2.Effects of Porphyromnonas gingivalis lipopolysaccharide on the expression of RANTES and fractalkine in human urnbilical vein endothelial cells.
Xiaoling QI ; Lei ZHAO ; Shanshan CHEN ; Shu MENG ; Yafei WU
West China Journal of Stomatology 2016;34(2):194-199
OBJECTIVEA study was conducted to investigate the effects of Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) on the expression of regulated upon activation normal T-cell expressed and secreted (RANTES) and fractalkine in human umbilical vein endothelial cells (HUVECs).
METHODSHUVECs were incubated with different concentrations of Pg-LPS (200, 500, and 1000 ng x mL(-1)) for 1, 6, 12, and 24 h, respectively. Then real time quantitative polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent method (ELISA) were adopted to detect the protein levels and mRNA levels of RANTES and fractalkine.
RESULTSThe RANTES protein levels and mRNA levels, as well as fractalkine mRNA levels, were significantly higher in all experimental groups of 1, 6, and 12 h than in the control group (P<0.05), except the expression of RANTES mRNA in 200 ng x mL(-1) group of 12 h and RANTES protein in 200 ng x mL(-1) group of 1 h. The expression levels of RANTES mRNA and fractalkine mRNA were highest in 1000 ng x mL(-1) group of 6 h and were 4.88- and 6.20-fold higher, respectively, than those in the control group. The expression levels of RANTES protein, mRNA, and fractalkine mRNA decreased 6 h after stimulation, and were significantly higher than those in the control group (P<0.05) in the RANTES and fractalkine in HUVEC, and such expression is important in the development of atherosclerosis 500 ng x mL(-1) group of 24 h. There was a significant difference between the expression of fractalkine mRNA in 1000 ng x mL(-1) group of 6 and 12 h than in the control group (P<0.05).
CONCLUSIONPg-LPS infection might up-regulate the expression of RANTES and fractalkine in HUVEC, and such expression is important in the development of atherosclerosis.
Atherosclerosis ; Cells, Cultured ; Chemokine CCL5 ; genetics ; metabolism ; Chemokine CX3CL1 ; analysis ; genetics ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Human Umbilical Vein Endothelial Cells ; metabolism ; Humans ; Lipopolysaccharides ; pharmacology ; Porphyromonas gingivalis ; immunology ; isolation & purification ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Up-Regulation
3.Expression of MMP-1 and MMP-8 in periodontal tissues in rats with periodontitis
Shu MENG ; Yu CHEN ; Yafei WU ; Xiaoqin ZHAO
Journal of Practical Stomatology 2000;0(06):-
Objective:To investigate the expression of MMP-1 and MMP-8 in periodontal tissues in rats with periodontitis at different stages of inflammation with varied severity. Methods:Periodontitis was induced by silk ligature. P.gingivalis(Pg) or Pg with F.nu were used to induce the varied severity of periodontitis in 40 rats. 14 and 28 days after periodontitis induction the animals were sacrificed, periodontal tissues were immunohistochemically stained by antibody of MMP-1 and MMP-8 respectively. Results:MMP-1 and MMP-8 were both strongly positive in gingival epithelia cells and fibroblasts in periodontal ligament in rats with periodontitis.Higher expression of MMP-1 and MMP-8 was observed at 14 d than at 28 d(P0.05). Conclusion:The expression of MMP-1 and MMP-8 varies in different stage of periodontitis. MMP-1 and MMP-8 may play an important role in development of periodontitis.
4.Effects of crocetin on VCAM-1 expression in human umbilical vein endothelial cells and monocyte-endothelial cell adhesion.
