In order to establish an UPLC-MS method for determination of twelve active compounds in Qili Qiangxin capsules including astragaloside, calycosin-7-0-glucoside, ginsenoside Rb1, ginsenoside Re, ginsenoside Rd, ginsenoside Rg1, ginsenoside Rf, periplocin, periplocoside H1, hesperidin, narirutin, isoquercitrin, the chromatographic separations were performedon a Phenomenex UPLC Kinetex C18 column (2.1 mm x 100 mm, 2.6 microm) with gradient elution of acetonitrile and 0.1% aqueous formic acidat a flow rate of 0.4 mL x min(-1). The temperature was set as 40 degrees C and injection volume was 5 microL. The monitoring of all analytes was achieved under the negative ionization mode with TOF-MS and TOF-MS/MS method. The twelve analytes showed good linearity (R2 > 0.9990) within the test ranges, the average recoveries were 98.0%-102%, respectively, and the RSD were less than 3.9%, respectively. The established method is simple, rapid, and sensitive, and can be used for quality control of Qili Qiangxin capsules.
Capsules ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry ; methods

Objective To investigate the clinical value for treatment of diabetic foot with PTA and PTA combined cinepazide maleate.Methods In 24 cases of diabetic associated vascular disease of lower limb,12 cases were treated with PTA and other 12 cases were treated with PTA combined einepazide maleate,We analysed and compared clinical effects before and after the procedure,together with 3 months follow up.Results In patients treated with PTA,the clinical symptom scores of posttreatment and follow-up decreased;ABI and TcPO_2 increased significantly.The clinical symptom score and ABI of follow-up remained,stable,but TcPO_2 decreased significantly.Control angiography showed improvement in degree of vascular stenosis and peripheral staining of 11 patients after treatment.The vascular patency remained in 12 patients and the peripheral staining decreased in 7 patients on follow-up.In patients treated with VIA combined cinepazide maleate,the clinical symptom score,ABI and TcPO_2 after treatment and on follow-up showed no signifcant changes compared with those in patients treated by PTA.F,Control angiography showed that the degree of vascular stenosis and peripheral staining were improved in 12 patients after treatment.The vascular pateney was maintained and peripheral staining was improved on follow-up.Before and after treatment,there were no significant differences in clinical symptom score.ABI and TcPO_2 between patients treated with PTA and PTA combined cinepazide maleate,however,there were significant differences in clinical symptom score and TcPO_2 on follow-up.Conclusion PTA can significantly improve clinical symptom of diabetic foot and the application of cinepazide maleate is a benefitial and necessary supplement.PTA combined cinepazide maleate can be taken as one of the conventional treatment plans for diabetic foot.(J Intervent Radiol,2007,16:811-815)

To study the chemical constituents of Periplocae Cortex, the separation and purification of 70% alcohol extract were carried out by column chromatographies on AB-8 macroporous resin, silica gel and preparative HPLC. The structure of the compounds were identified by NMR and TOF-MS. A new compound was isolated and identified as 21-O-methyl-Δ5-pregnene-3β, 14β, 17β, 21-tetraol-20-one-3-O-β-D-oleandropyranosyl(1-->4)-β-D-cymaropyranosyl-(1-->4)-β-D-cymaropyranosyl (1), named as periplocoside P.
Glycosides ; chemistry ; isolation & purification ; Periploca ; chemistry ; Pregnenes ; chemistry ; isolation & purification ; Saponins ; chemistry ; isolation & purification

Factor VII Deficiency ; diagnosis ; genetics ; Female ; Humans ; Middle Aged ; Pedigree ; Phenotype

Complete mannanase gene with two introns was cloned from Trichoderrna reesei by PCR. The two introns were then removed by overlap extension PCR. The gene encoding the mature mannanase protein was inserted into the expression vector pPIC9K, downstream of a alpha-factor signal peptide sequence. The resultant recombinant vector was named pM242. After linearized with Sac I , pM242 was transformed to Pichia pastoris GS115 by electroporation. After screening, the recombinant strain Gpmf25 that expresses the secretory protein at high level was obtained. The activity of the recombinant mannanase reached 12.5 IU/mL. Optimum pH and temperature for the recombinant enzyme were 5.0 and 80 degrees C, respectively. The enzyme was stable at pH 5.0-6.0 and maintained over 50% of original activity after incubation at 70 degrees C for 30 min.
Fungal Proteins ; biosynthesis ; genetics ; Hydrogen-Ion Concentration ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Temperature ; Trichoderma ; enzymology ; genetics ; beta-Mannosidase ; biosynthesis ; genetics

