With the advance of drug development and research techniques, the drug metabolic processes and mechanism can be more deeply achieved. As the drug metabolism and pharmacokinetics process are mediated by drug metabolizing enzymes and transporters, study of drug metabolizing enzymes and transporters has become an important part for drug development. The traditional immunoassays with low sensitivity and poor specificity can not reflect the accurate expression level of drug metabolizing enzymes and transporters. We now give a brief review on the quantitative study of drug metabolizing enzymes and transporters by mass spectrometry-based proteomic approach.
Enzymes ; chemistry ; Humans ; Inactivation, Metabolic ; Mass Spectrometry ; Membrane Transport Proteins ; chemistry ; Pharmacokinetics ; Proteomics

The fragmentation pathways of five estrogens (estradiol, estrone, equilin sulfate, 17 a-dihydroequilin sulfate and equilenin sulfate) have been studied with high resolution and high mass accuracy using electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF/MS) in the negative ion mode. Molecular weights were obtained from [M-H](-) ions in the product ion spectra. The results indicate that the five structurally similar estrogens have similar fragmentation pathways. Using their stable isotope forms as internal reference compounds, the accurate mass and composition of the fragment ions were determined. During collision-induced dissociation (CID), cleavage is initiated by loss of oxygen atoms from carbon-17, after which D and C rings cleave sequentially and rearrange to finally form stable conjugate structures with highly abundant characteristic fragment ions at m/z 183 (accompanied by m/z 181), m/z 169 and m/z 145 (accompanied by m/z 143). Understanding these characteristic fragmentation pathways of estrogens will be helpful in identifying the structures of steroid hormones in general.
Chemical Fractionation ; methods ; Equilenin ; chemistry ; Equilin ; analogs & derivatives ; chemistry ; Estradiol ; chemistry ; Estrogens ; chemistry ; Estrone ; chemistry ; Ions ; Spectrometry, Mass, Electrospray Ionization

The fragmentation pathways of two taxanes drugs have been studied in positive ion mode by Q-TOF with the advantages of high mass accuracy and high resolution analysis. The [M+H] + ions were observed by ESI-MS, from which the molecular weights were obtained. Using the protonated pseudo-molecular ions [M+H]+ as internal reference compounds, the accurate mass and element composition of the fragment ions were determined. The collision induced dissociation (CID) data of the [M+H] ions provided fragmentation pathways of related compounds. Results showed that the major cleavage pathways of paclitaxel and docetaxel were the same that the cleavage of C-O bond between the side chain and taxol skeleton easily occurred, then stripping of the functional groups on the parent ring. Some common fragments were formed, such as m/z 105.033 7, 291.137 3, 309.148 5, 327.159 7, 387.181 2 and 509.217 4, which would provide a basis for future qualitative and quantitative analysis of taxanes in vitro and in vivo.
Paclitaxel ; chemistry ; Peptide Fragments ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Taxoids ; chemistry

The bevacizumab and its glycoforms were analyzed by matrix-assisted laser desorption ionization-time of flight-time of flight-mass spectrometry (MALDI-TOF-MS), sodium dodecyl sulfate polyacrylamide gel electrophoresis ( SDS-PAGE) and short-gun strategy, with the sequence of unique peptide and seventeen glycoforms being characterized. The bevacizumab and its glycopeptides concentrations in mice plasma with different intravenous injection doses of bevacizumab were detected and the concentration-time curves were obtained by parallel reaction monitoring ( PRM) method based on liquid chromatography-tandem mass spectrometry (LC-MS / MS) technique. First, standard curves were created for quantification of mAb in mice plasma, which showed good linearity, with the correlation coefficient (R2 ) value of 0. 998 and the lower limit of quantification of 66 fmol. Detection results of high and low doses of the drug in the mice plasma samples showed that the drug concentration-time curve trend was consistent, e. g. the concentration was decreasing. However, the results of quantitation of seventeen glycoforms demonstrated that the metabolism of different glycoforms was different. The concentrations of most glycoforms increased first, whereas the metabolism afterwards differed by different glycoforms.

