Objective To study the in vitro effects of different doses and different kinds of LMWH on CR, and to determine whether the CR test could be used to monitor LMWH. Methods The CR value was measured with different reagents ( glass beads, celite and kaolin ) in blood samples from twenty volunteer donors, which were spiked with increasing concentration of LMWH ( dalteparin, 0-1.8 IU/ml ). Then the CR test was performed again on the same blood samples spiked with the same concentration ( 0. 8 IU/ml ) but different LMWH ( dalteparin, enoxaparin and nadroparin ). Regression analysis was performed to establish a regression equation from corresponding LMWH levels. Results With the increasing of dalteparin dose, CR values were reduced gradually for all three reagents. When the concentration of dalteparin was 0-1.8 IU/ml,the value of CR was 20. 0-4. 5 IU/min for glass beads, 26. 1-6.6 IU/min for celite and 27. 2-7. 5 IU/min for kaolin. An exponential relationship was observed between the CR value and dalteparin concentration for three reagents( R2 = -0.796, -0.884, -0.921 ,P ＜0.01 ). All three kinds of LMWH with the same concentration (0.8 IU/ml ) induced a different change in CR. The value of CR was 7.4 IU/min with dalteparin,8. 5 IU/min with enoxaparin and 8.5 IU/min with nadroparin. Compared with the control group ( CR was 17.6 IU/min ), three kinds of LMWH had statistical significance ( t = 18.45, 12. 33, 14. 93, P ＜ 0.01 ).Compared with the enoxaparin and nadroparin, dalteparin induced a higher CR value ( t = 2. 552,2. 924,P＜0. 05 ). Conclusions There is an exponential relationship between CR value and dalteparin concentration for three reagents. Three kinds of LMWH can significantly reduce the value of CR. CR test can be used to monitor the anticoagulant effect of LMWH.

Animals ; Comet Assay ; DNA Damage ; drug effects ; Lymphocytes ; drug effects ; Male ; Organometallic Compounds ; toxicity ; Rats

Background Hypertension is a multigenetic inheritable disease.Gene therapy with long-term effects and less side effects by regulating gene expression has been shown to be a potential and exciting prospect. Objective To investigate the effects of RNA interference(RNAi)targeting angiotensin-converting enzyme(ACE)on the blood pressure and ACE expression in kidney of spontaneously hypertensive rats(SHR).Methods SHR were randomly to receive placebo(n=12)or control adenovirus Ad5-EGFP)or a single injection of recombinant adenovi- ral vectors,Ad5-EGFP-ACE-shRNA(n=12,iv).Normotensive Wistar-Kyoto rats(WKY)were served as normal control group.SBP was measured before and after the intervention.Aorta,lung,myocardium and kidney were studied using fluorescence microscope to identify the sites of Ad5-EGFP-ACE-shRNA.Expressions of ACE mRNA and protein in kidney were evaluated by RT-PCR and Western blot.Results SBP of the treat group was effectively reduced by 19.0?3.2 mmHg at the 3rd day,and 22.1?3.3 mmHg at the 13th day of the experiment.The anti- hypertensive effect significant remained at least for 14 days.On the contrary,increase in BP was shown in placebo and the adenovirus control group.Compared with placebo or adenovirus control rats,ACE mRNA expression level in kidney of the treated rats was lower by 61.1% and 62.3% respectively,with ACE protein expression level lower- ing by 56.2% and 53.30% as well(ail P0.05). Conclusion RNA interference targeting ACE gene inhibits the expressions of ACE mRNA and protein.A single dose injection resulted in a prolonged decrease in BP.The evidence of strong antihypertensive effect by genetic therapy justifies efforts for further investigation.

OBJECTIVETo study the impact of total flavones from Artemisia anomala (TFAS) on activation of macrophages, cell oxidative stress, auto-nitration of CuZn-SOD, platelet aggregation and isolated vascular tension.

METHODLPS and IFN-gamma induced activation of macrophages and oxidative stress in rats; H2O2 and nitrite induced auto-nitration of CuZn-SOD; ADP, AA and collagen induced platelet aggregation in vitro in mice; PE stimulates isolated vascular tension; nitrite content of macrophages was measured by Griess assay; MTT assay and FRAP assay was applied for cell viability and total cell antioxidant capacity; auto-nitration of CuZn-SOD was measured by Western blot and colorimetric methods; platelet aggregation was detected by turbidimetry; and aorta ring relaxation was recorded by isolated vascular function experience devices for rats.

