To establish a LC-MS/MS method for quantification of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid in rats plasma and study its pharmacokinetics after administration of Mailuoning injection at a single dose to rats. Plasma samples were acidified with hydrochloric acid and extracted with ethyl acetate. The analytes were determined by LC-MS-MS using a ZOBAX SB C18 column with a mobile phase of methanol-water (containing 2 mmol x L(-1) ammonium acetic) (60:40)at a flow rate of 0.5 mL x min(-1) and detected using ESI with negative ionization mode. Ions monitored in the multiple reaction monitoring (MRM) mode were m/z 353.1/191.0 [M-H]- for chlorogenic acid, m/z 178.9/134.9 [M-H]- for caffeic acid, m/z 515.2/353.0 [M-H]-for 3,4-DCQA, m/z 193.0/133.9 [M-H]-for ferulic acid, m/z 146.9/102.9 [M-H]- for cinnamic acid and m/z 246.0/125.8 [M-H]- for tinidazole (IS). After administration of Mailuoning injection at a single dose to eight Sprague-Dawley rats, the concentrations of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid in plasma were determined by LC-MS/MS method. The main pharmacokinetics parameters of measured data were caluculated by using DASver 1.0 software. The linear concentration ranges of the calibration curves for chlorogenic acid, caffeic acid, 3,4-DCQA and cinnamic acid were 2.006-1,027 microg x L(-1) (r = 0.999 6), 1.953-1,000 microg x L(-1) (r = 0.999 7), 28.51-1.459 x 10(4) microg x L(-1) (r = 0.998 9), 1.836-940.0, g x L(-1) (r = 0.997 7) and 4.780-2,447 microg x L(-1) (r = 0.998 6) respectively. The inner and inter-days relative standard deviations were both less than 5.0%, indicating legitimate precise and accuracy to the requirement of biological sample analysis. For chlorogenic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (49.78 +/- 12.81) min, (123.55 +/- 14.82) mg x min x L(-1) and (0.004 3 +/- 0.000 5) L x min(-1), respectively. For caffeic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (36.65 +/- 10.59) min, (91.67 +/- 11.77) mg x min L(-1) and (0.005 7 +/- 0.000 7) L x min(-1), respectively. For 3,4-DCQA, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (50.08 +/- 13.78) min, (278.34 +/- 31.82) mg x min x L-1 and (0.001 6 +/- 0.000 2) L x min(-1), respectively. For ferulic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (51.39 +/- 15.52) min, (34.72 +/- 4.67) mg x min x L(-1) and (0.000 4 +/- 0.0001) L x min(-1), respectively. For cinnamic acid, the pharmacokinetic parameter t1/2, AUCo-t, and CL were (74.42 +/- 18.32) min, (34.63 +/- 4.82) mg x min x L(-1) and (0.007 7 +/- 0.001 1) L x min-', respectively. The assay method is proved to be sensitive, accurate and convenient. It can be applied to the pharmacokinetic study of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid.
Animals ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Female ; Hydroxybenzoates ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

Objective To evaluate effects of interleukin-36α (IL-36α) on psoriasiform skin lesions and C-C motif chemokine ligand 20 (CCL20) expression in mice.Methods Totally,30 BALB/c female mice were randomly and equally divided into 3 groups:control group treated with topical vaseline cream on the shaved back and intracutaneous injection with phosphate buffer saline (PBS),model group treated with topical imiquimod cream on the shaved back and intracutaneous injection with PBS,experimental group treated with topical imiquimod cream on the shaved back and intracutaneous injection with IL-36α solution.Psoriasis area severity index (PASI) was used to evaluate changes of psoriasiform skin lesions in mice,and light microscopy to observe morphological changes of skin lesions and to measure the thickness of the epidermis.Real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis were performed to determine the expression of IL-36α in skin lesions in the control group and model group,and qRT-PCR,Western blot analysis and immunohistochemical study to evaluate changes of CCL20 levels in skin lesions.Results The model group showed significantly increased mRNA (△ Ct value:0.0195 ± 0.0059) and protein expression (3.922 ± 0.248) of IL-36α compared with the control group (mRNA:0.0012 ± 0.0004,P ＜ 0.05;protein:0.690 ± 0.025,P ＜ 0.05).The mRNA and protein expression of CCL20 were significantly higher in the experimental group than those in the model group (mRNA:2.152 ± 0.793 vs.0.999 ± 0.178;protein:0.397 ± 0.033 vs.0.145 ± 0.030;both P ＜ 0.05),and higher in the model group than those in the control group (mRNA:0.378 ± 0.075;protein:0.025 ± 0.009;both P ＜ 0.05).Immunohistochemical study showed that the expression intensity of CCL20 in skin lesions significantly increased in the experimental group compared with that in the model group (Z =2.294,P ＜ 0.05).Conclusion IL-36α may aggravate psoriasiform skin inflammation in mice by promoting CCL20 expression.

