AIM: To observe the expression of transforming growth factor ? 1 (TGF-? 1), MAPK 1/3 and fibronectin (FN) in the development of renal tubulointerstitial disease. METHODS: Wistar male rats were randomly divided into normal control group, diabetic group of 1week, 2 weeks, 4 weeks and 8 weeks. Diabetic model was induced by peritoneal injection of streptozotocin. Immunohistochemistry was employed to detect the expression of TGF-? 1, MAPK 1/3 and FN in the kidney. TGF-? 1 protein in the renal cortex was checked by Western blot. BG, Scr and UP were analysed by biochemical methods, and the morphological changes in renal tubulointerstitium were also examined under microscopy on sections stained with HE and PAS. RESULTS: The expression of MAPK 1/3 and FN was observed, but not the expression of TGF-? 1 in normal renal tissue. Positive staining of TGF-? 1 was observed in the renal tubulo-interstitium in 1-week diabetic group and thereafter it increased in the course of diabetes. A continuous increase in the expression of MAPK 1/3 and FN was also observed in two - week diabetic rats. Chronologically the expression of TGF-? 1,MAPK 1/3 and FN and the ratio of KW/BW were positively correlative with each other in diabetic animals except one -week diabetic rats. There was also a positive correlation between MAPK 1/3 and FN in l -week diabetic rats. CONCLUSION: Our data suggest that TGF-? 1 appears in the renal tubulointerstitium in early period of diabetes and then its signal is mediated by MAPK 1/3 cascades to accelerate production of FN ,and in turn leads to renal hypertrophy and tubulointerstitial fibrosis. [

Objective To explore the effect of the simvastatin administration on the expression of connective tissue growth factor （CTGF） in fihrotic lungs of rats. Methods The cats were treated with single intratracheal instillation of bleomycin （BLM） or instillation of the same volume of normal saline （NS） as a control. The administration of simvastatin（20 mg/kg）began once a day immediately or 7 days later after intratracheal BLM instillation respectively with the same volume of NS was given as a vehicle control. The rats were killed on day 7,14 and 28 respectively. Pathological alteration of lung tissue was observed hy HE staining and Masson staining. Hydroxyproline（HYP）content in lung tissue was used to determine the severity of pulmonary fibrosis. The expression of CTGF in lung tissue was exanfined by immunohistochem- istry staining and photodeusitometry. Results Histopathological changes of pulmonary fibrosis emerged gradually after the instillation of BLM. The expression level of CTGF was increased in lungs of rats after intratracheal instillation of BLM, compared with the control. The administration of simvastatin immediately or 7 days after intratracheal instillation of BLM, attenuated the histopathological changes of bleomycin- induced puhnonary fibrosis and prevented the increased expression of CTGF in lung tissue on day 28. Conclusion The adntinistration of simvastatin, immediately or 7 days after intratracheal BLM instillation, prevented the up-regulation of CTGF in fibrotic lungs of rats, which ntight be one of the mechanisms of the anti-fihrosis of simvastatin in lungs.

Reuse of waste oil in cooking has been the social concern for long time, but until now there is no reliable method to identify the gutter oil.In this work, based on the generation of long-chain aliphatic aldehydes in the refinement of gutter oil, gutter oil and adulterate cooking oil were identified by determining hexanal, heptanal, octanal, nonanal and decanal in oil sample with high perfomance liquid chromatography (HPLC) method using O-(3-(9H-carbazol-9-yl)propyl)hydroxylamine as the fluorescent derivative reagent.At first, 10 μL of oil sample was dissolved in 200 μL of isopropanol, then reacted with O-(3-(9H-carbazol-9-yl)propyl)hydroxylamine to form the stable derivatives, extracted with acetonitrile and injected into HPLC for analysis.The derivatives were separated on a C18 column within 15 min with ACN-H2O (90∶10, V/V) as mobile phase.The fluorescence detector was set at λex/λem=292 nm/348 nm.The linear range of 5 kinds of long-chain aliphatic aldehydes was 0.01-1.00 μmol/L with limit of detection (LOD, S/N=3) of 0.05-0.10 nmol/L, and the spiked recoveries were 95.6%-101.4%.The results showed that the edible oils obtained from supermarket almost had no hexanal, heptanal, octanal, nonanal, decanal, but these aldehydes increased significantly in gutter oil.Thus the 5 kinds of long-chain aldehydes could be used as the indictor of gutter oil.Taking the advantage of good specificity, higher sensitivity, good accuracy and simplicity, the method was suitable for the rapid identification of gutter oil.

