This study aims to investigate the effect of total flavones of Fructus Chorspondiatis (TFFC) on the mRNA and protein expression of collagen type I and III of rat cardiac fibroblasts (CFs) induced by angiotensin II (Ang II), and explore its anti-myocardial fibrosis molecular mechanism. Neonatal rat CFs were prepared from Sprague-Dawley rats (1-3 d after birth). The expression of collagen type I and III mRNA and protein were measured by RT-PCR and Western blotting, respectively. The study showed that stimulation of neonatal rat CFs with 100 nmol.L-1 of Ang II for 72 h resulted in a significant increase of the expression of collagen type I and III mRNA and protein. The changes on the expression level were blocked by TFFC. The results demonstrated that TFFC can inhibit myocardial fibrosis induced by Ang II in rats, which is probably associated with the collagen type I and III mRNA and protein levels up-regulated by Ang II, and TFFC was shown to decrease the expression levels of collagen type I and III mRNA and protein.

Adolescent ; Adult ; Automobile Driving ; China ; Electrocardiography ; Female ; Heart ; physiopathology ; Humans ; Male ; Middle Aged ; Noise, Occupational ; adverse effects ; Occupational Exposure ; adverse effects ; Stress, Psychological ; etiology ; physiopathology ; Vibration ; adverse effects

OBJECTIVETo observe the clinical efficacy of Tiaozhi Jiangtang Tablet (TZJT) on patients suffering from diabetes mellitus with blood stasis syndrome.

METHODSSixty patients were randomly assigned into 2 groups, the TZJT group (n = 30) treated with TZJT tablet and asprin, the control group (n = 30) treated with asprin alone.

RESULTSThe improvement of symptoms was more significant in the TZJT group than that in the control group (P < 0.05). Levels of serum endothelin (ET), nitric oxide (NO) content and blood viscosity were decreased in both groups after treatment, and the effect of TZJT plus asprin was superior to that of asprin alone.

CONCLUSIONTZJT combined with asprin is effective in improving the serum content of ET and NO and reducing blood viscosity.

Adult ; Aged ; Aged, 80 and over ; Coronary Artery Disease ; drug therapy ; etiology ; Diabetes Mellitus, Type 2 ; complications ; drug therapy ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy

This study aims to investigate the effect of total flavones of Fructus Chorspondiatis (TFFC) on the mRNA and protein expression of collagen type I and III of rat cardiac fibroblasts (CFs) induced by angiotensin II (Ang II), and explore its anti-myocardial fibrosis molecular mechanism. Neonatal rat CFs were prepared from Sprague-Dawley rats (1-3 d after birth). The expression of collagen type I and III mRNA and protein were measured by RT-PCR and Western blotting, respectively. The study showed that stimulation of neonatal rat CFs with 100 nmol.L-1 of Ang II for 72 h resulted in a significant increase of the expression of collagen type I and III mRNA and protein. The changes on the expression level were blocked by TFFC. The results demonstrated that TFFC can inhibit myocardial fibrosis induced by Ang II in rats, which is probably associated with the collagen type I and III mRNA and protein levels up-regulated by Ang II, and TFFC was shown to decrease the expression levels of collagen type I and III mRNA and protein.
Anacardiaceae ; chemistry ; Angiotensin II ; pharmacology ; Animals ; Animals, Newborn ; Cells, Cultured ; Collagen Type I ; genetics ; metabolism ; Collagen Type III ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Fibroblasts ; cytology ; metabolism ; Flavones ; administration & dosage ; isolation & purification ; pharmacology ; Fruit ; chemistry ; Myocardium ; cytology ; metabolism ; Plants, Medicinal ; chemistry ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley

AIM: To study the clinical efficacy, complications and visual related quality of life(VRQoL)of Ozurdex in the treatment of macular edema secondary to retinal vein occlusion(RVO-ME).

METHODS: Totally 30 patients with ME, which had developed secondary to either CRVO(13 eyes)or BRVO(17 eyes), were monitored for 6mo after treatment with Ozurdex in the Department of Ophthalmology of our Hospital. We measured the best corrected visual acuity(BCVA), intraocular pressure(IOP), central macular thickness(CMT)at different time after treatment(1wk, 1mo, 2mo, 3mo, 4mo, 5mo, and 6mo after treatment), and we also measured the Chinese version vision related quality of life questionnaire-25(CVRQoL-25)at 3mo after treatment and compared them separately with the ones measured before treatment to evaluate the efficacy, adverse reactions and the visual related quality of life.

