Objective:To detect the degree of oxidative stress in the process when Porphyromonas gin-givalis ( P. gingivalis) stimulates human vascular endothelium, And to investigate the effect of peroxi-some proliferator-activated receptor(PPAR)γ on oxidative stress during this process. Methods:Human vascular endothelial cells ( HVECs) line EA. hy926 ( American Type Culture Collection ,United States) was cultured in high glucose Dulbecco' s modified eagle medium ( DMEM) . Four groups were designed:control group, P. gingivalis infected group, PPARγactivated group and PPARγblocked group. In con-trol group HVECs were cultured with only DMEM. In P. gingivalis infected group, HVECs were time-dependently stimulated by P. gingivalis W83 from 0 to 12 h. In PPARγ activated group or PPARγblocked group, PPARγ was pre-activated or blocked by a representative PPARγ agonist(15d-PGJ2 10μmol/L) or antagonist ( GW966210μmol/L) 30 minutes before the cells were stimulated by P. gingiva-lis. At 0, 0. 5, 1, 1. 5, 2, 4, 8, and 12 h, the culture medium was collected individually and centri-fuged, and the supernatant was stored for assay. Glutathione peroxidase (GSH-PX) and malondialdehyde( MDA) were analysed by enzyme-linked immunosorbent assay. Cellular reactive oxygen species ( ROS) were detected through 2',7'-dichlorofluorescin diacetate (DCFA-DA) fluorescent probe at various time points of the different groups. Results:In P. gingivalis infected group, the levels of GSH-PX [(5. 56 ± 0. 97) μmol/L] and MDA [(0. 84 ± 0. 18) nmol/L] were significantly higher than those in control group [GSH-PX(4. 71 ± 0. 64) μmol/L, MDA (0. 59 ± 0. 18) nmol/L)]. The levels of GSH-PX and MDA in PPARγactivated group [GSH-PX (5. 38 ± 0. 84) μmol/L, MDA (0. 84 ± 0. 22) nmol/L] and in PPARγblocked group [GSH-PX (5. 37 ± 0. 76) μmol/L, MDA (0. 85 ± 0. 14) nmol/L] were signi-ficantly higher than those in control group (P <0. 05). In the PPARγ activated group, the levels of GSH-PX at 0 . 5 and 8 h were significantly higher than those from 1 . 5 h to 4 h ( P<0 . 05 ) , while no difference was observed on the MDA levels at different time points. There was no significant difference at various time points for the levels of GSH-PX and MDA in PPARγ blocked group. The level of cellular ROS detected by DCFH-DA in P. gingivalis infected group was significantly higher than that in control group (10 108. 65 ± 1 805. 18 vs. 6 049. 06 ± 1 199. 19,P<0. 05). No difference was observed be-tween PPARγ activated group (7 120. 94 ± 1 447. 30) or PPARγblocked group (6 727. 35 ± 1 483. 68) and control group. Conclusion:Oxidative stress happens when P. gingivalis stimulates human vascular endothelium. PPARγ may involve in modulating oxidative stress during this process.

Objective To observe the protective effects of FTY720 on the Con A-induced mouse hepatic fibrosis injury,and to find the possible mechanisms of protective effects.Methods The pathologic models of hepatic fibrosis injury in the mice caused by Con A were set up.Forty mice were randomly divided into control group, model group,high dose of FTY720 (4 mg·kg-1 )group and low dose of FTY720 (1 mg·kg-1 )dose group (n=10).The serum alanine aminotransferase (ALT)and asparate aminotransferase (AST)activities,hepatic index and pathological changes of hepatic tissue were detected .Results Compared with model group,the serum ALT and AST activities in low and high doses of FTY720 groups were decreased significantly (P < 0.05 or P < 0.01).The optical microscope results showed that there were inflammatory cells and hepatocellular necrosis in model group. The masson staining results showed that there were surrounding fiber bundle and hepatic lobule fusion in model group;compared with model group,the damage degree in low and high doses of FTY720 groups was reduced.The protective effects of FTY720 on hepatic injury showed linear relation to the drug dose.Conclusion FTY720 could decrease the levels of ALT/AST,thus FTY720 alleviate hepatic damage degree and delay the process of hepatic fibrosis.The protective effects of FTY720 on hepatic injury in experimental hepatic fibrosis mice may be related to the mechanisms mentioned above.

