BACKGROUND: As a common anticoagulant, heparin is widely used in clinic, but it has remarkable side effects such as severe bleeding and heparin-induced thrombocytopenia, and it cannot inactivate fibrin-bound thrombin. Annexin Ⅴ derivative (AND) is inosculated with C-terminal of hirudin and annexin Ⅴ, and its anticoagulation and anti-thrombosis effects are compared with those of heparin. OBJECTIVE: To investigate the relationship between quantitative effectiveness and time effectiveness of AND on coagulation and thrombosis, and study its reliability. DESIGN: Completely randomized grouping design and controlled study. SETTING: Cardiac Department of amunicipal hospital. MATERIALS: The experiment was conducted at the Animal Laboratory of Jiangsu Provincial People's Hospital from July 2000 to April 2001. Totally 32 male New Zealand white rabbits were randomly divided into 4groups, namely, high dosage AND group, low dosage AND group, common heparin group and saline group with 8 in each group. METHODS: Heparin and AND were diluted with saline.①High dosage AND group: 0.7 mg/kg AND was injected intravenously and followed by intravenous dripping of 0.35mg/(kg ·h)for 2 hours.Low dosage AND group: 0.3 mg/kg AND was injected intravenously and followed by intravenous dripping of 0.15 mg/(kg·h) for 2 hours. Heparin group: 75 IU/kg heparin was injected intravenously and followed by intravenous dripping of 37.5 IU/(kg·h) for 2 hours. Saline group: The same volume of saline and medication were used as those in drug groups.② Blood sample was collected from the femoral vein before administration so as to test blood routine, activated partial thromboplastin time(APTT)and prothrombin time (PT) after 15-, 30- and 60-minute administration and 2-hour withdrawal.③Saccule was separated from endothelium of femoral artery to measure blood pressure of distal femoral artery at 15 minutes after administration.Time of pulse pressure equal to 0 mmHg was recorded when the vessel was occluded completely by a thrombus.Finally the injured femoral arteries whose vessel was stripped were collected to measure its length, wet weight and dry weight. ④Observation of AND toxicity and sideeffects:During the experiment,vital signs of the animals were measured,such as blood pressure,heart rate and breath;in addition,bowelhemorrhage was observed and the number of leucocytes was counted after dissection of some of the animals. MAIN OUTCOME MEASURES:①Effect of AND on blood coagulation system and arterial thrombosis.②AND toxicity and side effects. RESULTS: All the 32 white rabbits entered the final analysis. ① Anticoagulant effect: APTT: Fifteen minutes after administration, APTT in AND group was the longest,which was(136.86±39.46)s in high dosage AND group and (122.90±34.19) s in low dosage ANDgroup.Moreover, APTT was longer than that in saline group [(95.14±24.64) s], but shorter than that in common heparin group [(180.00±0.00) s, P ＜ 0.05, 0.01]. At 30 minutes after administration,AND in high dosage group still had coagulation,and APTT was (124.61±40.19) s in high dosage group, which was longer than that in saline group [(85.57±27.67) s], but APTT was (112.94±43.17) sin low dosage group,which was shorter than that in common heparin group [(85.57±27.67)s,P ＜ 0.05].APTT was shorter in high and low dosage groups than in common heparin group at 60 minutes after administration (P ＜ 0.05),and longer than that in saline group 2 hours after drug withdrawal,but there was not significant difference(P ＞ 0.05).PT:PT in common heparin group was longer than that in high and low dosage groups at 15,30 and 60 minutes after administration (P ＜ 0.05).② Effect on arterial thrombosis:Wet weight of thrombus:It was lighter in AND group than in common heparin group(P ＜ 0.05). Dry weight of thrombus:Thrombus was lighter in high and low dosage groups than in common heparin group, and was lighter in high dosage group than in low dosage group (P ＜ 0.05).Thrombus length:It was shorter in low dosage group than in saline group (P ＜ 0.05), and shorter in high dosage groupthan in common heparin group (P ＜ 0.05). Time of complete occlusion: It was longer in high and low dosage groups than in saline group(P ＜ 0.05).③ AND toxicity and side effects:The behavior of rabbits in high and low dosage groups was similar to that in other two groups. Obvious hemodynamic changes were not found, and bowel hemorrhage was not observed, either. CONCLUSION: AND is an effective anticoagulant and anti-thrombosis agent; the highest anticoagulation effect occurs at 15 minutes afteradminis tration. However, the anticoagulant effect is poor as compared to heparin.The effect is poorer after 60-minute administration. Effect of AND on thrombus is stronger than that of heparin,but the size of thrombus is smaller than that of heparin, and the dosage-dependence manner was found. In addition, the anti-thrombus effect of AND is stronger in high dosage group than in low dosage group.

