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Phosphocreatine ( PCr), a natural high energy phosphate, plays a pivotal role in maintaining energy homeostasis of the body. Exogenous PCr has been developed as a cardio-protective drug and extensively used in treatment of cardiovascular diseases.PCr has special chemical structure,which confers much bioinformation on it,and becomes a multitarget-directed drug.Since the 21st century,with rapid development of molecular biology,the multiple target action mechanisms of PCr have continually gained elucidation,including energy-related and non-energy-related mechanisms,intracellular and extracellular mechanisms,which are leading to its extensive clinical applications in cardiovascular diseases.Based on author' s research and published literatures,this article reviews the research progress in multiple target action mechanisms of PCr,including energy supply,membrane stabilization, anti-platelet aggregation, electrophysiology, enzyme inhibition and protection of mitochondria, antiapoptotic effect,etc.
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Objective To investigate effects of diabetes mellitus (DM) on pharmacokinetics (PK) of cephradine (CED).Methods DM model was induced by iv.alloxan 60 mg·kg-1.A reversed phase HPLC internal standard method was developed for measurement of CED plasma concentration.After blood collection,rats were sacrificed to collect kidneys for calculating kidney index(KW/BW).DM and normal control (CTL) rats were randomly assigned to receive iv.or ig.CED at a dose of 180 or 90 mg·kg-1.The 3p97 program was used to calculate PK parameters.Results The developed HPLC method was validated to have high specificity,precision,recovery and good storage stability,and met requirements for PK study of CED.The CED in rats of both DM and CTL groups showed the iv.two-compartment PK and ig.one-compartment PK and followed the first-order kinetics.Following iv.dosing,a remarkably decreased t1/2β and MRT,increased CLt were evident in DM group as compared with CTL group (P ＜ 0.05).After ig dosing,a significant decrease in t1/2k and t a remarkable increase in CLt and Cm=were observed for DM group as compared with CTL group (P ＜ 0.05).The DM rats showed a trend of decreased t1/2ka vs CTL rats.There was no significant difference in the oral bioavailability between the two groups (P ＞ 0.05).KW and KW/BW in DM group were increased remarkably compared with CTL group (P ＜ 0.05).Conclusion The DM vs CTL results in faster absorption and elimination of CED in rats,but does not have significantly affect in oral bioavailability.The compensatory hypertrophy and hyperfunction of early-stage diabetic kidneys may constitute one of causes of quick elimination of CED in rats with DM.
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With the deepening of modernization of traditional Chinese medicine (TCM) and continuing emergence of new theories, methods and techniques, a very rapid and significant development has been achieved in the pharmacokinetics (PK) of TCM. This paper reviews the main research progresses of PK of TCM, including integrated PK of multiple effective components of TCM, fingerprint PK of TCM, novel dosage form PK of TCM, polysaccharide PK of TCM and drug interactions of TCM; and further sets up the prospects.
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LC-MS/MS method was used to simultaneously determine anti-oxidative active catechins EGCG, ECG, EGC and EC in plasma of rats treated with tea polyphenols (TP). The integrated plasma concentration (C') of TP was calculated by means of self-defined weighing coefficient based on percent AUC of individual components, thereby assessing integrated pharmacokinetic (PK) parameters of TP via log C'-T curve. The anti-free radical effects of TP were estimated using inhibitory rate of drug-containing serum collected at different times from rats against in vitro lipid peroxidation of mouse liver homogenate. The obtained E-T curves were used to calculate anti-free radical pharmacodynamic (PD) parameters of TP. E-logC and E-log C' plots and linear regression were carried out in order to obtain the correlation coefficient (R2). The results indicated that the log C'-T curves of TP, which could be best described by three-compartment model, corresponded to elimination rule of iv administration of drugs. The integrated PK parameters showed that TP was distributed in body rapidly and widely, and eliminated from deep compartment slowly. From comparison of R2 values and consistence of C'-T course and E-T course, it was evident that TP integrated PK behaviors correlated much better with its PD behaviors than individual active components, and thus demonstrated that integrated PK parameters could characterize to maximal extent holistic disposition of Chinese herbal drugs and reflect residence properties of holistic effective substances in biological body.
