To establish neonatal rat cardiac fibroblast inflammatory secretion model by using LPS 100 microg x L(-1) combined with ATP 5 mmol x L(-1), in order to study the inhibitory effect of quercetin on the secretion of inflammatory factors TNF-alpha, IL-1beta and IL-6 of cardiac fibroblasts, further investigate the effect of quercetin on the protein expression of p-NF-kappaB p65 (S276) and p-Akt (S473) by western blot, and discuss the inhibitory effect of quercetin on the inflammatory secretion of cardiac fibroblasts. According to the findings, quercetin with the concentrations between 51.74 micromol x L(-1) and 827.81 micromol x L(-1) had no significant effect on the activity of cardiac fibroblasts. Quercetin with the concentrations of 82.78, 41.39, 20.70 micromol x L(-1) could notably inhibit the increase of TNF-alpha and IL-1beta induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 36 h (P < 0.01 or P < 0.05). Quercetin with the concentrations of 82.78, 41.39 micromol x L(-1) could notably inhibit the increase of IL-6 induced LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 36 h (P < 0.05), without any notable effect of quercetin with the concentration of 20.70 micromol x L(-1). Quercetin with the concentrations of 82.78, 41.39, 20. 70 micromol x L(-1) could notably inhibit the NF-kappaB p65 (S276) activation induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 15 min, with the most significant effect in 20.70 micromol x L(-1). Quercetin with the concentrations of 82.78, 41.39, 20.70 micromol x L(-1) could notably inhibit the increase of p-Akt(473) expression induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 240 min (P < 0.05). Therefore, this study believes that quercetin could attenuate the secretion of inflammatory factors TNF-alpha, IL-1beta and IL-6 of cardiac fibroblasts by inhibiting the activation of NF-kappaB p65 (S276) and Akt (473).
Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; Endomyocardial Fibrosis ; drug therapy ; genetics ; immunology ; Female ; Fibroblasts ; drug effects ; immunology ; Heart ; drug effects ; Humans ; Interleukin-6 ; genetics ; immunology ; Male ; Proto-Oncogene Proteins c-akt ; genetics ; immunology ; Quercetin ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; genetics ; immunology

AIM : To construct and analyze eukaryotic expression plasmid inserted by Cx50 with V64G mutation through bioinformatics software.METHODS: The full coding domain sequence of Cx50 with V64G mutation was acquired from the blood of patients with cataract and was cloned into pcDNA3.1 /Amp (+).The constructed plasmid was identified with PCR , enzyme digestion and sequencing. The analysis of Cx50 with V64G mutation was performed with bioinformatics software.RESULTS : Cx50 with V64G mutation was successfully amplified and its eukaryotic expression plasmid was constructed. Valine-64 is well conserved in the first extracellular loop of connexin 50 in different species and also in different human α -type gap junctional proteins.CONCLUSION : The successive reconstruction and verification of eukaryotic expression plasmid containing Cx50 with V64G mutation established the foundation for further studying the mechanism of cataract.

Objective To review the clinical diagnosis and treatment of acute focal renal infarction. Methods Three cases of focal renal infarction were reported and the literature was reviewed.The patients aged from 45 to 63 years with mean age of 54. Two cases had low back pain, 1 case with abdominal pain. Based on clinical history, B-ultrasonography and CT scan, focal renal infarction was diagnosed in 3 patients. There were 2 cases on left kidney and 1 case right. All cases were applied digital subtraction angiography (DSA) and thrombolytic anticoagulant therapy. Results Two cases received DSA and thrombolytic therapy. The other one case received pethidine 50 mg, progesterone 20 mg treatment, the salvia infusion and low molecular heparin 6000 U anticoagulant therapy. All patients had symtoms relieved after 1 d. A week later CT scan, 3 cases of renal infarction were apparently disappeared. Serum creatinine and urea nitrogen were normal. Three patients were followed, mean follow-up time was 1. 5 (0. 5-2) years. Conclusions The diagnosis of acute focal renal infarction mainly depends on B-ultrasound and CT. Early diagnosis and treatment is important for achieving recovery of the compromised renal function. Renal infarction should be suspected in the presence of abdominal pain of sudden onset.

Objective To study the decomposition kinetics of omethoate in blood.Methods The acetonitrile precipitated protein was added into the blood, with the chromatographic column of a Waters BEH C18column (2.1 mm×50 mm, 1.7μm), the mobile phase of 5 mmol/L ammonium acetate aqueous solution-methanol, and the gradient elution with a flow rate of 0.3 mL/min and injection volume of 2μL.With electrospray ionization (ESI) source and positive ion detection, qualitative and quantitative analyses were taken using multi-reaction monitoring mode.Omethoate standard was added into blank human blood to the mass concentrations of 0.78, 1.40, 2.30, 4.50, and 7.20μg/mL, and each mass concentration was preserved at 3 temperatures of-20℃, 4℃, and 20℃, respectively.The content of omethoate was detected at different time points (0, 1, 3, 4, 7, 11, 15, 24, 32, 40, 48, 64, 80, 96, and 120 d).Results Different concentrations of omethoate all showed a descended trend in human blood under different temperature conditions.The decomposition in storage environment of-20℃, 4℃, and 20℃was fit to a one-compartment open model with a first-order kinetic process, which could be expressed as Ct=Coe-αt, with the calculated theoretical values of omethoate concentration close to the measured values.Conclusion All concentrations of omethoate are decomposed in the blood, which vary a lot in different preservation conditions.It is suggested that blood samples should be frozen and detected timely in suspected omethoate poisoning cases.

