ObjectiveTo compare the contents and relative molecular weight distributions of proteins and polypeptides in Naoxuekang dropping pills， Huoxue Tongmai capsules and Maixuekang capsules of Hirudo single medicinal preparations， to evaluate the in vitro anticoagulant， antiplatelet and fibrinolytic activities of the three preparations， and to investigate the effects of temperature， pH and digestive enzymes on the anticoagulant activities of the three preparations. MethodsThe contents of soluble proteins and polypeptides in the three preparations were determined by bicinchoninic acid assay（BCA） and Bradford method， and the relative molecular weight distributions of the three preparations were determined by electrophoresis combined with gel chromatography. The antithrombin activity of the three preparations was evaluated by fibrinogen-thrombin time（Fibg-TT） method， and their anticoagulant activities were further assessed by the elongations of activated partial thromboplastin time（APTT）， prothrombin time（PT） and thrombin time（TT）. The antiplatelet aggregation activities of the three preparations were measured by turbidimetry and the fibrinolytic activities were measured by fibrin plate method. Relative TT was used as index to investigate the effects of temperature， pH and digestive enzyme buffer on anticoagulant activities of the three preparations. ResultsAt the lowest single dosage， the contents of proteins and polypeptides were in the order of Maixuekang capsules>Huoxue Tongmai capsules>Naoxuekang dropping pills. Both Huoxue Tongmai capsules and Maixuekang capsules had 11 electrophoretic bands between 4.0 kDa and 90 kDa， the bands of Maixuekang capsules were more clear in the range of >25 kDa， and there was 1 obvious band at 14 kDa for the two capsules. Huoxue Tongmai capsules had one specific band at 9.0 kDa and Maixuekang capsules had one specific band at 48.0 kDa. Naoxuekang dropping pills only had 2 electrophoretic bands at 6.5 kDa and 8.5 kDa， primarily containing peptides below 2 kDa， most of which were oligopeptides. The anticoagulant activity concentrations of the three preparations exhibited a certain dose-dependent effect. At the lowest single dosage， The anticoagulant activity concentrations were ranked as Naoxuekang dropping pills>Huoxue Tongmai capsules>Maixuekang capsules. The prolongation effect of the three preparations on coagulation time was dose-dependent. At the same concentration， the prolongation effect of Naoxuekang dropping pills and Huoxue Tongmai capsules was APTT prolongation rate>TT prolongation rate>PT prolongation rate， whereas for Maixuekang capsules， the sequence was TT prolongation rate>APTT prolongation rate>PT lengthening rate. At the single minimum dosage， the order of APTT prolongation rate was Maixuekang capsules>Huoxue Tongmai capsules≈Naoxuekang dropping pills， the order of PT prolongation rate was Naoxuekang dropping pills≈Maixuekang capsules>Huoxue Tongmai capsules， and the order of TT prolongation rate was Maixuekang capsules>Huoxue Tongmai capsules>Naoxuekang dropping pills. The three preparations showed dose-dependent effects on platelet aggregation induced by adenosine diphosphate（ADP） and arachidonic acid（AA）， and the effect induced by ADP was stronger than that induced by AA. The anti-platelet aggregation effect of Naoxuekang dropping pills was significantly stronger than that of Maixuekang capsules（P<0.01）， whereas Huoxue Tongmai capsules had the effect of promoting platelet aggregation. None of the three preparations had the ability to dissolve fibrin. The anticoagulant activity of Naoxuekang dropping pills was least affected by heating， while the activities of the two capsules decreased significantly within 5 min above 80 ℃， and continued to decrease within 2 h. Compared with pure water， the anticoagulant activities of the three preparations could be increased by 1-3 times under strong acidity（pH 1-3）. In the pepsin buffer， the anticoagulant activity of Naoxuekang dropping pills could be increased by 1-3 times， while the anticoagulant activities of Huoxue Tongmai capsules and Maxuekang capsules were significantly decreased， the lowest levels were about 60% and 20%， respectively. In trypsin buffer， the anticoagulant activities of Naoxuekang dropping pills， Huoxue Tongmai capsules and Maixuekang capsules decreased significantly， and the lowest levels decreased to about 41%， 41% and 35%， respectively. ConclusionThe contents of proteins and polypeptides and relative molecular weights of the preparations derived from lyophilized fresh Hirudo powder， dried Hirudo powder and reflux extract of Hirudo decrease sequentially， and the anticoagulant activity decrease gradually， but the anticoagulant pathway is different. And the anti-platelet aggregation activity of the reflux extract is significantly enhanced. The heat resistance and gastrointestinal stability of the three preparations increase successively， and the first two are suitable for enteric-soluble preparations， while the latter is suitable for routine oral administration. The above results can provide data reference for the rationality of different preparation methods， active substances， pharmacodynamics and mechanism of Hirudo preparations.