Shu-guo ZHENG ; Meng-qiu ZHAO ; You-nan REN ; Jie-ren YANG ; Zhi-yu QIAN
Acta Pharmaceutica Sinica 2015;50(1):34-38
Crocetin, a naturally occurring carotenoid, possesses antioxidant and antiatherosclerotic properties, of which the underlying mechanism remains unclear. In the present study, we examined the effects of crocetin (0.1, 1, 10 μmol·L(-1)) on angiotensin II (Ang II, 0.1 μmol·L(-1)) induced expression of vascular cell adhesion molecule-1 (VCAM-1) in human umbilical vein endothelial cells (HUVECs) and monocyte-endothelial cell adhesion. The effects of crocetin on the activation of nuclear factor kappa B (NF-κB) and intracellular reactive oxygen species (ROS) were also observed. The results demonstrated that crocetin notably suppressed Ang II induced NF-κB activation (P<0.01) and VCAM-1 expression (P<0.05, P<0.01) in HUVECs, accompanied by a markedly reduced monocyte-endothelial cell adhesion (P<0.05, P<0.01). In addition, preincubation with crocetin resulted in a significant enhancement of cellular antioxidant capacity (P<0.05, P<0.01), while Ang II induced intracellular ROS decreased markedly (P<0.05, P<0.01). These results indicated that crocetin was capable of suppressing Ang II induced VCAM-1 expression and monocyte-endothelial cell adhesion by suppression of NF-κB activation, which might be derived from the enhancement of antioxidant capacity and subsequent reduction of intracellular ROS.
Angiotensin II
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metabolism
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Antioxidants
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pharmacology
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Carotenoids
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pharmacology
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Cell Adhesion
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drug effects
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Human Umbilical Vein Endothelial Cells
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cytology
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drug effects
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metabolism
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Humans
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Monocytes
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cytology
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NF-kappa B
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metabolism
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Reactive Oxygen Species
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metabolism
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Vascular Cell Adhesion Molecule-1
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metabolism
5.Treatment of stage 3b diabetic kidney disease patients with macroalbuminuria by qizhi jiangtang capsule: a multicenter randomized control clinical study.
Zhao-An GUO ; Chun-Jiang YU ; Gang LIU ; Fan-Chen MENG ; Yue LI ; Shu-Ling PENG
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(9):1047-1052
OBJECTIVETo observe the efficacy and safety of Qizhi Jiangtang Capsule (QJC) in treating stage 3b diabetic kidney disease (DKD) patients with macroalbuminuria.
METHODSPatients who conformed to the diagnostic criteria of stage 3b DKD were randomly assigned to two groups according to random digital table, the experiment group and the control group, 84 in each group. All patients received a two-week elution period, and then were treated with basic Western therapy. Patients in the experiment group took QJC, 5 pills per time, 3 times a day, while those in the control group took Valsartan Capsule 160 mg each time, once daily. The observation period of follow-ups was limited within 6 months, and the time points were set as the baseline, 1st month, 3rd month, and 6th month. Systolic blood pressure (SBP), diastolic blood pressure (DBS), 24 h urine protein quantitative (24 h UPQ), plasma albumin (ALB), and serum creatinine (SCr) were detected and recorded, and estimated glomerular filtration rate (eGFR) was calculated. The occurrence of hypoglycemic reaction, coagulation disorder, gastrointestinal tract reaction, allergy, hyperkalemia, doubling of creatinine, and overall adverse events were observed and recorded at same time.