AIM:To investigate the effects of simvastatin on the expression of Toll-like receptor 2 ( TLR-2 ) , interferon-γ(IFN-γ) and monocyte chemoattractant protein-1 (MCP-1) in lung tissues of mice with mouse cytomegalovirus ( MCMV) pneumonia and to explore the possible mechanism .METHODS:Male BALB/c mice (6~8 weeks old, n=40) were randomly divided into 5 groups: normal control (NC) group, MCMV infection group, simvastatin group 1 (SMV1 group), simvastatin group 2 (SMV2 group), and simvastatin group 3 (SMV3 group).The mice in SMV1, SMV2 and SMV3 groups were gavaged with simvastatin (50 mg· kg-1 · d-1 for 7 d) 7 d before, on the same day of and 3 d after in-traperitoneal injection of MCMV , while the mice in normal control group and MCMV infection group were gavaged with the same volume of normal saline .HE staining was used to observe the pathological changes of lung tissues in mice .Total tis-sue protein was extracted from the lung homogenates to detect the expression of TLR-2 by Western blot and immunohisto-chemical staining .Real-time PCR was used to analyse the content of MCMV DNA .The levels of IFN-γand MCP-1 were measured by enzyme-linked immunosorbent assay (ELISA).RESULTS:Compared with NC group, the pathological chan-ges of the lung tissues of the mice in MCMV group showed alveolar interstitial edema , alveolar wall widening and a large number of inflammatory cells .The expression of TLR-2 in the lung tissues of the mice in model group was increased signifi-cantly.The content of MCMV DNA was increased , and the expression of IFN-γand MCP-1 was also increased significant-ly.Compared with the mice in MCMV group , the pathological changes of the lung tissues of simvastatin groups showed that the inflammatory cells were decreased .The expression of TLR-2 was down-regulated.The content of MCMV DNA was de-creased, and the levels of IFN-γand MCP-1 were also decreased significantly .At the same time, the expression of TLR-2 and the content of MCMV DNA in SMV1 group were less than those in SMV2 and SMV3 groups (P<0.05), and no statis-tically significant difference between SMV 2 and SMV3 groups was observed .CONCLUSION:Simvastatin down-regulates the TLR-2 signaling pathway , and reduces the expression of TLR-2 and replication of MCMV DNA , thus attenuating the pathological damage of the lung tissue .Early intervention with simvastatin plays an important role in preventing the infection of MCMV and reducing the inflammation .

This study was amid to construct the pharmacophore model of L-type calcium channel antagonist in the application of screening Drugbank and TCMD. This paper repositions the approved drugs resulting from virtual screening and discusses the relocation-based drug discovery methods, screening antihypertensive drugs with L-type calcium channel function from TCMD. Qualitative hypotheses wre generated by HipHop separately on the basis of 12 compounds with antagonistic action on L-type calcium channel expressed in rabbit cardiac muscle. Datebase searching method was used to evaluate the generated hypotheses. The optimum hypothesis was used to search Drugbank and TCMD. This paper repositions the approved drugs and evaluates the antihypertensive effect of the chemical constituent of traditional Chinese medicine resulting from virtual screening by the matching score and literature. The results showed that optimum qualitative hypothesis is with six features, which were two hydrogen-bond acceptors, four hydrophobic groups, and the CAI value of 2.78. Screening Drugbank achieves 93 approved drugs. Screening TCMD achieves 285 chemical constituents of traditional Chinese medicine. It was concluded that the hypothesis is reliable and can be used to screen datebase. The approved drugs resulting from virtual screening, such as pravastatin, are potentially L-type calcium channels inhibitors. The chemical constituents of traditional Chinese medicine, such as Arctigenin III and Arctigenin are potentially antihypertensive drugs. It indicates that Drug Repositioning based on hypothesis is possible.
Animals ; Antihypertensive Agents ; chemistry ; pharmacology ; Calcium Channel Blockers ; chemistry ; pharmacology ; Calcium Channels, L-Type ; genetics ; metabolism ; Drug Repositioning ; methods ; Molecular Structure ; Myocardium ; metabolism ; Rabbits