OBJECTIVE To characterize paclitaxel nanoparticles （PTX-PLGA-NPs） and evaluate their in vitro inhibitory effect on Lewis lung cancer cells. METHODS The PTX-PLGA-NPs prepared by the emulsion-solvent evaporation method were characterized in terms of particle size， polydispersity index （PDI）， Zeta potential， microscopic morphology， encapsulation efficiency， drug loading， ultraviolet-visible absorption characteristics and stability. Using mouse Lewis lung cancer cells as the subjects and paclitaxel reference substance as the control， the cytotoxicity and in vitro killing activity of PTX-PLGA-NPs were detected using CCK-8 method and Calcein-AM/PI double staining method， respectively. The effects of PTX-PLGA-NPs on cell apoptosis and cell cycle were assessed by Annexin Ⅴ-FITC/PI staining method and PI staining method， respectively. RESULTS PTX-PLGA-NPs were spherical with an average particle size of （172.03±0.95） nm， PDI of 0.098±0.012， and Zeta potential of （－1.76±0.02） mV. The encapsulation efficiency and drug loading were （52.32±0.66）% and （7.07±0.18）%， respectively， and the ultraviolet-visible absorption characteristics were not affected by the carrier polylactic-co-glycolic acid. When stored in the dark at 4 °C for 7 days， no significant change was noted in particle size， and the average PDI （after 1， 2， 4 and 7 days of storage） was under 0.3. Compared with the paclitaxel reference substance group， the PTX-PLGA-NPs group had more cells in a state of death， the survival rate （at the PTX concentration of 11.2 μg/mL） was significantly decreased， and both the apoptosis rate and the proportion of G2 phase cells were significantly increased （P＜0.05）. CONCLUSIONS The prepared PTX-PLGA-NPs indicate homogeneity in particle size， uniform dispersion， stable properties， and stronger in vitro killing effect on lung cancer cells than PTX.

Objective To identify the content of tubeimoside I in Paniculate Bolbostemma from 16 differ-ent locations and determine the high-containing locations and influencing factors.Methods The quanti-tative determination method of tobeimoside I in Tubeimu(Paniculate Bolbostemma, Rhizoma Bolbostem-atis)in Chinese Pharmacopoeia(2015 Edition)was used to determine the content of tobeimoside I in 16 batches of Tubeimu for deciding producing areas with higher content of Tubeimu and analyzing the influ-encing factors.The origin producing areas was classified by using group-average clustering analysis meth-od taken content of tobeimoside I as the index.Results The results showed that the content of tobeimo-side I was in a range from 1.234%to 3.174%in different producing areas,which was accorded with the requirement of Chinese Pharmacopoeia.The clustering analysis classified 16 producing areas into 4 groups,and content of tobeimoside I was the highest in Tubeimu material produced from Shangluo and Hanzhong.Conclusion The temperature and humidity had influence on Tubeimu material, and content of tobeimoside I was higher in the producing areas with higher temperature and humidity.

OBJECTIVE: To explore the relationship among perceived organizational support,job burnout and depressive tendency in nursing staffs. METHODS: A total of 807 nurses from 7 municipal hospitals in Zhengzhou City,Henan Province were selected as the study subjects by multi-stage cluster random sampling method. The questionnaires of Perceived Organizational Support,Maslach Burnout Inventory-General Survey and Center for Epidemiological Survey-Depression Scale were used to conduct the survey. RESULTS: The total scores of perceived organizational support and job burnout were( 69. 3 ± 18. 5) and( 36. 3 ± 13. 7) respectively. The median of the total score of depression tendency was 17. 00. The total score of nurses' perceived organizational support was negatively correlated with the total scores of job burnout and depression tendency( P < 0. 01). The total score of job burnout was positively correlated with the total score of depression tendency( P < 0. 01). The degree of explanations for the change of perceived organizational support and job burnout on depression tendency were 9. 1% and 13. 1%,respectively. CONCLUSION: Perceived organizational support and job burnout play important roles in predicting depression tendency. Job burnout plays a mediating role in the relationship between perceived organizational support and depression tendency.