RESULTSFinally 81 patients in the experiment group and 80 patients in the control group were effectively included. Compared with the baseline level, SBP and DBS obviously decreased in the control group at month 1 of treatment (P < 0.05), and more significantly decreased at month 6 of treatment (P < 0.01). SBP at month 1, 3, and 6 of follow-ups; DBS at month 6 of follow-ups was lower in the control group than in the experiment group (P < 0.05). At month 1, 3, and 6 of follow-ups, 24 h UPQ of the experiment group was significantly lower than the baseline level (P < 0.01). It was also significantly lower than the level of the control group at the same time point (P < 0.05). There was no significant difference in 24 h UPQ at month 1, 3, and 6 of follow-ups between the control group and the baseline level (P > 0.05). ALB of the experiment group showed an increasing trend. It was significantly higher than the baseline level at month 6 (P < 0.05), which was also higher than that of the control group at same period (P < 0.05). There was no significant difference in the ALB level in the control group (P > 0.05). SCr of two groups showed an increasing trend. SCr of the experiment group was significantly higher at month 1, 3, and 6 follow-ups than the baseline level (P < 0.05). But the increment of SCr was higher in the control group than in the experimental group, and obviously higher than the baseline levels (P < 0.05). eGFR of both groups showed a decreasing trend. The decrement was higher in the control group than in the experimental group (P < 0.05). The proportion of progression of renal functions at month 1, 3, and 6 of follow-ups in the experimental group was 0.0% (0 case), 9.55% (8 cases), and 21.4% (18 cases), while they were 8.3% (7 cases), 21.4% (18 cases), and 40.5% (34 cases) in the control group. There was no statistical difference in the proportion of progression of renal functions between the two groups at month 3 and 6 of follow-ups (P < 0.05). There was no statistical difference in the incidence of adverse reactions between two groups (P > 0.05).
CONCLUSIONQJC could effectively reduce urinary protein of patients with stage 3b DKD, and delay the progression of renal functions.
Adult ; Albumins ; analysis ; Albuminuria ; drug therapy ; Blood Pressure ; drug effects ; Creatinine ; blood ; Diabetic Nephropathies ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerular Filtration Rate ; Humans ; Male ; Middle Aged ; Tetrazoles ; therapeutic use ; Treatment Outcome ; Valine ; analogs & derivatives ; therapeutic use ; Valsartan
6.Inhibition of invasive and chemotactic abilities of SKOV3 cells bv human epithelial growth receptor-2 small interfering RNA
Shu-Lan ZHANG ; Yan-Ming LU ; Li-Rong MENG ; Yan-Yan ZHAO ;
Chinese Journal of Obstetrics and Gynecology 2001;0(01):-
Objective To investigate the effects of RNA interference(RNAi)targeting human epithelial growth receptor-2(HER-2)gene on the invasive and chemotactic ability of SKOV3 cells. Methods Glyeeraldehyde-3-phosphate dehydrogenase(GAPDH)was used as the positive control. Lipofectamine 2000 mediated transient transfection was conducted to transmit the siRNA into SKOV3 cells. Three pairs of specifically targeted(HER-2 siRNA Ⅰ,HER-2 siRNA Ⅱ,HER-2 siRNA Ⅲ)sequence were selected in the coding region of HER-2 mRNA.Transfection of HER-2 siRNA was conducted with lipofeetamine 2000 in ovarian carcinoma cell line SKOV3.The HER-2 gene expression was assessed by real- time PCR and western blot assays.Changes of invasive and chemotaetie capacity of SKOV3 cells were measured by polycarbonates coated with or without matrigal.Results Western blot results showed that the expression of GAPDH protein was decreased in specifically transfected cells and with the increase of siRNA dose,the expression of GAPDH protein was decreased.GAPDH protein gray value in control group, different doses(0.5,1.0,1.5,2.0 ?g)GAPDH siRNA interference groups were 0.6855?0.0259,0.5698 ?0.0275,0.4542?0.0296,0.3341?0.0178 and 0.1816+0.0180,respectively.There was a significant difference in each group(F=198.126,P
7.Evaluation of Tourette's syndrome by 99Tcm-TRODAT-1 SPECT imaging
Feng, DONG ; Hong, LIU ; Zhao-wei, MENG ; Jian, TAN ; Ben-shu, ZHANG