This study aimed to explore the impact of depression caused by chronic unpredictable mild stress (CUMS) on in vivo activity of six kinds of CYP450 isoforms in rats. According to 'Katz' method, the model of CUMS was established. Tolbutamide, chlorzoxazone, theophylline, midazolam, omeprazole and dextromethorphan were chosen as probe substrates of CYP2C6, CYP2E1, CYP1A2, CYP3A2, CYP2D1 and CYP2D2 of rats. Plasma concentration of six kinds of CYP450 in control group and model group were determined by LC-MS/MS and computed pharmacokinetic parameters. Consequently, metabolism of theophylline and chlorzoxazone accelerated significantly (P < 0.01), but tolbutamide, dextromethorphan, omeprazole and midazolam had no significant difference. The present study proved that depression caused by CUMS had strong induction to CYP1A2 and medium induction to CYP2E1.
Animals ; Chlorzoxazone ; metabolism ; Chromatography, Liquid ; Cytochrome P-450 Enzyme System ; metabolism ; Depression ; Dextromethorphan ; metabolism ; Liver ; enzymology ; Midazolam ; metabolism ; Omeprazole ; metabolism ; Rats ; Stress, Physiological ; Tandem Mass Spectrometry ; Theophylline ; metabolism ; Tolbutamide ; metabolism

OBJECTIVETo observe the clinical effect and safety of circumcision stapler in the treatment of phimosis and redundant prepuce.

METHODSWe treated 120 patients with redundant prepuce or phimosis using circumcision stapler and another 60 by conventional dorsal-incision circumcision. We observed intraoperative blood loss, operation time, postoperative pain, wound healing time, cosmetic appearance of the penis, and postoperative complications and compared them between the two groups of patients.

RESULTSStapler circumcision showed obvious advantages over the conventional method in intraoperative blood loss ([2. 3 ± 1. 3] vs [15.6 ± 2.9] ml), operation time ([7.1 ± 1.4] vs [22.6 ± 4.6] min), wound healing time ([12.0 ± 2.9] as [16.3 ± 3. 1] d), postoperative pain score (1. 9 ± 1. 3 vs 5. 2 ± 1. 7), incision edema, and cosmetic appearance of the penis (all P <0. 05). Besides, stapler circumcision exempted the patients from stitch-removal pain. However, the incidence rate of postoperative local ecchymosis was significantly higher in the circumcision stapler group than in the conventional circumcision group (20. 8% vs 8. 3% , P <0. 05).

CONCLUSIONCircumcision stapler, with its advantages of easier manipulation, shorter operation time, better cosmetic penile appearance, less pain, and fewer complications, is superior to conventional circumcision in the treatment of phimosis and redundant prepuce.

Blood Loss, Surgical ; Circumcision, Male ; instrumentation ; methods ; Ecchymosis ; etiology ; Humans ; Male ; Pain, Postoperative ; Penis ; abnormalities ; Phimosis ; therapy ; Postoperative Complications ; Postoperative Period ; Surgical Staplers ; adverse effects ; Wound Healing

Objective To explore the impact factors for post-thaw embryo survival rate and clinical pregnancy rate in frozen-thawed embryo transfer program. Methods The clinical data of 573 cycles of frozen-thawed embryo transfers were retrospectively analysed. Groups were divided according to the pre-freeze embryo quality, pre-freeze embryonic developmental stage, frozen-thawed embryo quality and cryopreservation technique, respectively, and post-thaw embryo survival rates and/or clinical pregnancy rates were compared among groups. Results The clinical pregnancy rate of high quality pre-freeze embryo was significantly higher than that of low quality pre-freeze embryo (31.8% vs 20.0%) (P< 0.05). There was no significant difference in the post-thaw survival rates and clinical pregnancy rates between embryos frozen at day 2 of ferrtilization and those frozen at day 3 of ferrtilization(79. 1% vs 82.9% and 25.5% vs 31.2%, respectively) (P>0.05). The clinical pregnancy rates of the transfer cycles only with fully intact embryos and with mixed embryos were significantly higher than that only with partially damaged embryos(36.7% vs 24.1% and 29.2% vs 24.1%, respectively)(P<0.05). The post-thaw survival rate and post-thaw high-quality embryo rate were significantly higher in those processed with modified cryopreservation technique than in those processed with original cryopreservation technique (82.0% vs 66.3% and 50.0% vs 27.5%, respectively)(P<0.05). Conclusion Pre-freeze embryo quality, post-thaw embryo survival rate and post-thaw embryo quality have a positive correlation to subsequent clinical pregnancy rate. Favorable cryopreservation technique may ensure the success of post-thaw embryo recovery and transfer.