RESULTTFAS demonstrated dose dependence (25, 50, 100, 200 mg x L(-1)) on inhibiting induced macrophages NO production from generating, while increasing cell viability and total anti-oxidant capacity. Auto-nitration of CuZn-SOD was suppressed by TFAS in dose dependence (0.5, 5, 50 mg x L(-1)). TFAS showed an inhibitory effect on collagen-induced platelet aggregation at 50 mg x L(-1) and an endothelium-dependent relaxation effect on PE-induced vasoconstriction at 1 g x L(-1).

CONCLUSIONTFAS shows effect on anti-inflammation, anti-oxidation, anti-nitration, anti-platelet aggregation and vasodilatation in experiment in vitro, which may inhibit vascular inflammatory by regulating multiple target points. It is among material bases for promoting blood circulation and removing blood stasis.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Aorta ; drug effects ; immunology ; physiology ; Artemisia ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Flavones ; administration & dosage ; Humans ; Macrophages ; drug effects ; immunology ; Mice ; Oxidative Stress ; drug effects ; Rats ; Vasodilation ; drug effects

BACKGROUND:Sepsis-induced myocardial injury is one of the major predictors of morbidity and mortality of sepsis. The cytoprotective function of erythropoietin (EPO) has been discovered and extensively studied. However, the cardioprotective effects of EPO on sepsis-induced myocardial injury in the rat sepsis model has not been reported.METHODS:The rat models of sepsis were produced by cecal ligation and perforation (CLP) surgery. Rats were randomly (random number) assigned to one of three groups (n=8 for each group):sham group, CLP group and EPO group (1000 IU/kg erythropoietin). Arterial blood was withdrawn at 3, 6, 12, and 24 hours after CLP. cTnI, BNP, CK-MB, LDH, AST, TNF-α, IL-6, IL-10, and CRP were tested by the ELISA assay. Changes of hemodynamic parameters were recorded at 3, 6, 12, 24 hours after the surgery. Histological diagnosis was made by hematoxylin and eosin. Flow cytometry was performed to examine cell apoptosis, myocardium mitochondrial inner membrane potential, and NF-κB (p65). Survival rate at 7 days after CLP was recorded.RESULTS:In the CLP group, myocardial enzyme index and inflammatory index increased at 3, 6, 12 and 24 hours after CLP compared with the sham group, and EPO significantly blocked the increase. Compared with the CLP group, EPO significantly improved LVSP, LV +dp/dtmax, LV -dp/dtmin, and decreased LVEDP at different time. EPO blocked the reduction of mitochondrial transmembrane potential, suppressed the cardiomyocyte apoptosis, inhibited the activation of NF-κB, and reduced the production of proinflmmatory cytokines. No difference in the survival rate at 7 days was observed between the CLP group and the EPO group.CONCLUSION:Exogenous EPO has cardioprotective effects on sepsis-induced myocardial injury.

Dental age plays an important role in age estimation. It has often been used together with skeletal age to improve the accuracy of age estimation abroad, but seldom performed in China. As a noninvasive technology, dental radiological imaging has been widely used in age estimation. By observing the age-related changes such as the pulp cavity and development of crown and root on radiographs. Gleiser and Hunt, as well as Demirjian have developed different methods to determine the age of human. Demirjian's method has been proved to be more accurate but with limitation when used in persons of eighteen and above. The accuracy and reliability of the measurements on pulp cavity could be improved as the development of computed tomography with its high resolution and intelligent software. As a convenient and accurate method, age estimation from dental computed tomographs would be more promising in the future for forensic scientists and anthropologists.
Age Determination by Teeth/methods* ; Dental Pulp/diagnostic imaging* ; Forensic Dentistry ; Humans ; Radiography, Panoramic/methods* ; Tomography, X-Ray Computed/methods*

Oculocerebrorenal syndrome of Lowe (OCRL) is an X-linked recessive disorder. This study investigated the history of a Chinese family with OCRL and used direct DNA sequencing to screen all exons of OCRL gene for mutations. A missense mutation (1736 A→G) in exon 15 was revealed, which resulted in the change of His (H) 507 to Arg (R). The patient's mother was the carrier of the heterozygous mutation in X-chromosome. To our knowledge, H507R mutation in OCRL gene has not been reported in Chinese people.
DNA Mutational Analysis ; Humans ; Infant ; Male ; Mutation, Missense ; Oculocerebrorenal Syndrome ; genetics ; Phosphoric Monoester Hydrolases ; genetics

OBJECTIVETo investigate the effects of RNA interference (RNAi) targeting angiotensin-converting enzyme (ACE) on the blood pressure and myocardial remodeling in spontaneously hypertensive rats (SHRs).

