1.Effect of rosiglitazone on the secretion of chemokines in renal tubular epithelial cells stimulated by lipopolysaccharide and the possible mechanism
Ying LU ; Qiao ZHOU ; Fang ZHONG ; Xu HAO ; Cong LI ; Weiming WANG ; Nan CHEN
Chinese Journal of Nephrology 2010;26(12):909-914
Objective To investigate the inhibitory effect and mechanism of rosiglitazone on chemokines secretion in renal tubular epithelial cells (HK-2) stimulated by lipopolysaccharide (LPS). Methods Cells were divided into four groups: control (CON), LPS (1 mg/L),rosiglitazone (10 μmol/L), rosiglitazone (10 μmol/L) +LPS (1 mg/L). MCP-1 and IL-8 expression was measured using real time PCR and ELISA. PPARγ was knockdown by RNAi to investigate whether the inhibitory effect of rosiglitazone was PPARγ-dependent or -independent. The NF-κB in nucleus was detected by Western blotting. The DNA binding activity of NF-κB was determined by electrophoretic mobility shift assay. Results Compared with CON group, the expressions of IL-8 and MCP-1 were increased by (4.30±0.45) and (4.80±1.29) times in mRNA level, (1.39±0.18)and (2.11 ±0.47) times in protein level, respectively, in LPS-stimulated HK-2 cells (P<0.05).Application of rosiglitazone followed by LPS significantly reduced IL-8 and MCP-1 secretion compared with LPS group (decreasing by 66.37% and 71.88% in mRNA levels, while 41.68% and 47.87% in protein levels) (P<0.05). In pcDNATM 6.2-GW/EmGFP-miPPARγ transfected cells, IL-8and MCP-1 only were decreased by 18.16% and 16.83% in mRNA level, while 11.39% and 11.86%% in protein level in rosiglitazone pretreated group, showing no significant difference compared with LPS group. Rosiglitazone did not block NF-κB nuclear translocation while significantly inhibiting the DNA binding activity of NF-κB. Conclusions Rosiglitazone inhibits the expressions of MCP-1 and IL-8 via a PPARγ-dependent mechanism in HK-2 cells, resulting from inhibition the DNA binding activity of NF-κB.
2.Effect of lifestyle quantilization based-weight management on overweight or obesity occupational groups
Yujiang LU ; Yan WANG ; Nan WANG ; Junjun HAO ; Di WU ; Hui FU ; Yueling HUANG
Chinese Journal of Health Management 2013;(1):14-17
Objective To study the effects of lifestyle quantilization based-weight management on overweight or obesity police officers.Methods One hundred and seven overweight or obesity police officers received lifestyle quantilization based-weight management (i.e.Jinbi weight management) and were then assigned to the excellent performance group (group A,n =50),good performance group (group B,n =42)and loss to follow-up group (group C,n =15).Dietary habits,body weight,height,waist circumference (WC),blood pressure (BP),total cholesterol (TC),triglyceride (TG),low-density lipoprotein cholesterol (LDL-C),high-density lipoprotein cholesterol (HDL-C) and fasting plasma glucose (FPG) were measured before and after the intervention.After 8 weeks' intervention,the changes of above parameters were recored.Student's t test was used for data analysis.Results Of group A and B,83 reported weight loss (group A t =13.31,group B t =5.04 ; both P < 0.05).In group A,body weight,body mass index (BMI),WC,body fat and visceral fat index were significantly decreased,in contrast to body water (t values were 13.31,13.72,10.8,8.59,6.83 and-6.62,respectively; all P < 0.05).However,there were no significant changes of BP,FPG,TC,TG,LDL-C and HDL-C in group A.Following intervention,daily dietary energy intake of group A was reduced by 74.1 k J,fat intake was decreased by 11.6 g,energy ratio of dietary fat was decreased by 1.8%,energy ratio of cereal was increased by 4.2%,and sodium chloride and cooking oil was decreased by 1.3 g and 10 g,respectively.Conclusion Lifestyle quantilization based-weight management shows effectiveness among overweight or obese police officers,and thus may be recommended for other functional communities.
