To investigate the effect of losartan on the axis of prolylcarboxypeptidase (PRCP)--kallikrein of the two-kidney, one-clipped (2K1C) hypertensives rats, and explore the novel protection mechanism of losartan on the kidney. Sprague-Dawley (SD) rats were used to develop the 2K1C hypertensive rats. Then, the rats were treated with prazosin (5 mg x kg(-1) x d(-1)) or losartan (5, 15 and 45 mg x kg(-1) x d(-1)) or vehicle, separately. At the same time, the blood pressures were observed. After treated for four weeks, the ratio of right kidney weight and body weight, the change of glomerular morphology, and K+, Na+, creatinine and blood urea nitrogen (BUN) of the serum were used for evaluation of kidney. The expressions of PRCP mRNA in the kidneys were determined by RT-PCR. The protein levels of PRCP, tissue kallikrein, plasma kallikrein, TGF-beta1 in kidney or plasma were measured by Western blotting. Results showed that the changes of body weight and kidney weight ratio, glomerular fibrosis degree and the biochemistrical index of serum induced by hypertension were relieved when the hypertensive rats treated with losartan for four weeks. Meanwhile, treatment of losartan also significantly decreased expression of TGF-beta1 and increased expressions of PRCP, plasma kallikrein and tissue kallikrein. The protective effects of losartan on the kidney of 2K1C hypertensive rats are activation of the axis of PRCP-kallikrein and reducing the expression of TGF-beta1.

AIM: To elucidate the effects of losartan on the expression ofmatrix metalloproteinases-2, JNK1/2 and proliferation in cardiac fibroblast. METHODS: Neonatal rat cardiac fibroblasts were cultured. The cells proliferation was determined by MTT. To determine effects of AngⅡ on JNK1/2 activity, cells were incubated (for 0, 2, 5, 10, 30, 60, 120 min) in serum-freemedia with AngⅡ, and the other group fibroblasts were exposed to serum-free media with or without AngⅡ and losartan (AngⅡ 100 nmol/L, AngⅡ 100 nmol/L+losartan 100 nmol/L, losartan100 nmol/L, losartan for 45 min before). Cells protein was collected with MBST buffer. The relative abundance of MMP-2, JNK1/2 and p-JNK1/2 in cells was determined by immunoblotting. The secretion of MMP-2 in media of cell culture was determined by ELISA. RESULTS: AngⅡ increased the proliferation of CFB in a dose-dependent manner, whereas losartan decreased the proliferation of CFB stimulated by AngⅡ in a dose-dependant manner, too (P＜0.05). The relative abundance of JNK1/2 was highest in AngⅡ of the 2-min-stimulated group. AngⅡincreased expression of JNK1/2 and MMP-2 protein (P＜0.05), on the contrary, losartan inhibited JNK1/2 and MMP-2 protein expression.CONCLUSION: AngⅡ induce the increase of proliferation of CFB, expression of JNK1/2 and MMP-2 in CFB, and losartan inhibits these effects of AngⅡ.

Aim To compare the effects of the non-peptide angiotensin Ⅱ receptor type Ⅰ antagonist,Losartan,and the active vascular peptide,calcitonin gene-related peptide(CGRP),on the proliferation of vascular smooth muscle cells induced by angiotensin Ⅱ,and to explore the mechanism of depressor effect of Losartan and CGRP in vivo.Methods MTT,Thymidine incorporation and flow cytometry,were used to determine the ability of proliferation of VSMC induced by angiotensin Ⅱ in the presence or absence of Losartan or CGRP,Western blotting was used to determine the activity of ERK1/2.Results Losartan or CGRP inhibited the viability,DNA synthesis,cell proliferation index,and the activity of ERK1/2 in a dose-dependent manner.Conclusion Losartan or CGRP significantly inhibits the proliferation of VSMC induced by angiotensin Ⅱ;the inhibitory effect of CGRP is stronger than that of Losartan.The signaling path way is involved in ERK1/2.

