A rapid, sensitive and convenient LC-MS/MS method was developed for the determination of γ-aminobutyric acid (GABA) in human plasma. d2-γ-Aminobutyric acid (d2-GABA) was synthesized as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all analytes were separated on a Luna HILIC column (100 mm x 3.0 mm, 3 μm) using an isocratic mobile phase of water: acetonitrile: formic acid (20 : 80 : 0.12) with a flow rate of 0.5 mL x min(-1). Acquisition of mass spectrometric data was performed in multiple reaction monitoring mode (MRM) in positive electrospray ionization using the transitions of m/z 104 --> 69 for GABA and m/z 106 --> 71 for d2-GABA. The method was linear in the concentration range of 5.00 to 1 000 ng x mL(-1). The intra- and inter-day precisions were within 9.9%, and accuracy ranged from 99.1% to 104%, within the acceptable limit across all concentrations. The method was successfully applied to a pharmacokinetic study of GABA tablets in healthy Chinese volunteers.
Chromatography, Liquid ; Humans ; Tandem Mass Spectrometry ; gamma-Aminobutyric Acid ; blood

By means of preparative HPTLC and column chromatography over silica gel and Sephadex LH-20, ten sesquiterpenes were isolated and purified from the whole plants of Solanum septemlobum Bunge. Based on the physico-chemical properties and spectral data, their structures were elucidated and identified as: lyratol D(1), solajiangxin B(2), 1 ,2-dehydrocyperone(3), solanerianone A (4), dehydrocarissone(5), ligucyperonol(6), nardoeudesmol A(7), solajiangxin F(8), and lyratol B(9), solajiangxin D(10). For the first time, compounds 1-10 were isolated from Solanum septemlobum, and compounds 5-7 were obtained from the genus Solanum.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Sesquiterpenes ; chemistry ; isolation & purification ; Solanum ; chemistry

Objective To construct a eukaryotic vector which contains avian H5N1 influenza virus hemagglutinin (HA) antigen and the cholera toxin B subunit (CTB) and to investigate its expression in COS7 cells,and the ability to induce specific immune responses in vivo in different periods.Methods After cloned by polymerase chain reaction (PCR),CTB and HA genes were digested with BamH Ⅰ and connected into CTB-HA gene with T4 ligase.The connected gene was referred to as CH.After double digestion,CH gene was inserted into a eukaryotic recombinant plasmid pCI-neo.The pCI-CH plasmid was then transfected into COS7 cells.Western blot was used to detect the expression of HA antigen.After New Zealand white rabbits were immunized,the titer of HA antigen-specific antibody in serum and its specificity with other strains such as H1N1,H9N1,H3N2 and influenza B virus were determined by indirect enzyme-linked immunosorbent assay.Results The pCI-CH vector (DNA vaccine) was successfully constructed,which could be efficiently expressed in COS7 cells and induce specific antibodies against pCI-CH in rabbits.Cross reactions indicated that DNA vaccine pCI-CH specific antisera could not only react with H5N1 strain (P/N＞2.1),but also H1N1,H9N1 and H3N2 strains,but did not cross react with influenza B virus.Conclusion The newly constructed avian H5N1 influenza virus nucleic acid vaccine has good immunogenicity.

