Objective To investigate the effects of p-cresol on human umbilical vein endothelial cells.Methods The effects of p-cresol on endothelial cell growth,cell cycle,cell morphological change and p21 protein were detected by the CCK-8 assay,flow cytometry assay,inverted microscope and Western blotting.Results P-cresol could inhibit the growth of human umbilical vein endothelial cells in dose-and time-dependent manners (all P ＜ 0.05).The human umbilical vein endothelial cells treated with p-cresol became elongated processes,cloudy cytoplasm,and irregular shapes.The p-cresol stopped human umbilical vein endothelial cells at cell cycle G1 and had no effect on cell apoptosis.The p-cresol could increase protein expression of p21 in a dose dependent manner (P ＜ 0.05).Conclusion P-cresol can increase protein expression of p21,induce cell cycle arrest at G1 stage and inhibit the proliferation of human umbilical vein endothelial cells.

Animals ; Antibiotics, Antineoplastic ; pharmacology ; Blotting, Northern ; Cells, Cultured ; Dexamethasone ; pharmacology ; Doxorubicin ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Gene Expression Regulation ; drug effects ; Glucocorticoids ; pharmacology ; Kidney Glomerulus ; cytology ; drug effects ; metabolism ; Mice ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Aged ; Alzheimer Disease ; drug therapy ; physiopathology ; Brain ; physiopathology ; Chitosan ; pharmacology ; therapeutic use ; Electroencephalography ; Female ; Humans ; Male ; Phospholipids ; pharmacology ; therapeutic use

Background Studies determined that rapamycin has not only the antibiosis but also suppressing the auto-immunology.The treating effect of rapamycin on experimental autoimmune uveoretinitis (EAU) is still concerned.Objective This work was to investigate the therapeutic effect of rapamycin on EAU and study the effect on the expression of inflammatory cytokines which were secreted by T lymphocyte subgroup in EAU.Methods EAU was induced in 20 SPF male Lewis rats by subcutaneous injection of interphotoreceptor retinoid binding protein (IRBP) R16 peptide emulsified in adjuvant.The rats were randomized into model control group and rapamycin injection group and 10 rats for each group.The rapamycin of 0.2 mg/( kg · d) 0.4 ml was intraperitoneally injected for the consecutive 7 days immediately after modeling,and the equal volume of normal saline solution was used at the same fashion in the model control group and 5 normal matched rats( normal control group).The ocular manifestation of the rats were examined under the slit lamp regularly.The retinal sections of the rats were prepared in the 14 days after modeling for the histopathological examination with hematoxylin and eosin staining.The ocular signs of inflammation and histopathological severity were scored based on the criteria of Caspi.Expression of interferon-γ (IFN-γ) and interleukin-17 (IL-17) in rat retinas were detected by immunohistochemistry.This experiment followed the Regulation for the Administration of Affair Concerning Experimental Animals by Tianjin Municipal Government.Results The scores of ocular signs elevated gradually from 6 through 12 days after modeling and slowly declined after that in the model control group.A same tendency was seen in the rapamycin injection group.The significant differences were seen in the scores of ocular signs between the two groups from 6 to 14 days( P＜0.01 ).The disorder of retinal structure and infiltration of inflammatory cells were seen in the model control group,but the retina layers were normal in the rapamycin injection group.The pathological score was evidently declined in the rapamycin injection group ( 0.90 ± 0.45 ) in comparison with model control group( 3.30±0.48 ) ( t =16.541,P＜0.01 ).The expressions of the IFN-γ and IL-17 in retina located in the outer nuclear layer,inner nuclear layer,internal plexiform layer and retinal ganglion cell layer with the weakened levels(A values) in rapamycin injection group compared with model control group,showing a considerably difference between them ( IFN-γ:21.16±4.23 vs 62.14 ±7.32; IL-17:49.86±6.59 vs 124.85 ±6.33 )(q=33.334,q=56.923,P＜0.01 ).Conclusions Rapamycin down-regulates the expressions of IFN-γ and IL-17 in retina and further eliminates the inflammatory response in the rat with EAU by the suppression of Th1 and Th17 cells function in EAU.