Objective To study the apoptosis of K562 cells induced by a new immunosuppressive agent FTY720 and its mechanism,and to provide experimental basis for the treatment of leukemia in clinic.Methods The K562 cells were cultured in vitro and divided into blank control group and FTY720 treatment group.The K562 cells in FTY720 treatment group were treated with 6μmol·L-1 FTY720 for 3,6,and 12 h,or treated with different concentrations of FTY720 (2,4,6,8μmol·L-1)for 24 h.The apoptosis,level of reactive oxygen species (ROS),mitochondrial membrane potential(MMP)and cell cycle were measured by flow cytometry.The inhibitory rate of proliferation of K562 cells after treated with FTY720 was detected by WST-1 reducting assay.Results The results of flow cytometry showed that the percentages of apoptotic cells were increased after treated with 6μmol·L-1 FTY720 for 3,6,and 12 h with the prolongation of time compared with blank control group(P<0.01).The percentages of apoptotic cells were also increased after treated with different concentrations of FTY720 for 24 h compared with blank control group(P<0.01).Compared with blank control group,the ROS levels were increased with the increasing of FTY720 concentration,while the MMP was decreased(P<0.01).Compared with blank control group,the percentage of cells at G0/G1 phase was increased,while those at S and G2/M phases were decreased with the increasing of FTY720 concentration (P<0.05).The WST-1 reduction assay results indicated that the inhibitory rates of proliferation of K562 cells after treated with 2,4,6,and 8μmol·L-1 FTY720 for 72 h were (24.0±4.1)%,(46.4±3.9)%,(67.0±4.8)%,and (88.2±5.6)%,respectively,compared with blank control group.The concentration of FTY720 to result the inhibitory rate of 50% (IC50 )on K562 cells was 5.5μmol·L-1 .Conclusion FTY720 could inhibit the proliferation of K562 cells by blocking cell cycle and inducing apoptosis through provoking ROS.

OBJECTIVETo detect the expression of vascular endothelial growth factor (VEGF), epidermal growth factor receptor (EGFR), p16 protein in lip cancers and oral squamous cell carcinoma (OSCC) as well as their clinicopathological significance.

METHODSImmunohistochemisty for expression VEGF, EGFR, P16 were carried out in 69 cases of lip cancers and OSCC.

RESULTSExpression of VEGF, EGFR, p16 protein in OSCC and lip cancers was respectively 71.01%, 46.37%, 28.98% and there were no significance between their positive expressions (P > 0.05) as well as in different sites of them (P > 0.05). Expression of VEGF was respectively 71.01% in cancers and 10.00% in non-tumor tissues, there was statistic significance among those (P < 0.05).

CONCLUSIONSThe results show that there is no correlation to the expression of VEGF, EGFR and P16 protein in OSCC and lip cancers. It is suggests that the expression of VEGF might become one of the useful markers for OSCC.

Adult ; Aged ; Carcinoma, Squamous Cell ; chemistry ; pathology ; Cyclin-Dependent Kinase Inhibitor p16 ; analysis ; Endothelial Growth Factors ; analysis ; Female ; Humans ; Immunohistochemistry ; Intercellular Signaling Peptides and Proteins ; analysis ; Lip Neoplasms ; chemistry ; pathology ; Lymphokines ; analysis ; Male ; Middle Aged ; Mouth Neoplasms ; chemistry ; pathology ; Receptor, Epidermal Growth Factor ; analysis ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVETo investigate the expression of NOEY2 gene in breast cancer tissue and its relation to clinicopathological and other molecular parameters.

METHODSThe mRNA expression of NOEY2 gene was monitored in benign and malignant breast lesions by RT-PCR and in situ hybridization (ISH) with transcripted antisense RNA probes. The protein expression of estrogen receptor (ER), Ki67, p27 and p21(WAF1) in the 60 breast cancer lesions was detected by immunohistochemical method.