Objective To study the efficacy and security of postoperative radiotherapy alone and post-operative radiotherapy combined with chemotherapy in patients with high-grade cerebral glioma.Methods Fifty-nine patients with high-grade cerebral glioma confirmed by pathology, including 35 with grade Ⅲ and 24 with grade Ⅳ, admitted to our hospital from June 2004 to June 2008, were chosen in our study; these patients were randomly divided into 3 groups (A, B and C). All the patients underwent three-dimensional conformal radiation therapy (3D-CRT) with a dose of 54-66Gy/27-33f/5-7w by 6/10MV-X ray; and the median total dose was 60 Gy; group A (n=21, including 13 with grade Ⅲ and 8with grade Ⅳ) received chemotherapy with termozolomide (TMZ) at a dosage of 75 mg/m2·d during radiotherapy, following by 150-200 mg/ (m2·d) for 5 d, and enjoying 28 d per cycle for a total of 3-5cycles; group B (n=26, including 14 with grade Ⅲ and 12 with grade Ⅳ) received chemotherapy with the regimen of nimustine (ACNU) with a dosage of 90 mg/m2 on the 1st d and teniposide (VM-26) at a dosage of 60 mg/m2 on the 1st, 2nd and 3rd d, enjoying 6-8 weeks per cycle for a total of 4-6 cycles; group C (n=12,including 8 with grade Ⅲ and 4 with grade Ⅳ) received radiotherapy alone. Clinical evaluations of tumor response and adverse effects were performed periodically. The primary end points were disease progression-flee survival (PFS) and overall survival (OS) Results The effective rate in the group A, B and C was 81.0％, 71.4％ and 33.3％, respectively, indicating that significant differences exited between group C and both group A and B (P＜0.05). The incidence of gastrointestinal side effects and toxic response in the group B was obviously higher than that in the group A (P＜0.05). The 1-, 3- and 5-year survival rates in group A were 66.7％, 19.0％ and 9.5％, respectively, which were the highest among the 3groups; those in group B were 53.8％, 15.4％ and 3.8％, respectively; those in group C were 25％, 8％ and 0％, respectively. The PFS in all the patients was 8 months and OS was 15 months; log-rank test indicated that significant differences of PFS and OS existed between patients received concomitant chemoradiotherapy and radiotherapy (X2=10.710, P=0.005; X2=7.185, P=0.028); the incidence of PFS and OS in group A and B was significantly higher than that in group C (P＜0.05). Conclusion To post-operative patients with high-grade cerebral glioma, concomitant chemoradiotherapy can improve the effective rate and extend the PFS and OS. TMZ is recommended as the concomitant chemotherapy regimen, having similar therapy effect with ACNU plus VM-26, but enjoying less adverse effects.

OBJECTIVETo investigate the Churg-Strauss syndrome (CSS) associated lung involvement, concentrating on clinical characteristics, pathological findings of lung involvements, response to treatment, and prognosis.

METHODSWe retrospectively analyzed the characters of the clinical manifestations, thin-section CT and pathological findings of CSS. The study involved 16 patients. Clinical data were obtained by chart review. All patients underwent transbronchial lung biopsy (TBLB). Six of them underwent surgical lung biopsy as well.

RESULTSThe patients included 7 men and 9 women, aged from 14 to 61 years (median, 47.5 years). Extrathoracic organs involved included nervous system (7/16) and skin (5/16). Respiratory symptoms included cough (12/16), exertional dyspnea (11/16), hemoptysis (4/16), and chest pain (3/16). CT findings included bilateral ground-glass opacities (12/16), bilateral patchy opacities (12/16), and centrilobular nodules (6/16). The pathological findings of TBLB demonstrated increased eosinophils (3/16), vasculitis (3/16), and interstitial pneumonia (16/16). The pathological findings of surgical lung biopsy of 6 cases showed necrotizing vasculitis in 4 cases, capillaries in 5, eosinophilic pneumonia in 3, granulomas in 2, and airway abnormalities in 3. All patients improved in symptoms after therapy during the study period (range, 3 to 51 months; median, 15 months).

CONCLUSIONSAsthma may be present in CSS patient when there is bronchial involvement. Ground-glass opacities and consolidation seen on high-resolution CT reflect the presence of eosinophilic pneumonia, vasculitis, and pulmonary alveolar hemorrhage. TBLB has significant limitations for the diagnosis of CSS. Early diagnosis and therapy can result in satisfactory prognosis.