3.Suppression of MDR1 gene expression and reversal of cisplatin resistance in renal carcinoma cells by RNA interference
Yixin HAO ; Zhengwen HE ; Nan DU ; Qiong LU ; Wenhua XIAO ; Qian SHEN
Tumor 2010;(2):115-118
Objective:To investigate the inhibitory effect of RNA interference (RNAi) on the expression of multidrug resistance (MDR1) gene and analyze the altered sensitivities of human renal carcinoma cell line to cisplatin.Methods:Three small interfering RNA (siRNA) sequences targeted MDR1 gene were synthesized and transfected into renal carcinoma A498 cells. The expression level of MDRl mRNA was measured by RT-PCR to identify the most effective siRNA sequence. The recombinant plasmid was packed by lentivirus and transfected into A498 cells. RT-PCR was used to screen the A498 cells with the optimal silencing efficacy. The MDR1 protein expression level in the cloned cells was verified by Western blotting. The inhibitory effect of cisplatin on the proliferation of A498 cells was assessed by MTT assay and the IC_(50) value was calculated. Results:The 3 siRNA sequences suppressed MDR1 gene expression at different degrees. The siRNA 1 sequence silenced MDR1 gene more effectively with a significant reduction of 67%. The MDR1 protein expression greatly decreased in screened A498 cells compared with non-transfected cells (P<0.01), and the IC_(50) value of cisplatin on screened A498 cells was significantly decreased by 83.37% (P<0.01). Conclusion: The RNAi could effectively inhibit the expression of MDR1 gene and increase the sensibility to cisplatin in human renal carcinoma A498 cell line, which make it possible to reverse the resistance of renal carcinoma to chemotherapy.
4.Hernsubanine E, a new hasubanan alkaloid from Stephania hernandifolia.
Nan ZHANG ; Li-Jia TANG ; Lu-Lu DENG ; Huan-Yu GUAN ; Jian-Xin ZHANG ; Xiao-Jiang HAO ; Yuan-Hu ZHANG
China Journal of Chinese Materia Medica 2014;39(6):1020-1023
A new hasubanan alkaloid, hernsubanine E (1), as well as two known compounds p-hydroxybenzaldehyde (2) and (-)-syringaresinol (3) have been isolated from the whole plants of Stephania hernandifolia by various column chromatographic methods. Their structures were identified by physicochemical properties and spectral analyses. Compounds 2 and 3 were isolated from the genus of Stephania for the first time.
Alkaloids
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chemistry
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Heterocyclic Compounds, 4 or More Rings
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chemistry
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isolation & purification
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Stephania
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chemistry
5.Construction of eukaryotic recombinant expression plasmids with glyceraldehydes-3-phosphate dehydrogenase and cysteine protease inhibitor gene of periodic Brugia malayi and its expression in HeLa cells
Xiao-jun, LIU ; Xiao-feng, GUO ; Sai-nan, ZHANG ; Shi-juan, LU ; Hao, FANG ; Bang-sheng, XU ; Zheng, FANG
Chinese Journal of Endemiology 2011;30(4):371-375
Objective To construct the eukaryotic expression plasmid containing glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and cysteine protease inhibitor ( CPI ) gene from periodic Brugia malayi (Bm) and to lay foundation for studying multivalent vaccines. Methods Total RNA was extracted from periodic Bin. The BmGAPDH and BmCPI genes were amplified by RT-PCR. The PCR product was cloned and then subeloned into eukaryotic recombinant plasmid vector pcDNA3.1 (+). pcDNA3.1 (+)/BmGAPDH/BmCPI was constructed. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and were transformed into HeLa cell subsequently. The transient expression of BmGAPDH and BmCPI were examined by RT-PCR. The expressed protein was identified by sodium dodeeylsulphate-polyacrylamide gel electrophoresis(SDS-PAGE). Results Two specific bands of around 877 bp of BmGAPDH and 621 bp of BmCPI were amplified, consistent with the expected value. The same bands were obtained by double restriction enzyme digestion of recombinant plasmids or PCR using recombinant plasmid as template. BmGAPDH and BmCPI mRNA were highly expressed in transfeeted HeLa cell. The relative molecular mass (Mr) of the recombinant protein was about 54 × 103. Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1 (+)/BmGAPDH/BmCPI has been constructed successfully and the protein is expressed correctly in mammalian cell.