Objective To investigate the effects of dibutylphthalate (DBP) on the proliferation and apoptosis of HL-60 leukemic cells and to study its mechanism to purge leukemic cells.Methods The effects of DBP on proliferation of HL-60 leukemic cells were measured by cell culture method. The effects on apoptosis were measured by the percentage of the apoptotic cells in morphology and of the DNA fragmentation and by DNA gel electrophoresis. The intracellular free Ca~(2+) concentration ([Ca~(2+)]i) of leukemic cells were measured by Fura-2AM method. The protein expressions of c-myc and bcl-2 genes of leukemic cells were measured by immunohistochemical assay.Results DBP could suppress the proliferation of HL-60 leukemic cells in a dose-dependent and time-dependent manner and induce them to die via apoptosis.It could elicit a intracellular Ca~(2+) redistribution and a potent extracellular calcium influx. It also down-regulated the protein expressions of c-myc and bcl-2 genes of HL-60 leukemic cells.Conclusion DBP can purge (HL-60) leukemic cells in vitro by increasing [Ca~(2+)]i of cells to initiate apoptosis and down-regulating bcl-2 and c-myc proto-gene expressions to promote cell apoptosis.

To observe the effects of total flavones of Metasequoia glyptostroboides Hu et Cheng (TFM) on volume-overload cardiac hypertrophy and the expression of c-Fos protein in rat. Methods　Volume-overload cardiac hypertrophy of rat was induced by aortocaval shunts. The rats were given ig TFM (400, 40 and 4 mg/kg/d). c-Fos protein in the ventricles were measured by immunocytochemical study. Results　TFM at the above dosage decreased heart weight and contents of RNA and protein in the myocardium, inhibited the expression of c-Fos protein in the ventricles. Conclusion　TFM can prevent volume-overload cardiac hypertrophy in rats. The inhibitory effects on the expression of c-Fos protein may be its mechanism in the molecular level.

OBJECTIVETo explore the mechanism of Astragalus polysaccharides (APS) for improving the cardiac function of Sjogren's syndrome (SS) model rats based on Keapl-Nrf2/ARE signaling pathway.

METHODSTotally 48 male Wistar rats were randomly divided into four groups by random digit table, i.e., the blank control group,the model control group,the APS group, and the hydroxychloroquine group, 12 in each group. Except those in the blank control group, 0. 1 mL mixed antigen protein of sufficiently emulsified Freund's complete adjuvant and submandibular gland protein was injected from two feet plantar to induce SS model. The intervention was started from 19th day after inflammation induction. Equal volume of normal saline was given to rats in the blank control group (1 mL/100 g), APS was administered to those in the APS group (1 mg/100 g), and hydroxychloroquine (0.03 125 g/kg) was administered to those in the hydroxychloroquine group. All rats were intervened once per day for 30 consecutive days. Changes of rats' body mass and drinking water quantity, submandibular gland index, spleen index, histological changes of glands were observed. Changes of the heart function were monitored using invasive hemodynamics. Serum reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), total antioxidant capacity (TAC), tumor necrosis factor alpha (TNF-alpha), and interleukin-35 (IL-35)were detected using ELISA method. The pathological changes were observed using HE staining. The protein expression of ROS, reactive nitrogen species (RNS), glutathione (GSH), and thioredoxin (TRX) were observed by immunohistochemical staining. The mRNA expression of Keap1, Nrf2, and ARE was detected using real time fluorescent quantitative PCR. The protein expression levels of gamma-glutamic acid and a half long glycine synthetase (gamma-GCS) and heme oxygenase 1 (HO-1) in the myocardial tissue were determined by Western blot method. Results Compared with the blank control group, the quantity of drinking water, submandibular gland index, spleen index, heart rate (HR), cardiac index (HI), left ventricular systolic pressure (LVSP), left ventricular diastolic pressure (LVEDP), MDA, ROS, TNF-alpha, ROS protein expression, RNS protein expression, Keap 1 mRNA expression, Maf mRNA expression, Nfr2 mRNA expression, and HO-1 protein expression, and gamma-GCS protein expression significantly increased (P <0.01); body mass, +/-dp/dtmax, SOD, TAC, IL-35, GSH, and TRX significantly decreased (P <0.01) in the model group. Compared with the model group, the quantity of drinking water, submandibular gland index, spleen index, LVEDP, MDA, ROS, TNF-alpha, ROS protein expression, RNS protein expression, Keap1 mRNA expression, Maf mRNA expression, Nfr2 mRNA expression, and HO-1 protein expression, and gamma-GCS protein expression significantly decreased (P<0.05); body mass, +/-dp/dtmax, SOD, TAC, IL-35, GSH protein expression, and TRX protein expression significantly increased (P < 0.05, P <0.01) in the AR group and the hydroxychloroquine group. In the hydroxychloroquine group HR increased (P <0.05). In the AR group HR and LVSP decreased (P <0. 05, P <0. 01). Compared with the hydroxychloroquine group, HR, LVEDP, - dp/dtmax, y-GCS protein expression significantly decreased (P <0. 05, P <0. 01); SOD, TAC, GSH, TRX, HO-1 protein expression increased (P <0.01 )in the AR group. HI was positively correlated with ROS (P <0. 05). LVSP and LVEDP were positively correlated with Keap1 -Nrf2/ARE signaling pathways (P <0. 01) , and negatively correlated with TAC (P <0. 05, P <0. 01 ). +/-dp/dtmax was negatively correlated with Keap1-Nrf2/ARE signaling pathways(P <0.05), and positively correlated with TNF alpha (P <0. 05).