Adult ; Air Pollutants ; adverse effects ; Humans ; Male ; Military Medicine ; Monitoring, Physiologic ; Noise, Occupational ; adverse effects

BACKGROUND:Acellular nerve scaffolds have the three-dimensional structure of natural nerves and low immunogenicity,but their effect on long nerve defects is still not ideal.Therefore,it is necessary to construct tissueengineered nerve using acellular nerve and seed cells in order to improve the therapeutic effect.OBJECTIVE:To systemically review the efficacy of combination of acellular nerve grafts (ANGs) and mesenchymal stem cells (MSCs) or Schwann cells (SCs) transplantation in the treatment of sciatic nerve defects in a rat model.METHODS:Randomized controlled trials (RCTs) about the effects of combination of ANGs and MSCs or SCs transplantation for sciatic nerve defects in rats were searched in PubMed,The Cochrane Library,EMbase,CNKI,WanFang and VIP from inception to July 2016.Three reviewers independently screened literature according to the inclusion and exclusion criteria,extracted data,and assessed the risk of bias of included studies.Then,a Meta-analysis was performed using Review Manger5.3 software.RESULTS AND CONCLUSION:A total of 10 RCTs involving 252 rats were included.The results of meta-analysis showed that:compared with the control group (simple acellular nerve scaffold group),the sciatic functional index (SFI) of the combined group (combination of ANGs and MSCs or SCs transplantation) were superior at 2 weeks [SMD=2.73,95％ CI (1.92,3.45),P ＜ 0.000 01],4 weeks [SMD=4.57,95％ CI (3.43,5.70),P ＜ 0.000 01],6 weeks [SMD=1.62,95％CI (0.18,3.06),P=-0.03],8 weeks [SMD=4.90,95％ CI (2.96,6.84),P ＜ 0.000 01] after surgery.The nerve conduction velocity [SMD=1.39,95％ CI (0.99,1.78),P ＜ 0.000 01),latency period (MD=-0.98,95％ CI (-1.19,-0.76),P ＜ 0.000 01],and amplitude [SMD=1.23,95％ CI (0.62,1.85),P ＜ 0.000 1] were superior at 12 weeks after surgery.The myelin sheath thickness was superior at 8 weeks [MD=0.14,95％ CI (0.07,0.21),P ＜ 0.000 1],12 weeks [SMD=1.85,95％ CI (1.63,2.08),P ＜ 0.000 01] and the number of myelinated nerve fibers were superior at 12 weeks [SMD=3.59,95％CI (2.63,4.55),P ＜ 0.000 01] after surgery.The gastrocnemius wet weight was superior at 8 weeks after surgery [SMD=4.22,95％ CI (2.40,6.03),P ＜ 0.000 01].Current evidence indicates that the combination of ANGs and MSCs or SCs can promote the regeneration and functional recovery of the peripheral nerve.Due to the limited quality of the included studies,the above conclusion should be verified by conducting high-quality and large-scale RCTs.

OBJECTIVETo investigate the expression of zinc finger E-box binding homebox 1 (ZEB1) in the prepuce of hypospadias children and its relationship to the incidence of hypospadias.

METHODSPrepuce tissues were collected from 37 children aged 6-15 months undergoing hypospadias repair and 11 age-matched controls receiving circumcision. Based on the position of the urethral meatus, the hypospadias cases were classified as severe (n = 13) and mild-moderate (n = 24). The mRNA and protein expressions of ZEB1 were determined by immunohistochemistry and RT-PCR.

RESULTSThe expression of the ZEB1 protein was remarkably higher in the severe (100% [13/13]) and mild-moderate hypospadias patients (75.0% [18/24]) than in the controls (9.1% [1/11]), with statistically significant differences between any two groups (P < 0.05). RT-PCR showed the integrated density value (IDV) of the ZEB1 mRNA expression to be (0.67 ± 0.21), (0.81 ± 0.24), and (1.55 ± 0.29) in the control, mild-moderate, and severe hypospadias patients, respectively, significantly higher in the severe hypospadias than in the control and mild-moderate hypospadias groups (P < 0.05), but with no significant difference between the latter two (P = 0.64).

CONCLUSIONThe expression of ZEB1 is significantly increased in hypospadias patients, and its upregulation is positively correlated with the severity of hypospadias, which suggests that the overexpression of ZEB1 may contribute to the development of hypospadias.

