Objective To investigate the role of plasma tissue factor (TF) and tissue factor pathway inhibitor-1 (TFPI-1) level and to observe the effect of extrinsic TFPI-1 on no-reflow (NR) in a rabbit model of ischemia/reperfusion. Methods Rabbits were randomized into four groups (n = 10 each): ischemic- reperfusion group (IR, subjected to 120 minutes of coronary artery occlusion and followed by 60 minutes of reperfusion); ischemic- reperfusion TFPI-1 group (100 ng/kg bolus and 1 ng · kg~(-1) · min~(-1) infusion during reperfusion) ; ischemic group (subjected to 180 minutes of coronary artery occlusion) and sham group. The NR area and ischemic area were determined by thioflavin S and Evan's blue staining in vivo. Plasma TF and TFPI-1 levels were measured before operation, before and at 120 minutes post coronary artery ligation, 10 and 60 minutes after reperfusion by ELISA. Results Plasma TF and TFPI-1 levels before and at 120 minutes post coronary artery ligation were similar among the four groups (all P > 0.05). At 10 and 60 minutes after reperfusion, the plasma TF levels in the IR group was significantly higher than those in ischemic group and sham group [10 minutes: (20.7 ±4. 1) pg/ml vs. (13.9 ±2. 2)pg/ml(P <0. 001), (20.7±4. l)pg/ml vs. (13.2±2.6) pg/ml(P<0. 001); 60 minutes; (15.8±2.6) pg/ml vs. (13.5± 1.6) pg/ml(P<0.05), (15.8 ±2.6) pg/ml vs. (12.1 ±0.7) pg/ml (P < 0. 001)] while the plasma TFPI-1 levels were similar among IR, ischemic and sham groups at 10 minutes after reperfusion and at 60 minutes after reperfusion (all P >0. 05). TFPI-1 level [(9.7 ± 1. 6) ng/ml] was significantly lower in the IR group than in the ischemic group [(11.6 ±1.6) ng/ml, P < 0. 05] and sham group [( 10. 1 ±1.3) ng/ml, P < 0. 01] . TF mRNA expression in the NR area in IR group was significantly up-regulated compared to the ischemic group (P<0. 05) and sham group (P <0. 001 ) while TFPI-1 mRNA expression was similar between IR group and ischemic group ( P > 0. 05 ) . NR severity in the ischemic-reperfusion TFPI-1 group was significantly attenuated compared to IR group (0. 39 ±0. 11 vs. 0.54±0.06, P<0.01). Conclusion Upregulated TF mRNA expression in the NR area and increased plasma TF level during reperfusion period, reduced plasma TFPI-1 level during reperfusion period as well as attenuated NR severity by extrinsic application of human rTFPI-1 in this model suggested an important role in the pathogenesis of the NR phenomenon.

Objective To observe the protein and gene expression of immunoglobulin binding protein (BiP) in the femur of fluoride-treated rats, and preliminarily study the possible role of endoplasmic reticulum stress in the pathogenesis of skeletal fluorosis. Methods Sixty Wistar rats were divided into 4 groups according to body weight, n =15. The control and low-calcium groups were fed with normal diet(0.79％ calcium) and low-calcium diet(0.063％ calcium), respectively, and both drank tap water(fluoride concentrations ＜ 1 mg/L). High-fluoride and coexpesure to low-calcium groups were fed with conventional feed (0.79％ calcium) and low-calcium diet (0.063％ calcium), respectively, and both drank tap water containing sodium fluoride (sodium fluoride concentration of 221 mg/L). During experimental period, rats were measured body weight once a week with a stand diet and water available ad libitum. The experiment lasted for 12 weeks. The immunohistochemical and reverse transcription polymerase chain reaction(RT-PCR) techniques were used to detect the protein and gene expression of BiP in the femur of fluoride-treated rats and control subjects. Results The bone mineral contents of high fluoride, lowcalcium and coexposure groups[(0.131 ± 0019), (0.097 ± 0.011 ), (0.083 ± 0.007)g/cm] were lower than those of the control group[(0.159 ± 0.029)g/cm, all P ＜ 0.05]; the bone mineral density of low calcium and coexpesure to fluoride group[(0.243 ± 0.018), (0.223 ± 0.022)g/cm2] was lower than that of the control group[(0.296 ± 0.046)g/cm2, all P ＜ 0.05]. The immunohistochemical staining showed that the anti-BiP antibody positive osteoblasts were significantly increased in the low calcium diet and coexposure to fluoride groups than that in the control, and coexposure to fluoride elevated the positive cells than that in only low calcium diet group. The mRNA expression of osteopontin(OPN) and osteocalcin(OCN) in coexposure to fluoride with low-calcium group(1.36 ± 0.20, 1.31 ±0.11 ) was higher than that of the control groups (0.82 ± 0.16, 0.85 ± 0.15, all P ＜ 0.05) ; moreover, OPN expression significantly increased in this group than that of the only high fluoride group (0.97 ± 0.29, P ＜ 0.05). The mRNA expression of BiP in the low calcium and coposure to fluoride group (1.38 ± 0.24,1.35 ± 0.12) was significantly higher than that of the control group ( 1.14 ± 0.06, all P ＜ 0.05 ). Conclusions Higher fluoride or coexposure to low calcium diet stimulates the gene and protein expression in rat femur BiP, indicating that varying degrees of endoplasmic reticulum stress is likely involved in the pathogenesis of rat skeletal fluorosis.