METHODSSaline (control), adenovirus (Ad5) and recombinant adenoviral vectors (Ad5-ACE-shRNA expressing ACE gene-specific shRN) were randomly administered by caudal intravasation to SHRs (n = 12 each group) at day 1 and day 16. Normotensive Wistar-Kyoto rats (WKY) served as normal controls. Systolic blood pressure (SBP) of the caudal artery was measured daily. Expressions of ACE at mRNA and protein levels in myocardium and aorta were evaluated by RT-PCR and Western blot respectively, ACE serum concentration was measured by ELISA at day 3 (n = 6 each group). The ratio of left ventricular to body weight (LVW/BW), myocardial collagen content were measured and myocardial ultrastructure observed under transmission electron microscope at the study end.

RESULTSAd5-ACE-shRNA injection significantly reduced SBP (-22 mm Hg, 1 mm Hg = 0.133 kPa) and the antihypertensive effect could last at least 14 days post each injection. SBP was not affected by saline and Ad5 injections. ACE expressions at mRNA and protein levels at myocardium and aorta as well as serum ACE were significantly decreased in Ad5-ACE-shRNA treated SHRs compared to that in saline and Ad5 groups (all P < 0.05) and was comparable to that in WKY group (P > 0.05). The LVW/BW ratio (2.24 +/- 0.19) and myocardial collagen content [(1.283 +/- 0.019) microg/mg] in Ad5-ACE-shRNA treated SHRs were also significantly lower than those in saline treated [3.21 +/- 0.13 and (1.686 +/- 0.013) microg/mg, both P < 0.05] and Ad5 treated SHRs [3.13 +/- 0.12, (1.682 +/- 0.009) microg/mg, both P < 0.05] but still higher than those of WKY group [2.06 +/- 0.11, (1.257 +/- 0.019) microg/mg, both P < 0.05]. Myocardial ultrastructure was also significantly improved in Ad5-ACE-shRNA treated SHRs compared to saline and Ad5 treated SHRs.

CONCLUSIONRNAi targeting ACE gene significantly inhibited the expressions of ACE at mRNA and protein levels and resulted in prolonged antihypertensive effects and myocardial ultrastructure improvements in this SHR model.

Animals ; Blood Pressure ; Disease Models, Animal ; Heart Rate ; Hypertension ; genetics ; physiopathology ; Male ; Peptidyl-Dipeptidase A ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Ventricular Remodeling

OBJECTIVETo study the effects of the change in transient receptor potential vanilloid 1 (TRPV1) channel activity on the degree of airway inflammation in asthmatic mice.

METHODSBALB/c mice were randomly divided into control, asthma, capsaicin (TRPV1 agonist), capsazepine (TRPV1 antagonist), and dexamethasone groups. The asthmatic mouse model was established by intraperitoneal injection of mixed ovalbumin-aluminium hydroxide solution and ultrasonic atomization with OVA for sensitization and challenge. The capsaicin, capsazepine, and dexamethasone groups were given intraperitoneal injection of capsaicin (30 μg/kg), capsazepine (10 μmol/kg), and dexamethasone (2 mg/kg) respectively, at 30 minutes before challenge. Hematoxylin and eosin staining was used to observe the degree of pulmonary inflammation. ELISA was used to measure the content of interleukin-8 (IL-8) and interleukin-13 (IL-13) in bronchoalveolar lavage fluid (BALF). Real-Time PCR was used to measure the relative content of TRPV1 mRNA in lung tissue.

RESULTSCompared with the asthma group, the capsazepine and dexamethasone groups showed reduced pulmonary inflammation, while the capsaicin group showed aggravated pulmonary inflammation. Compared with the control group, the asthma and capsaicin groups showed increases in the content of IL-13 and IL-8 in BALF and the mRNA expression of TRPV1 in lung tissue (P<0.05). Compared with the asthma group, the capsazepine and dexamethasone groups showed reductions in the content of IL-13 and IL-8 in BALF and the mRNA expression of TRPV1 in lung tissue (P<0.05). The capsaicin group showed increases in the content of IL-13 and IL-8 in BALF (P<0.05).

