Using brain microdialysis and LC-MS/MS to detect acetylcholine in rat brain to investigate the effects of ligustrazine. A liquid chromatography-tandem mass spectrometry method has been developed and validated for the determination of acetylcholine in rat brain dialysate sampling by microdialysis. The results indicated that ligustrazine administration by subcutaneous injection significantly increased Ach release in rat medial prefrontal cortex and nucleus accumbens in a dose-related manner. The drug' s effect on Ach release in rat brain could be directly detected by microdialysis combined with HPLC-MS/MS and this method is selective and sensitive.
Acetylcholine ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Dose-Response Relationship, Drug ; Ligusticum ; chemistry ; Male ; Microdialysis ; Nucleus Accumbens ; metabolism ; Plants, Medicinal ; chemistry ; Prefrontal Cortex ; metabolism ; Pyrazines ; administration & dosage ; isolation & purification ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

OBJECTIVETo investigate the effect and mechanism of BSDW on the model of allergic rhinitis and the model of guinea pigs by histamine shocking in guinea pigs.

METHODUsing the model of allergic rhinitis in guinea pigs caused by 10% TDI, we observed the effect of BSDW on physiological and pathological symptoms of allergic rhinitis in guinea pigs, the effect of the levels of serum IgE and serum and nasal histamine. Using the model of guinea pigs by histamine shocking, we observed the effect of BSDW on physiological symptoms in guinea pigs.

RESULTBSDW significantly relieved the pathological symptoms of allergic rhinitis in guinea pigs, alleviated the hyperplasia of columnar epithelium, decreased the number of monocyte and eosinocyte compared with the model group. It also reduced the levels of serum IgE, and decreased the release of serum and nasal histamine. BSDW significantly prolonged the occurent time of gasping, eclampsia and death caused by shock, reduced the times of gasping in the model of guinea pigs by histamine shocking.

CONCLUSIONBSDW has significant effect against allergy. The mechanism relates to its effects of decreasing the levels of serum IgE and inhibiting the release of serum and nasal histamine.

Administration, Intranasal ; Animals ; Anti-Allergic Agents ; pharmacology ; Asarum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Female ; Guinea Pigs ; Histamine ; blood ; Immunoglobulin E ; blood ; Lamiaceae ; chemistry ; Male ; Nasal Mucosa ; immunology ; Plants, Medicinal ; chemistry ; Rhinitis, Allergic, Perennial ; immunology ; Scutellaria ; chemistry ; Toluene 2,4-Diisocyanate

OBJECTIVETo explore the effect of rosiglitazone on the activity of signal transducer and activator of transcription 1 in rats with severe acute pancreatitis.

METHODSFifty-four male Wistar rats were randomly allocated into three groups (n = 18). SO group: sham-operated animals served as control, operation was executed and sodium chloride but not sodium taurocholate was injected. SAP group: SAP was induced by the retrograde injection of 5% sodium taurocholate into the biliopancreatic duct. ROSI group: same as SAP group, but rosiglitazone (6 mg/kg) was administered intravenously 30 min before operation. Rats in each group were sacrificed at 3,6 and 12 h after operation. The levels of serum amylase and histologic scores of pancreatic tissue were measured. The expression of TNF-alpha mRNA in pancreatic tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR). The expression of phosphorylated STAT1 in pancreatic tissue was assayed by immunohistochemistry.

RESULTSCompared to SO group, the levels of serum amylase and phosphorylated STAT1, TNF-alpha mRNA and histologic scores of pancreatic tissue were significantly elevated at the same time points after SAP (P < 0.01). The levels of these detection in ROSI group were lower than those of the SAP group at the same time points (P < 0.05), but higher than SO group (P < 0.05).

CONCLUSIONSSTAT1 was activated in severe acute pancreatitis. Rosiglitazone has a protective effects in rats with severe acute pancreatitis. The mechanism of its protective effects maybe that it inhibits the activation of JAK/STAT pathway, which can down-regulate the expression of TNF-alpha mRNA and block the the inflammatory cascade partially.