OBJECTIVE To study the mechanism of loganin,the effective componet of Cornus officinalis,on protecting the glomerulus mesangial cell (GMCs) injured by advanced glycation end products (AGEs).METHODS GMCs were divided into control,model (AGEs: 200mg/L),loganin (0.1,1,10μmol/L) groups.The cell proliferation,FN and COL-Ⅳ secretion,microstructure lesions,RAGE,SphK1,S1P and TGF-β protein expressions were measured.RESULTS AGEs induced cell proliferation,FN and COL-Ⅳ secretions,microstructure lesions,RAGE,SphK1,S1P and TGF-β protein expressions were suppressed by loganin.CONCLUSION Loganin can improve the AGEs induced GMCs injury by regulating the AGEs/RAGE/SphK1 signal pathway,and then ameliorate the diabetic nephropathy.

Objective To explore the distribution of arsenic speciafion and to estimate the effect of arsenic on glutathione(GSH)levels in the blood and liver of mice exposed to different concentrations of inorganic AsⅢ through drinking water.Methods Mice drank water containing arsenite at concentrations of iAsⅢ of 0(contr01),25,50,100 ms/L for 6 weeks.Blood and liver were sampled to asses$the levels of inorganic arsenic(iAs),monomethylarsenic acid(MMA),dimethylarsenic acid(DMA)by the method of hydride generation trapping and ultra-hypothermia coupled with atomic absorption spectrometry,and the level of GSH by the method of 5,5'-Dithio-bis (2-Nitrobenzoic acid).Results Leveh of iAs.MMA and DMA in blood and in liver increased along with the increase of iAs concentrations in drinking water.Primary methylated index(PMI)and secondary methylation index (SMI)of liver and blood were significantly higher in exposed groups than those in control group(P＜0.05).SMI of liver in 50 mg/L exposed group[(50.45±2.94)%]was significantly higher than those in 25 mg/L and 100 mg/Lgroups[(41.68±7.09)%and(41.19±8.87)%,respectively],the difference being statistically significant(P＜0.05).The ratio of iAs.MMA and DMA in blood and liver in exposed group were 2:3:5 and 4:3:3,the percentage of level of organic arsenic(MMA+DMA)were 80%and 60%.GSH in blood and liver in exposed group decreased along with iAs concentrations in drinking water and had significant differences compared with those in control group (P＜0.05).However,levels of GSH in liver and blood did not differ significantly between exposed groups and control group(P＞0.05).Conclusions Membolism of iAs in liver is maximized when the iAs concentrations in drinking water increases to a certain level.However,the percentage of arsenic speciation in blood is different from that in liver,suggesting that other organs and tissues may be capable of methylation of inorganic arsenic.The level of GSH in liver and blood in mice is a good mark tO reflect the toxicity of arsenic.