Betulinic acid is a naturally occurring pentacyclic triterpenoid, which has antiretroviral, antimalarial, and anti-inflammatory properties. The purpose of this study is to investigate the HBV DNA replication inhibition in the mouse model with betulinic acid. Hydrodynamic injection method via the tail vein with the Paywl. 3 plasmid was used to establish the animal mode (n = 15), and the mice were randomly divided into the PBS control group (n = 5), Betulinic acid treatment group (n = 5) and lamivudine control group (n = 5). The day after successful modeling , the mice would have taken Betulinic acid (100 mg x kg(-1)), lamivudine (50 mg x kg(-1)), PBS drugs orally, once daily for 7 days, blood samples were acquired from the orbital venous blood at 3, 5, 7 days after the administering, HBsAg and HBeAg in serum concentration were measured by ELISA and the mice were sacrificed after 7 days, HBV DNA southern detections were used with part of mice livers. The results showed that betulinic acid significantly inhibited the expression of HbsAg in the mice model at the fifth day compared with the control group, and there was no significant differences between the effects of lamivudine and the PBS control group; both the betulinic acid and lamivudine groups had no significant inhibition for the HBeAg expression; the HBV DNA expressions of the liver tissue from the betulinic acid and lamivudine groups were inhibited compared with the control group. Taken together, these results reveal betulinic acid can inhibit the HBsAg expression and replication of the liver HBV DNA in the mouse model.
Acute Disease ; Animals ; Antiviral Agents ; pharmacology ; DNA Replication ; drug effects ; DNA, Viral ; biosynthesis ; Hepatitis B ; blood ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; drug effects ; genetics ; immunology ; physiology ; Male ; Mice ; Plasmids ; genetics ; Triterpenes ; pharmacology ; Virus Replication ; drug effects

OBJECTIVETo explore the diagnostic value of the dual-energy technique with dual-source computed tomography (DSCT) for anterior cruciate ligament injuries.

METHODSThe clinical data of 8 patients with arthroscopic results were retrospectively reviewed. All patients underwent two- and three-dimensional imaging by multiplanar reconstruction, volume rendering, and tendon mode on DSCT. Dual-energy characteristics were compared with arthroscopic results.

RESULTSSix patients who were arthroscopically diagnosed as anterior cruciate ligament injuries, all of them were also correctly diagnosed by DSCT. Two patients who were arthroscopically diagnosed as normal, one was also diagnosed as normal by DSCT and the other was misdiagnosed. The overall agreement rate was 87.5% (7/8) . Under the dual energy tendon mode, the dual energy staining of the injured anterior cruciate ligament was lower than that of the contralateral normal cruciate ligament of the patient.

CONCLUSIONThe staining diminution in DSCT imaging may be a new feature that can be used to effectively diagnose anterior cruciate ligament injury.

Adolescent ; Adult ; Anterior Cruciate Ligament Injuries ; Female ; Humans ; Knee Injuries ; diagnostic imaging ; Male ; Tomography, X-Ray Computed ; methods ; Young Adult

OBJECTIVETo explore the changes in the threshold of auditory brainstem response (ABR) and [Ca(2+)]I and calmodulin (CaM) in cochlear nucleus of newborn mice infected by murine cytomegalovirus (MCMV) in the brain.

METHODSSixty-nine newborn mice were randomized into model group and control group. The model group (54 mice) was established by intracranial injection with MCMV viral suspension 20 l and the same volume of 0.9% sodium chloride was injected in the control group (15 mice). After 1 month, the ABR was tested in a sound-electric screen environment and the threshold was recorded. Then intracellular free calcium [Ca(2+)]i and the mRNA level of CaM in the cochlear nucleus were assayed by flow cytometry and RT-PCR.

RESULTSCompare to the control group [(64.0 ± 1.3) dBSPL], the threshold of ABR in the model group [(84.5 ± 2.7) dBSPL] was increased (F = 2.789,P = 0.000). Moreover, in the model group the intracellular free calcium [Ca(2+)]i and the mRNA level of CaM in the cochlear nucleus were increased (F = 1.290, P = 0.000; F = 4.252, P = 0.023), and the differences were statistically significant.

CONCLUSIONSThe intracranial injection of MCMV can lead to abnormal changes in the threshold of ABR in mice, and the change of [Ca(2+) ]I/CaM in cochlear nucleus may be the important pathological basis of sensorineural hearing loss induced by MCMV infection.

3T3 Cells ; Animals ; Auditory Threshold ; Calcium ; metabolism ; Calmodulin ; metabolism ; Central Nervous System Viral Diseases ; metabolism ; virology ; Cochlear Nucleus ; metabolism ; Cytomegalovirus ; Cytomegalovirus Infections ; metabolism ; Evoked Potentials, Auditory, Brain Stem ; Female ; Male ; Mice ; Mice, Inbred BALB C