Adolescent ; Adult ; Asthma ; physiopathology ; Biopsy ; Churg-Strauss Syndrome ; diagnosis ; diagnostic imaging ; drug therapy ; pathology ; Cyclophosphamide ; Female ; Humans ; Immunosuppressive Agents ; therapeutic use ; Lung ; diagnostic imaging ; pathology ; physiopathology ; surgery ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Tomography, X-Ray Computed ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo study serum levels of brain natriuretic peptide (BNP) in children with Kawasaki disease (KD) and the correlation between BNP levels and the heart function.

METHODSForty-three children with KD and thirty healthy children were enrolled. Serum levels of BNP were measured using ELISA. KD children received an echocardiographic examination, including measurements of left ventricular ejection fraction (LVEF), left ventricular shorten fraction (LVSF), cardiac index (CI) and left ventricular inflow velocity through the mitral annulus.

RESULTSMean serum level of BNP at the acute stage in children with KD was significantly higher than that at the recovery stage as well as the control group (p<0.01). The LVEF, LVSF and CI levels at the acute stage were significantly lower than those at the recovery stage in children with KD (p<0.05). The linear regression analysis showed that the BNP level was negatively correlated with the levels of LVEF, LVSF and CI (r=-0.63, -0.52, -0.53, p<0.05).

CONCLUSIONSThe serum BNP levels increase significantly in KD children at the acute stage, and are negatively correlated with the levels of LVEF, LVSF and CI. Measurement of serum BNP level is useful for the early diagnosis of KD.

Child ; Child, Preschool ; Female ; Heart ; physiopathology ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; blood ; diagnosis ; physiopathology ; Natriuretic Peptide, Brain ; blood

OBJECTIVETo explore the effect of arterial perfusion of marrow multifunctional stem cells (MFSC) in treating femoral head necrosis and its mechanism.

METHODSThe rabbit model of femoral head necrosis was established by large dose of methyl-prednisone through Shwartzman response. Bone marrow was extracted from femoral bone of model rabbit and isolated in vitro for culturing and proliferating MFSC. The experimental rabbits were randomly divided into 4 groups, treated with normal saline (A), Salvia + urokinase (B), MFSC (C) and MFSC + Salvia + urokinase (D), respectively, they were sacrificed in batches at 2 and 4 weeks after treatment, and changes in various parameters, including molybdenum target roentgenogram, routine pathology with HE staining, tetracycline labeled fluorescent microscopy and ultrastructure alteration by scanning electron microscope (SEM), were observed.

RESULTSTypical appearance of femoral head necrosis was shown in the successfully modeled rabbits. Two and 4 weeks after treatment by high selective drug via medial and lateral femoral circumflex arterial perfusion, the X-ray examination showed significant improvement of bone density; pathohistologic manifestation showed decrease of empty bone lacuna, increase of osteoblast and new bone formation; tetracycline fluorescent labeled microscopic picture showed bright fluorescent band of increased osteoblasts in necrosis repairing region with widened border; SEM displayed irregularly arranged fibrosis in necrosis region, abundant organelles in osteoblasts with few empty bone lacuna. The above-mentioned improvement was more significant in rabbits treated by MFSC.

CONCLUSIONHigh selective femoral drug arterial perfusion in treating femoral head necrosis could accelerate the process of revascularization and re-ossification in rabbits. As compared with Salvia, MFSC showed quicker and more potent effect.

Animals ; Bone Marrow Transplantation ; methods ; Femur Head ; blood supply ; Femur Head Necrosis ; chemically induced ; physiopathology ; therapy ; Infusions, Intra-Arterial ; Male ; Methylprednisolone ; Osteogenesis ; Rabbits ; Stem Cell Transplantation ; methods

To observe the serum samples and the anti-inflammatory effect of Tripterygium wilfordii in treating RA by using the pharmacokinetic-pharmacodynamic model, make a correlation analysis on concentration-time and effect-time curves, and explore RORγt, IL-17, STAT3, IL-6 mRNA transcriptional levels in rats by PCR. Methotrexate, tripterine and high-dose T. wilfordii could down-regulate RORγt, IL-17, STAT3, IL-6 mRNA transcriptional levels in AA rat lymph nodes. The study on PK-PD model showed correlations between inflammatory factors and blood concentration of T. wilfordii. T. wilfordii and its main active constituent tripterine could show the inflammatory effect and treat RA by inhibiting IL-17 cytokine.
Animals ; Arthritis, Rheumatoid ; drug therapy ; immunology ; Biomarkers ; Female ; Interleukin-17 ; antagonists & inhibitors ; genetics ; Interleukin-6 ; genetics ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Tripterygium ; Triterpenes ; pharmacokinetics ; pharmacology