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This article is report the study of the pharmacokinetics and metabolic disposition of exogenous phosphocreatine (PCr) in rats by means of an ion-pair HPLC-UV assay. PCr and its metabolite creatine (Cr) and related-ATP in rat plasma and red blood cell (RBC) were simultaneously determined. A blank plasma and RBC were initially run for baseline subtraction. Plasma and RBC samples were deproteinized with 6% PCA prior to HPLC. Following i.v. administration of PCr 500 mg x kg(-1) and 1 000 mg x kg(-1) the C-T curve could be described by the two-compartment model with t1/2beta 22.5-23.3 min, V(d) 0.956 4-0.978 6 L x kg(-1), CL 0.029 L. kg(-1) x min(-1). The Cr as PCr degraded product appeared as early as 2 min post i.v. dosing with t(max) 20 min, t1/2kappa (m) 40.6-42.7 min and f(m) 60%-76%. After po administration of PCr, the parent drug in plasma was undetectable, but the metabolite Cr was detected with t(max) 65-95 min, t1/2kappa (m) 56.0-57.7 min, metabolite-based bioavailability F(m) 55.02%-62.31%. PCr i.v. administration resulted in significant elevation of ATP level in RBC but not in plasma, the related-ATP in RBC was characterized by t(max) 68-83 min, t1/2kappa 49-52 min. In RBC no exogenous PCr was found but Cr was detected following i.v. administration of PCr, with the t(max) 120 min and t1/2k (m) 70 min for Cr. The above results indicate that PCr eliminates and bio-transforms in body very rapidly; K > K(m) confers ERL, instead of FRL, type upon the metabolic disposition of Cr. Following po administration of PCr, the degraded product Cr is absorbed but not the parent drug PCr. The formed Cr can be accounted for by most of i.v. and po PCr. Intravenous dosing leads apparently increased and sustained Cr and related-ATP concentration in RBC.
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Objective To develop a liquid chromatography technique coupled with tandem mass spectrometry(LC-MS/MS)for simultaneous determination of four active catechins EGCC,ECG,EGC,and EC of tea polyphenols(TP)in rat plasma in order to further study its multi-component pharmacokinetics.Methods Following a single step liquid-liquid extraction of plasma samples with ethyl acetate,the four catechins were separated on a Hypersil ODS C18 column using an isocratic mobile phase composed of methanol-water(30:70).The detection using a mass spectrometer was performed under negative ESI in the MRM mode.The analytes were identified by reference to both MRM and tR values and quantified using peak area internal standard method.Results The method was shown to be specific without interference from matrix,metabolites,and impurities present in TP raw material and to be sensitive with LOD and LOQ of 1.5 and 10 ng/mL(EGCG)as well as 0.75 and 5 ng/mL(ECG,EGC,and EC).A good linearity was obtained over a wide range of 10-10000 ng/mL for EGCG and 5-5000 ng/mL for other three catechins(r > 0.996).The method was validated to be reproducible and reliable,as evidenced by intra-batch and inter-batch precision of less than 10% and 11%,accuracy of 97.13%-106.05% and 99.22%-103.14%,respectively.The recovery of extraction ranged from 72.74% to89.13%,matrix effect from 88.76% to 105.97% for four cateckins.The method was successfully used to study the pharmacokinetics of TP iv administered to rats at a dose of 100 mg/kg.Conclusion This method is shown to completely meet requirements for the multi-component pharmacokinetic study of TP in rats.
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OBJECTIVE:To investigate the neuropotective effect and anticoagulation effect of total Saponins of Radix Liriopes on focal cerebral ischemia in rats.METHODS:Chemical reagent Fecl3 was locally applied on the injured vessels to establish middle cerebral artery thrombosis model,and the effects of total Saponins of Radix Liriopes on rats' behavioral disturbance,brain infarct size,the histopathological changes in brain and the expression rate of nNOS immunoreactive positive neurons were measured,and the bleeding time and coagulation time were also detected with glass tube method and tail transection method.RESULTS:Due to the use of total Saponin of Radix Liriopes(10 and 40mg? kg-1),the brain infarct size was significantly decreased,the behavioral disturbance were improved and the expression rate of the nNOS immunoreactive positive neurons in rats were decreased.At doses of 20 and 60mg? kg-1,total Saponin of Radix Liriopes significantly prolonged the coagulation time and bleeding time.CONCLUSION:Total Saponin of Radix Liriopes has nuroprotective effect on focal cerebral ischemia induced by middle cerebral artery thrombosis and significant anticoagulation effect.
