ObjectiveTo evaluate a new cross-linking reagent—polyepoxy compound(PC)in vivo. MethodsTo reconstruct abdominal aota with jugular homografts of SD rats (diameter 0.8～1.1mm) pret4reated by glycero polyglycidyl ether(a kind of PC) or glutaraldehyde(GA). Grafts were explanted at different elapsed time, and their histological changes were studied. ResultsThere was no difference between the patencies of the PC and GA prepared homografts ( P >0.05), but the inci-dence of aneurysms in GA is higher than that in PC group ( P < 0.05). The healing procedures of endothelial cells in two experimental groups were basically same at the end of second month after transplantation. The endothelial cells cover the whole inner wall of the grafts. ConclusionCompaired PC graft with GA graft, it shows that PC are more unbreakable and compliant, lower rate of aneurysms and less calcification. PC might be a potential vessel-crosslink-reagent.

Objective To evaluate the clinical effectiveness of extended profundaplasty for treatment of critical leg ischemia. Methods Twenty-five legs (stage Ⅲ and Ⅳ in Fontaine′s classification) of 23 patients with critical leg ischemia caused by extensive occlusion of the distal arteries were reviewed during past 12 years. All patients underwent extended profundaplasty and were evaluated by clinical observation of API, tcPO 2, PSaO 2, Treadmill test, NIRS and ultrasound perioperatively. Results Fourteen legs in 13 patients had a favorable outcome, seven legs in 6 patients had an improvement on their symptom and sign, three legs in 3 patients underwent below knee and one case medio-tarsal amputation during the periods of 7 months to 7 years follow-up. Conclusion The extended profundaplasty is simple, less invasive, safe and effective for the treatment of limb-threatening ischemia.

Objective To evaluate the therapeutic effect of mini-incision surgery for the treatment of varicose vein of lower extremities. Methods 106 limbs of 95 paitnets with varicose vein underwent saphenectomy combined with ligation of communicating veins by way of mini-incision from November 1997 to April 2001. The ascending venography and StethoDoppler examination were performed preoperatively in all of the patients. Results Venous reflux in deep vein of lower extremities were at grade 0 in 62 limbs, at grade Ⅰ in 28 limbs and at grade Ⅱ in 16 limbs demonstrated by ascending venography with Valsalva test. StethoDoppler examination showed that valve insufficiency of great saphenous vein at saphenofemoral junction was in 99 limbs, the insufficiency of small saphenous vein at saphenopopliteal in 5 limbs and incompetent perforating vein in all of the limbs. Postoperative course was uneventful, the average hospitalization days after operation were 2.8 days and no recurrent varicose veins occurred in all the patients during the follow-up period lasting 1-30 months. Conclusions With the help of preoperative StehoDoppler examination, the mini-incision surgery for the treatment of varicose vein of lower extremities is safe, simple, effective and lower costs.

Objective To evaluate the effect of Ji Tang Zhi on glucose metabolism in insulin resistance (IR) HepG 2 cell line,and to explore the related mechanism.Methods The HepG2 cells were incubated in culture medium addition of 10-7 mol/L insulin for 24 h to establish the IR cell model.Effect of Ji Tang Zhi on rate of glucose absorption in HepG2 cell was detected by the method of glucose oxidase-peroxidase (GOD-POD).We performed an MTT assay to determine cytotoxicity effects of Ji Tang Zhi on HepG2 cell line.The expression of p-IRS-1 Ser307,PI3K and GLUT-4 were detected by Western blotting.Results Incubated with 10-7 mol/L insulin for 24 h,the insulin resistance cell model had been built.Compared with model group,the rate of glucose absorption of cell treated with JTZ (30 ～ 120 μg/mL) was significantly improved.According to model cells,the expression of GLUT-4 and PI3K decreased significantly compared to control cells.While the expression of p-IRS-1 Ser 307 was inhibited and GLUT-4 and PI3K expression were increased in IR cells after treated with JTZ (30 ～ 120 μtg/mL).Conclusion JTZ exert beneficial effects on hyperglycosemia in IR cell line possibly through regulating the levels of GLUT-4,p-IRS-1 Ser307 and PI3K in HepG2 cell.

