Objective To investigate the expression of vascular endothelial growth factor (VEGF) in colon cancer and its relation with the microvessel density (MVD) and p53 gene. Methods The expression of VEGF, p53 gene and MVD of different region of colon cancer were studied by immunohistochemical method from 68 by surgical resected specimens. Results The positive rates of VEGF, p53 and MVD in colon cancer were significantly higher than those in peritumor and normal tissue. The expression of VEGF and p53 positively correlated with depth of invasion, lymph node metastasis, distant metastasis, vessel invasion and Dukes' stage, but their relation to the histologic types was not significant. The MVD in p53(+) ( 34.6 ?12.2 ) and VEGF(+) (31.2?12.6) tumors was significantly higher than that in p53(-) and VEGF(-) tumors (15.0?7.9; 12.7?6.3, respectively, P

Objective To investigate whether ultrasound-mediated microbubble destruction can effectively deliver pEGFP-N1 plasmid into human gingival fibroblasts (HGFs).Methods The primary cultured HGFs were divided into 4 groups,that is,plasmid,microbubble+plasmid,ultrasound+plasmid,and ultrasound+microbubble+plasmid groups.pEGFP-N1 plasmid was used to transfect to HGFs as a gene marker with ultrasound-mediated microbubble destruction.The last group was further divided into subgroups to optimize the transfection conditions.After 48 h,phase-contrast fluorescent microscopy was employed to evaluate the expression of green fluorescent protein (GFP).The cell vitality was measured by the MTT assay.Results The transfection efficiency of the ultrasound+microbubble+plasmid group was higher than other experiment groups.Optimal gene expression was found when ultrasound was radiated at 1.5 W/cm2 for 60 s,microbubble was at a concentration of 10% and plasmid was at a concentration of 6.67 ?g/ml,and the transfection efficiency was highest under this condition.Conclusion Under specific conditions,ultrasound mediated microbubble destruction enhances the reporter gene transfection and expression in HGFs.

ObjectiveTo observe the effects of bifidobacterial sectory adhesin on NF-κB DNA binding activity,cytoplasm IκB,nuclei NF-κBp65,TNF-α,IL-1β,and IL-8 expression in intestinal epithelial cell post stress response.MethodsThe cultured intestinal epithelial cell line Lovo cells were divided into 5 groups,which included directly stimulated by lypopolysaccharide (LPS) (100 ng/ml) or H2O2 (200 μmol/L) for 3 hours groups,bifidobacterial sectory adhesin (30μg/ml) pre-incubation for 30 minutes and then treated by LPS (100 ng/ml) or H2O2 (200 μmol/L) for 3 hours groups and control group with no treatment.NF-κB DNA binding activity was evaluated by electrophoretic mobility shift assay (EMSA).The cytoplasm IκB and nuclei NF-κBp65 expression were determined by Western blot.The expressions of TNF-α,IL-1β and IL-8 at mRNA level were detected by semi-quantatitive assay of RT-PCR.ResultsThe expressions of NF-κB DNA binding activity [(6.20±0.35) times and (4.16 ± 0.52) times of that of non treated control group,respectively] and nuclei NF-κBp65 [ (0.64±0.05) and (0.67±0.06)] were higher in cells after LPS or H2()2 stimulated,however the cytoplasm IκB expression [ (0.28 ± 0.10) and (0.39 ± 0.12) respectively] was weaker.After bifidobacterial sectory adhesin pre-incubation,the expressions of NF-κB DNA binding activity and nuclei NF-κBp65decreased obviously,but cytoplasm IκB expression increased.After treated with LPS or H2O2,the mRNA expressions of TNF-α,IL-1β and IL-8 were significantly increased [LPS treated group2 (0.92±0.10)、(0.38±0.03)、(1.44±0.25),H2O2 treated group:(0.89±0.13)、(0.36±0.06)、(1.42±0.18)].After bifidobacterial sectory adhesin pre-incubation,their expressions decreased.The mRNA expressions of TNF-α,IL-1β and IL-8 have significantly positive correlation with NF-κB DNA binding activity.ConclusionsThere is a significant inhibition role of bifidobacterial sectory adhesin in LPS and H2O2 induced NF-κB DNA binding activity in intestinal epithelia cells.The activation of NFκB may associate with the regulation of TNF-α,IL-1β and IL-8 proinflammatory cytokines expression.