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Objective To develop a method for the determination of Hesperidin,magnolol from "He-Wei-Yin" Mixture,and for its quality control,high-performance liquid chromatography (HPLC) is used.Method After plenty of study about the condition,the final result choosed was as following:an analytical column of HypersiC18(200mm?4.6mm,5?m ) maked by HP was utilized. The column number was 13164421196.The mobile phase was adapted gradient with methanol-1% acetum solution,according to the following profile:0～30min,methanol concentration from 30% to 70%;30～40min,from 70% to 80%;40～50min,from 80% to 30%. The flow rate was 1.0ml/min with the column kept at ambient temperature. All the components had come out in 45 min with the detection wavelength at 294nm. It followed the order of Heperidin,Magnolol according to the retention times in the chromatography.Results In the result,the regression equations were as following:Hesperidin was Y=139.718X+4.511 with a good linearity (r=0.99994). Magnolol was Y=1456.342X+106.506 with a good linearity (r=0.99957) between the peak area and the mass of the standard.The recoveries of the method were as following:in the "He-Wei-Yin" Mixture,for hesperidin,precision of the method RSD=1.87%(n=5),recovery of the method 96.53%(RSD=3.36%,n=3);for Magnolol,precision of the method RSD = 0.34% (n=5); recovery of themethod 93.17%(RSD=4.69%,n=3).At the same condition as above,the determination of content was carried out for six different dose and batch No.Conclusion The method is simple and reliable,easy to operate,suitable for the quality control of Hesperidin and Magnolol in "He-Wei-Yin" Mixture.
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Objective To optimize the extraction procedures of compound Three-Flower-Tea (CTFT) by orthogonal design. Methods After having established the methods of quantifying the chlorogenic acid by TLC-UV, the orthogonal design was used to select the best extraction procedures, using chlorogenic acid content and extract yield as the measuring index, through controlling four different factors, naming extraction temperature (A), extraction time (B), watervolume added (C) and extraction number of times (D), and three levels at each factor. Result The effective factors which influenced the extraction procedures were A, C and D. Conclusion The optimum extraction procedures for CTFT should be 15-fold water volume, three-time decocting with 20 minutes per time at temperature of 100 ℃.
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Arg-Gly-Asp (RGD) is a unique minimal integrin-binding sequence found within several glycoprotein ligands and also in snake-venom disintegrins. Adinbitor, a protein with 73 amino acid residues including 12 cysteins and an RGD motif, was cloned from Gloydius blomhoffi brevicaudus in my laboratory. As a new member of disintegrin family, adinbitor can inhibit both human platelet aggregation induced by ADP and angiogenensis in vivo and in vitro, the typical characters of disintegrin family. To separate the effect of inhibiting platelet aggregation from that of inhibiting angiogenensis, the motif KGD was introduced into adinbitor cDNA to replace RGD by site-directed and PCR-based mutagenesis. The recombinant protein (recombinant adinbitor (KGD)) was expressed in E. coli BL21 and purified through the His· Bind affinity chromatography. Recombinant adinbitor (KGD) could inhibit ADP-induced platelet aggregation with IC50 value of 85 nmol/L. Considerably, it was a more effective inhibitor on platelet aggregation than recombinant adinbitor (RGD), which has an IC50 of 150 nmol/L. Interestingly, recombinant adinbitor (KGD) has no potency in inhibiting angiogenesis in vivo compared with recombinant adinbitor (RGD). These findings showed that KGD containing adinbitor was more suitable for inhibiting ADP-induced human platelet aggregation as a potential and specific inhibitor of human platelet aggregation, which might have promising therapeutic potential as an antithrombotic agent.
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Objectives To investigate the infection and colonization of Mycoplasma genitalium and Ureaplasma urealyticum in different male populations, to explore the association of M. genitalium and U. urealyticum with nongonococcal urethritis (NGU) respectively. Methods A case-controlled, cross sectional study of four different male populations was performed, namely: NGU patients (G1), non-NGU subjects attending STD clinic (G2), men who had sex with men participating in a health education program (G3), and healthy volunteers (G4). Nested PCR and culture were used to detect U. urealyticum. Nested PCR and PCR product sequencing were applied to detect M. genitalium. Results The prevalence rates of M. genitalium in the four study populations were 25.0%(25/100), 6.4%(6/94), 5.5%(6/110) and 0% respectively. Significant difference was found between each two groups except G2～G3 with a p value of 0.80. By multivariate regression analysis, controlling for the age of first sex, new sexual partners, urethritis and condom use in the previous 3 months, M. genitalium was only associated with urethritis (P= 0.004, OR = 6.754, 95% CI 1.833～24.893). The direct sequencing of PCR products showed gene mutations, in comparison with the reference sequence in GenBank, in 3 samples. The prevalence rates of U. urealyticum by PCR in 4 groups were 40.0%, 44.7%, 22.7% and 46.9% respectively, and there was no significant difference between G1～G2, G1～G4 or G2～G4 with a p value of 0.419, 0.325, 0.868 respectively, but the prevalence rate of U. urealyticum in G3 was significantly lower than that in other groups. Conclusions M. genitalium is strongly associated with NGU and the prevalence rate is significantly higher in groups with high risk sexual behaviors than that in general population. There is no association between the colonization of U. urealyticum and NGU.
