Objective To observe the effects of fosinopril(FOS),a new generation angiotensin-converting enzyme inhibitor(ACEI),on protein and mRNA expression of transforming growth factor-?_1(TGF-?_1) of rat glomerular mesangial cell(GMC) induced by lipopolysaccharide(LPS);to demonstrate the preventive mechanism against glomerular sclerosis by applying FOS.Methods The cultured GMC in classic way were divided into 3 groups:control group;LPS group;LPS+FOS group.TGF-?_1 concentration in GMC supernatant fluid was detected by ELISA;TGF-?_1 mRNA expression was determined by semiquantitative real-time RT-PCR.Results LPS group was obviously higher than control groups in TGF-?_1 secretion and mRNA expression,while LPS+FOS group decreased distinctively in TGF-?_1 secretion and mRNA expression compared with LPS group.Conclusions FOS has obviously inhibited on TGF-?_1 expression of rat GMC both at protein level and mRNA level,which reveals that it may be an important mechanism by FOS on restraining the development of glomerulosclerosis.

OBJECTIVEIdiopathic nephrotic syndrome (INS) is a common glomerular disease. The pathogenesis of the disease remains unclear. Recent studies indicate that transforming growth factor beta (TGF beta) is the main cytokine involved in glomerular disease. It plays an important role in the development of INS and in occurrence of glomerulosclerosis. The present study aimed to study changes and significance of TGF beta in children with idiopathic nephrotic syndrome (INS).

METHODSTotally 35 cases with INS (13 males, 22 females) were studied. The age of onset was between 2 years and 1 months and 14 years with an average of 8 years and 3 months. The active stage group had 35 cases and the remission stage groups had 25 cases. The cases in active stage group had first onset of the disease with obvious clinical symptoms and abnormal laboratory findings without use of corticosteroids. The cases in remission stage group were asymptomatic without abnormal laboratory findings. Protein in urine was negative over 4 weeks after oral administration of prednisone for 8 weeks. Twenty five cases were steroid responsive and 10 cases were steroid non-responsive among the 35 cases. Thirty healthy young children were enrolled as control. TGF beta was detected by ELISA in peripheral blood mononuclear cell (PBMC) culture medium. The TGF beta mRNA gene expression was measured by in situ PCR in PBMC.

RESULTS(1) Concentration of TGF beta(247 +/- 26) ng/L and TGF beta mRNA expression (0.57 +/- 0.18) in active stage of simple type or nephritis type INS were higher than those of remission stage and control (P < 0.01). Concentration of TGF beta[(125 +/- 16) ng/L] and TGF beta mRNA expression (0.30 +/- 0.12) in remission stage were higher than that of control (P < 0.05). (2) The level of TGF beta protein in nephritis type [(275 +/- 26) ng/L] was significantly higher than that in simple type [(220 +/- 18) ng/L] in active stage INS (t = 6.45, P < 0.01). No significant difference in TGF beta mRNA expression was found between the nephritis type (0.58 +/- 0.15) and simple type (0.55 +/- 0.16) in active stage INS, either (P > 0.05). But these two types were different from the control (P < 0.01). (3) Concentration of TGF beta and TGF beta mRNA expression after therapy was clearly lower than that before therapy in steroid responsive group (P < 0.01). Whereas no significant change was seen in steroid non-responsive group. Both indicators were higher in steroid non-responsive group than in steroid responsive group whether before or after therapy.

CONCLUSIONTGF beta may play an important role in the mechanism of INS and its level in PBMC can be used as an immunological indicator for the illness state, therefore, determination of TGF beta level and mRNA may be of some clinical significance.

Adolescent ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Male ; Nephrotic Syndrome ; blood ; drug therapy ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; genetics ; metabolism

OBJECTIVETo observe the effects of fosinopril (FOS) on proliferation and secretion of extracellular matrix of rat glomerular mesangial cell induced by LPS.

METHODSIn vitro culture method for glomerular mesangial cells (GMC) of rat was established and passages 3 - 10 of the cells were used in the experiment after identification. The experiment included the following 5 groups: control group (Ctrl), LPS group (LPS), high, medium and low dose FOS groups (FOS1, FOS2 and FOS3 groups, respectively). GMC proliferation was detected by methyl thiazolyl tetrazolium (MTT) incorporation method at 24 and 48 h; the changes of laminin (LN), fibronectin (FN) and ColIV protein secretion was detected by the enzyme-linked immunosorbent assay (ELISA). The changes of LNbeta(2) mRNA expression was detected by semi-quantitative real-time RT-PCR.