Chinese Journal of Nuclear Medicine 2011;31(2):104-107
Objective To observe dopamine transporter (DAT) binding capacity using 99Tcm-TRODAT-1 in drug-naive patients with Tourette's syndrome (TS) on SPECT imaging, and explore possible correlations between 99Tcm-TRODAT-1 uptake ratio and TS patient's age, disease duration, and tic severity.Methods Eighteen drug-naive TS patients, male 14, female 4, as well as 8 age- and gender-matched healthy subjects were recruited. Brain SPECT imaging was performed 2. 5 h after intravenous injection of 11.1 - 14.8 MBq/kg 99Tcm-TRODAT-1. ROI was drawn on the striatum including its sub-regions of caudate and putamen, with cerebellum as the background. Striatum/cerebellum ratio was calculated. Comparisons of the ratios between TS patients and controls were carried out by independent-sample t-test. Pearson correlation analysis was performed between DAT uptake ratios of striatum and patients' age, disease duration, tic severity. Results Compared with the control, higher symmetrically striatum uptake of 99Tcm-TRODAT-1 in TS patients was observed (2.17±0.23 vs 1.87±0.24, t =2.957, P<0.05). Age (r= -0.320, P>0.05)and tic severity(r = 0. 345, P > 0.05) scores were not significantly correlated with specific uptake ratios measured in the striatum. But there was significant negative correlation between disease duration and the specific uptake ratios (r = - 0. 483, P < 0. 05). Conclusions 99 Tcm-TRODAT-1 SPECT imaging may play an adjuvant role for initial evaluation of untreated TS.
8.Expression of recombinant cytolethal distending toxin of Actinobacillus actinomycetemcomitans.
Shu MENG ; He YANG ; Lei ZHAO ; Ya-Fei WU
Chinese Journal of Stomatology 2009;44(7):409-412
OBJECTIVETo examine the expression of recombinant cytolethal distending toxin (CDT) produced by Actinobacillus actinomycetemcomitans (Aa).
METHODSCDT encoding gene cdtABC was amplified by PCR. Through TA clone and restriction endonuclease digestion, gene cdtABC and vector pQE60 were ligated to form pQE60-cdtABC expression system which transformed into competent cells. Protein expression was induced by IPTG and examined by SDS-PAGE and Western-blotting.
RESULTSRandom colony PCR of pQE60-cdtABC transformed cells demonstrated that all strains contained cdtABC gene. The DNA sequence was blast with cdtABC gene from GenBank and 99% homology was obtained. SDS-PAGE and Western-blotting confirmed that recombinant CDT was obtained.
CONCLUSIONSCDT protein expression system was reconstructed and recombinant protein was obtained. Actinobacillus actinomycetemcomitans;
Aggregatibacter actinomycetemcomitans ; genetics ; metabolism ; Bacterial Toxins ; genetics ; metabolism ; Genetic Vectors ; Recombinant Proteins ; genetics ; metabolism
9.Status of plasma folate in the third trimester of pregnant women and newborn babies in the northern rural areas of China
Shu-Hua ZHAO ; Lin-Lin WANG ; Wen-Jie XU ; Lei JIN ; Le ZHANG ; Jian-Meng LIU
Chinese Journal of Epidemiology 2012;33(7):654-657
Objective To study the plasma folate concentrations in the third trimester of pregnant women and newborn babies so as to assess the association between them.Methods Pregnant women in Yuanshi and Laoting counties in Hebei province from May to June in 2009 were recruited with related information collected at enrollment.Those pregnant women being enrolled were followed up until delivery.Maternal blood was collected before delivery,and cord blood was collected after the expulsion of the placenta.Data from 437 pairs of women and newborns were analyzed.Plasma folate concentration was measured by Microbiological assay,with maternal plasma folate concentration <6.8 nmol/L defined as folate deficiency.Neonatal plasma folate concentration below 10% was defined as relative deficiency.Student t-test and ANOVA were used to compare the plasma folate concentrations between the groups and x2 test was used to compare the situation of folate deficiency.In order to assess the association between maternal and newborn folate levels,logistic regression analysis was used to estimate the odds ratio of the neonatal plasma folate relative deficiency between the maternal folate deficient