Background and purpose: Cervical conization is a common operation to treat precancerous tissues performed under non-intubated anesthesia. As common opioid analgesics have side effects of inhibiting respiration and circulation, other kinds of analgesic drugs should be coordinated to improve the anesthetic effect, without interfering the respiration and circulation. This study aimed to evaluate the effects of dezocine or flurbiprofen combined with propofolremifentanil in cervical precancerosis conization. Methods: Sixty patients who underwent cervical conization were equally randomized into dezocine group (group D), flurbiprofen group (group F) and 0.9％ natural saline (group N) with 20 patients in each group, and received dezocine 0.1 mg/kg, flurbiprofen 1 mg/kg or 0.9％ natural saline in 5 mL respectively before anesthesia induction. During the anesthesia induction, the targeted control infusion of remifentanil in effect concentration was set at 1.5 ng/mL, and the plasma concentration of propofol was set at 2 μg/mL. Heart rate (HR), respiratory rate (RR), surplus pulse O2 (SPO2) and mean arterial pressure (MAP), MAP were monitored before the anesthesia induction (T0) and after (T1), at the start of cervical conization (T2), and at the end of operation (T3). The incidence of respiratory depression and body movements during surgery were observed. The satisfaction degree of the surgeon to the opening status of cervix was evaluated. The post-operative recovery time, visual analogue scale (VAS) scores, nausea and vomiting in the following 12 hours were also recorded. Results: The HR, RR, SPO2 and MAP in three groups did not have any significant change (P＞0.05) at T0, T1 and T3. At T2 the HR and MAP decreased significantly in group D and group F compared with group N (P＜0.05), and there was no significant difference between group D and group F (P＞0.05). The surgical satisfaction degree of "Good" in group D was 80％, significantly higher than that in group N (30％) and group F (50％), indicating a better cervix opening in group D. The recovery time in three groups had no significant difference, and the VAS scores in group D and group F were lower than those in group N (P＜0.05) after operation, and patients did not have nausea or vomiting in the following 12 hours. Conclusion: Both the dezocine and flurbiprofen could improve the anesthetic effect in cervical conization and post-operative comfort, with less respiratory or circulation depression. Dezocine showed better improvement than flurbiprofen in cervix opening and the inhibition of stress response and body movements during surgery.

Objective To explore the correlations of biochemical factors as gamma-aminobutyric acid (GABA) level with amino acid metabolism level in the blood, levels of intestinal immunoglobulin A (IgA)+complement factor 3 (C3), feeding intolerance and fatiguability in children with hypotonia cerebral palsy. Methods Ninety-six children with hypotonia cerebral palsy,admitted to our hospital from January 2009 to January 2012,were chosen in our study; questionnaire was used to understand the clinical symptoms of the children; the blood ammonia level,hepatic function and IgA+C3 level were obtained from routine blood chemistry testing, and amino acid metabolism was detected by blood tandem mass spectrometry; the correlations of GABA level with amino acid metabolism level in the blood, levels of IgA+C3, feeding intolerance and fatiguability were statistically analyzed. Results In all the 96 children with hypotonia cerebral palsy,63 (65.63％) had low arginine; 52 had both decreased arginine and elevated blood ammonia levels, enjoying negative correlation (r=-0.776,P=0.000); 42 had decreased arginine and reduced levels of IgA+C3 enjoying positive correlation (r=0.351,P=0.000); both decreased arginine level and feeding intolerance were noted in 47 with positive correlation (r=0.372,P=0.000).In these 96 children,30 (31.25％) had carnitine metabolism abnormality,including decreased propionyl carnitine/free carnitine or propionyl carnitine/acetylcarnitine levels in 21 (21.88％),increased hydroxyl palmitoyl carnitine/hydroxyl Palm enoyl carnitine level in 9 (9.37％),and decreased cysteine content in 3 (3.12％). Conclusion Metabolic abnormalities of arginine,carnitine and cysteine are noted in children with hypotonia cerebral palsy; a lot of exercise will consume arginine,carnitine and cysteine,which causes fatigue; children with low blood arginine content might also have increased blood ammonia level,reduced IgA+C3 level,trends of vomiting,susceptibility to infection and feeding difficulties,and therefore,children's mental state,immune function and exercise tolerance ability are affected.

