This research is to study the relationship between HPLC fingerprints of Moutan Cortex, Paeoniae Radix Rubra and Paeoniae Radix Alba and their activity on lipopolysaccharide-induced acute lung injury. HPLC fingerprints of each extract of Moutan Cortex,Paeoniae Radix Rubra and Paeoniae Radix Alba were established by an optimized HPLC-MS method. The activities of all samples against protein and tumor necrosis a factor were tested by the model of lipopolysaccharide-induced acute lung injury. The possible relationship between HPLC-MS fingerprints and the activitieswere deduced by the Partial least squares regression analysis method. Samples were analyzed by HPLC-MS/MS to identify the major peaks. The results showed that each sample had some effect on acute lung injury. Four components with a lager contribution rate of efficacy were calculated by the research of spectrum-effect relationship. Moutan Cortex exhibited good activity on acute lung injury, and gallic acid, paeoniflorin, galloylpaeoniflorin and paeonol were the main effective components.
Acetophenones ; chemistry ; pharmacology ; Acute Lung Injury ; chemically induced ; drug therapy ; Animals ; Bridged Bicyclo Compounds, Heterocyclic ; chemistry ; pharmacology ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gallic Acid ; chemistry ; pharmacology ; Glucosides ; chemistry ; pharmacology ; Lipopolysaccharides ; pharmacology ; Male ; Monoterpenes ; chemistry ; pharmacology ; Paeonia ; chemistry ; Plant Roots ; chemistry ; Rats ; Rats, Wistar ; Tandem Mass Spectrometry ; methods

OBJECTIVETo study the relation on overfilling with gutta-percha point or paste and acute periapical periodontitis.

METHODSCollected sixty cases of acute periapical periodontitis which had been filled with gutta-percha point and paste within 1 week, and took dental radiographs. The cases that dental radiographs showed only guttapercha point was overfilling were assigned to group A (34 cases), and the cases that dental radiographs showed only paste was overfilling were assigned to group B (26 cases). The cases that dental radiographs showed both gutta-percha point and paste were overfilling were excluded. Sixty cases were divided into light group and severe group according to clinical sign. Measured gutta-percha point length or paste areas over apex. Took out the ohturation material cornpletely, adjusted occlusion when necessary and changed root canal medicament every day until clinical sign disappeared completely. Recorded the time of clinical sign disappeared completely.

RESULTSIn group A, gutta-percha point length over apex averaged 1.01 mm on light cases, and 1.79 mm on severe cases. In group B, the paste areas over apex averaged 2.45 mm2 on light cases, and 8.26 mm2 on severe cases. Group A had 13 light cases and 21 severe cases, and group B had 18 light cases and 8 severe cases. In group A, the average time of clinical sign disappeared completely was 3.56 days, and in group B the average time was 6.19 days. The statistical test showed there were significant differences among these four couples.

CONCLUSIONThe more overfilling, the more severe clinical sign was. Clinical sign caused by gutta-percha point overfilling was more severe. The time of clinical sign which caused by gutta-percha point overfilling disappeared completely was shorter.

Gutta-Percha ; Humans ; Periapical Periodontitis ; Root Canal Filling Materials ; Root Canal Preparation

OBJECTIVETo evaluate the time-effect of silica on the expression of lung tissue nitric oxide synthase (NOS) in early inflammatory damage stage of silicotic rat.

METHODSAnimal models were established by direct tracheal instillation of silica into rat lungs. Total NOS and induced NOS (iNOS) activities in bronchoalveolar lavage fluid (BALF) were assayed. The expression of iNOS protein in paraffin-embedded lung sections with Streptavidin/peroxidase (SP) immunohistochemistry were measured by tissue microarray and Image-Pro Plus.

RESULTSMost of the expression of iNOS was in the cytoplasm of macrophages and neutrophils. iNOS integrated optical density (IOD) of lung tissue increased 1.47 x 10(5) and 2.73 x 10(5) more respectively in silicatreated rats 3, 7 days after exposure than in control rats (P < 0.05), and decreased 1.11 x 10(5) more 28 days after exposure (P < 0.01). The activities of iNOS in BALF increased by 0.86, 1.89 and 0.92 U/ml respectively 3, 7, 14 days after exposure (P < 0.05 or P < 0.01). The activities of total NOS in BALF increased by 1.43, 2.05, 2.61 and 2.19 U/ml respectively 1, 3, 7, 14 days after exposure (P < 0.05 or P < 0.01).