CONCLUSIONSDeclined heart function exists in SS rats. The mechamechanism of APS for improving the heart function might be closely correlated with activating Keap1-Nrf2/ARE signaling pathway.

Animals ; Astragalus Plant ; Blotting, Western ; Carboxylic Ester Hydrolases ; metabolism ; Heme Oxygenase-1 ; metabolism ; Hydroxychloroquine ; Male ; Malondialdehyde ; metabolism ; Myocardium ; enzymology ; NF-E2-Related Factor 2 ; metabolism ; Plant Extracts ; therapeutic use ; Polysaccharides ; metabolism ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Signal Transduction ; Sjogren's Syndrome ; Submandibular Gland ; Superoxide Dismutase ; metabolism ; Thioredoxins ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

A 23-year-old man in remission from acute myeloblastic leukemia after allogeneic peripheral blood stem cell transplantation developed peripheral neuropathy presenting as sciatic and peroneal nerve deficits. Electrophysiological tests localized the lesions to the left sciatic and common peroneal nerve. Magnetic resonance imaging revealed nerve thickening and enhancement, while a positron emission tomography-computed tomography scan demonstrated increased fluorodeoxyglucose uptake tracking along the nerve, suggesting peripheral nerve infiltration. This report demonstrates an unusual presentation of acute leukemia relapse presenting as focal neuropathy

Objective To explore the effective method of treating childhood acute aplastic anemia(AAA) by overviewing the curative effect of therapeutic alliance to the children diagnosed as AAA with immunodepressants.Methods Retrospective analysis was performed on 13 patients diagnosed as AAA,who were treated by high dose of methyllprednisolone,antilymphocyte globulin,ciclosporin A,large dose of gamma globulin,granulocyte colony-stimulating factor and blood transfusion in 4 years.Results Among 13 cases,9 cases were healing well on the whole,one was relieved,one was improving,two were ineffective(one of them was dead).In 9 cases who were healing,WBC took(24.60?8.86) days to reach 4?10~9/L,hematoglobin took(152.22?68.88) days to reach normal and platelet took(125.55?55.86) days to reach 80?10~9/L.During the procedure of treating,infection appeared in 7 cases,the ALT and AST was high in 2 cases,hypoproteinemia and edema in 1 case,gross hematuria in 2 cases,gingival hyperplasia obviously and dedentition in 1 case.Conclusion Therapeutic alliance with immunodepressants to childhood AAA is a safe and effective method.

Obstructive sleep apnea-hypopnea syndrome(OSAHS) is a common sleep-related breathing disorder,which has a series of impact on the cardiovascular system.The dctection of some biochemical indicators plays an important role in predicting this kind of cardiovascular damage.The role of inflammatory predicting factors such as TNF-?,IL-6,CRP,IL-10,MMPs and ICAM-1 is reviewed in this paper.

To investigate the biological role of snake venom cystatin(sv-cystatin) in tumor progression, the cDNA of sv-cystatin amplified by PCR from pUC18-cystatin plasmid was cloned into methanol-inducible expression vector pPICZ?A. The linearized recombinant plasmid pPICZ?A-cystatin was transfered into Pichia pastoris, strain GS115 by electrophoration. Transfermants with phenotype Mut+ selected were identified by PCR analysis and induced in 1.0% methanol. The reombinant sv-cystatin protein was examined by SDS-PAGE, Western blot analysis. The molecular mass of expression product was about 14 kDa and approximately 16 mg/L of recombinant sv-cystatin was produced from one of GS115-cystatin transformants. The chromatography purified protein could reduce the activity of papain. The ability of B16F1 cells treated with recombinant sv-cystatin to invade the reconstituted basement membrane decreased significantly (P