ObjectiveTo determine whether there is correlation between the pathological classification and urinary level of transforming growth factor(TGF)-β1,macrophage inflammatory protein (MIP)-1α,and Neutrophil gelatinase-associated lipocalin (NGAL) of lupus nephritis.MethodsELISA was used to test the levels of urinary level of TGF-β1,MIP-lα and NGAL.The correlation between these three urinary markers and the pathological classifications,Austin score,histological semi-quantitative score and clinical data were analyzed.Comparisons between groups were performed with t-test,ANOVA and X2 test.Correlation analysis was performed with Pearson analysis.Results①There was significant difference in urinary TGF- β1,MIP1α,and NGAL levels when compared the lupus nephritis group,SLE patients without renal damage group and the normal control group [TGF-β1(351±219),(92±60),(74±29) pg/ml],[MIP-1α(18.0±15.5),(8.5±2.3),(7.1±1.9) pg/ml],[NGAL (1.104±0.519),(0.181±0.030),(0.146±0.024) ng/ml],while there was no significant difference between lupus nephritis (LN) group and patients with other glomerulonephritis.② There was significant difference in the three urinary markers when stratified the LN patients based on the pathological classifications,however,there was no significant difference between the LN group and other glomerulonephritis group.③ Analysis of the correlation between the three urinary markers and semiquantitative histopathological score of the LN patients showed that the urinary level of TGF-β1 and MsMI,GCI,TCI,VCI was closely related to each other,the urinary MIP-1α was related to and endol,and the dGAI and VCI was closely related, while the urinary NGAL was closely related with endol,dGAI and MsHI.The correlation analysis between the three urinary markers and acute and chronic pathological index score of the LN patients showed that TGF- β1 was correlated with the chronic index(r=0.89,P＜0.01 ),the MIP-lα and NGAL were significantly correlated with the activity index(r=0.71,P＜0.01 ; r=0.60,P＜0.01 ).Conclusion There is a positive correlation between the urine TGF-β1 level and chronic kidney disease.The urinary MIP-lαand NGAL are associated with active renal damage.

Objective To investigate the effects of myricetin (Myr) on A549 lung cancer cell proliferation and its mechanism. Method The inhibitory effects of Myr on the growth of A549 cells were investigated and its value of IC50 after 72 h incubation was measured by 3-(4,5-dimethyl-thiazolyl -2)-2,5-diphenyl tetrazolium bromide (MTT) method. The phosphorylation level of Akt protein was studied by Western blotting. Results Myr inbibited A549 cell proliferation in dose-dependent way. The IC50 value of Myr treatment for 72 h was 41.7 ?g/ml. Treatments with 32 ?g/ml Myr for 12 h significantly reduced the protein kinase B (Akt Ser473) phosphorylation. Conclusion Myr could inhibit the growth of A549 cell in a dose-dependent way. Akt phosphorylation may be the mainly molecular target of Myr in A549.

OBJECTIVETo investigate the antagonistic effects of three species of oral Streptococcus on the growth of oral Saccharomyces albicans in vitro.

METHODSDirect inoculation method, reverse inoculation method and mixed culture methods were respectively chosen to observe the changes of Saccharomyces albicans colony formation on the effects of Streptococcus mutans, Streptococcus sanguis and Streptococcus salivarius.

RESULTS1) No clear inhibition zone was observed in each of the groups by direct inoculation method. 2) Compared with the control groups, Saccharomyces albicans colony formation on soft agar of Streptococcus sanguis decreased significantly (P < 0.05). 3) Mixed culture method results showed that Streptococcus mutans could inhibit the growth of Saccharomyces albicans significantly at different time points (P = 0.001). 4) Under the action of bacteria culture supernatant, the count of Saccharomyces albicans in experiment groups showed statistical significance when compared with the control groups at 24, 48, 72 h (P = 0.001); The differences among the experimental groups were of no statistical significance at majority times (P > 0.05).

CONCLUSIONStreptococcus mutans, Streptococcus sanguis, and Streptococcus salivarius could obviously inhibit the growth of Saccharomyces albicans in vitro. However, it is still unclear that among which the inhibition effects is stronger. The antagonistic effects is weakened gradually.

In Vitro Techniques ; Saccharomyces ; Streptococcus ; Streptococcus mutans ; Streptococcus sanguis

OBJECTIVEThe aim of this study was to investigate whether the three species of oral Actinomyces have inhibitory effects on the growth of oral Candida albicans in vitro.

METHODSStraight o'clock method was used to observe the bacteriostasis circle. Reverse o'clock and mixed culture method were used to study the quantitative changes of Candida albicans colony respectively.

RESULTS(1) None of the groups had been viewed the bacteriostasis circle. (2) Compared with control groups, there was a significant decrease of Candida albicans colony on Actinomyces viscosus TPY soft agar (P < 0.05). Actinomyces naeslundii and Actinomyces odontolyticus TPY soft agar were both devoid of obvious Candida albicans colony (P < 0.01). The former group (Actinomyces viscosus) and the two latter groups (Actinomyces naeslundii and Actinomyces odontolyticus) showed a striking contrast (P < 0.01). (3) Compared with control groups, a decrease of Candida albicans showed up in the mixed culture, and the difference was significant (P < 0.05). The discrepancies among the three experimental groups were of no statistical value (P > 0.05).

