OBJECTIVETo discuss the effect of taurine (Tau) on the proliferation of hypoxia-induced pulmonary artery smooth muscle cells (PASMCs), and study whether the extracellular signal-regulated kinase 1/2 (ERK1/2) signal pathway participated in the Tau-inhibited PASMC proliferation process and the possible molecular mechanism.

METHODThe primary culture was performed for PASMCs in rats. The second to fifth generations were adopted for the experiment. The Tau concentration was 80 mmol x L(-1). The concentration of ERK1/2 blocker (PD98059) was 50 micromol x L(-1). The drug administration time was 24 h. The effect of Tau on the PASMC proliferation was detected by MTT assay, immunofluorescence staining method and western blot under different conditions. The PASMCs were growing were divided into four groups: the normoxia group, the normoxia + Tau group, the hypoxia group and the hypoxia + Tau group. The Western blot was adopted to detect whether the ERK1/2 signal pathway participated in the Tau-inhibited PASMC proliferation process. Subsequently, the PASMCs were divided into five groups: the normoxia group, the hypoxia group, the hypoxia + Tau group, the hypoxia + Tau + PD98059 group and the hypoxia + PD98059 group.

RESULTHypoxia could induce the PASMC proliferation. Under the conditions of normoxia, Tau had no effect on the PASMC proliferation. Under the conditions of normoxia and hypoxia, Tau had no effect on the expression of the tumor necrosis factor-alpha (TNF-alpha) among PASMCs. Tau could reverse the expression up-regulation of hypoxia-induced proliferative cell nuclear antigen (PCNA) (P < 0.01) and Cyclin A (Cyclin A) (P < 0. 05). Under the conditions of normoxia, Tau had no effect on the expression of phosphoryl extracellular signal-regulated kinase 1/2 (p-ERK1/2). Hypoxia could up-regulate the p-ERK1/2 expression (P < 0.01). Tau could reverse the up-regulation of the hypoxia-induced p-ERK1/2 expression(P < 0.01). Both PD98059 and Tau could inhibit the up-regulated expressions of PCNA, Cyclin A and p-ERK1/2. According to the comparison between the single addition of Tau and PD98059 under conditions of hypoxia, the hypoxia + Tau + PD98059 group showed more significant down-regulation in the expressions of PCNA, Cyclin A and p-ERK1/2.

CONCLUSIONTau could inhibit the hypoxia-induced PASMC proliferation, and may regulate it through ERK1/2 pathway.

Animals ; Cell Hypoxia ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; MAP Kinase Signaling System ; drug effects ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Oxygen ; metabolism ; Pulmonary Artery ; cytology ; drug effects ; metabolism ; physiopathology ; Rats ; Rats, Wistar ; Taurine ; pharmacology ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

PKS gene was screened by PCR from thirty strains of spone-associated bacteria including twenty-one actinomycetes isolated from Craniella anstrialiensis and nine bacillus isolated from Dysidea avara in the South China Sea.As a result,a 669 bp KS domain gene was successfully amplified from Bacillus C89.BLAST analysis showed that the KS domains were most closely related to the KS sequences of Bacillus subtilis subsp.subtilis str.168 with 96% similarity.Phylogenetic analysis demonstrated the KS domain belong to trans-AT KS domains.This study demonstrated the existence of PKS gene in bacteria associated with sponge Dysidea avara for the first time,and provided proof for the hypothesis that sponge-associated bacteria are perhaps the true producers of many novel bioactive compounds in sponge.Meanwhile,this study lays a basis for the microbial screening for polyketide compounds production.