BackgroundStudies determined that Th17 cells are important inflammatory cell group in experimental autoimmune uveoretinitis ( EAU ).Interleukin-17 ( IL-17 ),as a marker of Th17,is involved in the occurrence and development of EAU.Mesenchymal stem cells (MSCs) play an immunomodulatory role,mainly by inhibiting the expression of Th17 in a variety of self-autoimmune disease.This is one of the current research focuses.ObjectiveThe present study was to investigate the therapeutic effect of MSCs in EAU and their impact on IL-17 expression in the retina.MethodsMSCs were isolated from the bone marrow of the femurs from 10 SPF Wistar rats and cultured and passaged.The third to fifth generations of cells were used in this experiment.EAU models were induced in 48 6-8 week-old SPF Lewis rats by subcutaneous injection of interphotoreceptor retinoid binding protein (IRBP) R16 peptide emulsified in adjuvant.EAU rats were randomly assigned to the model control group and the MSCs group.MSCs suspension (5×106) of 1 ml was injected via the rat tail vein once a day for 3 consecutive days after immunization,and the same amount of PBS was injected in the model control group in the same manner.Six matched normal Lewis rats were used as the normal control group.The inflammatory response was clinically examined under the slit lamp biomicroscope daily,and the histopathological changes of the retina were examined by hematoxylin and eosin staining on days 9,12,15 and 20.The clinical and histopathological scoring was performed according to the Caspi criteria.Expression of the IL-17 protein in the retina was detected by immunohistochemistry on 9,12,15 and 20days following molding.The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by Tianjin Municipality Science and Technology Commission.Results MSCs showed the fusiform in shape and vortex-like growth.Flow cytometry verified that presented the positive expression for CD29 and CD44 but absent expression for CD45 and CD34.The scores of the anterior segment were significantly lower ( U=2.815,P =0.005 ; U =2.768,P =0.006 ; U =2.900,P =0.004 ; U =2.855,P =0.004 ),and the retinal inflammation scores were lower in the MSCs group than the model control group at various time points ( U =2.345,P =0.019 ; U =2.559,P =0.011 ; U =2.166,P =0.030 ; U =2.373,P =0.018 ).Im mnunochemistry showed that the expressions levels of the IL-17 protein (A value) in the rat retina were 26.47±5.68,77.78± 9.65,47.02±6.68 and 26.59±5.94 in the MSCs group on days 9,12,15 and 20,and those in the control group were 45.34±4.63,105.95± 10.74,64.11 ±9.76 and 37.02±6.51,showing a significant reduction ( t =6.305,P =0.000 ; t =4.799,P =0.001 ; t=3.540,P=0.005;t=2.900,P=0.016). ConclusionsMSCs can inhibit the aggravation of EAU and suppress the expression of IL-17 in ocular tissue.