and normal groups after adjusting factors as age,BMI,region,career and education.Linear regression was used to test the trend by quintiles of maternal plasma folate concentration.Pearson' s test was used to test the relationship between the ratio of neonatal and maternal plasma folate level and the level of maternal plasma folate.Results The geometric mean of maternal plasma folate concentration was 8.0(95%CI:7.6-8.5) nmol/L and the deficiency was 29.3%,but in newborn babies,they were 24.0(95%CI:23.1-25.0) nmol/L and 0.9% respectively.The plasma folate level in newborn babies was 3.0 times as high as in maternal (t=32.519,P<0.01 )but the neonatal plasma folate deficiency status was higher than in matemal ( x2=137.2,P<0.01 ).When compared with the normal plasma folate level group,the risk on neonatal plasma folate relative deficiency in the maternal folate deficiency group was significantly higher aiter adjusted for confounders (OR=1.96,95%CI:1.02-3.80).The neonatal plasma folate level significantly increased along with the maternal plasma folate level (Ptrend<0.05).The ratio of neonatal and maternal plasma folate level was significantly inversely correlated with the maternal folate level (r=-0.810,P<0.001 ).Conclusion Folate status in newborns was much better than in their mothers',in the northern rural areas of China.The maternal folate status was positively correlated with their offspring' s.Active placental transport for folate was significantly increasing when the maternal plasma folate level decreased.
10.Study on the characteristics of temporal distribution and the epidemic trend of autumn-winter type scrub typhus under time series analysis
Lei DING ; Shu-Jun DING ; Meng ZHANG ; Xian-Jun WANG ; Zhong LI ; Zhong-Tang ZHAO
Chinese Journal of Epidemiology 2012;33(7):698-701
Objective To explore the characteristics of temporal distribution and epidemic trend of autumn-winter type scrub typhus using the time series analysis.Methods Based on the data of scrub typhus collected from Shandong Diseases Reporting Information System from 2006 to 2011,both spectral analysis and moving average analysis were used to analyze the annual data of scrub typhus while scrub typhus incidence in 2012-2014 was forecasted.Seasonal decomposition analysis was applied to analyze the monthly data from January of 2006 to October of 2011,followed by Autoregressive Integrated Moving Average Model (ARIMA) which was constructed to forecast case number in November and December of 2011 and compared to the actual incidence.Results The results of spectral analysis showed that the prevalence of autumn-winter type scrub typhus had a feature of ‘3-year-periodicity’.A long-term up-trend was confirmed by method of moving average analysis,with annually case numbers of 310,337 and another number of 366 forecasted for 2012 to 2014,respectively,with the annual increase rate as 9% per-year.Data from analysis of monthly data of scrub typhus showed that through multiple seasonal decomposition analysis,the results indicated that the prevalence of this disease possessed a typical autumn-winter type.The seasonality indexes for scrub typhus in October and November were 8.454 and 2.230,respectively,while others were less than 1.000.The ARIMA (0,1,1 ) (0,1,0)12 model of ( 1 -B) ( 1 -B12)X,=( 1 -0.811B)u,that was used to forecast the prevalence of autumn-winter type scrub typhus and was constructed with the residual error of 16 lags as white noise.The Box-Ljung test statistic for the model was 3.116,giving a P value of 0.999.The model fitted the data well.Good accordance was achieved between the observed values and the forecasted values of scrub typhus in November and December of 2011 which was produced by the ARIMA model,and all observed values were within the forecasted 95% CI.Conclusion The prevalence of autumn-winter type scrub typhus showed a 3-year-periodicity,with a long-term up-trend,and the case numbers of 2012 to 2014 were forecasted,rising on the end with an increasing rate of 9% per year,which occurred seasonally with October as the peak time in every year.The ARIMA (0,1,1 ) (0,1,0) 12 model seemed to be quite appropriate in predicting the autumn-winter type scrub typhus.