Objective To explore the value of MRI-R2 * and to compare clinical effect of two iron chelators(deferasirox and deferoxamine) in iron-overloaded patients.Methods By completely randomized balanced design,24 iron-overloaded patients were randomly divided into 2 groups,which consisted of 12 patients treated with deferasirox and 12 patients treated with deferoxamine.The planned deferasirox dose was 40 mg· kg-1 · d-1,and the deferoxamine dose was no less than 50 mg · kg-1 · d-1 All patients underwent quantitative MRI at the time points of the primary screening,6 months and 12 months.Pair Wilcoxon rank sum test was used to compare the differences of liver R2 * values of the 2 groups at various time points respectively.Wilcoxon rank sum test was used to compare the differences of change rate of liver R2 * values between the two groups at the time point of 6 months,12 months,respectively.Results Deferasirox group's liver R2 * values of primary screening,6 months and 12 months were 1081,889 and 712 Hz,while deferoxamine group's liver R2 * values were 1042,838 and 488 Hz.There was no statistically significant difference between liver R2 * values of two groups at primary screening (Z =-0.029,P ＞ 0.05).The change rate of liver R2 * of deferasirox group at 12 month was-32％,while it was-58％ for the deferoxamine group,and there was statistically significant difference between the two groups (Z =-3.060,P ＜0.01).The change rate of serum ferritin of deferasirox group at 12 month was-15％,while it was -55％ for the deferoxamine group,and there was statistically significant difference between the two groups (Z =-2.945,P ＜ 0.01).Conclusion By using MRI-R2*,it suggest that both deferasirox and deferoxamine can effectively remove liver iron and deferoxamine is superior to deferasirox.

To study the inhibitory effect of RNA interference (RNAi) on MMP-9 gene expression in THP-1 cell line.To investigate the application of RNAi on the therapy of leukemia.Methods:Small interfering RNA ( siRNA) for MMP-9 gene was designed and transfected into THP-1 cells.MMP-9 mRNA expression was assessed by RT-PCR, and MMP-9 protein expression was tested by Western blot.MTT and trypan blue staining were used to observe the effect on the proliferation of THP-1 cells after RNAi.The changes in cell morphology were observed under the microscope.Results:The expressions of MMP-9 mRNA and protein were inhibited in THP-1 MMP-9 siRNA-transfected cells ,significantly lower than those of control cells.The results of MTT and trypan blue staining in-dicated that the proliferation ability of THP-1 cells obviously decreased after siRNA-transfected 48h and 72h.The growth of cells was in-hibited and the cells survival rate was significantly lower than that of control group ( P<0.05 ).The cells of control groups grew semi-quote wall under inverted microscope.The outline of cells was clear and the shape was uniform.The cells grew vigorously.While the growth of cells in siRNA group was inhibited.The morphology of siRNA group cells changed obviously by the Wright staining.Most cells expressed changes of apoptosis.Conclusion: siRNA for MMP-9 gene can not only reduce the expressions of MMP-9 mRNA and protein,but also inhibit the proliferation and induce apoptosis of THP-1 cells.

Objective To investigate the effects of antihypertension and reversing left ventricular hypertro- phy by carvedilol or bisoprolol in patients with mild to moderate essential hypertension.Methods 40 cases of mild to moderate essential hypertension patients were selected for this random single-blind,paralleling controlled clinical study.Results Patients were randomized to take 12.5～25mg carvedilol tablet orlce daily or bisoprolol 2.5～5mg once daily if DBP was still in the range of 12.0～14.6kPa(90～110mmHg)after 2 weeks' placebo baseline. Carvedilol group included 20 cases,bisoprolol group included 20 cases,and the course was 24 weeks.Blood pressure and heart rate were measured and symptoms and signs were recorded.At the end of placebo and in 24 weeks heart ultrasound,blood routine,serum glucose,blood lipid,hepatic function and renal function were examined.SBP,DBP and heart rate of patients in two groups decreased obviously.There were significant differences between the two groups.Ventricular hypertrophy of carvedilol group improved than that in pretherapy.There were significant differ- ences between the two groups.Conclusion Carvedilol was well-tolerated with less side effects such as mild headache,tiredness,dizziness,slightly elevating of serum glucose.Carvedilol could well treat the mild moderate essen- tial hypertension effectively and safely by 12.5～25mg once daily.

AIMTo explore the role of endogenous and exogenous hydrogen sulfide (H2S) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats and the underlying mechanisms.

METHODS120 Sprague-Dawley rats were randomly divided into four groups: control, LPS (instilled intratracheally to induce ALI), NaHS (H2S donor) + LPS, and propargylglycin (PPG) + LPS. Animals were sacrificed at 4 h or 8 h after agent administration. Lung weight/body weight ratio (LW/BW) was measured and calculated. Morphological changes of lung tissues were observed. H2S concentration, NO concentration (NO) and carbon monoxide (CO) level in plasma were tested. Malondialdehyde (MDA) content, CSE activity, inducible nitric oxide synthase (iNOS) activity and hemeoxygenase (HO) activity of the lung were determined. PMN and protein content in BALF were also tested. Immunohistochemisty technique was performed to examine the expression of iNOS and HO-1 protein in lung tissues. The correlation of H2S content with the above indices was analyzed.

