Adult ; Ascitic Fluid ; metabolism ; Endometriosis ; blood ; metabolism ; Female ; Humans ; Interleukin-18 ; analysis ; blood ; Middle Aged

Objective: To observe the regulating action of Zushima Gancao Tablet on abnormal metabolites by analyzing the changes of endogenous metabolites in plasma of rheumatoid arthritis (RA) rats, and to explore the mechanism of Zushima Gancao Tablet in treating RA from the perspective of metabonomics. Methods: Rats were divided into normal group (NG), model group (MG), positive medicine group (PMG), and Zushima Gancao Tablet group (ZGG). Adjuvant arthritis (AA) rats were established with freund complete adjuvant, rats in PMG and ZGG groups were ig administered with tripterygium glycosides and Zushima Gancao Tablet respectively for a month. Plasma samples were collected on days 1, 10, 20, and 28 before and after administration. UPLC/LTQ-Orbitrap-MS was applied to analyzing the metabolic profile changes of each group in different durations of the disease and mechanism of drug intervention. Results: The related pathways of AA mainly involved metabolisms of amino acids, lipids, nucleic acids, and vitamins. Zushima Gancao Tablet had great effect on AA by regulating 12 biomarkers. Conclusion: Small molecule metabolites in AA rats are deviated from the normal ones. The effects of Zushima Gancao Tablet and positive medicine (tripterygium polyglycoside) on AA are similar, but the mechanisms were different. Zushima Gancao Tablet down-regulates the level of LPC in secondary lesions while tripterygium polyglycoside tablets could effectively inhibit the metabolisms of a variety of amino acids during acute inflammation.

The hexane extract from marine actinomycetes 124092 showed potent inhibition on B16 cell line by MTT assay. The hexane extract was fractionationed on silica gel column by vacuum liquid chromatography to afford 6 fractions(Fr1~Fr6), and Fr6 showed cytotoxic activity. To determine the bioacitve components of hexane extract, Fr6 was analyzed by GC/MS. The main components were identified as palmitic acid (11.76%), oleic acid (12.16%), linoleic acid (14.77%), and lactobacillic acid (61.31%). It have been reported that palmitic acid, oleic acid, and linoleic acid possess cytotoxic activity on rat ascites tumor cells and linoleic acid have suppressive effect on human lung adenocarcinoma cells.

The objective of the present study was to establish a method based on principal component analysis (PCA) for the study of transdermal delivery of Chinese medicinal formulae, and to choose the best penetration enhancers for Qingfei Xiaocuo gel depend on this method. Using improved Franz type diffusion cell and excised rat skin in vitro as transdermal barrier, the receptive solution fingerprint was established by HPLC, harvesting the areas of the common peaks in the fingerprint, then the total factor scores of the concentrations at different times were calculated using PCA and were employed instead of the concentrations to compute the cumulative amounts (Q12) and enhancement ratio (ER), the latter of which were considered as the indexes for optimizing penetration enhancers. Compare to the control group, the ER of the other groups increased significantly and furthermore, 2.5% azone with 2.5% menthol manifested the best effect. PCA represent most information in the receptive solution, the method above could choose the best penetration enhancers, it could be a reference for the study of transdermal delivery of Chinese medicinal formulae.
Administration, Cutaneous ; Animals ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Gels ; In Vitro Techniques ; Male ; Medicine, Chinese Traditional ; Mice ; Principal Component Analysis ; Skin ; metabolism

Objective To explore the role of white matter injuries in the schizophrenia induced by the NMDA re-ceptor antagonist. Methods Adult male C57BL/6J mice (8 week old) were equally divided into four groups. One group was sub-chronically treated with saline solution, and the other three groups were intraperitoneally treated with MK-801 at dose of 0.025 mg/mL (M1), 0.050 mg/mL (M2) and 0.100 mg/mL (M3) in a volume 10 ml per kilogram body weight. All ani-mals were tested using Morris water maze at the 9th-15th day and using the Hole Board exploration as well as Rota Rod performance tests on the 16th day. The myelin basic protein (MBP) and the ultrastructure of the myelin sheaths in the cor-pus callosum were then examined using immunohistochemical methods, transmission electron microscope technique and stereological methods. Results The repeated sub-chronic MK-801 treatment did not induce impairment of spatial learning and memory in Morris water maze. The MK-801 treatment at 0.25 mg/kg and 1.00 mg/kg but not at 0.50 mg/kg resulted in less exploration to a new environment. The myelin staining with anti-MBP antibody was less intense in all three schizo-phrenic groups when compared to saline control group (P<0.01). Furthermore, MK-801 treatment caused pathological al-terations of the myelin sheaths including segmental demyelination of myelinated fibers and splitting of myelin sheath lamel- lae in schizophrenic groups. The ratio of the injured myelinated nerve fibers in the corpus callosum of MK-801 treated mice [M3 group, (22.42 ± 4.24)%] was significantly higher when compared to the control mice [(3.84 ± 1.35)%,P<0.01)]. Conclusions The present study demonstrated the white matter damages, mainly low MBP expression and segmental demye-lization in the corpus callosum in the mice sub-chronic treated with MK-801, indicating that the white matter changes might be involved in the schizophrenia induced by NMDA antagonist.