Objective To investigate the relationship between the patient-based questionnaires and the computed tomography (CT) staging in patients with chronic rhinosinusitis (CRS). Methods Quantitative data of 121 preoperative recruits with CRS were collected by using the Lund-Mackay CT staging system, a visual analogue scale ( VAS), sino-nasal outcome test-20 ( SNOT-20), and the medical outcome study short-form 36 items (SF-36). The patients were classified into several subgroups according to whether CRS was associated with nasal polyps (NP) or not, sex, duration of disease, and educational background.Correlation between the patient-based questionnaires and the CT staging were analyzed in the total cohort patients and subgroups. Results In the total cohort patients, there were significant correlations between SNOT-20 and SF-36 ( r = -0. 561, P ＜ 0. 01 ), SNOT-20 and VAS ( r = 0. 743, P ＜ 0. 0l ), and SF-36 and VAS ( r = - 0. 504, P ＜ 0. 01 ), however, the CT staging did not correlate with the patient-based questionnaires (P ＞ 0. 05 ). Significant but weak correlations were found between the CT staging and the patient-based questionnaires in the C RS with NP subgroup (CT vs SNOT-20, r = 0. 318, P = 0. 005; CT vs SF-36, r = - 0. 358, P = 0. 002; CT vs VAS, r = 0. 358, P = 0. 002). Compared between CRS with NP and without NP subgroup, there were statistic differences on the Lund-Mackay CT stage and the SNOT-20 and VAS scores (t value was 3.249, -2.409, -2.957, respectively, all P＜0.05). Conclusions The patient-based questionnaires correlate well with each other, but CT staging correlated significantly but weakly with the patient-based questionnaires only in the CRS with NP subgroup. Nasal polyps do not appear to be responsible for the adverse effects of CRS on quality of life.

Objective To evaluate the optimal administration regimen of flomoxef for the bacterial infection based on pharmacokinetic/pharmacodynamic (PK/PD) models.Methods A literature search was conducted in PubMed to capture the pharmacokinetic data of flomoxef.Minimum inhibitory concentrations (MICs) were determined using two-fold agar dilution method.The percent time that drug concentration exceeds the minimum inhibitory concentration (％fT＞MIC) was used as the PK/PD indicator correlated with efficacy.Monte Carlo simulation was employed to determine the appropriate regimens of flomoxef based on the probability of target attainment (PTA) against the clinical isolates of strains.Results The regimens of 1 g q6 h,1 g q8 h and 1 g q12 h with 1 hour infusion at 70％ of ％ fT＞MIC achieved 93.1％,89.1％ and 66.8％ of PTA against Escherichia coli (ESBL +),respectively.For the Klebsiella pneumoniae (ESBL+),these regimens achieved 81.8％,78.7％ and 62.3％ of PTA at ％fT＞MIC =70％.The regimen of 2 g q12 h achieved the similar PTA as 1g q6 h and 1 g q8 h at 50％ and 70％ of ％fT＞MIC,but not at higher ％ fT＞MIC.Furthermore,flomoxef also showed potent bactericidal activity against Escherichia coli (ESBL-),Klebsiella pneumoniae (ESBL-),methicillin-susceptible S.aureus (MSSA),methicillin-susceptible S.Epidermidis (MSSE) and Moraxella catarrhalis,etc.with all dosing regimens according to the PK/PD analysis.Conclusion As a time-dependent antibiotic,the clinical outcome of flomoxef can be improved by shortening dosing interval,extending infusion time and/or increasing dose.The first two strategies played more significant roles.