Objective To determine the diversity of Porphytomonas gingivalis lipopolysaccharide (LPS)induced IL-1β,TNF-αand IL-6 levels in THP-1 cells and the associated Toll-like receptors(TLR).Methods P.gingivalis strain ATCC33277 LPS (Pg-LPS)was prepared using phenol-water method and then identified by both infrared spectrometry and limulus test.The levels of IL-1β,TNF-α and IL-6 secreted by THP-1 cells under inducement of Pg-LPS were quantitatively detected using commercial ELISA kits.The bloc-king test using TLR2 or TLR4 monoelonal antibody(McAb)plus the ELISA were used to determine the types of Pg-LPS binding TLR on the surface of target cells.In this study.a commercial LPS from E.coli strain O111:B4(E-LPS)was used as the contr01.Results When 1彬ml ofPg.u,s induced THP.1 ceHs for0.5,6 and 6 h.or l∥rnl ofE-LPs induced for 6,24 and 24 h,respectively,tIIe levels ofTNF.a,IL-1B and IL广6 were in.creased obviously(P<0.01).However,the maximal concentration of tlle t11ree cytokines induced by Pg.LP$ were similar to that induced by E-LPS(P>0.05).tLR2 or TLR4 McAb could block the effects of Pg-LPS in-ducing THP-1 cells to secrete IL-1β and IL-6(P<0.05),where as only TLR2 McAb displayed the inhibition of TNF-α secretion(P<0.05).On the contrast,only TLR4 McAb showed the effects blocking the three cytokines secretion in the THP-1 cells under inducement of E-LPS(P<0.05).Conclusion Pg-LPS shows a slight high-er activity inducing THP-1 cells to secrete IL-1β,TNF-α and IL-6 than E-LPS.TLR2,but not TLR4,is the major receptor of Pg-LPS on the target cells to mediate the secretion of the three cytokines.

: To assess the () recombinant gingivalis gingipain R2 (rRgpB)-induced Ca mobilization in human gingival fibroblast (HGF) mediated by protease-activated receptor (PAR) and its downstream signal transduction pathways. : Flow cytometry was used to detect the expression of PAR in HGF. The proliferation of HGF was measured by CCK-8. The dynamic changes of intracellular Ca concentration in HGF induced by rRgpB and the blocking effect of PAR-1 antagonist were observed by laser confocal microscopy. Western blot was performed to determine the phosphorylation levels of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) 1/2, p38 mitogen-activated protein kinase (p38 MAPK) and p65 in HGF. : PAR-1 and PAR-3 were expressed in HGF, and the rRgpB could promote the proliferation of HGF. rRgpB caused a transient increase in [Ca], which could be completely suppressed by vorapaxar, a PAR-1 antagonist. The phosphorylation levels of JNK, ERK1/2 and p65 were significantly up-regulated after the induction of rRgpB for and (all <0.05), which was completely inhibited by vorapaxar. However, the phosphorylation level of p38 MAPK had no significant change after rRgpB stimulation. : rRgpB causes an increase in [Ca] in HGF mediated by PAR-1. JNK, ERK1/2 and nuclear factor-κB may be involved in intracellular signal transduction after PAR-1 activation.
Fibroblasts ; Humans ; JNK Mitogen-Activated Protein Kinases/metabolism* ; MAP Kinase Signaling System ; Phosphorylation ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases/metabolism*

Objective To construct a 5-lipoxygenase (5-LO) transgenic mouse model of atherosclerosis.Methods Purified 5-LO fragment was injected into male pronucli and the firtilized eggs were transplanted into pseudopregnant mice.PCR and Southern blot were used to detect the genotype of DNA separated from the newborn mouse tail tissues.RT-PCR and Western blot analysis were used to detect the gene transcription and expression.Results PCR and Southern blot results showed that 7 of 25 mice were transgenic mice.Expression of 5-LO and FLAP was found in the bone marrow,spleen,kidney,and peritoneal cells.Results of RT-PCR and Western blot showed that No.9,20,24transgenic mice expressed a higher level of 5-LO and FLAP than those in the wild type C57BL/6 mice.The expression levels in bone marrow and peritoneal cells were significantly different(P<0.05).Conclusion A 5-LO transgenie mouse line has been established in this study and may be used for future study on the function of 5-LO gene.