BACKGROUND: Studies have shown that bone marrow mesenchymal stem cell (BMSC) transplantation can effectively improve cardiac function after myocardial infarction. However, few reports have been issued on myocardial electrophysiology after BMSCs transplantation. OBJECTIVE: To observe the effects of BMSCs transplantation on voltage-gated K+channel protein and myocardial infarction-related cytokines, thereby providing basic evidence for further exploration on the mechanism underlying arrhythmia in myocardial infarction due to BMSCs transplantation. METHODS: Forty male Wistar rats, SPF grade, were randomly divided into four groups: sham group, model group, cell culture medium group and BMSCs group. The myocardial infarction model was created in rats by permanent ligation of the left descending coronary artery. At 15-20 minutes after surgery, BMSCs (100 μL, 1×106) or cell culture medium (100 μL) was injected at four sites in the peri-infarct zone. Four weeks after cell therapy, cardiac samples were taken, the pathological morphology of the infarcted myocardium was observed by hematoxylin-eosin staining, and the infarct size was calculated; the expression levels of voltage-gated K+channel proteins Kv1.2 and Kv1.5 and cardiac troponin T (cTnT) were measured by western blot assay; and the expression levels of apoptotic factor (Caspase-3), autophagy factor (Bcl-2), nitric oxide and superoxide dismutase were tested by immunohistochemistry. RESULTS AND CONCLUSION: Compared with the model group and cell culture medium group, the infarct size decreased in the BMSCs group (P < 0.05); the expression levels of cTnT, Kv1.5 and superoxide dismutase increased (P < 0.05), and the expression levels of Caspase-3, Bcl-2 and Kv1.2 decreased (P < 0.05) in the BMSCs group. In summary, BMSCs transplantation can promote the expression of voltage-gated K+channel proteins, and improve anti-oxidation capacity of the myocardium and decrease apoptosis and autophagy.

Objective To investigate the difference of late-phase of limb ischemia preconditioning (L-LIP) verse early-phase (E-LIP) on patients with percutaneous coronary intervention (PCI).Methods A total of 160 patients with unstable angina pectoris who were planned to undergo PCI were divided equally into two groups at random.The late-phase of limb ischemia preconditioning group (80 patients) were provided with L-LIP (three 5-minute inflations up to 200mmHg by applying the sphygmomanometer cuff around the right upper arm,followed by 5-min intervals of reperfusion,twice a day) 3 days before PCI.The Earlyphase of limb ischemia preconditioning group (80 patients) were provided with E-LIP (method as above)2 hours before PCI.Comparison of procedural parameters during PCI and the levels of cTnT,CK-MB,hs-CRP were made 24 hours after PCI.Estimation of the rate of adverse events at 1 year between the two groups was evaluated by Kaplan-Meier analysis.Results Compared to the E-LIP group,the rates of angina,arrhythmia and TIMI flow ≤ 2 during PCI were significantly lower in the L-LIP group (all P ＜ 0.05).At 24 hours after PCI,the levels of cTnT and CK-MB were declined more significantly in the L-LIP group[(11.52±2.41) pg/ml vs.(27.53±4.78)pg/ml,P =0.021;(14.11±2.87)Iu/L vs.(30.23±5.17)Iu/L,P =0.032].There was no difference in the level of hs-CRP between the 2 groups [(128±0.71)mg/dl vs.(1.33±0.69)mg/dl,P =0.742].The Kaplan-Meier survival curve showed that the incidence rate of adverse events in the L-LIP group at l year was lower than the E-LIP group (3.75％ vs.13.75％,P =0.024).Conclusions L-LIP is more effective to in protecting myocardial cell in patients with unstable angina pectoris undergoing elective PCI and may reduce the rate of future adverse event.

Coronary Artery Disease ; physiopathology ; Fistula ; physiopathology ; Heart Ventricles ; Humans ; Iatrogenic Disease ; Male ; Middle Aged ; Percutaneous Coronary Intervention ; adverse effects

Objective To study the non-fastidous antibiotic-resistance bacteria pollution in tap water from a neighborhood in Tianjin.Methods Tap water samples were collected and bacteria were isolated by R2A agar,and 16S rRNA gene sequence for the identification of bacteria isolates was used.Antibiotic susceptibility testing was performed using the Kirby-Bauer disc diffusion method.Results Among the 39 non-fastidous bacteria isolates,the resistance rate of antibioticresistance bacteria was 79.49％,including Enterococcus,Staphylococcus,Acinetobacter and Pseudomonas.Some bacteria (28.21％) showed resistance against only one antibiotic.The others (51.28％) were multiple resistant bacteria.Trimethoprim/sulfamethoxazole resistance was most prevalent (53.85％),and sulfamethoxazole resistance was also widely distributed (28.21％).Conclusion Tap water from this neighborhood is heavily polluted by non-fastidous antibioticresistantance bacteria,which deserves more attention.

OBJECTIVETo establish a new culture system for mouse tooth germs in chick embryo.

METHODSThe mandibular first molar germ fragments of 15 embryonic days' Kunming mouse embryo were implanted into the lateral mesenchyme of 4-5 days' chick embryo wing buds in ove. Eggs were reincubated and implanted tissues were examined by histochemistry.

RESULTSThe cultured tooth germ development continued from cap stage to latest bell stage. The ameloblast and the odontoblast all differentiated maturely and secreted matrix.

CONCLUSION4-5 days' wing buds chick embryo could serve as developing the mouse tooth germs and demonstrate well physiological process of differentiation and morphogenesis.