RESULTSCompared with control conditions, severe injuries of lung tissues and a raised LW/BW, MDA content, PMN and protein content in BALF were observed in rats treated with LPS. LPS also lead to a drop in plasma H2S concentration and lung CSE activity. The enzyme activity of iNOS and HO, the protein expression of them and plasma NO, CO level increased after LPS instillation. Administration of NaHS before LPS could attenuated the changes induced by LPS. Pre-administration of PPG exacerbated the injuries induced by LPS, increased PMN and protein content in BALF, the plasma NO level, lung iNOS activity and its protein expression, but there was no prominent variation in CO level, HO activity and HO-1 protein expression compared with those of LPS group. The H2S content was positively correlated with CSE activity, CO content and HO-1activity (r = 0.945-0.987, P < 0.01), and negatively correlated with the other indices (r = -0.994 - -0.943, P < 0.01).

CONCLUSIONDownregulation of H2S/CSE was involved in the pathogenesis of acute lung injury induced by LPS. Endogenous and exogenous H2S provided protection against the lung injuries, which might be explained by its anti-oxidative effects, attenuating inflammatory over-reaction in lung induced by PMN,the downregulation NO/iNOS system and the upregulation of CO/HO-1 system.

Acute Lung Injury ; chemically induced ; physiopathology ; Animals ; Antioxidants ; metabolism ; pharmacology ; Bronchoalveolar Lavage Fluid ; Carbon Monoxide ; metabolism ; Heme Oxygenase (Decyclizing) ; metabolism ; Hydrogen Sulfide ; metabolism ; pharmacology ; Lipopolysaccharides ; antagonists & inhibitors ; toxicity ; Male ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the relationship between B-type natriuretic peptides (BNP) and subclinical target organ damage in essential hypertensive (EH) patients.

METHODSA total of 317 EH patients were divided into 3 groups according to BNP levels, namely normal (BNP<600 ng/L) group (n=102), moderate (600-883.5 ng/L) group (n=116), and elevated BNP (>883.5 ng/L) group (n=99). The blood pressure, left ventricular mass index (LVMI), the intima media thickness (IMT) of the common carotid artery, the plaque size in the coronary artery (CS) and microalbuminuria levels were analyzed in these patients.

RESULTSThe EH patients with moderate and elevated BNP showed significantly higher LVMI, IMT, CS and microalbuminuria levels than those with normal BNP level (LVMI: 102.8∓23.12 and 123.9∓26.47 vs 91.09∓18.71 g/m2; IMT: 0.95∓0.32 and 1.16∓0.37 vs 0.84∓0.28 mm; microalbuminuria: 31.36∓20.55 and 36.73∓22.07 vs 23.21∓18.68, P<0.01). After adjustment, BNP was positively correlated to LVMI, IMT, CS and microalbuminuria level (r=0.45, 0.43, 0.39 and 0.41, respectively, P<0.01). Multivariate logistic regression analysis showed that age, systolic blood pressure, BNP, FPG, and microalbuminuria, LDL-C, and BMI were all related to the occurrence of subclinical target organ damages.

CONCLUSIONBNP is positively correlated to subclinical target organs damages in EH patients.

Adult ; Aged ; Aged, 80 and over ; Albuminuria ; pathology ; Carotid Artery, Common ; pathology ; Carotid Intima-Media Thickness ; Female ; Humans ; Hypertension ; blood ; pathology ; Hypertrophy, Left Ventricular ; pathology ; Male ; Middle Aged ; Natriuretic Peptide, Brain ; blood