CONCLUSIONAfter silica instillation, the iNOS-positive cells in rat lung tissue were mostly macrophages and neutrophils. There is a parabolic changing trend in the level of expression of lung iNOS during 1 - 28 day exposure to silica.

Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; cytology ; Female ; Immunohistochemistry ; Lung ; drug effects ; enzymology ; Male ; Models, Animal ; Nitric Oxide Synthase ; analysis ; metabolism ; Rats ; Rats, Wistar ; Silicon Dioxide ; toxicity

This study was purposed to detect the expression of transforming growth factor β1 (TGF-β1) and its receptors (TGF-βR) and to investigate their roles in pathogenesis of immune thrombocytopenic purpura (ITP). The expressions of TGF-β1 and their receptors TGF-βRI, TGF-βRII and TGF-βRIII in the peripheral blood of patients with ITP and healthy persons were detected by the real-time PCR, and differences of their expression levels were analysed. The results showed that the expression of TGF-β1 and TGF-βRII mRNA in ITP patients was significantly higher than that in the healthy controls, while the TGF-βRI mRNA expression in ITP patient was significantly lower than that in the controls. The expression of TGF-βRIII was not statistically different between the two groups. It is concluded that TGF-β1 and its receptors including TGF-βRI and TGF-βRII express abnormally in the peripheral blood of ITP patients, which suggests that the TGF-β signaling pathway probably play a vital role in the pathogenesis of the ITP.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Child ; Female ; Humans ; Male ; Middle Aged ; Protein-Serine-Threonine Kinases ; metabolism ; Purpura, Thrombocytopenic, Idiopathic ; metabolism ; pathology ; Receptors, Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Young Adult

Objective To study the bone biomechanics of the rabbit osteoporosis models induced by dexamethasone sodium phosphate injection (DX) using a combined treatment modality of 99Tc-MDP and GuKangLing.Methods Rabbits were intramuscularly injected with DX (2 mg/kg) twice a week for 6 weeks.The animal osteoporosis model group (Group C) and normal group (Group A) were compared to confirm the model was available.Another control group (Group B),the osteoporosis control group (Group D) were set for the comparison at the end of the experiment.The 99Tc-MDP therapy group (Group E),GuKangLing therapy group (Group F) and 99Tc-MDP plus GuKangLing therapy group (Group G) were included in the study.The treatment lasted for 16 weeks.The bone biomechanics,cytopathology bone histomorphology,bone mineral density (BMD),X-ray,CT,bone scintigraphy and serum bone alkaline phosphatase (BALP)and P (bone gla protein) were chosen as the markers or methods to evaluate the treatment results (excellent,effective and invalid).The analysis of variance (ANOVA) and t-test were used for group comparison analysis.Results Cytopathology result indicated that there was no bone trabecula destruction in Group A.However,there was distinct bone destruction in Group C.The bone biomechanics (left femur head (265.914 ±52.773) N,L4(369.671 ±94.919) N),BMD(left femur (0.238 ±0.016) g/cm2,L4(0.236 ±0.016) g/cm2)and bone histomorphology ( (66.230 ± 10.848) ％ ) in Group C reduced clearly as compared with Group A ((405.343±55.410) N,(750.870±53.718) N,(0.294±0.017) g/cm2,(0.302±0.023) g/cm2,( 131.500 ± 21.846) ％ ) ( t ≥4.550,all P ＜ 0.01 ).Radionuclide bone scan also showed that the uptake of tracers was higher by the main arthrosis in Group C than that in Group A.Vertebra was not clearly visualized on bone scan image.There were significant differences between Group A and Group C in serum BALP and P ((45.000±7.303) vs (12.485 ±1.512) U/L,(0.168±0.018) vs (0.115 ±0.017) μg/L,t =4.126,5.476,both P ＜ 0.01 ),which indicated that the animal osteoporosis model was available.The pathological results showed an improved recovery of bone structure and trabecular in Groups E and G,but a worse recovery in Group F.Biomechanics result in Groups E and G (left femur head (386.457 ±77.077) N and (432.771 ± 17.525) N,L4(649.550 ± 126.859) N and (655.443 ±76.555) N) improved apparently,which were similar to Group B.The radiotracer uptake in Group F was lower than that in group D.The bone biomechanics,bone histomorphology,BMD,serum BALP and P after the treatment showed significant differences in Groups E,F and G (F:8.556 - 31.608,all P＜0.01 ),and the bone biomechanics result in Group G was a little better than that in Group E (t =2.625,P ＜ 0.05 ).The results of Group G and E were considered as excellent,and Group F was considered as effective.Conclusions The treatment of 99Tc-MDP combined with GuKangLing could improve the bone biomechanics of rabbit osteoporosis models and may be a potential method to increase the bone strength for resisting external force.