Objective:To observe the clinical effect of mild moxibustion for knee osteoarthritis (KOA) and to explore the role of mild moxibustion in relieving pain,reducing stiffness and improving joint dysfunction in patients with KOA.Methods:Eighty patients with KOA were randomly allocated into either a moxibustion group or a medication group by the random number table,with 40 cases in each group.The moxibustion group used mild moxibustion at Neixiyan (EX-LE 5),Dubi (ST 35),Xuehai (SP 10) and Liangqiu (ST 34),30 min each time,3 times a week;the medication group was given celecoxib capsule (celebrex),0.2 g each time,once a day.Both groups were treated for 4 weeks.The visual analog scale (VAS) and Western Ontario and McMaster Universities osteoarthritis index (WOMAC)scores were evaluated before and after treatment.The efficacy of the two groups was compared after treatment.Results:After treatment,the overall efficacy of the moxibustion group was significantly different from that of the medication group (P＜0.05).The VAS and WOMAC scores of the two groups were lower than those before treatment (both P＜0.01).The changes in the VAS and WOMAC scores after treatment in the moxibustion group were significantly different from those in the medication group (both P＜0.05).After treatment,in single item of WOMAC,the changes in pain and joint dysfunction in the moxibustion group were more statistically significant than those in the medication group (both P＜0.05).Conclusion:Mild moxibustion and oral celebrex can reduce the VAS and WOMAC scores of patients with KOA.Mild moxibustion is superior to oral celebrex in relieving pain and improving joint function.

A simple, sensitive, selective, and reproducible liquid chromatography-tandem mass spectrometric method was developed for the simultaneous determination of repaglinide and metformin in human plasma using d5-repaglinide and d6-metformin as internal standards (ISs). After a simple protein precipitation using acetonitrile as the precipitation solvent, both analytes and ISs were separated on a Venusil ASB C 18 (150 mm x 4.6 mm, 5 microm) via gradient elution using acetonitrile--10 mmol x L(-1) ammonium acetate as the mobile phase. A chromatographic total run time of 7.5 min was achieved. Mass spectrometric detection was conducted with atmospheric pressure chemical ionization under positive-ion and multiple-reaction monitoring modes. The method was linear over the 0.2 to 60.0 ng x mL(-1) concentration range for repaglinide and over the 4 to 1 000 ng x mL(-1) range for metformin. For both analytes, the intra- and inter-accuracies and precisions were within the +/- 15% acceptable limit across all concentrations. The validated method was successfully applied to a clinical bioequivalence study.
Administration, Oral ; Adolescent ; Adult ; Area Under Curve ; Carbamates ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, Liquid ; Drug Stability ; Female ; Humans ; Hypoglycemic Agents ; administration & dosage ; blood ; pharmacokinetics ; Male ; Metformin ; administration & dosage ; blood ; pharmacokinetics ; Middle Aged ; Piperidines ; administration & dosage ; blood ; pharmacokinetics ; Tandem Mass Spectrometry ; Therapeutic Equivalency ; Young Adult

A simple and rapid method was developed based on high performance liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) to determine sivelestat and its metabolite XW-IMP-A in human plasma. After a simple protein precipitation, the samples and internal standards were analyzed on a C18 column by a gradient elution program. The mobile phase consisted of 30% acetonitrile in methanol and 5 mmol · L(-1) ammonium acetate at a flow rate of 0.7 mL · min(-1). The mass spectrometric data was collected in multiple reaction monitoring mode (MRM) in the negative electrospray ionization. The standard curves were linear in the range of 10.0-15,000 ng · mL(-1) for sivelestat, and 2.50-1000 ng · mL(-1) for XW-IMP-A. The low limits of quantitation were identified at 10.0 and 2.50 ng · mL for sivelestat and XW-IMP-A, respectively. The intra- and inter-day precision were within 11.3% and 13.1% for sivelestat and XW-IMP-A, and accuracy was 0.3% and 0.6% for sivelestat and XW-IMP-A, within the acceptable limits across all concentrations. The method was successfully validated in the pharmacokinetic study of sivelestat in healthy Chinese volunteers.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Glycine ; analogs & derivatives ; blood ; Humans ; Inosine Monophosphate ; blood ; Reproducibility of Results ; Sulfonamides ; blood ; Tandem Mass Spectrometry

OBJECTIVETo evaluate the efficacy and safety of domestically manufactured recombinant human thrombopoietin (rhTPO) in the treatment for chemotherapy-induced thrombocytopenia in patients with solid tumors.