The paper is aimed to provide a novel index, named as multidimensional spatial triangular area, for the evaluation of the release-absorption correlation of multiple component traditional Chinese medicines (TCMs). The applicability of the method was demonstrated by the example data. The method and standard practice for evaluation of the release-absorption correlation for western medicines with single compound could not be applied to TCMs with multiple components. The release percentage or absorption percentage of the multiple components for TCMs at the sampling time was a point in the multidimensional space. The area of the triangle formed byt the sequential three points represented the changing characteristics of the components' release and absorption kinetics. The side lengths of the triangle could be calculated from the spatial distances between each two of the sequential three points. Then the triangle area could be obtained by the side lengths. The in vitro release-in vivo absorption correlation of the multiple components could be represented by the correlation between the integrating values of the release triangle areas and that of the absorption triangle areas. The results of the examples indicated that the multidimensional spatial triangular area method could treat the multiple components in a holistic way, in line with the holism the hi he TCMs. Therefore, the multidimensional spatial triangular area method provided new methodology for the release-absorption correlation of the TCMs with multiple components.
Algorithms ; Chemistry Techniques, Analytical ; Drug Combinations ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; pharmacokinetics ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry

OBJECTIVETo evaluate and analyze the mechanism of Pizhen for the treatment of patients with scapular muscle fasciitis.

METHODSOne hundred and sixteen patients were divided into two groups according to random number table method, 58 cases in the treatment group and 58 cases in the control group. After treatment, 109 patients were followed up, and 7 patients lost follow-up (2 cases in the treatment group and 5 patients in the control group). In the treatment group, there were 15 males and 41 females, with a mean age of (28.02 +/- 4.85) years; the course of disease ranged from 0.25 to 3 years; and the patients were treated by Pizhen once a week,and 2 weeks were one treatment course. In the control group, there were 19 males and 34 females, with a mean age of (27.23 +/- 4.54) years old; the course of disease ranged from 0.25 to 1 year; and the patients were treated by acupuncture massage instrument, 20 minutes at a time,once a day for 2 weeks. Soft tissue displacement and banana area were tested by a soft tissue tension tester, pressure value was measured by pressure measuring instrument, and VAS was recorded by pain visual analog scale record.

RESULTSThere were significant differences in tissue displacement, banana area, pressure value and VAS between two groups before and after treatment. The tissue displacement and pressure value in the treatment group were higher than those in the control group,while banana area and VAS were lower than those in the control group. In the treatment group, 30 patients healed, 13 patients got marked improvement, 12 good and 1 poor;while 16 patients healed, 19 patients got marked improvement,11 good and 7 poor in the control group. The therapeutic effects in the treatment group was better than that in the control group.

CONCLUSIONPizhen is an effective method to treat pain in patients with scapular muscle fasciitis by relaxing tendon of soft tissue around pain point of scapular and relaxing compress and stimulation of nerves.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Adult ; Case-Control Studies ; Fasciitis ; therapy ; Female ; Humans ; Male ; Treatment Outcome ; Young Adult

Due to the diversity of components within the traditional Chinese medicines (TCMs), the release profiles of the components in the TCM dosage forms vary dramatically and no quantification method is available to determine the variance yet. Based upon the principles of Kalman filter method, the authors defined a new parameter, relative chemomic error (epsilon), to evaluate the asynchronous nature of the components in TCMs, and a derivative parameter as synchronization factor (SF) to quantify the synchronicity of the chemome of the TCMs. The average synchronization factor (SF(av)) was accordingly derived to simultaneously quantify the release/dissolution profiles of the multi-components in TCMs. Randomly generated simulation data were processed to demonstrate the chemomic data processing and the methodology. The results indicated that the novel parameter epsilon was well correlated (r = 0.996 8) with the coefficient of variation from the conventional release profiles of all the components. As the asynchronicity was the intrinsic characteristics of the multi-component TCMs, the synchronicity might be a new target of quality control of TCMs. The methods established by this report can be used a quantitative tool for the evaluation of the chemomic release synchronization of TCMs.
Algorithms ; Chemistry Techniques, Analytical ; Drug Combinations ; Drugs, Chinese Herbal ; analysis ; chemistry ; Evaluation Studies as Topic ; Medicine, Chinese Traditional

BACKGROUNDThe urokinase plasminogen activator system is believed to play an important role in degradation of the extracellular matrix associated with cartilage and bone destruction; however its precise roles in temporomandibular disorders have not yet been clarified. The aims of this study were to investigate the gene expression of fibrinolytic factors urokinase plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) in the articular cartilage of rabbit temporomandibular joint (TMJ) with disc displacement (DD) and to probe the relationship between fibrinolytic activity and cartilage remodeling.