6.Prediction of the Th/B cell epitopes on HA of influenza virus(H1N1)and antigenicity analysis
Kaiyan WANG ; Taiyuan LI ; Huijun LU ; Lei TAN ; Wenlong NAN ; Mingyao TIAN ; Jinshuang ZHANG ; Hao LIU ; Ningyi JIN
Chinese Journal of Immunology 2010;26(1):8-12
Objective:To predict Th/B cell epitopes in HA of influenza virus(H1N1)and analyze antigenicity of the candidate epitopes in order to develop epitope-bacterin by the way of bioinformatics.Methods:The HA amino acid sequences of infiuenza virus(H1N1),which the viral infection was prevalent recently,were downloaded from Genbank.The Th/B cell epitopes were predicted and analyzed by bioinformatics methods.Then,specificity and conservation of the candidate epitopes were estimated.Finally,antigenicity of the candidate epitopes was identified by influenza virus(H1N1)positiVe serum samples of mice.Results:Three Th/B cell epitopes containing HA_(73-87),HA_(125-139),HA_(188-205) were acquired Two of the candidate epitopes were in a relatively conserved domain of HA1,and a deal of 2006-2009 influenza virus(H1N1)isolates contained the sequences.Moreover,the candidate epitopes were showedin a distinct antibody combining reactivity with the influenza virus (H1N1)positive serum of mice,which inferred the predicted epitopes to be functional ones.Conclusion:The selected epitopes are able to be functional HA Th/B cell epitopes of influenza virus(H1N1).Our study also establish the foundations for the further research of influenza virus infectlon and immunity mechanism,the recognition of influenza virus(H1N1)functional epitope and the development of epitope vaccines.
7.Prediction of the Th/B cell epitopes on HA of influenza virus(H1N1) and antigenicity analysis
Kaiyan WANG ; Taiyuan LI ; Huijun LU ; Lei TAN ; Wenlong NAN ; Mingyao TIAN ; Jinshuang ZHANG ; Hao LIU ; Ningyi JIN
Chinese Journal of Immunology 1985;0(01):-
Objective:To predict Th/B cell epitopes in HA of influenza virus(H1N1) and analyze antigenicity of the candidate epitopes in order to develop epitope-bacterin by the way of bioinformatics.Methods:The HA amino acid sequences of influenza virus (H1N1),which the viral infection was prevalent recently,were downloaded from Genbank.The Th/B cell epitopes were predicted and analyzed by bioinformatics methods.Then,specificity and conservation of the candidate epitopes were estimated.Finally,antigenicity of the candidate epitopes was identified by influenza virus (H1N1) positive serum samples of mice.Results:Three Th/B cell epitopes containing HA73-87,HA125-139,HA188-205 were acquired.Two of the candidate epitopes were in a relatively conserved domain of HA1,and a deal of 2006-2009 influenza virus (H1N1) isolates contained the sequences.Moreover,the candidate epitopes were showedin a distinct antibody combining reactivity with the influenza virus (H1N1) positive serum of mice,which inferred the predicted epitopes to be functional ones.Conclusion:The selected epitopes are able to be functional HA Th/B cell epitopes of influenza virus (H1N1).Our study also establish the foundations for the further research of influenza virus infection and immunity mechanism,the recognition of influenza virus (H1N1) functional epitope and the development of epitope vaccines.
8.Clinical research of angle-splitting ostectomy based on three dimensional computed tomography true-up technique.
Yan-feng ZHAO ; Yun-fei HAO ; Ping LU ; Xiao-nan ZHOU ; Chang-feng QU
West China Journal of Stomatology 2009;27(5):516-520
OBJECTIVETo illustrate the morphological changes of mandible after angle-splitting ostectomy.
METHODSFrom January 2006 to April 2008, 10 cases had undergone mandibular angle-splitting ostectomy to reduce the width of the lower face. For each patient, CT datum of mandible at three stages (preoperative, immediate postoperative, 6 months postoperative) were collected. By the application software of reverse engineering (Surfacer V9) and true-up and dissection techniques based on three-dimensional spiral computed tomography (3D-CT), operative efficacy and bone regeneration at the operation area of angle-splitting ostectomy were evaluated 6 months postoperative.
RESULTS1) Concavity could be seen at the angle-splitting ostectomy area 6 months postoperative, especially at the mandibular external oblique line region. Average cup depth was (3.64 +/- 1.67) mm by contrasted to preoperative. Diminution of bone volume was 55% +/- 9% for the local operative area 6 months postoperative. 2) Bone regeneration could be seen at the area that mandibular outer cortex had been removed. Compared with immediate postoperative, ratio of neoformative bone was 84.6% +/- 7.3% 6 months postoperative. The main region of bone regeneration was mandibular angle.