RESULTS(1) LPS could induce the mesangial cell proliferation, FOS inhibited this effect of proliferation induced by LPS. (2) Mesangial cells could secrete some extracellular matrix (ECM) protein in normal culture medium, mesangial cell secreted ECM protein was significantly higher in LPS group than that in Ctrl group (P < 0.01), but significantly lower in all FOS groups than that in LPS group (P < 0.01). (3) Mesangial cell could express LNbeta(2) mRNA in normal culture medium, LNbeta(2) mRNA expression was significantly higher in LPS group than that in Ctrl group at all time points, but was significantly lower in FOS group than that in LPS group.

CONCLUSIONSLPS could induce increased secretion of the ECM, including LN, FN, ColIV; FOS could inhibit the secretion of ECM in GMC in a dose-dependent manner at mRNA and protein levels.

Animals ; Cell Proliferation ; Cells, Cultured ; Extracellular Matrix Proteins ; secretion ; Fosinopril ; pharmacology ; Gene Expression Regulation ; Lipopolysaccharides ; Mesangial Cells ; drug effects ; metabolism ; Rats

Objective:To study therapeutic effect of levosimendan combined milrinone on severe refractory heart failure (SRHF) and its influence on serum levels of brain natriuretic peptide (BNP) and uric acid (UA).Methods:A total of 156 SRHF patients were enrolled,randomly and equally divided into levosimendan group and milrinone group,both groups re-ceived corresponding medication based on routine treatment for 7d.Heart rate,blood pressure,serum BNP and UA levels,left ventricular end-systolic dimension (LVESd),left ventricular end-diastolic dimension (LVEDd) and left ventricular e-jection fraction (LVEF) before and after treatment,total effective rate and incidence of adverse reactions were observed and compared between two groups.Results:Compared with milrinone group after treatment,there were significant reduc-tions in heart rate [ (73.79 ± 7.61) beats/min vs.(70.39 ± 7.45) beats/min],systolic blood pressure [ (128.84 ± 13.11) mmHg vs.(121.86 ± 12.53) mmHg],scores of lung wet rales [ (2.05 ± 0.33) scores vs.(1.53 ± 0.21) scores],difficulty breathing [ (2.11 ± 0.36) scores vs.(1.60 ± 0.25) scores] and lower extremity edema [ (2.03 ± 0.34) scores vs.(1.50 ± 0.18) scores],serum levels of BNP [ (459.62 ± 46.27) μg/L vs.(248.73 ± 25.91) μg/L] and UA [ (355.97 ± 36.47) μmol/L vs.(282.75 ± 28.61) μmol/L],LVESd [ (41.62 ± 4.52) mm vs.(36.87 ± 3.71) mm] and LVEDd [ (51.89 ± 5.37) mm vs.(47.85 ± 4.83) mm],and significant rise in 24h urine volume [ (3204.59 ± 321.52) ml vs.(3695.78 ± 370.62) ml] and LVEF [ (42.36 ± 4.31)% vs.(47.85 ± 4.86)%] in levosimendan group,P＜0.01 all.Total effective rate of levosimendan group was significantly higher than that of milrinone group (89.74% vs.71.79%),and incidence rate of adverse reactions was significantly lower than that of milrinone group (5.13% vs.28.21%),P＜0.01 both.Conclusion:Levosimendan therapy can significantly reduce serum BNP and UA levels,improve cardiac function in SRHF patients.It possesses significant therapeutic effect,and it＇s safe and reliable,which is better than milrinone.