RESULTSAll 6 benign lesions, and 13 (72.2%) of the 18 breast cancers were NOEY2 positive by RT-PCR. By ISH, positive NOEY2 was obtained in all 10 benign lesions but only in 31 (51.7%) of the 60 breast cancer lesions. The difference was statistically significant (P = 0.025). NOEY2 positive rate tended to decrease with the increase of histological grade. However, NOEY2 expression was negatively correlated with the status of axillary lymph nodes. The positive NOEY2 rate was 75% in those without lymph node metastasis but only 26.7% in those with metastasis (P < 0.001). No correlation with other clinicopathological parameters including ER, Ki67, p27 or p21(WAF1) were found.

CONCLUSIONNOEY2 gene may be related with the pathogenesis of breast cancer. A link between NOEY2 loss expression and the spreading mechanism of breast cancer may possibly exist.

Breast Neoplasms ; metabolism ; Female ; Gene Expression ; Humans ; In Situ Hybridization ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; rho GTP-Binding Proteins ; biosynthesis ; genetics

Objective To explore the expression of zinc finger protein 217 (ZNF217) in non-small cell lung cancer (NSCLC) and its correlation with prognosis of patients. Methods A total of 120 NSCLC patients in Chongqing Three Gorges Central Hospital from January 2012 to October 2013 were selected. Immunohistochemical method was used to test the expression of ZNF217 in NSCLC tissues and paracancerous tissues. The correlation of ZNF217 expression with patient's clinicopathological features was analyzed. At the same time, the Kaplan-Meier method and Cox regression model multiple factor analysis method were used to explore the factors affecting the prognosis of patients after NSCLC radical operations. Results ZNF217 mainly existed in cell nucleus of NSCLC. The positive expression rate of ZNF217 in the cancer tissues was higher than that in the paracancerous tissues [52.5％ (63/120) vs. 20.1％ (25/120), χ 2 = 25.909, P < 0.05]. The positive expression rate of ZNF217 increased with the increase of tumor T stage (χ 2 = 7.333, P = 0.026), N stage (χ 2 = 7.782, P = 0.020) and TNM stage (χ 2 = 11.557, P = 0.003). The overall survival (P = 0.007) and progression-free survival (P = 0.004) of patients with positive ZNF217 were poorer than those of patients with negative ZNF217. Cox multiple factor analysis showed that ZNF217 was an independent risk factor affecting the prognosis of NSCLC. Conclusion ZNF217 is an independent risk factor affecting the prognosis of NSCLC, and it may be a potential target for accurate treatment of NSCLC.

Objective:To compare the efficacy and safety of ultrasound-guided percutaneous translumial septal myocardial ablation in dogs using laser and radiofrequency.Methods:Twelve healthy adult Beagle dogs (males or females) were randomly divided into two groups, namely, group laser and group radiofrequency (6 dogs each group). Under ultrasound guidance, laser fiber or radiofrequency ablation needle was respectively inserted into the basal and middle segments of the interventricular septa via the percutaneous transapical approach to perform ablation. The Beagle dogs received radiologic examination, laboratory tests and pathological detection before ablation, immediately after ablation, at 1 week after ablation, and at 1 month after ablation, respectively. The efficacy and safety of the two ablation procedures were compared.Results:All dogs survived after ablation. The peak gradient of LVOT decreased immediately after ablation using either laser or radiofrequency ( P<0.05), but it increased at 1 week after ablation than before ( P<0.05). At 1 month after ablation, no significant differences were found in the peak gradient of LVOT compared with that before surgery ( P<0.05). The interventricular septum thickness was increased immediately after ablation using either laser or radiofrequency than before ( P<0.05), but it decreased at 1 week and at 1 month after surgery than before ( P<0.05). The ablation zone using radiofrequency was slightly larger than that of using laser[(372.50±69.06)mm 3 vs (116.65±20.15)mm 3, P<0.001], and the surgical time of the former was significantly shorter than that of using laser [(56.00±3.22)s vs (260.00±65.39)s, P<0.05)]. Conclusions:Ultrasound-guided percutaneous translumial septal myocardial ablation is feasible, safe and effective using either laser or radiofrequency. Comparatively speaking, radiofrequency ablation is more simple and convenient.