ObjectiveTo study the relevant effect of proinflammatory cytokines interelenkin-17(IL-17), -6 and endothelin-1 (ET-1) on statins attenuating no-reflow phenomenon after myocardial ischemia-reperfusion in rats.MethodsEighteen healthy male Wistar rats were randomly divided into 3 groups according to body weight: sham operation, injury, preconditioning groups. The preconditioning group was given atorvastatin 2 mg·kg-1 ·d-1 and the other two groups were given the same volume of saline once. After 7 days, the rats were anesthetized with an intraperitoneal injection of chloral hydrate, and then the thoracic cavity was opened. The coronary artery of injury group and preconditioning group were ligated for 60 minutes, and then opened for 15 minutes, to establish the rat acute myocardial ischemia-reperfusion model. The sham operation group was was treated with a seam through the coronary artery without ligation. Eleetrocardiogram was checked before ligation, and ligation was carried out for 15, 30, 45 minutes and then reperfusion for 15 minutes. After reperfusion for 15 minutes, the thioflavine S and Even's were injected from femoral venous, then the heart and blood were obtained(keeping left ventricular only). Hearts were flushed with saline and sliced transversely into five to seven sections. Finally, observed at 365 nm wave length the existence of non-fluorescent areas, which was no-reflow zone. The level of serum IL-17, IL-6 and ET-1 was detected by ELISA. Results The electrocardiogram confirmed that the sham operation group had no ischemic damage and the model of myocardial ischemia- reperfusion was established in preconditioning group and injury group. The noreflow phenomenon could be observed under 365 nm wave length in preconditioning group and injury group. The ligated area[LA％, (57.34 ± 11.49)％, (53.08 ± 8.66)％] of injury group and preconditioning group was higher than that of sham operation group(0, all P ＜ 0.05); the area of no-reflow[ANF％, (48.96 ± 6.94)％, (21.37 ±3.35)％] of injury group and preconditioning group was higher than that of sham operation group(0, all P ＜ 0.05),and the ANF％ of preconditioning group was lower than that of injury group(P ＜ 0.05) ; the level of serum IL-17,IL-6 and ET-1[(151.67 ± 11.19) × 10-9, (167.89 ± 5.13) × 10-9, (322.37 ± 19.08) × 10-9 g/L] of injury group was higher than those of sham group and preconditioning operation group[(49.75 ± 14.06) × 10-9, (59.32 ± 5.26) ×10-9, (109.9 ± 12.12) × 10-9, (90.45 ± 11.63) × 10-9, (112.47 ± 10.40) × 10-9 and(198.91 ± 27.88) × 10-9 g/L,P ＜ 0.05], the level of serum IL-17, IL-6 and ET-1 of preconditioning group was higher than those of sham operation group(P＜ 0.05). Conclusionsno-reflow phenomenon is related with IL-17 and ET-1 which can promote the expression of IL-6, statins decreases the expression of IL-17 and ET-1, and then decreases the on-reflow phenomenon.

Objective To find methods to isolate and purify plasminogen activator (Pla) from artificial culture of Yersinia pestis. Methods Ultrasonication and urea extracting combined by ammonium sulfate salting-out were tried to extract Pla. High performance liquid chromatography(HPLC) was used to purify Pla. The first step was ion exchange and the second was gel filtration, Preparative electrophoresis was used to purify Pla, too. The enzyme activity of the isolated or purificated Pla was detected. Results Both 50% - 60% saturated ammonium sulfate deposition of supernatant of plague bacilli ultrasonication and 0 - 10% saturated ammonium sulfate deposition of supernatant of plague bacilli powder soaked by urea had three bands(Mr about 31×103, 35×103 and 37×103) and lysis rings were 6.5 and 7.2 mm in diameter respectively when the enzyme activity was detected. Pla purified by HPLC was mainly composed of three bands(Mr about 31×103, 35×103 and 37×103), occupying more than 80% of total protein weight and lysis ring was 5.0 mm in diameter. Pla purified by preparative electrophoresis mainly consisted of three bands(Mr about 31×103, 35×103 and 37×103) with other proteins of low concentration nearby, no lysis ring was detected. Conclusions Pla is collected by the methods of ultrasonication and urea extracting. Priliminary purification of Pla can be achieved by HPLC and preparative electrophoresis.

Objective To investigate the correlation between the expression of Ku70 and Ku80 in esophageal squamous cell carcinoma and chemotherapy sensitivity and prognosis.Methods The expression levels of Ku70 and Ku80 in esophageal squamous cell carcinoma were measured by immunohistochemistry.The relationship between the expression of Ku70 or Ku80 and clinical pathology parameters,recent curative effect of concurrent chemoradiation and prognosis of the patients were analyzed.Results The positive expression rate of Ku70 and Ku80 was 59.0％ (69/117) and 50.4％ (59/117) in esophageal squamous cell carcinoma.The expression of Ku70 and Ku80 was significandy related to recent curative effect of esophageal squamous cell carcinomas (P =0.017,0.012).The patients with complete response had lower positive expression rate of Ku70 and Ku80 [49.2％(32/65),40.0％(26/65)].But the expression had no relationship with the age,length of lesions in CT,lymph node metastasis of esophageal squamous cell carcinoma (P ＞0.05).There was no relationship between the expression of Ku70 and Ku80 neither (P =0.114).KaplanMeier analysis found Ku70 and Ku80 had worse prognosis in patients with positive expression (Ku70:P =0.004 ; Ku80:P =0.025).In Cox univariate analysis concluded the same result (Ku70:P =0.005,HR =1.971,95％ CI:1.230-3.159; Ku80:P =0.028,HR =1.659,95％ CI:1.057-2.605); but in Cox multivariate analysis,only Ku70 expression was the independent factor (P =0.033).Conclusions The upregulation of Ku70 and Ku80 is correlated to decreased concurrent chemoradiation sensitivity.In addition,the upregulation of Ku70 is correlated to worse prognosis.Ku70 and Ku80 can be a prediction index for predicting concurrent chemoradiation sensitivity in esophageal squamous cell carcinoma.