OBJECTIVETo establish a method for isolating and culturing human amniotic epithelial cells (hAECs) in a serum-free medium and investigate their transdifferentiation ability into islet-like cells.

METHODSThe culture condition of hAECs was optimized using DMEM with different supplements. The genetic stability of the tenth-passage cells was assessed by chromosome analysis and G-banding method. The stem cell characteristics of the cells were identified by examination of the surface markers using immunofluorescence methods. The endocrine-related genes and hormones of the cells were tested after induced differentiation into islet-like cells.

RESULTSThe hAECs allow stable passaging in the presence of 10 ng epidermal growth factor (EGF) in the culture medium. After 10 passages, the cells maintained a normal karyotype and G-banding profile. The hAECs expressed many multi-potent stem cell markers, including SSEA4, TRA-1-60, and TRA-1-81. After induced differentiation, the endocrine-related genes were expressed in the islet-like cells, including PDX1, ngn3, insulin and glucagon. Insulin secretion increased in the differentiated islet-like cells in response to high glucose exposure.

CONCLUSIONWe established a method for isolating and expanding the hAECs in a serum-free medium. hAECs possess stem cell characteristics and can be induced to differentiate into islet-like cells in vitro.

Amnion ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cells, Cultured ; Epithelial Cells ; cytology ; secretion ; Humans ; Insulin ; secretion ; Islets of Langerhans ; cytology ; Stem Cells ; cytology ; secretion

A capacitively coupled contactless conductivity detector for high performance liquid chromatography was developed. The detector was consisted of a signal generator, a signal amplifier, and a pair of tubular electrodes. A plastic connecting tube following the separation column was threaded through the two tubular electrodes. When the separated components passed the region between two tubular electrodes, they would be sensed. The instrumental parameters of this detector were investigated, including the internal diameter of connecting tube,the frequency and voltage of excitation signal, electrodes length, and the gap between two electrodes. The 0.5 mm of internal diameter of connecting tube, 70 kHz of excitation frequency, 60 V of excitation voltage,10 mm of electrodes length and 1.5 mm of gap between two electrodes were finally found to be the best working conditions for the detector. This HPLC-C4D(capacitively coupled contactless conductivity detector) was applied to the analysis of oleic and linoleic acids in Brucea javanica oil. The chromatographic peak area was linear with concentration for oleic and linoleic acids in 5-1000 μg/mL, and the limit of detection reached 2.5 μg/mL and 1.0 μg/mL,respectively. The results demonstrate that the present method is sensitive and accurate, proving this homemade contactless conductivity detector has a great application potential for pharmaceutical analysis in future development.

BACKGROUND AND OBJECTIVEc-Cbl and Cbl-b are two ubiquitous members of the Casitas B-lineage lymphoma (Cbl) family, which play important roles in the downregulation of epidermal growth factor receptor (EGFR) by acting as E3 ubiquitin ligases and multiadaptor proteins. This study investigated the expression of c-Cbl, Cbl-b, and EGFR in gastric carcinoma and its clinical significance.

METHODSThe expressions of c-Cbl, Cbl-b, and EGFR were detected by immunohistochemistry using tissue microarrays consisting of 124 specimens of gastric carcinoma and 16 specimens of normal gastric mucosa. The relationship between the expressions of c-Cbl, Cbl-b, and EGFR and clinicopathologic factors of gastric carcinoma were analyzed statistically.

RESULTSThe positive rates of c-Cbl, Cbl-b, and EGFR were higher in the gastric carcinoma group than in the normal group (71.0% vs. 18.0%, P<0.01; 82.3% vs. 25.0%, P<0.01; 56.5% vs. 12.5%, P<0.01, respectively). The expression of c-Cbl was positively correlated with depth of invasion (r=0.219, P=0.015), and TNM staging (r=0.266, P=0.003). The expression of Cbl-b was positively correlated with lymph node metastasis (r=0.190, P<0.034) and TNM staging (r=0.298, P<0.001). The expression of EGFR was positively correlated with depth of invasion (r=0.286, P<0.001) and TNM staging (r=0.362, P=0.000). The expression of both c-Cbl and Cbl-b was positively correlated with EGFR (r=0.241, P=0.007; r=0.183, P=0.042, respectively). Synchronous strong-positive expressions of c-Cbl, Cbl-b, and EGFR were observed in 27 specimens of gastric carcinoma, most of which were at advanced stage.