Biomarkers ; metabolism ; Case-Control Studies ; Circumcision, Male ; Foreskin ; metabolism ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Hypospadias ; classification ; etiology ; metabolism ; Immunohistochemistry ; Infant ; Male ; Penis ; RNA, Messenger ; metabolism ; Transcription Factors ; genetics ; metabolism ; Up-Regulation ; Urethra ; Zinc Finger E-box-Binding Homeobox 1

To establish a LC-MS/MS method for quantification of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid in rats plasma and study its pharmacokinetics after administration of Mailuoning injection at a single dose to rats. Plasma samples were acidified with hydrochloric acid and extracted with ethyl acetate. The analytes were determined by LC-MS-MS using a ZOBAX SB C18 column with a mobile phase of methanol-water (containing 2 mmol x L(-1) ammonium acetic) (60:40)at a flow rate of 0.5 mL x min(-1) and detected using ESI with negative ionization mode. Ions monitored in the multiple reaction monitoring (MRM) mode were m/z 353.1/191.0 [M-H]- for chlorogenic acid, m/z 178.9/134.9 [M-H]- for caffeic acid, m/z 515.2/353.0 [M-H]-for 3,4-DCQA, m/z 193.0/133.9 [M-H]-for ferulic acid, m/z 146.9/102.9 [M-H]- for cinnamic acid and m/z 246.0/125.8 [M-H]- for tinidazole (IS). After administration of Mailuoning injection at a single dose to eight Sprague-Dawley rats, the concentrations of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid in plasma were determined by LC-MS/MS method. The main pharmacokinetics parameters of measured data were caluculated by using DASver 1.0 software. The linear concentration ranges of the calibration curves for chlorogenic acid, caffeic acid, 3,4-DCQA and cinnamic acid were 2.006-1,027 microg x L(-1) (r = 0.999 6), 1.953-1,000 microg x L(-1) (r = 0.999 7), 28.51-1.459 x 10(4) microg x L(-1) (r = 0.998 9), 1.836-940.0, g x L(-1) (r = 0.997 7) and 4.780-2,447 microg x L(-1) (r = 0.998 6) respectively. The inner and inter-days relative standard deviations were both less than 5.0%, indicating legitimate precise and accuracy to the requirement of biological sample analysis. For chlorogenic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (49.78 +/- 12.81) min, (123.55 +/- 14.82) mg x min x L(-1) and (0.004 3 +/- 0.000 5) L x min(-1), respectively. For caffeic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (36.65 +/- 10.59) min, (91.67 +/- 11.77) mg x min L(-1) and (0.005 7 +/- 0.000 7) L x min(-1), respectively. For 3,4-DCQA, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (50.08 +/- 13.78) min, (278.34 +/- 31.82) mg x min x L-1 and (0.001 6 +/- 0.000 2) L x min(-1), respectively. For ferulic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (51.39 +/- 15.52) min, (34.72 +/- 4.67) mg x min x L(-1) and (0.000 4 +/- 0.0001) L x min(-1), respectively. For cinnamic acid, the pharmacokinetic parameter t1/2, AUCo-t, and CL were (74.42 +/- 18.32) min, (34.63 +/- 4.82) mg x min x L(-1) and (0.007 7 +/- 0.001 1) L x min-', respectively. The assay method is proved to be sensitive, accurate and convenient. It can be applied to the pharmacokinetic study of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid.
Animals ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Female ; Hydroxybenzoates ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

A polyphenolic compound, 1,2,4,6-tetra-O-galloyl-β-D-glucose (1246TGG), was isolated from the traditional Chinese medicine Phyllanthus emblica L. (Euphorbiaceae) and assayed for its potential as an anti-hepatitis B virus (HBV) agent. The cytotoxicity of 1246TGG on HepG2.2.15 as well as HepG2 cells was determined by observing cytopathic effects, and the effects of 1246TGG on secretion of HBsAg and HBeAg in HepG2.2.15 cells were assayed by enzyme immunoassay. Results indicates that treatment with 1246TGG (6.25 μg/mL, 3.13 μg/mL), reduced both HBsAg and HBeAg levels in culture supernatant, yet the inhibitory effects tend to decline with the assay time. This study provides a basis for further investigation of the anti-HBV activity and possible mechanism of action of 1246TGG.