Acute Disease ; Animals ; Disease Models, Animal ; Male ; Pancreas ; metabolism ; pathology ; Pancreatitis ; drug therapy ; metabolism ; pathology ; Random Allocation ; Rats ; Rats, Wistar ; STAT1 Transcription Factor ; metabolism ; Thiazolidinediones ; pharmacology

OBJECTIVETo investigate the effect of Helicobacter Pylori lipopolysaccharide (Hp-LPS) on expression of Gli and Ptch-1 proteins in sonic hedgehog (Shh) signaling pathway of gastric mucosa GES-1 cells.

METHODSThe LPS was extracted from Hp by hot phenol water method, and then the concentration of LPS was detected by the kinetic turbidimetric assay. GES-1 cells were stimulated by different concentrations of Hp-LPS (0, 1, 10, 20, 30 and 40 μg/ml). The inhibition rates of cell growth were measured by MTT assay after treated with Hp-LPS for 24 h. The expression of Gli and Ptch-1 proteins were determined by Western Blot.

RESULTSMTT assay showed that the inhibition rates of GES-1 cell growth after treatment by different concentrations of Hp-LPS (1, 10, 20, 30 and 40μg/ml) were 25.8% ± 2.7%, 34.2% ± 3.1 %, 46.3% 3.4%, 60.8% ± 2.1% and 82.9% ± 2.8% respectively (r=0.985, P<0.001). Western blot showed that the expressions of Gli and Ptch-1 proteins were decreased after Hp-LPS treatment (0, 1, 10, 20, 30 and 40 μg/ml): the relative expression values of Gli were 1.286 ± 0.180, 0.963 ± 0.067, 0.850 ± 0.085, 0.566 ± 0.058, 0.549 ± 0.056 and 0.377 ± 0.047, respectively (r=-0.945, P<0.001); those of Ptch-1 were 1.688 ± 0.088, 1.466 ± 0.061, 1.170 ± 0.065, 1.042 ± 0.064, 0.648 ± 0.057 and 0.482 ± 0.074, respectively (r=-0.985, P<0.001).

CONCLUSIONHp-LPS can decrease the related protein expression of Shh signaling pathway, which indicates that Hp may interfere with the function of Shh signaling pathway in gastric mucosa via the effect of its LPS.

Cells, Cultured ; Epithelial Cells ; drug effects ; Gastric Mucosa ; cytology ; Hedgehog Proteins ; metabolism ; Humans ; Lipopolysaccharides ; administration & dosage ; pharmacology ; Patched Receptors ; Patched-1 Receptor ; Receptors, Cell Surface ; metabolism ; Signal Transduction ; Transcription Factors ; metabolism ; Zinc Finger Protein GLI1

OBJECTIVETo test the dissolution rate of silymarin dropping pill as well as to be compared with other three commercial products of the silymarin.

METHODBy UV spectrophotometry, we studied the dissolution conditions of silymarin dropping pill and compared its dissolution rate with Yiganling tablets (film-coating, sugar-coating) and Legalon capsule which are available in the market.

RESULTThe dissolution parameters T50 and Td of silymarin dropping pill, Yiganling tablet (film-coating), Yiganling tablet (sugar-coating) and Legalon capsule are 6.78, 9.85 min, 51.01, 73.78 min, 74.35, 86.97 min and 53.10, 72.65 min.

CONCLUSIONThe dissolution rate of silymarin dropping pill is superior to that of two kinds of Yiganling tablets and Legalon capsule.

Capsules ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Silymarin ; administration & dosage ; chemistry ; Solubility ; Spectrophotometry, Ultraviolet ; Tablets

OBJECTIVETo study the remodeling of the anterior alveolar bone with parodontium under physiology loading using finite element method (FEM) and theory of bone remodeling.

METHODSA FEM model of the maxillary central incisor with parodontium was established, and the change of bone density during the remodeling of alveolar bone was investigated under physiology loading (60 - 150 N) based on the theory of bone remodeling about strain energy density (SED). The finite element analysis software Abaqus user material subroutine (UMAT) were used.