CONCLUSIONSOverexpressions of c-Cbl, Cbl-b, and EGFR are closely related to the invasion and progression of gastric carcinoma. c-Cbl and Cbl-b may serve as novel molecular markers for gastric carcinoma.

Adaptor Proteins, Signal Transducing ; metabolism ; Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; metabolism ; Disease Progression ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Proto-Oncogene Proteins c-cbl ; metabolism ; Receptor, Epidermal Growth Factor ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; Young Adult

OBJECTIVETo observe the effect of Zishen Huoxue Recipe (ZHR) on pathomorphology in coronary heart disease (CHD) rats with Shen deficiency blood stasis syndrome (SDBSS).

METHODSTotally 60 healthy Wistar rats were divided into the blank control group, the model group, high, middle, and low dose ZHR groups according to random digit table, 12 in each group. Myocardial ischemia SDBSS rat model was prepared by ligating the left anterior descending coronary artery and injecting hydrocortisone. ZHR physic liquor was administered to rats in high, middle, and low dose ZHR groups at the daily dose of 21.6, 10.8, 5.4 g/kg by gastrogavage for 7 successive days, equal volume of pure water was administered to rats in the blank control group and the model group by gastrogavage for 7 successive days. Rat heart was collected for pathomorphological observation under light microscope.

RESULTSIn the model group the heart muscle fiber was swollen and deformed with widened space, loose and dropsy tissues. Blood vessels in myocardial mesenchymal were dilated, infiltrated with more inflammatory cells. Myocardial cells were markedly swollen, degenerated, or necrotic, with caryolysis or disappearance of partial nuclear. A large amount of collagen fibrous tissue became hyperplasia. Endocardial blood vessels were swollen and degenerated with infiltration of few inflammatory cells. Epicardium tissue and structure were destroyed and got hyperplasia. Swollen, degenerated, or necrotic vessels could be seen, with infiltration of more inflammatory cells and collagen deposition. Pathomorphological injuries were alleviated in each ZHR group. The higher ZHR concentration, the milder the injury degree of myocardial tissue, the more limited range of damage.

CONCLUSIONZHR could attenuate pathomorphological injuries of myocardial ischemia rats with SDBSS and regulate myocardial function, thus improving myocardial ischemia in CHD rats with SDBSS.

Animals ; Coronary Artery Disease ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Medicine, Chinese Traditional ; Myocardial Ischemia ; Myocardium ; Rats ; Rats, Wistar

BACKGROUNDNonmyeloablative allogeneic bone marrow transplantation has been used since the 1990s as a new hematological stem cell transplantation strategy for treating hematological diseases. The purpose of this study was to explore the graft-versus-leukemia (GVL) effects of donor lymphocyte infusions (DLIs) after nonmyeloablative allogeneic bone marrow transplantations, while assessing the declines in treatment-associated morbidity, mortality, and graft-versus-host disease (GVHD).

METHODSA total of 615 (H-2k) mice were injected with L615 tumor cells and received 500 cGy (60Co gamma-ray) irradiation three days later, followed by an allogeneic bone marrow transplantation (allo-BMT). The allo-grafts consisted of 3 x 10(7) bone marrow cells and 1 x 10(7) spleen cells from BALB/C (H-2d) donor mice. Two days after the allo-BMT, the recipient mice were given 200 mg/kg of cyclophosphamide. Subsequently, recipient mice were infused with either donor spleen cells (2 x 10(7)) on day 14 or 21, or donor spleen cells (5 x 10(7)) pretreated with hydrocortisone and cyclosporin A (CsA) in vitro on day 14 post-BMT.

RESULTSThe median survival time of mice that received DLI on day 21 and pretreated DLI on day 14 post-BMT was longer than that of controls and the day 14 DLI group (P < 0.01). No evidence of severe GVHD was observed in the day 21 DLI group nor in the day 14 treated DLI group. Mixed chimerism was confirmed in the day 14 DLI group, the day 14 treated DLI group, and the day 21 DLI group on the thirteenth day post-transplantation; full donor chimerism was observed two weeks after DLI.

CONCLUSIONDonor lymphocyte infusion after nonmyeloablative bone marrow transplantation may reduce transplantation-associated morbidity and mortality while strengthening graft-versus-leukemia effects.

Animals ; Bone Marrow Transplantation ; immunology ; Cyclosporine ; pharmacology ; Female ; Graft vs Host Disease ; etiology ; Graft vs Leukemia Effect ; immunology ; Hydrocortisone ; pharmacology ; Lymphocyte Activation ; Lymphocyte Transfusion ; Male ; Mice ; Mice, Inbred BALB C ; Transplantation Chimera ; Transplantation, Homologous