METHODSDisc displacement of right joints was performed in 36 of 78 rabbits under investigation. The animals were sacrificed at 4 days and 1, 2, 4, 8 and 12 weeks after surgery, respectively. The right joints of these animals were harvested and processed for the examination of mRNA expression of uPA and PAI-1 in articular cartilage using in situ hybridization techniques.

RESULTSThe expression of uPA and PAI-1 was co-expressed weakly in the chondrocytes from transitive zone to hypertrophic zone and mineralized zone, while no hybridizing signals were shown in proliferative zone and superficial zone in control rabbits. The most striking was the up-regulation of uPA and PAI-1 mRNA in 4-day rabbits postoperatively at the onset of cartilage degeneration. The strongest hybridizing signals for uPA and PAI-1 were seen in 2-week rabbits postoperatively. After 2 weeks, the expression of uPA and PAI-1 began to decrease and reached nearly normal level at 12 weeks.

CONCLUSIONSThe expression of the uPA/PAI-1 system coincides with the pathological changes in condylar cartilage after DD. The uPA/PAI-1 system may be one of the essential mediators in articular cartilage remodeling.

Animals ; Cartilage, Articular ; metabolism ; Female ; Joint Dislocations ; metabolism ; pathology ; Male ; Mandibular Condyle ; metabolism ; pathology ; Plasminogen Activator Inhibitor 1 ; genetics ; RNA, Messenger ; analysis ; Rabbits ; Temporomandibular Joint ; metabolism ; Temporomandibular Joint Disc ; Urokinase-Type Plasminogen Activator ; genetics

OBJECTIVETo investigate the effect of anterior disc displacement on the expression of urokinase plasminogen activator and its inhibitor-1 (uPA/PAI-1) in synovial tissues.

METHODSForty Japanese white rabbits were used in this study. The animals were killed at 4 days, 1, 2, 4, 8 and 12 weeks postoperatively, respectively. In situ hybridization technology was applied to detect the expression of uPA/PAI-1 mRNA in synovial membrane.

RESULTSIn normal synovial tissues, synovial lining cells and a few fibrosblasts with mild positive staining were occasionally seen. More synovial lining cells and fibrosblasts with moderate postive signals were found 1 week after operation. Since then, the degree of staining for uPA/PAI-1 increased gradually. By the end of 12 weeks postoperatively, strong signals of uPA/PAI-1 mRNA were detected.

CONCLUSIONThere is a harmonized uPA/PAI-1 system existing in synovial tissues. The high expression of uPA and PAI-1 mRNA in synovial tissues indicates that the uPA/PAI-1 system may play an important role in the process of synovitis resulted from anterior disc displacement.

Animals ; In Situ Hybridization ; Plasminogen ; Plasminogen Activator Inhibitor 1 ; RNA, Messenger ; Rabbits ; Synovial Membrane ; Urokinase-Type Plasminogen Activator

Objective To explore the value of dual-phase contrast-enhancement multislice computed tomography (MSCT) in the assessment of acute myocardial infarction volume and perfusion in porcine models. Methods The distal left anterior descending coronary arteries of 5 pigs were balloon-occluded for 90 min and followed by reperfusion. MSCT was performed 1 min (early phase) and 5 min (delayed phase) after administration bolus of 100 mL of iodinated contrast material 30 min after reperfusion. On the same day, hearts were excised, sectioned in 8 mm short-axis slices, and stained with TTC. Infarction volume was defined as the sum of the hyper-enhanced area and surrounding hypo-enhanced area in all slices on delay enhanced phase of MSCT and the TTC-negative area on TTC staining slices. Infarction volume was expressed as percentage of total slice volume. Results Acute infarction detected by MSCT was characterized by early myocardial perfasion defects in the early phase of the contrast bolus (early defects) with surrounding residual defects and late enhancement observed in the late phase. Mean CT attenuation value of early defects was significantly different from CT attenuation value of remote myocardium [(213±55)HU vs (304±30)HU](P < 0.05), CT attenuation values of residual defects and late enhancement were also significantly different from those of remote myocardium [(360±75) HU vs (90±37) HU and (152±23) HU vs (190±37) HU, repectively](P < 0.01, P < 0.05). The mean infarction volume was (8.9± 1.0)% on MSCT and (9.2±1.4)% on TTC pathology images. The infarction volume assessed by MSCT compared well with TTC staining slices. Conclusion Acute reperfused myocardial infarction zone has specific enhancement pattens different to remote normal zone on dual phase MDCT, which is in good agreement with in vivo Trc pathology in the assessment of acute reperfused myocardial infarction shortly offer reperfusion.