RESULTS: Generalized estimation equation results showed that BCVA, CMT and IOP all had differences at different time points(P<0.001). A BCVA increase was achieved and CMT decreased in all patients at any time point after the onset of treatment(P<0.001). The changes of BCVA and CMT were the largest in the 2mo compared to the baseline(P<0.001). The score of CVRQol-25 at 3mo after treatment was significantly higher than that before treatment and then the central retinal thickness decreased and a BCVA increased compared to the baseline level(P<0.01). The score of CVRQoL-25 at 3mo was negatively correlated both with the LogMAR BCVA evaluated before treatment and at 3mo after treatment(rs= -0.717, -0.746, all P<0.001); Meanwhile, the score of CVRQoL-25 was also negatively correlated with CMT at 3mo after treatment(rs= -0.862, P=0.001). In 19 eyes(63%)of the patients with RVO-ME,a relapse was observed after a follow-up time of 1-3mo and the average recurrence time was(2.8±0.5)mo. In follow-up of 6mo, about(2.3±0.4)intravitreal Ozurdex injections per eye was observed. The increase in IOP was observed at 1wk, 1, 2, 3mo after pretherapy(P<0.05). The mean IOP values reached a peak at 2mo after injection, which rose(7.85±0.32)mmHg above the baseline level(P<0.05)and decreased to normal at 4mo after treatment. 10% of patients had an elevation in IOP above 25mmHg, which could be medically controlled and 4 eyes(13%)of patients had cataract formation, two of which needed to surgery.

CONCLUSION: Ozurdex proved to be efficacious with increase in visual acuity and reduction of central retinal thickness and improve the visual function-related quality of life of RVO-ME patients. After single injection of Ozurdex, visual acuity benefited for 2-3mo. 63% of the patients relapsed at about 3mo after treatment. Adverse reactions associated to the use of Ozurdex include the formation of cataracts and an increase in IOP.

OBJECTIVETo study the effect of phospholamban antisense RNA (asPLB) on sarcoplasmic reticulum Ca2+-ATPase activity and cardiac function in rats with diabetes mellitus (DM) mediated by recombinant adeno-associated virus (rAAV) vector.

METHODSSix weeks after the induction of DM by streptozotocin injected intraperitoneally, the rats were divided into three groups, namely: DM-rAAV-asPLB group, DM-saline group and DM group (control group). The rats in the DM-rAAV-asPLB group were intramyocardially injected with rAAV-asPLB, the rats in the DM-saline group were injected with saline, and those in the control group did not receive any treatment. Six weeks after gene transfer, the expressions of PLB protein and PLB phosphorylation were detected by Western-blot, while the activity of sarcoplasmic reticulum (SR) Ca2+-ATPase and left ventricular function were measured.

RESULTSThe PLB protein expression level was significantly higher whereas the PLB phosphorylation, SR Ca2+-ATPase activity and left ventricular function were significantly lower in the DM-saline group than in the control group. No significant difference was found in PLB protein expression level, PLB phosphorylation or SR Ca2+-ATPase activity between the DM-rAAV-asPLB group and the control group. The left ventricular function in the DM-rAAV-asPLB group was poorer than in the control group and was better than in the DM-saline group.

CONCLUSIONrAAV-asPLB can down-regulate PLB protein expression and up-regulate PLB phosphorylation and SR Ca2+-ATPase activity, thus contributing to the improvement of in vivo left ventricular function.

Animals ; Calcium-Binding Proteins ; genetics ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; Male ; Phosphorylation ; RNA, Antisense ; administration & dosage ; Rats ; Rats, Wistar ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; metabolism ; Ventricular Function, Left ; drug effects

This study was purposed to construct the recombinant hF9 minigene and its stable nonsense mutant cell lines, and to investigate its significance. Minigene hF9 was cloned into the mammalian expression vector pCMV-Tag3B; a nonsense mutant containing a premature termination codon (PTC) in the 121(st) amino acid residue was obtained by PCR site-directed mutagenesis; minigene hF9 and nonsense mutant were respectively transfected into HepG2 cells with G418 treatment to get stable HepG2-WT and HepG2-N cell lines. The results confirmed that the minigene hF9 and nonsense mutant were constructed successfully. The gene of interest was amplified by RT-PCR from the stable cell lines, and the minigene hF9 was expressed in the stable cell lines. It is concluded that the recombinant hF9 minigene and its stable nonsense mutant cell lines are constructed successfully. The cell lines can be used to screen the drugs treating the nonsense mutation-caused hemophilia according to PTC read-through approaches.
Cell Line, Tumor ; Codon, Nonsense ; Factor IX ; genetics ; Genetic Vectors ; Hemophilia B ; genetics ; Humans ; Recombinant Proteins ; genetics

OBJECTIVETo explore the feasibility of using plasma trypsinogen activation peptides (TAP) and serum interleukin-6(IL-6) as early markers for predicting the severity of experimental acute pancreatitis.