CONCLUSIONOral Actinomyces viscosus, Actinomyces naeslundii and Actinomyces odontolyticus could inhibit the growth of Candida albicans in vitro. However, which of them contributed more to the inhibitory effects was still not affirmed.

Actinomyces ; Actinomyces viscosus ; Candida albicans ; In Vitro Techniques

OBJECTIVETo observe the effect of matrix metalloproteinase-1 (MMP-1) from human host on degradation of dentin organic matrix of root dentin.

METHODSThe freshly extracted caries-free impacted teeth were selected. Teeth were cut transversely under the enamel-cementum junction into dentin sections with a thickness of about 5 mm. Then all sections with removal of cementum, pulp and predentin were randomly divided into four groups. In the first group, dentin sections were demineralized with acid solution for 21 days, and then incubated with MMP-1 solution for 7 days; the second group were only treated with acid solution for 21 days; the third group were only attacked by MMP-1 solution for 7 days; and the fourth group were untreated as a control. Then all sections were dehydrated in ascending strength of alcohol, critically dried, coated with platinum, and then observed under scanning electron microscope(SEM).

RESULTSThe dentin sections of root surface attacked by acid and MMP-1 showed that demineralization of dentin mineral and degradation of dentin matrix fibrae synchronously happened. The dentin matrix fibrae wasn't degradated in the groups treated with acid or MMP-1.

CONCLUSIONThe proteinases from human host may play an important role in the development of root surface caries. MMP-1 may distinctly degradate the organic matrix of demineralized dentin.

Dental Cementum ; Dental Enamel ; Dentin ; enzymology ; Humans ; Matrix Metalloproteinase 1 ; physiology ; Microscopy, Electrochemical, Scanning ; Root Caries ; enzymology ; Tooth Root ; enzymology

1 approximately 3 days old Piglet's primary preadipocytes in vitro were cultured and treated with 0micromol/L (control group), 10microlmol/L (lower dose group), 20micromol/L(middle dose group) and 50micromol/L, 100micromol/L (higher dose group) RES. Cell proliferation and viability were analyzed by MTT assay. The degree of differentiation and adipogenesis were measured by Oil Red O staining extraction assay and the expression of Sirt1 (sirtuin) mRNA were detected by RT-PCR. The results showed the optical density (OD) of MTT and Oil Red O staining were all decreased, especially treated by 50micromol/L, 100micromol/L RES at 72h and 96h (P < 0.01); the ratio of OD of the expression of Sirt1 mRNA to that of beta-actin mRNA were increased after treated by 100micromol/L RES (P < 0.01). RES can inhibit proliferation and differentiation of pig preadipocytes in certain degree. Higher dose of RES can markedly decrease adipogenesis and prevent preadipocytes differentiation into adipocytes, which may be in part associated with its effect on increasing the expression of Sirt1 mRNA.
Adipocytes ; cytology ; drug effects ; metabolism ; Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; RNA, Messenger ; analysis ; Sirtuin 1 ; genetics ; Stem Cells ; drug effects ; Stilbenes ; pharmacology ; Swine ; Transcription, Genetic

Objective:To investigate the effects of 3-methyladenine(3-MA)on extracellular matrix deposition in early diabetic nephropathy(DN)and its mechanism.Methods:A streptozotocin(STZ)-induced type 1 diabetes mouse model was used, and the mice were divided into vehicle control group, diabetes group(STZ group), 3-MA group, and chloroquine(CQ)group, 8 mice in each group. After 6 weeks of intervention, both kidneys were harvested, and the kidney-to-body weight ratio was recorded. Western blotting was performed to detect protein expressions of renal cortex fibronectin, α-smooth muscle actin(α-SMA), LC3, Beclin 1, p62, and vascular endothelial growth factor(VEGF). Immunohistochemistry was used to observe kidney fibronectin staining. Bioinformatics analysis was conducted on shared genes between diabetic nephropathy(DN)gene targets and 3-MA predicted gene targets.Results:Both 3-MA and CQ exhibited certain hypoglycemic effects in diabetic mice. Compared to the STZ group, the kidney-to-body weight ratio decreased in the 3-MA group( P<0.05). Western blotting showed that 3-MA reduced the expression of renal cortex matrix-related proteins fibronectin and α-SMA in diabetic mice( P<0.05 or P<0.01). Immunohistochemistry also revealed that 3-MA reduced fibronectin staining in the kidneys of diabetic mice. Both 3-MA and CQ inhibited the protein expression of renal cortex Beclin 1 in diabetic mice(both P<0.05), while 3-MA increased the expression of renal cortex p62( P<0.05). Bioinformatics analysis indicated a connection between shared genes of DN gene targets and 3-MA predicted gene targets with the VEGF signaling pathway. Western blotting results further showed that 3-MA reduced renal cortex VEGF expression in diabetic mice( P<0.01). Conclusion:3-MA can alleviate extracellular matrix deposition in the kidneys of early DN mice by inhibiting the VEGF signaling pathway.