Objective To investigate the effect of angiotensin Ⅱ-1 receptor antagonist losartan on expression of tumor necrosis factor-alpha (TNF-?) in the ventricular myocardium in the renovascular hypertension rats. Methods Renovascular hypertension model was obtained by clip left renal artery in Sprague-Dawley(SD) rats. After operation the rats were divided into 3 groups: sham group, two-kidney one clip (2K1C) group, and losartan treatment group(2K1C and losartan 20 mg/kg?d by drinking). Tail blood pressure was determined every week. Animals were euthanized after treatment with losartan for four weeks. Cardiac index(CI)was calculated by HW/BW, and TNF-? protein of ventricle myocardium was determined by ELISA and immunohistochemistry. Results Losartan significantly decreased blood pressure(P

This study is to investigate the impairment and possible mechanism of endothelium-dependent relaxation of mice mesenteric arteries induced by mmLDL. Wire myography was employed to examine endothelial function of mesenteric arteries. Ultramicrostructure of mesenteric vascular beds were detected by transmission electron microscope. The results showed that endothelium cell edema and peeling, vascular elastic membrane fracture traces in mmLDL group. Endothelium-dependent relaxation was decreased in a time-dependent and dose-dependent manner by using mmLDL, compared with normal arteries. In endothelium-derived hyperpolarizing factor (EDHF)-mediated relaxation, the Rmax and pIC50 were decreased from (63 +/- 5) % and 6.42 +/- 0.09 of normal saline control to (31 +/- 3) % and 5.67 +/- 0.07 in mmLDL group (P < 0.001, P < 0.001), respectively. In nitric oxide (NO)-mediated relaxation, the Rmax and pIC50 were decreased from (45 +/- 4) % and 5.93 +/- 0.08 in normal saline control to (32 +/- 4) % and 5.43 +/- 0.11 in mmLDL group (P < 0.05, P < 0.01), respectively. There is no significant alteration of prostacyclin I2 (PGI2) pathway between these two groups. In conclusion, mmLDL induced the impairment of the ultramicrostructure of mesenteric vascular endothelium cell as well as the endothelium-dependent relaxation. The latter includes the dysfunction of NO- and EDHF pathway mediated endothelium-dependent relaxation.

Objective To evaluate the clinical application of May-Grunwald-Giemsa staining followed by fluorescence in situ hybridization (MGG-FISH) technique in the differentiation diagnosis of Ph-chromosome positive acute lymphoid leukemia (Ph + ALL) from chronic myeloid leukemia in lymphoid blast crisis(CML-LBC). Methods The bone marrow smears of 4 patients with Ph+ ALL, 4 patients with CML-LBC, 1 patient with CML in myelocytic blast crisis complicated with lymphoma and 1 patient with CML in mixed blast crisis were assayed with the MGG-FISH technique in which the spectrum green labeled BCR and spectrum orange labeled ABL dual color dual fusion probes were used. Based on the morphological classification, the percentages of BCR-ABL positive cells were subsequently determined respectively in the erythroid, myeloid and lymphoid hneages for the 10 specimens. Results According to the MGG-FISH analysis, the erythroid lineage was not involved in the 4 Ph+ ALL specimens without BCR/ABL positive cells. While the BCR/ABL positive percentage of myeloid cells was 11% (1/9), 8% (1/12), 0% (0/8) and 10% (1/10) respectively and that of lymphoid cells was 97% (76/78), 98% (87/89), 98% (97/99) and 97% (75/77) respectively. On the other hand, the BCR/ABL positive percentage was 100% (8/8), 91% (10/11), 82% (9/11), 88% (7/8) in the erythroid lineage, 89% (8/9), 96% (94/98), 100% (47/47), 98% (40/41)in the myeloid lineage and 96% (78/81), 93% (52/56), 96% (68/71), 95% (58/61) in the lymphoid lineage respectively for the 4 CML-LBC specimens. The BCR/ABL positive percentages of the other 2 specimens were all above 80% and through MGG-FISH analysis we also identified the source of the malignant clones and ascertained the diagnosis of the 2 patients. Conclusions The MGG-FISH technique has proved useful in providing rapid and precise differentiation between Ph + ALL and CML-LBC. The source of the malignant clones can also be analyzed by this technique.

Aim To study the effect of Macrophage colony-stimulating factor(M-CSF) on MMP-9 in RAW 264.7 cell and explore the relationship between atherosclerosis caused by M-CSF and the activity of MMP-9. Methods Gelatin zymography analysis was used to investigate the effect of M-CSF and PD98059 on the activity of MMP-9 in cultured RAW 264.7 cell.Western blot was used to study the effect of M-CSF and PD98059 on the express of p-ERK1/2 in cultured RAW 264.7 cell. Results The enzyme activity of MMP-9 was significantly increased after 24-hour M-CSF treatment.Meanwhile, M-CSF upregulated the expression of p-ERK1/2. Pre-treatment with PD98059 blocked partly the increased expression of p-ERK1/2 and the activity of MMP-9 induced by M-CSF. Conclusion M-CSF can induce the secretion of MMP9 in RAW 264.7 cell, which may be mediated by the phosphorylation of ERK1/2.