OBJECTIVETo study the chemical constituents and the antibacterial activity from n-butanol extract of Sarcandra glabra.

METHODThe compounds were isolated by Diaion HP-20, Sephadex LH-20, MCI CHP-20 and silica gel column chromatographic methods. Their structures were identified on the basis of physicochemical and spectroscopic analysis. The antibacterial effect of the compounds were measured against Staphylococcus aureus by filterpaper slice method, finally the antibacterial ring in each group was recorded after 24 hours.

RESULTSeven constituents were isolated and elucidated as 5, 7, 3', 4'-tetrahydroxy-6-C-beta-D-glucopyranosylflavanone (1), kaempferol-3-O-beta-D-glucuronide (2), fraxidin (3), isofraxidin (4), isofraxidin-7-O-beta-D-glucopyranoside (5), kaempferol (6), pinostrobin (7). Diameters (in mm) of antibacterial ring in the compounds 2, 5, 6 were orderly recorded as follows: 14.67 +/- 0.08, 11.14 +/- 1.06, 8.26 +/- 1.26 and the compound 4 is not effective.

CONCLUSIONCompounds 1-3 and 5 were isolated from S. glabra for the first time.

Anti-Bacterial Agents ; chemistry ; isolation & purification ; pharmacology ; Butanols ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Magnetic Resonance Spectroscopy ; Magnoliopsida ; chemistry ; Staphylococcus aureus ; drug effects

Our previous studies showed that spinal neurons sensitization was involved in morphine withdrawal response. This study was to investigate the roles of spinal protein kinase C (PKC) alpha, gamma in morphine dependence and naloxone-precipitated withdrawal response. To set up morphine dependence model, rats were subcutaneously injected with morphine (twice a day, for 5 d). The dose of morphine was 10 mg/kg in the first day and was increased by 10 mg/kg each day. On day 6, 4 h after the injection of morphine (50 mg/kg), morphine withdrawal syndrome was precipitated by an injection of naloxone (4 mg/kg, i.p.). Chelerythrine chloride (CHE), a PKC inhibitor, was intrathecally injected 30 min before the administration of naloxone. The scores of morphine withdrawal symptom and morphine withdrawal-induced allodynia were observed. One hour after naloxone-precipitated withdrawal, Fos protein expression was assessed by immunohistochemical analysis and Western blot was used to detect the expression of cytosol and membrane fraction of PKC alpha and gamma in the rat spinal cord. The results showed that intrathecal administration of CHE decreased the scores of morphine withdrawal, attenuated morphine withdrawal-induced allodynia and also inhibited the increase of Fos protein expression in the spinal cord of morphine withdrawal rats. The expression of cytosol and membrane fraction of PKC alpha was significantly increased in the spinal cord of rats with morphine dependence. Naloxone-precipitated withdrawal induced PKC alpha translocation from cytosol to membrane fraction, which was prevented by intrathecal administration of CHE. During morphine dependence, but not naloxone-precipitated withdrawal, PKC gamma in the spinal cord translocated from cytosol to membrane fraction, and intrathecal administration of CHE did not change the expression of PKC gamma in the spinal cord of naloxone-precipitated withdrawal rats. It is suggested that up-regulation and translocation of PKC in the spinal cord contribute to morphine dependence and naloxone-precipitated withdrawal in rats and that PKC alpha and gamma play different roles in the above-mentioned effect.
Animals ; Male ; Morphine Dependence ; physiopathology ; Naloxone ; pharmacology ; Protein Kinase C ; metabolism ; physiology ; Protein Kinase C-alpha ; metabolism ; physiology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; metabolism ; physiopathology ; Substance Withdrawal Syndrome ; enzymology ; physiopathology

OBJECTIVETo search and identify the non-steroid receptor binding cis-acting elements in the L-plastin promoter in prostate cancer, and the correlative regulation pathway and transcription factors.