OBJECTIVETo investigate the effect of prostaglandins E2 (PGE2) in enhancing vascular endothelial growth factor (VEGF) expression in a rat macrophage cell line and the effect of the media from PGE2-inuced rat macrophages on angiogenetic ability of human umbilical vein endothelial cells (HUVECs) in vitro.

METHODSWestern blotting and qPCR were employed to investigate the expressions of VEGF protein and mRNAs in rat macrophage cell line NR8383 stimulated by PGE2 in the presence or absence of EP2 receptor inhibitor (AH6809) and EP4 receptor inhibitor (AH23848). Conditioned supernatants were obtained from different NR8383 subsets to stimulate HUVECs, and the tube formation ability and migration of the HUVECs were assessed with Transwell assay.

RESULTSPGE2 stimulation significantly enhanced the expression of VEGF protein and mRNAs in NR8383 cells in a dose-dependent manner. The supernatants from NR8383 cells stimulated by PGE2 significantly enhanced tube formation ability of HUVECs (P<0.05) and promoted the cell migration. Such effects of PGE2 were blocked by the application of AH6809 and AH23848.

CONCLUSIONPGE2 can dose-dependently increase VEGF expression in NR8383 cells, and the supernatants derived from PGE2-stimulated NR8383 cells can induce HUVEC migration and accelerate the growth of tube like structures. PGE2 are essential to corpus luteum formation by stimulating macrophages to induce angiogenesis through EP2/EP4.

Animals ; Cell Line ; Cell Movement ; Cells, Cultured ; Culture Media, Conditioned ; pharmacology ; Dinoprostone ; pharmacology ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; Humans ; Macrophages ; chemistry ; Neovascularization, Pathologic ; RNA, Messenger ; Rats ; Receptors, Prostaglandin E, EP2 Subtype ; metabolism ; Receptors, Prostaglandin E, EP4 Subtype ; metabolism ; Vascular Endothelial Growth Factor A ; Xanthones ; pharmacology

OBJECTIVETo investigate the role of endometrial macrophages in embryo implantation and in regulating the expression of vascular endothelial growth factor A (VEGFA) in mouse endometrium during the peri-implantation period.

METHODAt D3.5 (D0.5 defined as the morning when a vaginal plug was observed), pregnant mice were divided randomly into experimental group, control group and blank group. In the experimental group, the mice were subjected to intrauterine injection of clodronate liposomes on the left side of uterus to eliminate the macrophages, and PBS liposomes on the right side. PBS liposomes and PBS were administered in the control and blank groups, respectively. The uterine tissues were collected on D5.5 and stained with trypan blue to show the implantation sites. Flow cytometry was performed to examine the percentage of F4/80(+) CD11b(+) macrophages macrophages in the uterus. F4/80(+) macrophage population within the endometrium and ovary and changes in VEGFA expression at the implantation and non-implantation sites were examined using immunohistochemistry.

RESULTSEndometrial F4/80(+) CD11b(+) macrophages macrophages were significantly reduced by 74% following intrauterine injection of clodronate liposomes (P<0.05). The number of macrophages in the ovaries showed no significant difference among the 3 groups. In the experimental group, the left side of the uterine showed imcomplete cavity closure with a lower number of implantation site than the right side (2.20∓1.81 vs 5.10∓1.91, P<0.05). VEGFA expression at the implantation site were significantly decreased in the endometrium on the left side with macrophage suppression as compared with that on the right side (P<0.05).

CONCLUSIONEndometrial macrophages appear to modulate uterine receptivity by regulating the expression of VEGFA to affect embryo implantation, suggesting the important role of macrophages in embryo implantation.

Animals ; Embryo Implantation ; Endometrium ; physiology ; Female ; Immunohistochemistry ; Macrophages ; cytology ; Mice ; Ovary ; cytology ; Pregnancy ; Random Allocation ; Uterus ; cytology ; Vascular Endothelial Growth Factor A ; physiology

Objective: To explore the relationship betweenrheumatoid arthritis (RA) and chronic periodontitis (CP). Methods: A total of 48 RA patients were recruited from the Rhematology Department of The First Affiliated Hospital of Shantou University Medical College (SUMC). RA patients were matched on age and gender with healthy controls, who were recruited from the Stomatology Department. Dental parameters including unstimulated salivary flow rate(UWS), stimulated salivary flow rate (SWS), bleeding on probe (BOP), periodontal probing pocket (PD), clinical attachment level (CAL) and decayed, missing and filling (DMF) were recorded in all cases. Erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and Anticyclic citrullinated peptide antibody (Anti-CCP) were also recorded in RA patients. Results : There were significant difference between RA group and health control group on salivary flow rate, BOP, PD, CAL and DMF (P< 0.001). Higher percentage of RA patients were diagnosed as periodontal disease than those in control group (P< 0.001). There was relationship between CAL and Anti-CCP antibody (P< 0.001). Conclusion RA patients have higher risk of CP, and there might be relationship between RA and CP.