oralis.</p>
Subject(s)
Culture Media , Chemistry , Glucose , Hydrogen Peroxide , Metabolism , Hydrogen-Ion Concentration , Metals, Heavy , Streptococcus oralis , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To assess the long effect of atraumatic restorative treatment (ART) restorations in a school environment.</p><p><b>METHODS</b>Five middle-level dentists placed 294 ART restorations in the permanent teeth of 197 children. Standard instruments and procedures for ART were used. The restorative material used was a high-strength glass-ionomer. The restorations were evaluated annually by the same examiner who was not involved in the placement of the restorations. Occlusal wear of the restoration was assessed using a standard rheinberger scale. At the 5th year examination an independent external examiner evaluated the restorations using USPHS criteria.</p><p><b>RESULTS</b>The cumulative survival rates of small class I restorations in the sixth year it was 75%. The corresponding figures for large class I restorations were 61% (P < 0.05). The net wear rate for both the small and large class I restorations in the first year was about 48 microm, which was significantly higher than those of the later years, which were about 20 - 25 micro m (P < 0.05). In the fifth year 93% of the small restorations were assessed as being satisfactory for marginal integrity, as against 80% for the large ones by using USPHS criteria.</p><p><b>CONCLUSIONS</b>The ART approach for treating tooth decay in school children was appropriate, effective and acceptable. The 6-year survival rates of the restorations are high but are related to the size and type of the restoration.</p>
Subject(s)
Adolescent , Child , Humans , Dental Restoration, Permanent , Glass Ionomer Cements , Prospective StudiesABSTRACT
Objective To investigate the anti-fibrogenesis property of imatinib mesylate in a rat model of liver fibrosis induced by carbon tetrachloride/olive oil and its effect on the expression of trans- forming growth factor(TGF)-?1.Methods Rat liver fibrosis was induced by intraperitoneal administra- tion of carbon tetrachloride and olive oil mixture twice a week for eight weeks.Imatinib mesylate was given 20 mg/kg daily by oral lavage.The control rats received saline by oral iavage.Liver collagen depo- sition was evaluated by immunohistochemistry with Masson staining.The activation of hepatic stellate cells was detemined by the immunohistoehemistry staining of?-smooth muscle actin.The mRNA expres- sions of TGF-?1,c-Abl and TIMP-1 were measured by RT-PCR.While protein expressions of TGF-?1, phosphorylated platelet-derived growth factor receptor and c-Abl were detected by Western blot and im- munohistochemical staining.Hepatic hydroxyproline content was also quantified.Results The collagen deposition[(16.23?1.01)%vs(25.61?0.92)%]and the number of activated HSCs(10.52?1.33vs 13.10?1.21)were reduced in the imatinib mesylate treatment group compared with the control group by 35% and 20%,respectively(P