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Objective To study LI gene sequence of HPV cp6108 from 5 cases of condyloma acuminata. Methods T-A cloning and direct sequencing of PCR product were used. Results The LI gene sequences of HPV cp6108 from 5 specimens were presented with the homology of 99% to reference sequence in GenBank. A total of 3 gene mutations were found, including a nonsense mutation of G70A, a missense mutation of D77N, and a missense mutation of Tl16P. Conclusions In comparison with the sequence in GenBank, at least 3 gene mutations of HPV CP6108, i.e. one nonsense mutation of G70A and missense mutations of D77N and Tl 16P, are found in the present study.
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ObjectiveTostudytheincidenceandclinicalcharacteristicsoftheco-infectedsexuallytransmittedinfections(STI)inpatientswithgenitalherpes.MethodsTheclinicaldataof287caseswithherpeticlesionsorpatientswithsuspectedherpeslesionswerecollected,andthepathogensofsexuallytransmittedinfectionsweredetected.ResultsGenitalherpeswasconfirmedin64.8%(186/287)oftherecruitedcases.HIVantibodiesweredetectedin68cases,andnoHIVantibodywasdetected.Theco-infectionssuchascondylomaacuminatum,activeorlatentsyphilis,genitalcandidiasisandotherSTIswerediscoveredin23.1%(43/186)ofpatientswithgenitalherpes.Allgenitalherpescasesco-infectedwithotherSTIswerecausedbyHSV-2.ConclusionTheco-infectionsarecommoninpatientswithgenitalherpes,andthefeaturesofthelesionsmaybechangedbytheseco-infections.
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Objective To study the effects of verussurinine (VSRN), an alkaloid isolated from Veratrum nigrum var. ussuriense alkaloids (VnA), against thrombosis and its platelet aggregation inhibitory activity in rats in order to find out whether VSRN is the antithrombotic active ingredient of VnA. Methods The electrically induced rat carotid artery thrombosis and stasis-induced rat inferior vena cava thrombosis models were used to evaluate the anti-arterial and anti-venous thrombosis effect of VSRN, respectively. Borns turbidimetric method was used to examine the in vivo and in vitro anti-platelet effect so as to investigate the antiplatelet aggregation of VSRN. Results In comparison with saline, VSRN in five different doses (1.25—20.00 ?g/kg) showed significantly and dose-dependently prolonged occlusion time (OT) of carotid artery injured by electrical stimulation and reduced thrombus dry weight of inferior vena cava ligated for 4 h to cause stasis. Platelet aggregation was found to be inhibited by VSRN in the doses of 1.25—5.00 ?g/kg and at the concentration of 6.25—50 ?g/L in both in vivo and in vitro test. Conclusion VSRN has powerful arteriovenous antithrombosis and antiplatelet aggregation of rats. The antithrombotic effect of VSRN is related to its platelet aggregation inhibitory activity. The above findings indicate that VSRN is an antithrombotic active ingredient of VnA.
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Object To study the effects of Veratrum nigrum L. var. ussuriense Nakai alkaloids (VnA) on platelet aggregation in rats and coagulation time, bleeding time in mice. Methods The antiplatelet effect of VnA was examined by determining platelet aggregation rate in normal rats and blood stasis model rats by turbidimetric method developed by Born. Whole blood coagulation time (CT) in mice was measured by capillary glass tube method, bleeding time (BT) by hemorrhagic transection of mouse tail model. Results VnA (45, 30, and 15 ?g/kg, iv) significantly inhibited ADP-induced platelet aggregation in rats in a dose-dependent manner. VnA (12.5, 25, 50, and 100 ?g/kg, ip) markedly increased CT and BT in mice. VnA [49.3 ?g/kg, which was anticoagulantly equieffective to heparin (1.25 mg/kg), ip] prolonged BT. There was no statistically significant difference in BT between VnA and heparin, although BT increase induced by VnA was shorter than that induced by heparin. Conclusion VnA has significant antiplatelet effect in rats and can prolong CT and BT in mice. At equieffective dose VnA-induced BT increase does not exceed that heparin induced.