Objective:To explore the characteristics of Lowe syndrome, as well as OCRL1 gene mutation and its relationship with phenotype. Methods:Children diagnosed with Lowe syndrome during their visit to Nanfang Hospital of Southern Medical University (4 cases) and the First Affiliated Hospital of Sun Yat-sen University (3 cases) from January 2009 to January 2019 were included. The clinical data and peripheral blood samples were collected, and the sequence analysis of OCRL1 was performed after genomic DNA extraction. Then the clinical features of the children and the relationship between OCRL1 mutation and clinical phenotype were analyzed. Results:Seven patients from 6 families who presented with Lowe syndrome were included. All of them had different degrees of ocular-neural-renal symptoms. Six cases from 5 families had congenital cataract and neonatal hypotonia, one case from another family only had a thin lens without cataract. Four cases had nystagmus and 2 cases had glaucoma. Six cases from 6 families had psychomotor retardation and had proximal tubular impairment, included low-molecular-weight proteinuria (LMWP). Serum aspartate transaminase (AST), lactate dehydrogenase (LDH), creatine kinase (CK) and creatine kinase-MB (CK-MB) were increased in all 6 patients who were tested. Mutations of OCRL1 were detected in all the 6 families, which located in exon 10, 13, 16, 18, 22 and 23 respectively. The mutations of c.891 G>T, c.1682_1683insAA and c.2564_2567del are novel. Conclusions:Three OCRL1 novel mutations in 6 Chinese Lowe syndrome families are identified. The clinical manifestations in different mutations of OCRL1 are heterogeneous. The mutations of c.891 G>T in exon 10 without congenital cataract is rare in clinical.

Objective To explore the clinical efficacy of Shenkang Injection and its influence on C-reactive protein (CRP)and interleukin-6 (IL-6)levels in patients with diabetic nephropathy (DN).Methods A total of 120 patients with type 2 diabetes mellitus combined with DN admitted in our hospital from Jan.2012 to Jan.2014 were randomly divided into observation group and con-trol group,60 cases for each.Control group was treated with high-quality protein diets and insulin to control blood glucose and pressure,on which basis observation group was added with intravenous injection of Shenkang Injection.Clinical efficacy,fasting blood glucose (FBG),Triglyceride (TG), total cholesterol (TC),serum creatinine (SCr),24 h urinary protein (24 hUpro),CRP and IL-6 level changes before and after treatment in both groups were observed.Results Clinical efficacy was 90.00% in observation group,evidently higher than 75.00% in control group (P <0.05). Aboveindexeswere all improvedobviously after treatment than treatment before (P < 0 .0 5 ,P <0.01)and were markedly lower in observation group than in control group (P <0.01).Conclusion Shenkang Injection can effectively reduce IL-6 and CRP levels and decrease blood glucose and pressure,prolong disease progression and improve prognosis in DN patients.

Objective To explore the activation of tumor necrosis factor-α(TNF-α)signaling pathway and the expressions of the associated inflammatory factors in NPHP1-defective renal tubular epithelial cells.Methods A human proximal renal tubular cell(HK2)model of lentivirus-mediated NPHP1 knockdown(NPHP1KD)was constructed,and the expressions of TNF-α,p38,and C/EBPβ and the inflammatory factors CXCL5,CCL20,IL-1β,IL-6 and MCP-1 were detected using RT-qPCR,Western blotting or enzyme-linked immunosorbent assay.A small interfering RNA(siRNA)was transfected in wild-type and NPHP1KDHK2 cells,and the changes in the expressions of TNF-α,p38,and C/EBPβ and the inflammatory factors were examined.Results NPHP1KDHK2 cells showed significantly increased mRNA expressions of TNF-α,C/EBPβ,CXCL5,IL-1β,and IL-6(P<0.05),protein expressions of phospho-p38 and C/EBPβ(P<0.05),and IL-6 level in the culture supernatant(P<0.05),and these changes were significantly blocked by transfection of cells with siRNA-C/EBPβ(P<0.05).Conclusion TNF-α signaling pathway is activated and its associated inflammatory factors are upregulated in NPHP1KDHK2 cells,and C/EBPβ may serve as a key transcription factor to mediate these changes.