CONCLUSIONMandibular angle-splitting ostectomy is an effective technique for reducing the width of the lower face. Masseter muscular movement should be restricted postoperative to prevent hyperostosis at the angle area.
Adult ; Bone Regeneration ; Face ; Female ; Humans ; Mandible ; Masseter Muscle ; Osteotomy ; Tomography, X-Ray Computed
9.Feasibility of peptide mass fingerprinting for differential diagnosis of IgA and non-IgA nephropathy.
Jing GAO ; Yong WANG ; Xin-yu WEN ; Hong-hao LU ; Zhen-nan DONG ; Ya-ping TIAN
Journal of Southern Medical University 2011;31(8):1309-1313
OBJECTIVETo investigate the feasibility of peptide mass fingerprinting for non-invasive differential diagnosis of IgA nephropathy (IgAN) from the non-IgA nephropathy (IgAN).?
METHODSAccording to the results of renal biopsy, 56 patients were divided into IgAN group (n=28) and non-IgAN group (n=28), and peptide mass fingerprints were acquired from these patients using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).
RESULTSNine different peptides were identified between IgAN and non-IgAN. The two most distinctive differentially expressed peptides, with peptide peak values of 4476.46 and 1968.10, showed area under curve values of 86.18% and 79.77%. Principal component analysis demonstrated that the accumulated explained variance of the first 8 differential peptides reached 95%, suggesting the feasibility of differential diagnosis of IgAN from non-IgAN. Comparison with the Matrix protein database identified the peptide with a relative molecular mass of 5338.08 as a fragment of mucin 4 inform and the 2082.77 peptide as fragment of α1-II type collagen inform.
CONCLUSIONMALDI-TOF MS is feasible for differential diagnosis of IgAN and non-IgAN and also has great potentials in the classification of the subtypes of other systemic diseases.
Adult ; Diagnosis, Differential ; Feasibility Studies ; Female ; Glomerulonephritis, IGA ; diagnosis ; Humans ; Kidney Diseases ; diagnosis ; Male ; Middle Aged ; Peptide Mapping ; methods ; Peptides ; chemistry ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Young Adult
10.Normal reference values and predict equations of heart function.
Zhi-nan LU ; Sun XING-GUO ; Song-shou MAO ; M J BUDOFF ; W W STRINGER ; Wan-gang GE ; Hao LI ; Jie HUANG ; Fang LIU ; Sheng-shou HU
Chinese Journal of Applied Physiology 2015;31(4):332-336
OBJECTIVEFor heart functional parameters, we commonly used normal range. The reference values and predict formulas of heart functional parameters and their relationships with individual characteristics are still lack.
METHODSLeft ventricular (LV) volumes (end-diastolic volume and end-systolic volume), stroke volume (SV), ejection fraction (EF) and cardiac output (CO) were measured by cardiac CT angiography (CAT) in 1 200 healthy Caucasian volunteers, men 807 and women 393, and age 20-90yr. The results are analyzed by high-accuracy three-dimensional imaging technology, and then measured the dynamic changes of the volumes of each atriam and ventricule during their contractions and relaxations. The gender, age, height and weight were analyzed by multiple linear regression to predict LV functional parameters.
RESULTSExcept the LVEF was lower in man than in women (P < 0.001), all other LV functional parameters of EDV, ESV, SV, FE and CO were higher in man (P < 0.001). Multiple linear regression indicated that age, gender, height and weight are all independent factors of EDV, ESV and SV (P < 0.001). CO could be significantly predicted by age, gender and weight (P < 0.001), but not height (P > 0.05). The predict equation for CO (L x min(-1)) = 6.963+0.446 (Male) -0.037 x age (yr) +0.013 x weight (kg).
CONCLUSIONAge, gender, height and weight are predictors of heart functions. The reference values and predict equations are important for noninvasive and accurate evaluation of cardiovascular disease and individualized treatment.
Adult ; Age Factors ; Aged ; Aged, 80 and over ; Body Height ; Body Weight ; Cardiac Output ; Female ; Heart ; physiology ; Humans ; Male ; Middle Aged ; Reference Values ; Sex Factors ; Stroke Volume ; Ventricular Function, Left ; Young Adult