Liver fibrosis is a common pathological process for chronic liver injury caused by multiple etiological factors and an inevitable phase leading to liver cirrhosis. According to the previous studies, hesperidin (HDN) shows a very good protective effect on CCl4-induced chemical hepatic fibrosis in rats. In this experiment, based on the findings of the previous studies, a platelet-derived growth factor (PDGF)-induced HSC-T6 model was established to observe the inhibitory effect of HDN on HSC-T6 proliferation. The ELISA method was adopted to detect the content of collagen I in HSC-T6 supernatant. Transforming growth factor (TGF)-beta1, Smad2, Smad3, Smad7 and connective tissue growth factor (CTGF) mRNA expressions were measured by RT-PCR; TGF-beta1 and CT-GF protein expressions in HSC-T6 were determined by Western blot, in order to study HDN's effect on TGF-beta1 signaling pathway in HSC and its potential action mechanism. The results demonstrated that HDN could notably improve HSC-T6 proliferation, Collagen I growth and TGF-beta1, Smad2, Smad3 and CTGF mRNA.expressions. After being intervened with HDN, it could notably inhibit HSC-T6 proliferation and Collagen I growth, reduce TGF-beta1, Smad2, Smad3 and CTGF mRNA and TGF-beta1, CTGF protein expressions and increase Smad7 mRNA expression. HDN's antihepatic fibrosis effect may be related to the inhibition of HSC proliferation and activation by modulating TGF-beta/Smad signaling pathway.
Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Connective Tissue Growth Factor ; physiology ; Hesperidin ; pharmacology ; Platelet-Derived Growth Factor ; pharmacology ; Rats ; Signal Transduction ; drug effects ; Smad Proteins ; physiology ; Transforming Growth Factor beta1 ; physiology

Objective To investigate the role of oxidative stress in the epithelial-to-mesenchymal transdifferentiation (EMT) of peritoneal mesothelial cells in rat model of peritoneal fibrosis and the effect of probucol on peritoneal fibrosis. Methods The rat model of peritoneal fibrosis was induced by 4.25% high glucose peritoneal dialysis fluid (PDF). The rats were randomly divided into 4 groups:the control group, the saline group, the peritoneal fibrosis group, and the probucol group. A 4 hour peritoneal equilibration test (PET) was performed 4 weeks later. The peritoneal function and net ultrafiltration (UF) volume were determined. The level of malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in peritoneal tissue were examined. The histology of peritoneal membrane was evaluated by light microscopy. E-cadherin and α-smooth muscle actin (α-SMA) protein expression was evaluated by immunohistochemical method and Western blot.Results The mesothelial cells were detached from peritoneal membrane in peritoneal firbosis rats. Comparing with the control rats, the thickness of visceral peritoneum, the level of MDA, and the-SMA protein expression were increased while the net ultrafiltration volume, the level of GSH-Px and E-cadherin protein expression were decreased in peritoneal firbosis rats. All these changes were reversed in the rats treated with probucol.Conclusion Oxidative stress plays an important role in transdifferentiation of peritoneal mesothelial cell in the peritoneal fibrosis rats. Probucol can improve structure and function of peritoneum, and partially reverse the EMT by reducing the oxidative stress.

Objective To investigate the effects of connective tissue growth factor (CTGF) siRNA delivered by pRetro-Super (PRS) retrovirus vector on extracellular matrix and VEGF expression in human peritoneal mesothelial cells (HPMC). Methods Four pairs of oligonucleotides including 64 bp DNA were designed and synthesized in vitro according to siRNA target sequence and PRS retrovirus desire.PRS-CTGF-siRNA1-4 recombinant retrovirus vectors were constructed.The recombinant retrovirus vectors containing CTGF-siRNA were transferred into PT67 packaging cell lines with lipefectamine 2000,then infected HPMC.mRNA expression was determined by semi-quantitative RT-PCR and protein expression was determined by Western blot.Results Both mRNA and protein expressions of CTGF,FN,Col I,laminin (LN) and VEGF were significantly increased in HPMC with 5 μg/L TGF-β1 stimulation (P＜0.01,respectively).CTGF,FN,Col I,LN mRNA and protein and VEGF mRNA expression stimulated by TGF-β1 were significantly decreased in HPMC infected with PRS-CTGF-siRNA1～4 retrovirus vectors (P＜0.01,respectively).The inhibitory rates on CTGF were 69.3%,22.2%,27.4% and 38.8%,respectively (P＜0.01).At the same time,there was also a significant reduction of VEGF protein expression in HPMC infected with PRS-CTGF-siRNA1 vector (P＜0.01).There was no significant difference in HPMC infected with PRS void vector. Conclusion CTGF siRNA delivered by PRS retrovirus vector can effectively inhibit the enhancement of extracellular matrix and VEGF expression stimulated by TGF-β1 in HPMC.