METHODSA non-randomized controlled study was conducted. Seventy-two patients with platelet count < 75 x 10(9)/L after chemotherapy were enrolled into this study according to the standard criteria of NCI-CTC toxicity stratification. Thirty-five patients in the treatment-group received subcutaneous injection of rhTPO at a dose of 15,000 U/d, another 37 patients in the control group received subcutaneous injection of rhIL-11a at a dose of 3 mg/d as the paralleled control.

RESULTSThe mean minimal platelet count after rhTPO treatment was (46.2 +/- 20.3) x 10(9)/L in the treatment-group versus (37.2 +/- 16.7) x 10(9)/L in the control-group (P < 0.05), while the mean maximal platelet count was (250.2 +/- 159.0) x 10(9)/L versus (160.5 +/- 96.4) x 10(9)/L (P < 0.05). The incidence rate of adverse effects and duration of grade III and IV thrombocytopenia in the treatment-group was also significantly lower than those in the control-group. Furthermore, the patients receiving platelet transfusion were 4/35 in the treatment-group versus 11/37 in the control-group (P > 0.05). The side-effect rate in the treatment-group was significantly lower than that in the control-group (11.4% versus 78.4%, P < 0.01).

CONCLUSIONCompared with rhIL-11, administration of rhTPO after chemotherapy is safe and effective with mild adverse effects in reducing the degree and duration of thrombocytopenia.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Breast Neoplasms ; blood ; drug therapy ; Female ; Fever ; chemically induced ; Headache ; chemically induced ; Humans ; Lung Neoplasms ; blood ; drug therapy ; Male ; Middle Aged ; Platelet Count ; Recombinant Proteins ; adverse effects ; therapeutic use ; Thrombocytopenia ; chemically induced ; drug therapy ; Thrombopoietin ; adverse effects ; therapeutic use

OBJECTIVETo investigate the effect of Shenqi Fuzheng Injection (SFI) in radiation pneumonitis and its influence on the levels of plasma transforming growth factor beta (TGF-beta), tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) at various stages (pre-, mid- and post-radiation) of radiotherapy (RT).

METHODSFifty-eight patients enrolled in this study were randomized into two groups, all received conventional radio therapy, but the treatment group was intravenously dripped with SFI additionally, once daily, starting 3 days before radiotherapy to 1 week after the end of it. Levels of TGF-beta, TNF-alpha and IL-10 in blood samples collected and frozen at different time point of RT, before radiotherapy (Pre-RT), after received 40-50 Gy radiation (Mid-RT) and in time of terminating RT (Post-RT), were measured with enzyme linked immunosorbent assay (ELISA). And the occurrence of radiation pneumonitis was analyzed according to RTOG acute radiation pneumonitis criteria.

RESULTSLevel of TGF-beta increased in both groups after RT, either Mid- or Post-RT, but in comparing with that at Pre-RT, statistical significant (P < 0.05) only showed in the control group at Post-RT time when it raised to peak, while in inter-group comparing insignificance was shown. Level of TNF-alpha also increased in both groups, but statistical significance only showed in the control groups as compared with that of Pre-RT, accordingly, it was lower in the treatment group than that in the control group at corresponding time points (P < 0.05). Level of IL-10 decreased gradually in the process of RT, as compared with that at Pre-RT, significant difference was shown in both groups at Post-RT but not at Mid-RT, as compared between groups, no significance (P < 0.05) was shown, though the treatment group showed a high tendency. As for the ratio of IL-10/TNF-alpha, significant difference of lowering in the control group after RT was shown as compared with that at Pre-RT (P < 0.05), and also as compared with that in the treatment group at corresponding time point (P < 0.05). The occurrence of radiation pneumonia > or = grade 2 in the treatment group was significantly lower than that in the control group (chi2 = 8.7133, P < 0.01).