This study was aimed to search anti-platelet aggregation effectors from Gardenia jasminoides extract with the employment of platelet affinity extraction method coupled with HPLC, in order to provide pharmacological experi-mental evidences of the selected effectors to verify its feasibility. Under physiological conditions, washed rat platelets were added into G. jasminoides extract and then a mixture was gained. Consequently, some components from G. jas-minoides extract were combined to the platelets in the mixture while some were not owing to their special chemical structures and properties. Firstly, the uncombined components were washed off from the mixture. Secondly, the com-bined components in the leftover was washed down and collected, respectively, right after destroying the occupied platelets' structures. Thirdly, different collected eluents were analyzed, respectively, by HPLC established in the pre-vious work to search the effectors. Fourthly, pharmacological experiments were implemented for confirmation. The re-sults showed that dominant effective components from G. jasminoides extract acting on anti-platelet aggregation were identified as geniposide. Further evident was provided as well by pharmacological experiment that geniposide exhibit-ed significant inhibitory effect on anti-platelet aggregation in rats induced by ADP, rat tail collagen and thrombin(P< 0.01). It was concluded that the platelet affinity extraction-HPLC method proposed in this paper can be utilized to analyze the correlation of effectors from G. jasminoides extract and its pharmacological effects. Moreover, there are some correlations between screened chemical substances and their pharmacological effects.

OBJECTIVETo establish a method for determination of geniposide in Beagle dogs plasma by high performance liquid chromatography (HPLC), and study the pharmacokinetics and bioavailability of geniposide in Beagle dogs after oral administration Xingnaojing.

METHODTo determine the geniposide in Beagle dogs plasma by HPLC after oral administration or intravenous injection Xingnaojing, and the pharmacokinetic parameters were calculated by the software of Kinetica.

RESULTThe good linearity range of geniposide was 1.24 - 158.88 mg x L(-1). The main pharmacokinetic parameters after oral administration was as follows: Cmax (11.8 +/- 0.6) mg x L(-1), Tmax (52.0 +/- 4.5) min, AUC(1280.8 +/- 172.0) mg x min x L(-1), MRT(118.7 +/- 25.4) min, and these parameters after intravenous injection was follows: Cmax 107.4 +/- 6.3) mg x L(-1), AUC(7930.1 +/- 670.0) mg x min x L(-1), MRT(92.4 +/- 5.1) min. The bioavailability of geniposide in Beagle dogs after oral administration Xingnaojing was (6.46 +/- 0.87)%.

CONCLUSIONThe HPLC method had good applicability. The extract recovery, method recovery, intra-day precision and inter-day precision of the method were all met the requirements. The stability in conditions of room temperature and freeze-thaw cycle was good. The results indicated that the oral administration bioavailability of geniposide was in low degree.

Administration, Oral ; Animals ; Biological Availability ; Chromatography, High Pressure Liquid ; Dogs ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Iridoids ; administration & dosage ; pharmacokinetics

OBJECTIVETo study the transdermal absorbability of gentiopicroside, naringin and protosappanin B contained in Xuanbi gel plaster.

METHODThe Franz diffusing cells method was adopted for the in vitro model of rat belly skins. Three indexes, gentiopicroside, naringin and protosappanin B, residued in the accept liquid, skins and plaster were determined by HPLC.

RESULTThe penetration rates of gentiopicroside, naringin and protosappanin B were respectively 3.47, 1.59, 2.13 microg x cm(-2) x h(-1). After 24 h, their penetration rates were 25.42%, 11.73%, 17.78%, respectively. The residual quantities of gentiopieroside, naringin and protosappanin B in skin were 0.231, 0.593, 0.568 microg x cm(-2), ith the retention rates of 0.027%, 0.227%, 0.475%, respectively. The amount of residue of gentiopicroside, naringin and protosappanin B in plaster were 2179, 674, 278 microg, with the retention rates of 81.36%, 81.92%, 73.83%, respectively.

CONCLUSIONThe in vitro transdermal behavior of Xuanbi gel plaster is close to a zero-order process. The residual quantity the retention rate in skins is much lower than the penetration rate and the residual rate in plaster.