Objective To investigate the value of hs-CRP in the assessment of risk stratification,coronary artery disease severity and prognosis for patients undergoing PCI with coronary heart disease.Methods A total of 150 coronary heart disease patients were recruited in the study,who had undergone PCI,and before that all the patients had undergone coronary angiography,and the levels of hs-CRP,TG,LDL-C,HDL-C,NT-proBNP and hs-cTnT were measured.Patients were divided into three groups according to the serum concentration of hs-CRP.Gemini score was used to determine the degree of stenosis.After PCI,patients were followed up for 6 months,the major cardiovascular events(MACE)were recorded.The relationships between hs-CRP and other coronary risk factors,Gemini score,vasculopathy count,MACE events were analyzed.Results The serum concentrations of hs-CRP and HDL-C, hs-cTnT were significantly correlated(r=-0.17,0.42,P <0.05).The level of hs-CRP had increasing tendency with the increase of Gensini score,and the serum concentration of hs-CRP in severe vascular lesions group was significantly higher than both mild vascular lesions group and moderate vascular lesions group(P <0.05).The serum concentration of hs-CRP in single-vessel disease patients,double-vessel disease patients and multi-vessel disease patients were (4.64±5.39),(9.86±8.75),(14.93±10.34)mg/L, there was significant difference among them(P <0.05 ).The correlation coefficients of hs-CRP and Gemini score was 0.21 (P <0.05),while hs-CRP and vasculopathy count was 0.18(P <0.05).The incidence of MACE in three groups were 18.52%,43.75%and 58.66%,which were statistically significantly difference(χ2 =13.42,P =0.001).Logistic regression showed that hs-CRP was an independent risk factor for MACE(OR =2.05,95%CI :1.24 -3.39,P =0.005).Conclusion Hs-CRP is an independent risk factor for coronary heart disease in patients with MACE after PCI,and hs-CRP was correlated with PCI patient′s vascular disease. Hs-CRP can be used as an indicator in the assessment of patient′s condition,PCI risk stratification and prognosis.

ObjectiveTo investigate the protection effect of bifidobacterial adhesin for intestine ischemia/reperfusion (I/R) injury on gut barrier function in rat.MethodsSeventy-two male SD rats were randomly divided into sham operation group (n =24), I/R model group (n =24) and pretreatment group of bifidobacterial adhesin (pretreatment group, n = 24).Six rats were anatomized at 6 h, 1 d, 4 d and 7d after inducing I/R model in each group, respectively.The pathological changes of the terminal ilea and the blood levels of TNFα, IL-6, IL-10, diamine oxidase (DAO), and the activity and content of D-lactic acid were observed.ResultsThe blood levels of TNFα, IL-6, DAO and D-lactic acid in I/R model group were significantly higher than sham operation group at all time points (P ＜0.05) , while the blood level of IL-10 was no significantly change.The activity of IL-6 and DAO in pretreatment group was significantly lower than I/R model group at all time points (P ＜ 0.05), the blood level of TNFαt in pretreatment group was significantly lower than I/R model group at 1 d, the blood level of D-lactic was significantly lower than I/R model group at 4 d and 7 d (P ＜ 0.05). Intestinal pathological damages were obviously milder in pretreatment group than I/R model group at all time points (Chiu's pathological scores: 6 h, 3.22 ±0.22 vs 3.57 ±0.20;1 d,3.77 ±0.13 vs 3.90 ±0.12;4 d,2.93 ±0.23 vs 3.07 ±0.21;7 d,2.10 ±0.30 vs 2.22 ±0.17,all P ＜ 0.05).ConclusionThe pretreatment of bifidobacterial adhesin could protect the intestinal mucosa from I/R injury, and alleviate intestinal ischemic reperfusion injury.

Objective To observe glucose metabolism in C57 mice treated with different doses of fluoride.Methods Forty male C57 mice (body weight 20-24 g) were divided into four groups which were exposed to 0,50,100 and 150 mg/L sodium fluoride (NaF) by random number table according to body weight,each group had 10 mice.At 2,4,6,8,10 and 12 weeks after fluoride exposure,body weight was measured,blood glucose and glycosylated hemoglobin were detected by blood glucose meter and glycosylated hemoglobin meter,serum insulin and glucagon were detected by enzyme-linked immunosorbent assay (ELISA).Results At 10 and 12 weeks after fluoride exposure,the differences of fasting glucose between groups of C57 mice were statistically significant (F =35.12,21.92,all P ＜ 0.05),the fasting glucose of 100,150 mg/L fluoride groups [(7.7 ± 0.2),(7.3 ± 0.3),(8.6 ± 0.5),(9.1 ± 0.7)mmol/L] were higher than those of the control group [(5.4 ± 0.3),(5.0 ± 0.3)mmol/L,all P ＜ 0.01].The differences of glycosylated hemoglobin,glucagons between groups were statistically significant (F =3.85,8.74,all P ＜ 0.05).The glycosylated hemoglobin of 100,150 mg/L fluoride groups [(7.73 ± 0.76),(7.80 ± 1.15) mmo]/L] were higher than those of the control group [(5.43 ± 1.27) mmol/L,all P ＜ 0.05]; serum glucagon levels of 50,100,150 mg/L fluoride groups [(19.15 ± 11.84),(26.55 ± 15.97),(20.05 ± 7.29)ng/L] were lower than that of the control group [(48.35 ± 2.79)ng/L,all P ＜ 0.01].Conclusion Long term excess fluoride intake can reduce the function of sugar metabolism in C57 mice.