Objective To investigate the effects of tetramethylprazine (TMP) on left ventricular hypertrophy and the regulation of nicotinamide-adenine dinucleotide(NADPH) oxidases in renovascular hypertension of rat.Methods Hypertensive rats were obtained by using two-kidney-two-clip (2K2C) method.Rats were classified into sham-operated group,model group,low,high dose experimental groups (TMP,30,60 mg · kg-1 · d-1) and control group (candesartan,10 mg · kg-1 · d-1).The drugs were continuously administered for 2 weeks.Left ventricular (LV) hemodynamic parameters,serum superoxide ismutase (SOD),dmalondialdehyde (MDA) and hydrogen peroxide(H2O2)levels were measured.The ratio of LV weight to body weight (LVW/BW) was calculated.Hematoxylin-eosin(HE) staining was used for morphological analysis.RT-polymerase chain reaction and Western-blot were performed to investigate the protein expressions of brain natriuretic peptide (BNP),β-myosin heavy chain(β-MHC) and NADPH oxidases(Nox2,Nox4 and P47phox).Results Compared with model group,aortic systolic pressure (AoSP),aortic diastolic pressure (AoDP),LV end-systolic pressure (LVESP),LV end-diastolic pressure(LVEDP) significantly decreased in low,high dose experimental groups with siginificanly (all P ＜ 0.05) while (+ dp/dtmax) and (-dp/dtmax) increased in low,high dose experimental groups with siginificanly (P ＜ 0.05).Compared with model group on LVW/BW (2.8 ± 0.3) mg · g-1,LVW/BW of (1.9 ± 0.2),(1.7 ± 0.4) mg · g-1 in low,high dose experimental groups significantly decreased (P ＜ 0.05).Compared with model group SOD significantly increased while MDA and H2O2 decreased in low,high dose experimental groups with significandy (all P ＜ 0.05).The protein expressions of Nox2 in low,high experimental groups and model group were 0.82 ± 0.08,0.57 ± 0.05,0.30 ± 0.13;the protein expressions of Nox4 in above three groups were 1.00±0.10,0.79 ± 0.07,1.32 ± 0.15 and the protein expressions of P47phox in above three groups were 0.88 ± 0.08,0.65 ± 0.06,1.23 ± 0.15,compared with model group,the protein expression significantly decreased in low,high dose experimental groups with significantly (all P ＜ 0.05).Conclusion TMP attenuated cardiac hypertrophy possibly through decreasing NADPH oxidases (Nox2,Nox4,P47phox) expressions and oxidative stress in renovascular hypertensive rats.

Background It has been proven that ultrasonic destruction of microbubbles can enhance gene transfection efficiency into the noncardiac cells, but there are few reports about cardiac myocytes. Moreover, the exact mechanisms are not yet clear; whether the characteristic of microbubbles can affect the gene transfection efficiency or not is still controversial.This study was designed to investigate whether the ultrasound destruction of gene-loaded microbubbles could enhance the plasmids carried reporter gene transfection in primary cultured myocardial cell, and evaluate the effects of microbubbles characteristics on the transgene expression in cardiac myocytes.Methods The β-galactosidase plasmids attached to the two types of microbubbles, air-contained sonicated dextrose albumin (ASDA) and perfluoropropane-exposed sonicated dextrose albumin (PESDA) were prepared. The gene transfection into cardiac myocytes was performed in vitro by naked plasmids, ultrasound exposure, ultrasonic destruction of gene-loaded microbubbles and calcium phosphate precipitation, and then the gene expression and cell viability were analyzed.Results The ultrasonic destruction of gene-loaded microbubbles enhanced gene expression in cardiac myocytes compared with naked plasmid transfection ((51.95±2..41) U/g or (29.28±3.65) U/g vs. (0.84-0.21) U/g, P ＜0.01), and ultrasonic destruction PESDA resulted in more significant gene expression than ASDA ((51.95e2.41) U/g vs. (29.28±3.65)U/g, P ＜0.05). Ultrasonic destruction of microbubbles during calcium phosphate precipitation gene transfection enhanced 3-galactosidase activity nearly 8-fold compared with calcium phosphate precipitation gene transfection alone ((111.35±11.21) U/g protein vs. (14.13±2.58) U/g protein, P＜0.01). Even 6 hours after calcium phosphate precipitation gene transfection, ultrasound-mediated microbubbles destruction resulted in more intense gene expression ((35.63±7.65)U/g vs. (14.13±2.58) U/g, P＜0.05 ).Conclusions Ultrasonic destruction of microbubbles might be a promising method for the delivery of non-viral DNA into cardiac myocytes, and the gene tranfection is related to the characteristics of microbubbles.