Objective :To explore plasma expression levels of human maternal expression gene 3 (MEG3) and urotheli-al carcinoma antigen 1 (UCA1) in patients with acute myocardial infarction (AMI) and their clinical significance . Methods : A total of 90 AMI patients treated in our hospital were enrolled as AMI group ,and 50 healthy subjects un-dergoing physical examination in our hospital simultaneously were treated as healthy control group .Plasma expres-sions of MEG3 and UCA1 ,serum levels of creatine kinase isoenzyme MB (CK-MB) and cardiac troponin I (cTnI) were observed and compared between healthy control group and AMI group on 1h ,3h ,6h and 12h after onset . Spearman correlation analysis was used to analyze their correlation .Results : Compared with healthy control group , there was significant rise in plasma MEG3 expression [ (0-002 ± 0-001) vs .0-017 ± 0-003)] ,and significant reduc-tion in UCA1 expression [ (0-027 ± 0-005) vs .(0-017 ± 0-002)] in AMI group on 1h after onset , P＝0-001 both ;after 3h ,there were significant rise in serum levels of CK-MB [ (20-01 ± 3-05) IU／L vs.( (32-10 ± 4-40) IU／L] and cTnI [ (1-01 ± 0-87) ng／L vs.(2-10 ± 0-91) ng／L] in AMI group , P＝0-001 all.Spearman correlation analy-sis indicated that in plasma MEG3 expression was significant positively correlated with serum CK-MB and cTnI levels ( r＝0-351 ,0-368 , P＜0-05 both) ,and plasma UCA1 expression was significant inversely correlated with serum levels of CK-MB and cTnI (r＝ －0-416 ,－0-425 , P＜0-01 both) AMI patients .Conclusion : Plasma MEG3 level significant rises ,and UCA1 level significantly reduces in AMI patients during early onset period .Both are signifi-cantly correlated with CK-MB and cTnI levels ,which may be used as new markers diagnosing early AMI .

OBJECTIVETo explore the anti-tumor immunity in vitro induced by prostate cancer cell vaccine transfected with recombinant adenovirus encoding 4-1BBL in mice.

METHODSThe replication-deficient adenovirus AdEasy-1 system was used to construct recombinant adenovirus Ad-m4-1BBL and Ad-eGFP. The prostate cancer cell RM-1 of mice was transfected with Ad-m4-1BBL and Ad-eGFP, and treated with mitomycin (MMC) to produce TCV, TCV-Ad-eGFP and TCV-Ad-m4-1BBL, followed by co-culture with syngeneic murine spleen cells. Then the cytotoxic activity of the lymphocytes against RM-1 cells was analyzed with CCK-8 solution, and IL-2 and INF-gamma were detected by ELISA.

RESULTSThe 4-1BBL protein was highly expressed in the TCV-Ad-m4-1BBL of the 4-1BBL-transfected mice. TCV-Ad-m4-1BBL significantly increased the expressions of IL-2 ([180.24 +/- 2.22] pg/ml) and INF-gamma ([1512.46 +/- 23.64] pg/ml) as compared with TCV and TCV-Ad-eGFP (P < 0.05), and induced higher RM-1 cell specific cytotoxicity ([34.24 +/- 2.64]%) than the latter two ([9.82 +/- 1.48]%) and ([14.65 +/- 3. 21]%), (P < 0.05). But none of them exhibited significant cytotoxicity against hepatocellular carcinoma Hepal-6.

CONCLUSIONThe m4-1BBL-expressing prostate cancer cell vaccine can effectively induce anti-tumor immune responses.

4-1BB Ligand ; genetics ; immunology ; Animals ; Cancer Vaccines ; genetics ; immunology ; Cell Line, Tumor ; Coculture Techniques ; Cytotoxicity, Immunologic ; genetics ; Female ; Interleukin-2 ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Prostatic Neoplasms ; Transfection

OBJECTIVETo describe a HPLC method for assessing betaine in Fufang Guilu granule.

METHODThe content of betainephenaxcyl bromide in Fufang Guilu granule was determined by HPLC. The analytical column was a shim-pack CLC-ODS (6.0 mm x 150 mm) filling a 5 microm stationary phase; The mobile phase consisted of acetonitrile-water(35:65) with 0.1 mol x L(-1) NaClO4; The flow-rate was 1 mL x min (-1); The detector was set at 254 nm.

RESULTThe calibration curve was linear over the range of 0.09-0.585 microg (r = 0.9997). The average recovery of the method was 98.4%, RSD 2.5% (n = 5).

CONCLUSIONThe results showed that this method was reliable and accurate, and can be used for quality control of Fufang Guilu granule.