RESULTSWith the increase of physiology loading, the pressure stress on the buccal cervical margin increased gradually while the density was decreased gradually. The cortical bone was lower than its initial density 1.74 g/cm(3), which was 1.74 - 1.63 g/cm(3). The density of cancellous bone was 0.90 - 0.77 g/cm(3), which was lower than its intial density 0.90 g/cm(3). The lingual cervical margin was under tensile stress which also increased with loading, the density had no significant change. When the achieve to 120 N, the density of cortical bone was 1.74 - 1.73 g/cm(3). No significant change was found in the cancellous bone.

CONCLUSIONSThe simulation of the perodontium remodeling is achieved and proved to be effective by the relevant research based on the method of the study. And the result will be helpful to form the basis of analysis bone remodeling process and predict the results in the clinical work.

Alveolar Process ; physiology ; Bone Density ; Bone Remodeling ; physiology ; Computer Simulation ; Dental Stress Analysis ; methods ; Finite Element Analysis ; Humans ; Incisor ; physiology ; Maxilla ; physiology ; Periodontium ; physiology ; Stress, Mechanical

OBJECTIVETo construct a lentiviral-vector-mediated CyPA small interference RNA (siRNA) and study its function in non-small cell lung cancer.

METHODSFirst, four target sequences were selected according to CyPA mRNA sequence, the complementary DNA contained both sense and antisense oligonucleotides were designed, synthesized and cloned into the pGCL-GFP vector, which contained U6 promoter and green fluorescent protein (GFP). The resulting lentiviral vector containing CyPA shRNA was named Lv-shCyPA, and it was confirmed by PCR and sequencing. Next, it was cotransfected by Lipofectamine 2000 along with pHelper1.0 and pHelper 2.0 into 293T cells to package lentivirus particles. At the same time, the packed virus infected non-small cell lung cancer cell (A549), the level of CyPA protein at 5 d after infection was detected by Western Blot to screen the target of CyPA. A549 were infected with Lv-shCyPA and grown as xenografts in severe combined immunodeficient mice. Cell cycle and apoptosis were measured by FCM.

RESULTSIt was confirmed by PCR and DNA sequencing that lentiviral-vector-mediated CyPA siRNA (Lv-shCyPA) producing CyPA shRNA was constructed successfully. The titer of concentrated virus were 1 x 10(7) TU/ml. Flow cytometric analysis demonstrated G2-M phase (11.40% +/- 0.68%) was decreased relatively in A549/LvshCyPA compared with control groups (14.52% +/- 1.19%) (P<0.05). The apoptosis rate of A549/Lv-shCyPA (5.01% +/- 0.5%) was higher than control groups (0.35% +/- 0.17%) (P<0.05). Visible tumors were only detectable at 6th day after inoculated by A549/Lv-shCyPA. The xenograft tumors of A549/Lv-shCyPA remarkably delayed tumor growth and remained at a similarly small average size at 38th days after inoculation compared with the control group (P < 0.05).

CONCLUSIONLentiviral-vector-mediated siRNA technique effectively inhibits the expression of CyPA, induces the NSCLC cell apoptosis, inhibits the tumor growth. Elucidation of the precise role of CypA in these pathways may lead to new targeted therapies for non-small cell lung cancer.

Animals ; Carcinoma, Non-Small-Cell Lung ; genetics ; Cell Line, Tumor ; Cyclophilin A ; genetics ; Gene Silencing ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Lung Neoplasms ; genetics ; Mice ; RNA, Small Interfering

OBJECTIVEAnalyzing the relationships between peripheral blood CD4+ CD25hi regulatory T (Treg) cells and peripheral blood immune status or plasma HIV-lviral load in HIV-infected individuals,so as to determine whether Treg were related to the progression of HIV-infected disease.

METHODS116 HIV-infected patients in different stages and 21 healthy control individuals were included in this study. The CD4+ and CD8+ T cell counts were determined by a standard 4-color flow cytometry technique. The Treg cells were examined with 3-color immune staining flow cytometry. The plasma HIV-1 viral load was detected by real time PCR.