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Object To study the effects of WUZI SIWU GUASHITANG (WT) against multi glycosides of Tripterygium wilfordii Hook. f. (GTW) induced genital system toxicity in male rats and its mechanism. Methods Male rats in treatment groups were treated respectively with a 80 days oral administration of large , middle and small doses of WT plus a fixed dose of GTW. The rats as control were treated with GTW alone or saline. Thereafter, the changes in testis organ index, the number of spermatozoa, the rate of movable spermatozoa and the structure of spermatogenic cell epithlium were observed and the serum testosterone level was determined by IRMA. Then the comparison was made between treatment and control groups as to above variables. Results As compared with GTW group, large or middle dose of WT significantly increased the weight of testis, the number of spermatozoa and the rate of movable spermatozoa. Large dose of WT could relieve the damage of GTW to the spermatogenic cell epithelium completely and keep the serum testosterone at normal level. Conclusion The combined use of GTW with WT is very useful in relief of GTW induced adverse reactions of genital system in male rats. The mechanism by which WT exerts the counteractive effects is related to protecting the spermatogenic cell epithelium from being damaged by GTW, and keeping the serum testosterone at normal level.
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Objective To systematically investigate the molecular epidemiological profiles of human papillomavirus (HPV) in patients with condyloma acuminata(CA). Methods Two hundred and one samples of HPV DNA isolated from CA were PCR amplified by the PGMY09/11 primer system. The PCR products were simultaneously hybridized to 37 specific HPV probes immobilized on a nylon strip and then genotyped. All DNA templates were further PCR amplified using HPV 6 and 11 type specific primers for verification. Results All samples were HPV DNA positive consisting of totally 31 genotypes, the types of which were type 11(53.7%, 108/201), 6(43.8%, 88/201), 16(6.5%, 13/201), 52(6.0%, 12/201), 33(5.5%, 11/201), cp6108 (5.5%, 11/201) and 42 (5.0%, 10/201). The samples infected with a single and mixed types of HPV accounted for 60.2% (121/201) and 39.8% (80/201) respectively. Consistent results were found with the detection of HPV6 and 11 between hybridization assay and type-specific PCR. Conclusions At least 31 HPV genotypes are associated with CA. HPV 11 predominates while 68, 40, 54, 67, 73, 82, 35, 64 and 83 are rare in CA. Type cp6108 is detected in CA for the first time with a high prevalence. HPV26, 69, 70, 71,72 and IS39 might be not associated with CA. CA infected with a single and mixed HPV types accounts for 60.2% and 39.8%, respectively.
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Objective To detect and type herpes simplex virus (HSV) in genital lesions of the patients attending STD clinic. Methods Clinical data were collected and analyzed from patients with anogenital non-herpetic lesions including induration or furuncle, fissure, folliculitis, single ulcer and so on. HSV was detected and typed by culture and PCR with specimens taken from these lesions. Results One hundred and five cases were recruited in this study. Among them, 18 cases presented induration (furuncle), 15 fissure, 16 folliculitis, 7 abrasion, 12 single ulcer, 25 nonspecific erythema and 12 balanoposthitis with edema and exudation. HSV was found in 33.3%(6/18), 20%(3/15), 37.5%(6/16), 28.6%(2/7), 33.3%(4/12), 20%(5/25) and 50%(6/12) of these lesions, repectively, by PCR, while in 22.2%(4/18), 13.3%(2/15), 25%(4/16), 14.3%(1/7), 33.3%(4/12), 8%(2/25) and 41.7%(5/12), repectively, by viral culture. The positive rates of HSV from all these non-herpetic lesions were 30.5% (32/105) and 21% (22/105), respectively (? = 0.095, P = 0.114), by PCR and viral culture. The results of HSV typing were consistent between PCR and immunofluorescence with type-specific monoclonal antibodies. Among those with HSV infections, HSV-1 infection acounted for 9.4% (3/32), and HSV-2 90.6% (29/32). Conclusions The clinical manifestations of genital HSV infections vary, and HSV could be isolated from lesions of induration (furuncle), fissure, folliculitis, abrasion, single ulcer, nonspecific erythema and balanoposthitis with edema and exudation. HSV-2 is the predominant type.