Betaine ; analysis ; Chromatography, High Pressure Liquid ; methods ; Codonopsis ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; Lycium ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results

Objective :To explore influence of small and dense low density lipoprotein (sdLDL) on in-stent restenosis after percutaneous coronary intervention (PCI).Methods : A total of 631 CHD inpatients ,who received PCI in our department from Feb 2015 to Feb 2017were followed up for 12 months ,and 240 cases were successfully followed up . According to coronary angiography (CAG) results ,the vascular diameter stenosis ≥50% regard as in-stent resteno-sis ,all patients were divided into restenosis group (n＝105) and no stenosis group (n＝135).Another 35 healthy volunteers undergoing physical examination simultaneously were treated as healthy control groups .Serum levels of sdLDL ,serum lipid TC ,TG ,HDL-C and LDL-C were measured and compared among all groups .Correlation be-tween serum sdLDL level and Gensini score was analyzed in CHD patients .Results : There were no significant difference in levels of serumlipids between restenosis group and no stenosis group , P＞ 0. 05 all.Compared with healthy control group ,there was significant rise in serum sdLDL level [ (0.451 ± 0.135) mmol/Lvs .(0.673 ± 0.281) mmol/L] in CHD group , P＝0.001 ;compared with no stenosis group ,there were significant rise in serum sdLDL level [(0.606 ± 0.276) mmol/L vs.(0.695 ± 0.304) mmol/L] and Gensini score [(40.23 ± 9.24) scores vs. (58.42 ± 10.37) scores] in restenosis group (P＝0.019 ,0.001) respectively.Linear correlation analysis indicated that serum sdLDL level was significant positively correlated with Gensini score in CHD patients ( r＝ 0.514 , P＝ 0.032).Conclusion :Elevated serum sdLDL level suggests high risk of in-stent restenosis in CHD patients .It can provide reference for disease condition assessment and adverse event prevention after PCI .

A device to produce low temperature plasma ( LTP) was designed and constructed to serve as the ion source of a high resolution mass spectrometry, and was applied to qualitatively analyze the steroid samples. In comparison with conventional electrospray ionization mass spectrometry, low temperature plasma mass spectrometry ( LTP-MS) had some advantages such as simple sample pretreatment and less interference. Mass spectrometry and tandem mass spectrometry were used to characterize the steroid samples in this research, and it was found that the structural stability of each steroid sample was presented in its mass spectrum, while in the tandem mass spectra there were more fragments of H2 O lost. And then the fragmentation process of typical steroid samples in collision induced dissociation ( CID ) was discussed based on theoretical calculation. In addition, by comparing tandem mass spectrometry and the fragmentation process, a pair of isomers of testosterone and dehydroepiandrosterone could be distinguished successfully.

BACKGROUND: Edaravone (3-methyl-1-penyl-2-pyrazolin-5-one) is a potent free-radical scavenger and has the antioxidant ability to inhibit lipid peroxidation. The study aimed to examine the effect of edaravone on protecting the acute injury of human type Ⅱ alveolar epithelial cells (A549 cells) induced by paraquat (PQ) and the change of production of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD). METHODS: A549 cells were cultured and divided into PQ group (group P), edaravone-treated group (group E) and normal control group (group C). The cells in group P were exposed to paraquat (600 mol/L), and the cells in group E were treated with edaravone (100 mol/L) additionally, and no drug intervention was given to the cells in group C. Real-time monitoring by LSCM was used to detect the cell response and the intracellular dynamic change of ROS level in A549 cells after administration of PQ and edaravone. And the levels of SOD and MDA were detected respectively by biochemistry colorimetry. Data were expressed as mean ± standard error of the mean. Statistical analysis was carried out with the soft SPSS 16.0. RESULTS: The concentration of intracellular ROS significantly increased when PQ was given to A549 cells. But after administration of edaravone, the concentration of intracellular ROS was decreased. Compared to the PQ group, the levels of SOD in the edaravone group were significantly increased while the levels of MDA were markedly decreased. CONCLUSIONS: Paraquat can increase the oxidative stress, and induce the lipid peroxidation of A549 cells. Edaravone has the effect to scavenge reactive oxygen species, and to protect against the PQ-induced lung toxicity.