CONCLUSIONPlasma level of TNF-alpha and ratio of IL-10/ TNF-a could be the practicable indexes for estimating the susceptibility of acute radiation pneumonia; SFI can regulate the network of cytokine, and thus be effective in preventing and treating radiation pneumonitis.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; radiotherapy ; Carcinoma, Squamous Cell ; radiotherapy ; Cytokines ; blood ; Drugs, Chinese Herbal ; therapeutic use ; Esophageal Neoplasms ; radiotherapy ; Female ; Humans ; Interleukin-10 ; blood ; Lung Neoplasms ; radiotherapy ; Male ; Middle Aged ; Phytotherapy ; Radiation Pneumonitis ; blood ; prevention & control ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

OBJECTIVETo study the sensory integration function in children with primary nocturnal enuresis (PNE).

METHODSThe sensory integration function was assessed by the Childhood Sensory Integration Ability Development Checklist in 70 children with PNE and was compared with that in 74 normal children(control group).

RESULTSThe incidence of sensory integration dysfunction (76%) in the PNE group were significantly higher than that in the control group (35%; P<0.01). Severe sensory integration dysfunction occurred in more children in the PNE group compared with the control group (39% vs 18%; P<0.01). The scores of all sensory integration indexes revealed by sensory integration function testing in the PNE group were significantly lower than those in the control group (P<0.01).

CONCLUSIONSThe majority of children with PNE have sensory integrative dysfunction which presents in various aspects. It is necessary to assess the sensory integration function in children with PNE.

Brain ; physiopathology ; Child ; Female ; Humans ; Male ; Nocturnal Enuresis ; etiology ; physiopathology ; Sensation

The aim of the present study is to investigate the effects of Panax notoginseng saponins (PNS) on pneumocyte apoptosis and apoptosis-related protein, as well as c-Jun N-terminal kinase (JNK) in lung ischemia/reperfusion (I/R) injury. Thirty Wistar rats were randomly divided into control group, I/R group and PNS group. The unilateral lung I/R model was replicated by obstruction of left lung hilus for 30 min and reperfusion for 120 min in vivo. The rats in PNS group were given intraperitoneal injection of PNS at 60 min before ischemia and 10 min before reperfusion. Some lung tissues sampled at the end of the experiment were assayed for wet/dry weight ratio (W/T). The expressions of phosphorylated JNK (p-JNK) and JNK protein were detected by Western blot. The expressions of Bcl-2, Bax and Caspase-3 protein were detected by immunocytochemistry techniques. The pneumocyte apoptotic index (AI) was detected by terminal deoxynuleotidy1 transferase mediated dUTP nick end labeling (TUNEL). The morphological and ultrastructure changes were observed under light microscope and electron microscope, and the injured alveolus rate (IAR) was counted as well. The results showed that compared to control group, I/R group showed increased expressions of p-JNK, Bcl-2, Bax and Caspase-3 protein (all P < 0.01), decreased ratio of Bcl-2/Bax (P < 0.05), and increased values of AI, W/T and IAR (all P < 0.01). Moreover, light microscope and electron microscope showed serious morphological and ultrastructure injury in I/R group. Compared to I/R group, PNS group showed markedly decreased expressions of p-JNK, Bax and Caspase-3 protein (all P < 0.01), increased expression of Bcl-2 protein and ratio of Bcl-2/Bax (both P < 0.01), and lower values of AI, W/T and IAR (all P < 0.01). Meanwhile, light morphological and ultrastructure injury was found to be alleviated in PNS group. These results suggest that PNS can protect lung tissue from I/R injury, and the mechanism may correlate with suppressing JNK signal pathway, up-regulating the ratio of Bcl-2/Bax which results in inhibition of Caspase-3 dependent apoptosis.
Alveolar Epithelial Cells ; drug effects ; Animals ; Apoptosis ; drug effects ; Female ; Ischemia ; physiopathology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Lung ; blood supply ; metabolism ; pathology ; Male ; Panax notoginseng ; chemistry ; Rats ; Rats, Wistar ; Reperfusion Injury ; prevention & control ; Saponins ; isolation & purification ; pharmacology ; Signal Transduction ; drug effects