Objective To observe gene different expression of unfolded protein response signaling pathway in human osteoblasts under the excessive fluoride ,and explore the role of endoplasmic reticulum stress in fluorosis .Methods Human osteoblasts were cultured with fluoride ,intervening for 24 h .Cell viability and apoptosis were inspected by MTS assay and flow cytometer respective‐ly .The UPR signaling pathway was examined by real time PCR array ,and protein expressions were detected by Western blot .Re‐sults T he cell survival rates w ere (100 .678 5 ± 2 .830 3 )% ,(105 .393 4 ± 2 .538 4 )% ,(106 .125 7 ± 2 .048 3 )% ,(77 .977 3 ± 2 .544 3)% (P<0 .05) ,(30 .237 7 ± 0 .632 73)% (P<0 .05) treated with sodium fluoride at the concentration 0 ,5 ,10 ,20 ,40 ,80 mg/L respectively .Apoptosis rate inspected by flow cytometer was 4 .8% in 5 mg/L group ,13 .8% in 10 mg/L group ,37 .0% in 20 mg/L group ,58 .9% in 40 mg/L group ,63 .2% in 80 mg/L group (P<0 .05) .Only 1 gene was down regulated and 14 genes were up regulated .Western blot analysis showed BIP ,ATF4 ,CHOP and IRE1 both showed their protein expression gradually up regula‐ted with fluorine dose .XBP1 expression gradually increased in NaF 5-20 mg/L ,and its expression decreased at 40 and 80 mg/L . Conclusion Sodium fluoride can cause osteoblasts endoplasmic reticulum stress pathway through PTEN and IRE1 pathway ,and at high concentrations can cause apoptosis of osteoblast .

Objective To investigate the effect and potential mechanism of microRNA (miRNA)-377 on high glucose-induced proliferation and inflammation in human mesangial cells.Methods Cells were randomly divided into six groups:control group (5.5 mmol/L glucose),high glucose group (30.0 mmol/L glucose),negtive miRNA inhibitor transfection+high glucose group,negtive miRNA mimic transfection+high glucose group,miRNA-377 inhibitor transfection+high glucose group (miR-377i+high glucose group),miRNA-377 mimic transfection+high glucose group (miR-377m+high glucose group),miRNA-377 expression was detected by real-time PCR.Cell proliferation and cell cycle were detected by BrdU assay and flow cytometry,respectively.The release of tumor necrosis factor-α (TNF-α),interleukin (IL)-18,IL-6 and macrophages chemotaxis protein-1 (MCP-1) were evaluated by ELISA.The activations of NF-κB pathway,including the expressions of phosporylated (p)-IκBα,p-P65 and nuclear P65,were measured by Western blotting.Results Compared with those in control group,in high glucose group cell viability,miRNA-377 expression and cell proliferation rate increased (all P ＜ 0.05),proportions of S phase cell and G2/M phase cell in cell cycle increased (all P ＜ 0.05),the levels of TNF-o,IL-18,IL-6 and MCP-1 were higher (all P ＜ 0.05),as well as the expressions of p-IKBα/IκBα,p-P65/P65 and nuclear P65 were increased (all P ＜ 0.05).Compared with high glucose group,cell proliferation rate was restrained (P ＜ 0.05),proportions of S phase cell and G2/M phase cell in cell cycle was descreased (all P ＜ 0.05),the levels of TNF-α,IL-18,IL-6 and MCP-1 were lower (all P ＜ 0.05),as well as the expressions of p-IκBα/IκBα,p-P65/P65 and nuclear P65 were reduced (all P ＜ 0.05) in miR-377i+high glucose group.However,miR-377m+high glucose group presented opposite results (all P ＜ 0.05).Conclusions miRNA-377 knockdown can partially suppress high glucose-induced human mesangial cell proliferation and cell cycle transition,and restrain inflammatory molecules release.Its mechanism may be related to the inhibition of NF-κB pathway.

OBJECTIVETo study the molecular mechanisms of Fuyuan capsule serum containing, icariin and arasaponin R1 in the treatment of osteoarthritis from the urokinase-type plasminogen activator (uPA) system.