Background: As a traditional Chinese medicine, Cordyceps sinensis (CS) possesses a variety of immunoregulatory properties. This study aimed to explore the therapeutic potential of CS in a mice model of multiple sclerosis (MS)?experimental autoimmune encephalomyelitis (EAE). Methods: Female C57BL/6 mice were immunized with myelin oligodendrocyte glycoprotein35–55 to induce EAE, followed by an instant intragastric feeding with a low dosage of CS (low?CS group, n = 5), high dosage of CS (high?CS group, n = 5), or the same volume of normal saline (control group, n = 5).All the mice were observed for clinical assessment. Over the 30 days of CS treatment, flow cytometry was used to detect the frequency of helper T?cell (Th) subsets, Th1 and Th17, and CD4+ CD25+ regulatory T cells in the spleen and lymph nodes. Meanwhile, pathological changes in brain were determined using both hematoxylin?eosin and luxol fast blue staining. Data were analyzed using the one?way analysis of variance (ANOVA). Results: Over the 15 and 30 days of CS treatment, the clinical assessment for EAE demonstrated that both high?CS group (2.51 ± 0.31 and 2.26 ± 0.39 scores, respectively) and low?CS group (2.99 ± 0.40 and 2.69 ± 0.46, respectively) had lower disease severity scores than those of control group (3.57 ± 0.53 and 3.29 ± 0.53, all P < 0.01, respectively). Meanwhile, after 15 and 30 days, the high?CS group (19.18 ± 1.34 g and 20.41 ± 1.56 g, respectively) and low?CS group (18.07 ± 1.18 g and 19.48 ± 1.69 g, respectively) had a lower body weight, as compared with control group (16.85 ± 1.15 g and 18.22 ± 1.63 g, all P < 0.01, respectively).At 30 days post?CS treatment, there was a lower Th1 frequency in the lymph nodes (2.85 ± 1.54% and 2.77 ± 1.07% vs. 5.35 ± 1.34%, respectively; P < 0.05) and spleens (3.96 ± 1.09% and 3.09 ± 0.84% vs. 5.07 ± 1.50%, respectively; P < 0.05) and less inflammatory infiltration and demyelination in the brain of CS?treated mice than that of control group. Conclusions: Our preliminary study demonstrated that CS efficiently alleviated EAE severity and EAE?related pathology damage and decreased the number of Th1s in the periphery, indicating its effectiveness in the treatment of murine EAE. Thus, our findings strongly support the therapeutic potential of this agent as a new traditional Chinese medicine approach in MS treatment.

Adult ; Aged ; Antiviral Agents ; pharmacology ; DNA-Directed DNA Polymerase ; genetics ; Drug Resistance, Viral ; genetics ; Female ; Genes, Viral ; Hepatitis B virus ; drug effects ; genetics ; Humans ; Lamivudine ; pharmacology ; Male ; Middle Aged ; Nucleotides ; pharmacology ; Young Adult

OBJECTIVETo compare the singing formant differences between successful opera actors and non-successful opera actors during their adolescence period, and to compare the same index between adolescence and adult period of successful actors.

METHODSFrom 1985 to 1986, the author had 21 adolescent actors' voice recorded, all of them were from Beijing KunQu opera troupe. In 2000, all the 21 subjects had their voice recorded and singing formant (Fs) analyzed by using computer and sound spectrograph, 7 of them had become adult actors, others quitted their actors career after adolescents period.

RESULTSuccessful actors have obvious Fs, and stronger acoustic energy; successful actors had weaker Fs value during adolescence period than during adult period (t = 2. 9600, P < 0.05).

CONCLUSIONFs's presence and its acoustic energy were important to evaluate adolescent actors future locality potential.

Adolescent ; Child ; Humans ; Male ; Music ; Phonation ; Voice Quality

AIMTo explore the effect of hyperhomocysteinemia on vascular calcification and the underlying mechanism of it.

METHODSArterial calcification of Sprague-Dawley rats was induced by vitamin D3 plus nicotine. Hyperhomocysteinemia was established by feeding high methionine diet for six weeks and was assessed b y plasma total homocysteine (tHcy) level detected by HPLC method. Calcification was confirmed by von Kossa staining, measurement of calcium content, alkaline phosphatases (ALP) activity and osteocalcin (OC) concentration of vascular tissue. Lipid conjugated dienes formation were determined to reflecting the production of lipid peroxide.

RESULTSThe results showed that there were mass black granules deposited in aortic wall of the calcified rats by von Kossa staining. Calcium content, ALP activity, OC concentration in calcified rats increased by 8.09-fold, 45.57% and 2.81-fold compared with those of the control group (P < 0.01). Calcium content in calcified rats with high methionine diet increased by 34.29% compared with that of the calcified rats, while ALP activity and OC content decreased by 29.13% and 74.69% compared with that of the calcified rats. Lipid conjugated dienes formation in plasma of the rat with high methionine diet and of calcified rats with high methionine diet increased by 1.93 and 2.89-fold compared with those of the control group, respectively (P < 0.01), and in calcified rats with high methionine diet group was increased by 32.90% compared with that of high methionine diet group (P < 0.01).

CONCLUSIONHyperhomocysteinemia could promote vascular calcification, which might be mediated through the production of lipid peroxide.

Alkaline Phosphatase ; metabolism ; Animals ; Calcium ; metabolism ; Endothelium, Vascular ; Hyperhomocysteinemia ; metabolism ; pathology ; Lipid Peroxidation ; Male ; Methionine ; administration & dosage ; Osteocalcin ; analysis ; Rats ; Rats, Sprague-Dawley ; Vascular Calcification ; metabolism ; pathology