METHODSOn the basis of construction of the L-plastin promoter luciferase vectors which were removed the steroid hormone receptor AR and ER binding elements, the promoter on the vector was nest-deleted by Exonuclease III and the relative luciferase plasmids were constructed. Transfected these twelve plasmids into prostate cancer cell line LNCaP under dihydrotestosterone-stimulated situation or not and test the intensity of luciferase, then we got the regulation message of every 200 bp part of the promoter in prostate cancer. After the analysis of relative programme, we got the possible regu- lation pathway of non-steroid hormone transcription factors. After removing the possible transcription factors binding site sequence by site-specific mutagenesis, the changes luciferase of activities proved our reasoning.

RESULTSWe succeed in segmental deletion of the L-plastin promoter, and constructing the relative plasmids containing part L-plastin promoter on luciferase vector pGL3-basic. After testing the luciferase activities of constructed plasmids, we found the sequence from 206 to 1 of L-plastin promoter had significant luciferase activity. The software TRANSFECT showed that there were binding elements for transcription factors AP-4 at seq-198 to 192 and SP-1 at seq-54 to 41 on the short part promoter (206 to 1). The recombinant plasmids deleted the AP-4 and SP-1 binding elements had lower luciferase activity than the wild-type.

CONCLUSIONThere are some other non-steroid hormone pathway to regulate the expression of L-plastin except the steroid hormone pathway in prostate cancer. The main binding sites of the non-steroid hormone regulator lies in the sequence from 206 to 1. Transcription factors AP4 and SP-1 may up-regulated the expression of L-plastin by binding these sites.

Animals ; DNA-Binding Proteins ; physiology ; Gene Expression Regulation, Neoplastic ; Luciferases ; metabolism ; Male ; Membrane Glycoproteins ; Mice ; Microfilament Proteins ; Phosphoproteins ; biosynthesis ; genetics ; Promoter Regions, Genetic ; genetics ; Prostatic Neoplasms ; metabolism ; Response Elements ; Transcription Factors ; physiology ; Transfection ; Tumor Cells, Cultured ; Up-Regulation

OBJECTIVETo evaluate the relationship of chimerism status of cell subsets with engraftment, occurrence of acute graft versus host disease (aGVHD), graft rejection and disease relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSChimerism status in peripheral blood (PB) and bone marrow (BM) of 65 patients received allo-HSCT were monitored at regular intervals post-transplant. Fluorescence-activated cell sorter (FACS) was used to sort CD3(+)T lymphocytes in 65 cases, CD3(-)CD56(+)CD16(+)NK cells in 52 cases, CD15(+) granulocytes in 32 cases and CD19(+)B lymphocytes in 20 cases post transplants. The chimerism status of different lineage cells was analyzed by polymerase chain reaction amplification of short tandem repeats (PCR-STR).

RESULTSOn day +7, NK-cells donor chimerism (DC 55.5%) was higher than other cell subsets. T lymphocyte was the latest one to reach complete donor chimerism (CDC) with a median on day +21. Patients whose T lymphocytes donor chimerism was more than 70% on day +7 and more than 95% on day +14 had a high risk for acute aGVHD. In all cases except those with ALL, the decreased DC of T lymphocytes were observed before molecular or hematological relapse occurred.

CONCLUSIONSerial and quantitative T cell chimerism analysis provides a reliable and rapid screening method for the early detection of engraftment, graft rejection, disease relapse and occurrence of aGVHD, therefore, is a prognostic tool to identify patients at high risk of aGVHD and disease relapse following allo-HSCT.

Adolescent ; Adult ; Child ; Chimerism ; Female ; Graft Rejection ; immunology ; Graft vs Host Disease ; immunology ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Male ; Middle Aged ; Recurrence ; T-Lymphocytes ; immunology ; Young Adult

Objective:To explore the expression and clinical significance of ubiquitin-like with PHID and ring finger domains 2 (UHRF2) in intrahepatic cholangiocarcinoma (ICC).Methods:The expression of UHRF2 in 138 ICC was determined by immunohistochemistry,and the relationships between UHRF2 expression with prognosis and clinicopathological features of ICC were analyzed:Results:UHRF2 was highly expressed in ICC compared with paratumorous tissues.Level of UHRF2 was associated with high AFP level (P=0.010),poor tunor differentiation (P=0.026),lymph node metastasis (P=0.001),and advanced TNM stage (P=0.019).Hevated UHRF2 expression was suggested to be associated with poor prognosis and high recurrence rate in ICC patients (P＜ 0.001),and multivariable analysis indicated that UHRF2 was of theprognostic indicators of ICC (P＜0.05).Conclusions:High level of UHRF2 is related to the malignant phenotype of ICC,and can be a prognostic and recurrent indicator of postoperative patients.