Administration, Cutaneous ; Animals ; Flavanones ; metabolism ; Gels ; chemistry ; Iridoid Glucosides ; metabolism ; Male ; Phenols ; metabolism ; Rats ; Rats, Sprague-Dawley ; Skin Absorption

OBJECTIVETo study the characteristics of intestinal absorption of psoralen and isopsoralen of Xianlinggubao capsule, and compare the absorption of Xianlinggubao capsule prepared by different technologies.

METHODNon everted gut sac method was applied to investigate the influence of absorption sites and drug concentration on psoralen and isopsoralen absorption, which were determined by HPLC.

RESULTAlthough the absorption rate constants of psoralen and isopsoralen in duodenum were more than that in jejunum and ileum, there was no significance difference between them. The absorption rate constants of psoralen kept at the same level when the concentrations of drug solution were at middle and low level, while the absorption rate constant at high level was absolutely lower than them (P < 0.05). The results of isopsoralen were the same as psoralen's.

CONCLUSIONIntestinal absorption of psoralen and isopsoralen may be affected by the dissolution. The absorption rate constants of psoralen and isopsoralen in new Xianlinggubao capsules are higher. The absorptions of active components absorption has significant difference in different preparation processes of Xianlinggubao capsule.

Animals ; Capsules ; pharmacokinetics ; Drug Compounding ; Drugs, Chinese Herbal ; pharmacokinetics ; Duodenum ; metabolism ; Ficusin ; pharmacokinetics ; Furocoumarins ; pharmacokinetics ; Ileum ; metabolism ; Intestinal Absorption ; Intestines ; metabolism ; Jejunum ; metabolism ; Rats

OBJECTIVETo study pharmacokinetic of puerarin in rats following different methods of administration of Tongqiao Sanyu prescription.

METHODTongqiao Sanyu prescription was administered to rats by caudal vein injection, nasal administration and oral administration. Plasma samples were extracted with methanol and the plasma concentration of puerarin was analyzed by RP-HPLC. The pharmacokinetic parameters and bioavailability were calculated with Kinetica software.

RESULTThe main pharmacokinetic parameters were as follows: AUC(0-infinity) of caudal vein injection was (787.99 +/- 70.44) mg x min x L(-1); AUC(0-infinity) of nasal administration was (376.56 +/- 93.93) mg x min x L(-1); AUC(0-infinity) and oral administration (The dose was decuple higher than that of caudal vein injection and nasal administration) was (491.18 +/- 110.64) mg x min x L(-1). The absolute bioavailability of puerarin was 47.78% by nasal administration and 6.23% by oral administration.

CONCLUSIONThe bioavailability of nasal administration is higher than oral administration significantly, this result can provide some scientific foundantion for the method of administration and the reform of dosage form of Tongqiao Sanyu prescription.

Animals ; Biological Availability ; Drug Administration Routes ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Isoflavones ; administration & dosage ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo develop a GC-FID method for the determination of borneol concentration in rat plasma and to investigate the pharmacokinetics after injection of novel-Xingnaojing.

METHODNovel-Xingnaojing was injected via by caudal vein injection. The blood samples were collected by posterior orbital venous plexus approach at 0.5, 1, 3, 5, 8, 12, 20, 30, 45 min. The drug in plasma was extracted with ethyl acetate and then detected by GC-FID, octadecane was used as the internal standard. The pharmacokinetic parameters were calculated by the software of Kinetica.

RESULTThe calibration curve was good linear in the range of 1.67-16.67 mg x L(-1). The extraction recoveries of low, medium and high concentration were (92.81 +/- 1.11)%, (85.38 +/- 0.86)% and (84.58 +/- 0.58)%, respectivley. And the RSDs of within-day and between-day were below 3.00%. Plasma concentration of borneol was consistent with the two-compartment open model. The pharmacokinetic parameters were that the t1/2alpha was (1.18 +/- 0.20) min, the t1/2beta was (22.27 +/- 6.85) min, the C(max)(Calc) was (18.76 +/- 2.10) mg x L(-1), the MRT was (23.84 +/- 7.67) min(-1), and the AUC was (100.00 +/- 15.85) mg x min x L(-1).

CONCLUSIONThe GC-FID method developed can be applied to determination and pharmacokinetics. The borneol in novel-Xingnaojing is distributed and metabolized fast after being administrated.

Animals ; Bornanes ; pharmacokinetics ; Drugs, Chinese Herbal ; pharmacokinetics ; Flame Ionization ; methods ; Male ; Rats ; Rats, Sprague-Dawley