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.

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.

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.

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 ; chemistry ; Heterocyclic Compounds, 4 or More Rings ; chemistry ; isolation & purification ; Stephania ; chemistry

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.

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.

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.

OBJECTIVETo investigate the value of application of mandibular outer cortex as bone graft by comparing its bone absorption with cranial outer cortex.

METHODS8 minitype grown-up pigs at the age of 8 - 12 months underwent surgery of taking out the same size (2.5 cm x 1.0 cm) of outer cortex from mandible and craninium. The volume of the outer cortex was measured by volume-displacement method. Then the outer cortex of mandible and cranium were onlay grafted to the each side of the pig snout, respectively. 12 weeks later, 2 pigs were randomly selected for histological examination. The other 6 pigs were killed 24 weeks after surgery for measurement of the bone graft volume and histologic examination.

RESULTSThe bone graft absorption rate was (41 +/- 5)% for mandibular outer cortex and (46 +/- 12)% for cranial outer cortex, showing no significant difference between them (P = 0.51). The histologic examination results also had no marked difference in the bony healing and reforming between the two graft.

CONCLUSIONSMandibular outer cortex is a good donor site for onlay bone graft in craniofacial region.

Animals ; Bone Plates ; Bone Regeneration ; Bone Transplantation ; methods ; Female ; Male ; Mandible ; transplantation ; Skull ; transplantation ; Swine ; Swine, Miniature

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

OBJECTIVETo establish a tumor-bearing nude mouse model of human neuroblastoma in order to study the mechanisms of neuroblastoma invasion and metastasis, and to investigate potential therapeutic modalities in the experimental animal models.

METHODSA human neuroblastoma cell line was cultured in vitro. 1 x 10(7) cells undergoing exponential growth were collected in 0.1 ml of suspension and subcutaneously inoculated into the right flank next to the forelimb in nude mice. The biological characteristics of the developed tumors were observed, and histopathological and DNA microarray analyses were performed. The expressions of NSE in the subcutaneous tumor, metastatic tumor and the primary neuroblastoma tumor tissues from a pediatric patient were analyzed by immunohistochemistry.

RESULTSTumors successfully grew in 36 out of 48 injected mice, with a total tumor-formation rate of 75.0%. Metastasis occurred in 10 cases, and the metastatic rate was 20.8%. Tumors in five injected mice grew locally without metastasis. These tumors had large volume and the tumor weight reached up to half of the body weight of the host animal. Four mice exhibited systemic metastasis without tumor growth at the primary inoculation site. There were six mice with locally growing tumor accompanied by metastasis.

CONCLUSIONWe have successfully established a human neuroblastoma xenograft model in nude mice with high tumor growth and metastatic rates. This model depicting the natural cell growth, local infiltration and distant metastasis characteristics of human neuroblastoma, providing an ideal animal model for in vivo studies of neuroblastoma. In addition, the results of this study indicate the heterogeneous nature of neuroblastoma, it may play an important role in metastasis of this tumor.

Animals ; Cell Line, Tumor ; Child, Preschool ; Disease Models, Animal ; Female ; Gene Expression Profiling ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; secondary ; Male ; Mice ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Transplantation ; Neuroblastoma ; genetics ; metabolism ; pathology ; secondary ; Phosphopyruvate Hydratase ; metabolism ; Tumor Burden