BACKGROUND: Research has been reported that serum soluble CD146 (sCD146) expression was improved on the surface of endothelial cells and activated T cells by the stimulation of inflammatory factor. Therefore, it predicts that CD146 may participate in inflammatory reaction of tissue.OBJECTIVE: To investigate the expression and clinical significance of serum sCD146 in peripheral blood from patients with ankylosing spondylitis.METHODS: A total of 62 patients with ankylosing spondylitis were selected from the Sixth People's Hospital AffiUated to Shanghai Jiao Tong University. All patients were divided into two groups: active group (n=46) and inactive group (n=16); while, 20 healthy subjects were selected as the control group. Indicators including Bath Ankylosing SpondyUtis Disease ActivityIndex (BASDAI),Bath Ankylosing Spondylitis Functional Index (BASFI), patient's global assessment (PGA), night pain, visual analogue scale (VAS),morning stiffness time, erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP) were measured in all patients. The serum concentration of sCD146 from 62 patients with ankylosing spondlitis and 20 healthy controls was measured by enzyme-linked immunosorbent assay. Westergren method was used to measure ESR and immunoturbidimetry for CRP. Clinical data of the patients were collected as well.RESULTS AND CONCLUSION: sCD146 levels of patients with ankylosing spondlitis were significantly higher than normal control group (P ＜ 0.05). The sCD146 expression in the active group was significantly higher than inactive and normal control groups (P ＜0.05). Positive correlations were observed between sCD146 and BASDAI index of patients with ankylosing spondlitis (P ＜ 0.05).The sCD146 levels of ankylosing spondUtis patients with peripheral joint involvement were significantly higher than the patients with axial involvement alone or the normal controls (P ＜ 0.05).The expression level of sCD146 in peripheral blood was positively correlated with disease activities of patients with ankylosing spondlitis. It may play important roles in the pathogenesis in ankylosing spondlitis.

Aim To explore the effect of receptor component protein(RCP)in the signal transduction of vascular smooth muscle cell(VSMC)proliferation induced by static pressure.Methods The mouse-derived vascular smooth muscle cell line(A10VSMC)was employed in the experiment.Cells were exposed to static pressure,and MTT assay was used to detect the cell viability.Western blot was used to determine the expressions of PCNA,RCP and p-Akt,RCP mRNA was tested by RT-PCR,and co-immunoprecipitation was used to test the interaction between RCP and G proteins.Results The cell viability,expressions of PCNA and RCP increased with the elevation of static pressure and reached their peaks at 120 mmHg,and after 6 hours they got a plateau.The static pressure significantly increased the level of p-Akt,meanwhile,the binding of RCP and Gαs significantly decreased.However,the binding of RCP and Gβ increased in response to static pressure after stimuli of static pressure,but Gγ was obscure.Conclusion Static pressure can induce VSMC proliferation and expression of RCP,which may involve G protein signal transduction model.

Objective To observe the occurrence and risk factors of arrhythmia in chronic kidney disease (CKD) patients in different stages of renal function. Methods A total of 405 CKD patients were enrolled in this study and none of them received renal replacement therapy.The 24 h dynamic electrocardiogram (DCG) was performed,and baseline characteristics were compared.Multivariable Logistic regression analysis was used to examine the relationship between the severe arrhythmia and the potential risk factors,such as age,gender,CKD stage,diabetes,hypertension,hyperpotassaemia,left ventricular hypertrophy (LVH),etc. Results There were 69 patients (17.04％),79 patients (19.51％),82 patients (20.25％),88 patients (21.73 ％) and 87 patients (21.48％) in CKD stage 1,2,3,4 and 5,respectively.As high as 45.68％ of all the patients had severe arrhythmia,represented by 27.54％,29.11％,42.68％,57.95％ and 65.52％ in CKD stages 1-5 respectively.The occurrence of severe arrhythmia increased as the eGFR decreased in CKD stages 2,3,4 (p＜0.05).On multivariable Logistic regression analysis,the occurrence of severe arrhythmia was related to LVH,CKD stage,diaberes hyertension and hyperpotassaemia are signidicantly assoxiated with severe arrhythmia.