METHODChondrocytes were isolated, cultured and identified using type II collagens immunostaining. After stimulating with TNF-alpha 10 microg x L(-1), 1 h, then the chondrocytes were treatment with glucosamine hydrochloride 25 g x L(-1), 20% Fuyuan capsule serum containing, icariin 12.5 mg x L(-1), arasaponin R1 125 mg x L(-1), icariin 12.5 mg x L(-1) + arasaponin R1 125 mg x L(-1). After 2 h, expression of uPA and nuclear factor kappa B (NF-kappaB P65) mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR), the activities of NF-kappaB(P65) combine DNA were determined by electrophoretic mobility shift assays (EMSA), 1kappaBalpha were detected by Western blotting.

RESULTFuyuan capsule, icariin and arasaponin R1 could significantly reduce NF-kappaB (P65) activities and uPA mRNA expression, and increase expression of IkappaBalpha (P < 0.01), but no significant difference between all treatment groups.

CONCLUSIONFuyuan capsule and its two main active ingredients, icariin and arasaponin R1, could protect chondrocytes from damage through reducing the NF-kappaB (P65) activities, increasing the express of IkappaBalpha and then reducing uPA of chondrocytes.

Animals ; Capsules ; Cells, Cultured ; Chondrocytes ; drug effects ; metabolism ; DNA-Binding Proteins ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Flavonoids ; pharmacology ; therapeutic use ; Gene Expression Regulation ; Male ; NF-kappa B ; genetics ; metabolism ; Osteoarthritis ; drug therapy ; genetics ; metabolism ; RNA, Messenger ; genetics ; Rabbits ; Urokinase-Type Plasminogen Activator ; genetics ; metabolism

Objective: To explore the social functioning characteristics of children with co ocurrence of attention deficit hyperactivity disorder (ADHD) and oppositional defiant disorder (ODD) for intervention reference. Methods: The Chinese Version of Swanson Nolan and Pelham, Version IV Scale-Parent Form(SNAP-IV), the Chinese Version of Weiss Functional Impairment Scale-Parent(WFIRS-P), and the Questionnaire-Children with Difficulties (QCD) were applied to 192 children with ADHD, 243 children with co occurrence of ADHD and ODD, who firstly visited the Department of Children Psychological Health of Zhuhai Maternal and Child Health Care Hospital, and 118 healthy control children from a school in Zhuhai. Results: The scores of attention deficit factor in SNAP-Ⅳ scale of children in three groups were[1.9(1.7, 2.1), 1.8(1.6, 1.9), 1.0(0.6, 1.2)], the scores of hyperactive impulsivity were[1.8(1.4, 2.1), 1.6(1.1, 1.8), 0.7(0.2, 1.0)] the scores of oppositional defiant were[1.6(1.5, 1.9), 1.0(0.8,1.1), 0.8(0.5, 1.0)], the differences were statistically significant( H=268.44, 237.97, 418.66, P <0.01). The dimensions and total scores of the three groups of children s WFIRS-P scale were family[0.8(0.6, 1.1), 0.6(0.3, 0.8), 0.3(0.1, 0.6)]; learning and school[0.8(0.5, 1.1), 0.8(0.5, 1.0), 0.3(0.1, 0.5)]; life skills[1.0(0.7, 1.2), 0.8(0.6, 1.0), 0.6(0.4, 0.8)]; self management [1.0(0.3, 1.0), 0.7(0.3, 1.0), 0.3(0.0, 0.7)]; social activities [0.7(0.4, 1.0), 0.6(0.3, 0.9), 0.3(0.0, 0.4 )]; adventure activities[0.3(0.2, 0.5), 0.2(0.1, 0.4), 0.1(0.0, 0.2)]; the total score[0.8(0.6, 1.0), 0.6(0.5, 0.8), 0.4( 0.2 , 0.6)], the difference between the groups was statistically significant( H=108.82, 122.45, 60.17, 40.58, 96.17, 76.57, 138.30, P <0.01). The difference between the QCD scale scores of children in the three groups was statistically significant[30.0( 24.0 , 37.0), 32.0(27.0, 40.0), 47.0(37.0, 52.3), H=124.65, P <0.01). Multiple regression analysis showed that attention deficit, and oppositional defiant symptoms were associated with both the total WFIRS-P score and the QCD score of children( R 2= 0.40 , 0.25, P <0.05). Conclusion Children with co occurrence of ADHD and ODD have more severe deficits in all dimensions of social functioning than children with ADHD, which might be associated with attention deficit and oppositional defiant symptoms.