CONCLUSIONSTRPV1 channel agonist and antagonist can influence the degree of airway inflammation in asthmatic mice. Dexamethasone may reduce airway inflammation through regulating TRPV1 level.

Animals ; Asthma ; etiology ; Female ; Interleukin-13 ; analysis ; Interleukin-8 ; analysis ; Lung ; pathology ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; analysis ; TRPV Cation Channels ; genetics ; physiology

OBJECTIVETo investigate the effects of RNA interference (RNAi) targeting angiotensin II Type 1 receptor (ATlR) and angiotensin-converting enzyme (ACE) on blood pressure and myocardial remodeling in spontaneous hypertensive rats (SHRs).

METHODSSaline (control), adenovirus (Ad5) and recombinant adenoviral vectors (Ad5-ACE-shRNA, Ad5-AT1R-shRNA and Ad5-ACE-AT1R-shRNA expressing ACE, AT1R, ACE and AT1R gene-specific shRNA, respectively) were randomly administered by caudal intravasation to SHRs (n = 12 each group) at day 1 and 17. Normotensive Wistar-Kyoto rats (WKY) served as normal controls. Systolic blood pressure (SBP) of the caudal artery was measured daily. Expression of ACE and AT1R at mRNA levels in ventricle and aorta were evaluated by fluorescence quantitative PCR. Angiotension II serum concentration was measured by ELISA at day 3 (n = 6 each group). The ratio of left ventricular to body weight (LVW/BW) and myocardial collagen content were measured, myocardial ultrastructure observed under transmission electron microscope at the study end.

RESULTSThe caudal artery pressure of saline and Ad5 group was equally increased by about 26 mm Hg(1 mm Hg = 0.133 kPa) compared to baseline (both P < 0.05). Ad5-ACE-shRNA, Ad5-AT1R-shRNA and Ad5-ACE-AT1R-shRNA injection significantly reduced SBP (-24 mm Hg, -22 mm Hg and -26 mm Hg respectively, all P < 0.05 vs. baseline) and the antihypertensive effect could last at least 15 days post each injection. SBP was not affected by saline and Ad5 injections. ACE and AT1 mRNA expressions at ventricle and aorta were significantly decreased in Ad5-ACE-shRNA, Ad5-ACE-AT1R-shRNA and Ad5-AT1R-shRNA, Ad5-ACE-AT1R-shRNA treated SHRs compared to those in saline and Ad5 groups (all P < 0.05) and was comparable to that in WKY group (P > 0.05). The LVW/BW ratio [(2.22 +/- 0.18) microg/mg, (2.23 +/- 0.19) microg/mg, (2.17 +/- 0.16) microg/mg] and myocardial collagen content [(1.291 +/- 0.019) microg/mg, (1.298 +/- 0.019) microg/mg, (1.276 +/- 0.019) microg/mg] in Ad5-ACE-shRNA, Ad5-AT1R-shRNA and Ad5-ACE-AT1R-shRNA treated SHRs were also significantly lower than those in saline treated [(3.23 +/- 0.13) microg/mg and(1.683 +/- 0.013) microg/mg, both P < 0.05] and Ad5 treated SHRs [(3.25 +/- 0.12) microg/mg and(1.693 +/- 0.013) microg/mg, both P < 0.05], but still higher than those of WKY group [(2.06 +/- 0.12) microg/mg and (1.258 +/- 0.019) microg/mg, both P < 0.05]. Myocardial ultrastructure was also significantly improved in all SHRs underwent RNAi treatments compared to saline and Ad5 treated SHRs.

CONCLUSIONRNAi targeting ACE and AT1R gene significantly inhibited myocardial and aortic ACE and AT1R mRNA expressions and resulted in prolonged antihypertensive effects and myocardial ultrastructure improvements in SHRsl. The RNAi technology may be a potential new strategy of gene therapy for hypertension.

Animals ; Blood Pressure ; Gene Silencing ; Heart Rate ; Hypertension ; genetics ; physiopathology ; Male ; Peptidyl-Dipeptidase A ; genetics ; metabolism ; RNA Interference ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism ; Ventricular Remodeling