Objective:To investigate the mechanism of bladder cancer treatment by using Scutellaria barbata and Codonopsis pilosula drug pair through network pharmacology. Methods:The drug composition of the drug pair was screened using TCMSP, and their action targets were predicted using Swiss Target Prediction. GeneCards was used to obtain disease targets of bladder cancer, and venny 2.1 was used to obtain intersection targets. PPI analysis was performed using STRING, and a network diagram was constructed using Cytoscape. GO and KEGG analysis were conducted using Metascape. A drug-target-pathway network map was constructed using Cytoscape software. Nude mice were randomly divided into a model group and a treatment group to establish a bladder cancer mouse model. On the 8th day after model formation, the mice in the model group were administered intragastrically with a dose of 342.86 mg/kg, 0.2 ml, twice/day. On the 28th day after modeling, the tumor size of nude mice was measured. Prostaglandin G/H Synthetase 2 (PTGS2), PTGS1, Nuclear Receptor Coactivator 2 (NCOA2), Retinoic Acid X Receptor α (RXRA), Progesterone Receptor (PGR), Mitogen-Activated Protein Kinase 1 (MAPK1), Reticuloendothelial Proliferation virus oncogene homology A (RELA), and Akt1 levels were detected by enzyme-linked immunosorbent assay.Results:The results show that 45 active components of the drug pair directly acted on 187 disease targets through multiple pathways to treat bladder cancer, in which Quercetin, luteolin, wogonin, 7-Methoxy-2-methyl isoflavone, baicalein, beta-sitosterol, Stigmastero, and other core ingredients, as well as PTGS2, PTGS1, NCOA2, RXRA, PGR, MAPK1, RELA, and Akt1 are critical targets. The results of gene function annotation analysis show that the biological processes most likely related to crossover genes mainly involved responses to hormones, cell responses to lipids, responses to foreign stimuli, and responses to bacterial molecules. The cell components mainly involves transcription regulatory complexes, membrane rafts, membrane microregions, and RNA polymerase Ⅱ transcriptional regulatory complexes, etc. The molecular functions mainly involve transcription factor binding, DNA-binding transcription factor binding, RNA polymerase Ⅱ specific DNA-binding transcription factor binding, nuclear receptor activity, ligand-activated transcription factor activity, etc. The results of pathway enrichment analysis suggests that the main signaling pathways are AGE-RAGE, IL-17, PI3K-Akt, TNF, MAPK, HIF-1, apoptosis, p53, toll-like receptor, etc. Animal experiments show that the Scutellaria barbata and Codonopsis pilosula drug pair can significantly improve tumor size and also improve the expression levels of PTGS2, PTGS1, NCOA2, RXRA, PGR, MAPK1, RELA, and Akt1. Conclusions:The Scutellaria barbata and Codonopsis pilosula drug pair can regulate PTGS2, PTGS1, NCOA2, RXRA, PGR, MAPK1, RELA, and Akt1 and other diseases mainly through the regulation of AGE-RAGE, IL-17, PI3K-Akt, TNF, MAPK, HIF-1, apoptosis, p53, toll-like receptor, and other signaling pathways. Targeting enzyme activity and cell apoptosis can treat bladder cancer by regulating these biological processes.

OBJECTIVE To evaluate the safety and clinical efficacy of cefoperazone/sulbactam in the treatment of biliary tract infections.METHODS In this prospective multicenter study,159 hospitalized patients with biliary tract infections received cefoperazone/sulbactam,and the clinical and bacteriological efficacy as well as the side effects were evaluated.RESULTS The clinical effective rate of cefoperazone/sulbactam in the treatment of biliary tract infections was 86.78%.After treatment,the body temperature reduced to normal rapidly,the average time of defervescence was 3.09?1.81 days.Pathogen eradication rate was 85.71%.No adverse reactions were reported during the study period.CONCLUSIONS Cefoperazone/sulbactam can be used as one of antibiotics of choice in the initial empirical therapy for biliary tract infections.

OBJECTIVETo investigate the expression types of protease-activated receptors(PAR) in human gingival fibroblasts(HGF) and the functions of PAR in periodontitis.

METHODSPrimary HGF were cultured.Reverse transcription PCR(RT-PCR) was used to detect the expression of PAR in HGF. Recombinant gingipain R (rRgp) was applied to HGF. The change of PAR expression on the cell surface was analyzed by real-time quantitative RT-PCR, and enzyme-linked immunosorbent assay (ELISA) was used to detect the change of the interleukin (IL)-6 production from HGF. The results of RT-PCR and ELISA were statistically analyzed using the two independent samples t-test of SPSS10.0 software.

RESULTSHGF expressed PAR-1 and PAR-3. The expression of PAR-1 and PAR-3 changed after two rRgp treatment with HGF cells. The relative expression of PAR-1 was decreased from 1.04 ± 0.31 to 0.67 ± 0.11 and 0.31 ± 0.11. The relative expression of PAR-3 was decreased from 1.01 ± 0.44 to 0.79 ± 0.13 and 0.44 ± 0.12(P < 0.05). The level of IL-6 was increased after rRgp treatment for 8 h. The control group was (18.77 ± 4.09) µg/L, the rRgp treatment groups were (179.36 ± 15.81) and (320.56 ± 26.19) µg/L respectively.

CONCLUSIONSHGF expressed PAR-1 and PAR-3 and were involved in periodontal inflammation.

Adhesins, Bacterial ; Cell Membrane ; Cysteine Endopeptidases ; Enzyme-Linked Immunosorbent Assay ; Fibroblasts ; Gingiva ; metabolism ; Humans ; Interleukin-6 ; Periodontitis ; metabolism ; Receptors, Proteinase-Activated ; biosynthesis