RESULTSThe frequencies of Treg cells decreased in HIV-infected individuals with high CD4+ T cell counts( > 300/microl) compared with normal controls. With the progression of disease the frequencies of Treg cells were raised gradually, until were increased in HIV-infected individuals with low levels of CD4+ T cell counts ( < 100/microl). In addition, the frequencies of Treg cells were inversely related to CD4+ T cell counts and CD4+ /CD8+ ratio, data showed a statistically significant (respectively, r = -0.564, P < 0.001; r = -0.377, P < 0.001). Furthermore, the proportions of Treg cells were closely related to plasma HIV-1 RNA viral load (r = 0.514, P < 0.001).

CONCLUSIONCD4 CD25hi Treg cells should be a kind of important cells participating the immunopathogenesis of AIDS. It may play different roles in different stages of HIV-infected disease. The exact mechanism of Treg cells in the progression of the HIV-infected disease needs to be investigate further.

Adult ; Case-Control Studies ; Cells, Cultured ; Disease Progression ; Female ; HIV Infections ; immunology ; pathology ; virology ; HIV-1 ; genetics ; immunology ; isolation & purification ; Humans ; Interleukin-2 Receptor alpha Subunit ; immunology ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology ; Viral Load

OBJECTIVETo investigate the role of alginate-polylysine-alginate (APA) microcapsules in protecting rat islet cells in cryopreservation.

METHODPurified rat islet cells microencapsulated with APA and free islet cells were cryopreserved for one month and then thawed for culture in RPMI 1640 overnight. The morphology of the cells was observed and their function assessed by stimulated insulin release test.

RESULTAPA microcapsulation protected the fragile islets from freezing damage by increasing the recovery rate of the cells from 68.6%+/-2.9% to 94.7%+/-1.4% (P<0.05). After incubation with high glucose (16.7 mmol/L) solution, the insulin release from the encapsulated cells after cryopreservation significantly increased in comparison with that of the nonencapsulated cells (22.6+/-1.8 mU/L vs 11.7+/-1.5 mU/L, P<0.05). In high glucose solution containing theophylline, the calculated stimulation index of the encapsulated cells was about 3 times that of the nonencapsulated cells.

CONCLUSIONAPA microencapsulation may significantly increase the post-thaw recovery and improve the function for cryopreserved rat islets.

Alginates ; pharmacology ; Animals ; Capsules ; Cell Separation ; Cell Survival ; Cryopreservation ; methods ; Insulin ; secretion ; Islets of Langerhans ; cytology ; secretion ; Male ; Polylysine ; analogs & derivatives ; pharmacology ; Rats ; Rats, Wistar

OBJECTIVETo investigate the feasibility and benefits of co-culture of cryopreserved islets with small intestinal submucosa (SIS).

METHODSPurified rat islets cryopreserved for one month were divided into SIS group and control group, and after culture in standard islet culture media RPMI1640 for 1 week, the morphology and function of the islets were assessed.

RESULTSThe SIS protects the fragile islets from damage by cryopreservation, and increased the recovery from (60.6-/+3.3)% to (91.7-/+1.8) % (P<0.05). Compared with the control group, incubation of the islets of the SIS group in high-glucose (16.7 mmol/L) solution resulted in significantly enhanced insulin secretion (23.7-/+1.6 vs 12.5-/+1.1 mU/L, P<0.05). When the islets were incubated in high-glucose solution containing theophylline, the calculated stimulation index of SIS group was about 3-fold higher than that of the control group.

CONCLUSIONCo-culture of cryopreserved rat islets with SIS can increase the recovery of islet cells and improve their function.

Animals ; Coculture Techniques ; Cryopreservation ; methods ; Glucose ; pharmacology ; Insulin ; secretion ; Intestinal Mucosa ; cytology ; drug effects ; physiology ; Intestine, Small ; cytology ; drug effects ; physiology ; Islets of Langerhans ; cytology ; drug effects ; physiology ; Male ; Rats ; Rats, Wistar ; Theophylline ; pharmacology