OBJECTIVETo observe the effect of Ningdong Granule (NG) on serum levels of interleukin-12 (IL-12) and tumor necrosis factor-alpha (TNF-alpha) of children patients with Tourette's syndrome (TS).

METHODSTotally 90 TS children patients were randomly assigned to the NG group, the NG + Tiapride group (abbreviated as the combined treatment group), and the Tiapride group, 30 in each group. Besides,another 30 healthy children were recruited as the healthy control group. Patients in the NG group were treated with NG (consisting of all gastrodia rhizome, Codonopsis pilosula, Ophiopogon japonicus, white peony root, Rhinocerotidae, oyster, earthworm, licorice root, etc.), one dose daily, administered by dissolving it in boiled water, taken in two portions in the morning and in the evening respectively. Patients in the Tiapride group took Tiapride Tablet, 50 -100 mg each time, twice daily. The dosage was adjusted according to individual difference and changes of pathogenic conditions. The maximal dosage was 300 mg per day. Those in the combined treatment group were treated with equal dose of NG and Tiapride Tablet in combination. The treatment course was 3 months for all. Changes of pathogenic condition before and after treatment were assessed by Yale global tic severity scale (YGTSS). Serum levels of IL-12 and TNF-alpha were detected by enzyme-labeled immunosorbent assay (ELISA) before and after treatment.

RESULTS(1) The total effective rate of the NG group, the combined treatment group, and the Tiapride group was 79.3%, 83.3%, and 67.9%, respectively. It was the lowest in the Tiapride group (P < 0.05). It was significantly higher in the combined treatment group than in the NG group (P < 0.05). (2) The post-treatment YGTSS score was obviously lower in each group after treatment than before treatment (P < 0.05). The posttreatment YGTSS score was obviously lower in the NG group and the combined treatment group than in the Tiapride group (P < 0.05), but with no statistical difference between the fromer two groups (P > 0.05).(3) Compared with the healthy control group before treatment, serum levels of IL-12 and TNF-alpha (pg/mL) were 124.95 +/- 22.78 and 209.52 +/- 21.69 in the NG group, 126.14 +/- 25.65 and 208.97 +/- 22.46 in the combined treatment group, 123.00 +/- 24.26 and 205.10 +/- 26.16 in the Tiapride group, being higher than those in the healthy control group (64.56 +/- 27.59 and 78.13 +/- 33.42; P < 0.05). After treatment, serum levels of of IL-12 and TNF-alpha were 104.67 +/- 16.84 and 183.01 +/- 24.95 in the NG group, 109.04 +/- 16.81 and 179.87 +/- 23.45 in the combined treatment group, significantly lower than before treatment (P < 0.05). But there was no statistical difference in serum levels of IL-12 or TNF-alpha in the Tiapride group between before treatment (123.00 +/- 24.26 and 205.10 +/- 26.16) and after treatment (117.75 +/- 16.79 and 199.76 +/- 33.21; P > 0.05).

CONCLUSIONNG could modulate abnormal serum levels of IL-12 and TNF-alpha in TS children patients, which might be one of its pharmacodynamic mechanisms for treating TS.

Adolescent ; Child ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interleukin-12 ; blood ; Male ; Phytotherapy ; Tourette Syndrome ; blood ; drug therapy ; Tumor Necrosis Factor-alpha ; blood

Comparative analysis of characteristic of species and esterase-isoenzyme of isolates of Polyporus umbel-latus from different regions were processed. The results indicated that isolates of Jizhaoling ( Z) and Zhushiling (ZJ) have significant differences in characteristic, and enzymatic band types of the two species also have significant differences. The homology at genetics between the two isolates is 0% , and consanguinity between the two i-solates is the farthest.