METHODSNinety male adult Sprague-Dawley rats were equally randomized into five groups: edema pancreatitis group, treated with retrograde ductal infusion of 3% sodium taurocholate solution; necrosis pancreatitis group, treated with retrograde ductal infusion of 5% sodium taurocholate solution; treatment pancreatitis group, treated with retrograde ductal infusion of 3% sodium taurocholate solution and ulinastatin intravenous infusion half an hour later; control pancreatitis group, treated with 0.9% normal saline retrograde ductal infusion; and sham operation group, treated with sham operation. Rats in each group were equally randomized into three subgroups, which were killed by exsanguination 3, 6, or 24 hours after infusion, and blood specimens were obtained. Serum amylase, plasma TAP, and serum IL-6 were determined. The severity of pancreatitis was scored by two blinded pathologists under microscope.

RESULTSAt 3 and 6 hours after infusion, plasma TAP concentration of necrosis pancreatitis group [(4.798±0.169) and (3.999±0.299)nmol/L, respectively]were significantly higher than those of edema pancreatitis group [(2.416±0.148) and (3.356±0.211)nmol/L, respectively] (P<0.01); at 6 hours after infusion, serum IL-6 level of necrosis pancreatitis group [(1339.51±56.43)pg/ml]was significantly higher than that of edema pancreatitis group [(619.07±42.25)pg/ml] (P<0.01).

CONCLUSIONSIn this acute pancreatitis model, the peak levels of plasma TAP and serum IL-6 may appear earlier in rats with severer disease. Serum TAP level may be used as a marker for the accurate early prediction of the severity of acute pancreatitis.

Animals ; Biomarkers ; blood ; Disease Models, Animal ; Interleukin-6 ; blood ; Male ; Oligopeptides ; blood ; Pancreatitis, Acute Necrotizing ; blood ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate the antioxidant activities of different chemical constituents from Astragalus mongholicus Bunge and their protection against xanthine (XA)/xanthine oxidase (XO)-induced toxicity in PC12 cells.

METHODSThe compounds of Astragalus mongholicus Bunge were isolated by chromatography and the structures were elucidated on the basis of spectral data interpretation. Their antioxidant activities were detected by 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities in a cell-free system. Meanwhile, the effects against XA/XO-induced toxicity were assessed using MTT assay in PC12 cells.

RESULTSTen principal constituents were isolated and identified as formononetin (I), ononin (II), calycosin (III), calycosin-7-O-beta-D-glucoside (IV), 9,10-dimethoxypterocarpan-3-O-beta-D-glucoside (V), adenosine (VI), pinitol (VII), daucosterol (VIII), beta-sitoster (IX) and saccharose (X) from Astragalus mongholicus Bunge. The compounds I, III, and IV scavenged DPPH free radicals in vitro. Formononetin and calycosin were found to inhibit XA/XO-induced cell injury significantly, with an estimated EC50 of 50 ng/mL.

CONCLUSIONCompound II, VI, and VII are first reported in this plant. Calycosin exhibits the most potent antioxidant activity both in the cell-free system and in the cell system.

Animals ; Astragalus Plant ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Free Radical Scavengers ; chemistry ; pharmacology ; Free Radicals ; metabolism ; Isoflavones ; chemistry ; pharmacology ; PC12 Cells ; Rats ; Xanthine ; toxicity ; Xanthine Oxidase ; toxicity

OBJECTIVETo investigate the effect of ischemic postconditioning on the expression of rat myocardium matris metalloproteinase-2 (MMP-2) induced by ischemia/reperfusion (I/R) and relationship between its expression and interstitium and the effect on left ventricular function.

METHODSTwenty-four rats were randomly divided into 3 groups (n = 8): sham control (SC) group, ischemic/reperfusion (I/R) group and ischemic postconditioning (IPTC) group. The left ventricular peak systolic pressure and its derivate (+/- dp/dt) were calculated; The amount of myocardium collagenous were determined; The vitality of superoxide dismutase (SOD) and content of malondialdehyde (MDA) of plasma were detected; The activity of myocardium MMP-2 was measured by Western blot and RT-PCR.

RESULTSAs compared with I/R group, IPTC could lower the expression of MMP-2, ameliorate left ventricular function and increase the content of myocardium collagenous. In the meantime, the vitality of superoxide dismutase (SOD) of plasma were greatly enhanced and the content of malondialdehyde (MDA) of plasma were reduced in IFC group.

CONCLUSIONProtective effect of IPIC on myocardium may be due to reduce free radical, lower expression of MMP-2 and protect myocardial interstitium. MMPs plays an important role in the myocardial protection provided by IPTC.

Animals ; Collagen ; metabolism ; Ischemic Postconditioning ; Malondialdehyde ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Myocardial Reperfusion Injury ; enzymology ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism