OBJECTIVETo explore the effect of Jingui Shenqi pill (JGSQP) with various concentrations at different time points on pituitary adrencorticotropic hormone (ACTH) gene expression level in Shen-Yang deficiency rats.

METHODSThe Shen-Yang deficiency rats were randomly divided into the model control group and the high, medium and low dosage of JGSQP groups. Reverse transcriptase polymerase chain reaction was used to observe the effect of JGSQP on the ACTH mRNA of pituitary tissue in rats treated at different time points (10 d, 20 d, 30 d).

RESULTSAs compared with that in the model group, the ACTH gene expression level was significantly higher in the high dose JGSQP group (P < 0.05), and the increment in the medium dosage group was significantly higher in comparing with that in the high and low dosage groups (P < 0.05 or P < 0.01).

CONCLUSIONThrough up-regulation on ACTH gene expression is possibly one of the mechanisms of JGSQP in treating Shen-Yang deficiency.

Adrenocorticotropic Hormone ; biosynthesis ; genetics ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Hypothalamo-Hypophyseal System ; Kidney Diseases ; genetics ; metabolism ; Medicine, Chinese Traditional ; Pituitary Gland ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Yang Deficiency ; genetics ; metabolism

OBJECTIVETo investigated the effect of 50-Hz 3.6-mT sinusoidal electromagnetic fields (SEMFs) on the proliferation and differentiation of osteoblasts in vitro.

METHODSThe newborn rat calvarial osteoblasts were isolated by enzyme digestion and randomly divided into 6 groups after one passage. The treatment groups under 50-Hz 3.6-mT SEMFs and controls without SEMFs treatment. The cells were exposed in the SEMFs for 0.5 h, 1.0 h, 1.5 h, 2.0 h, and 2.5 h. They were observed under the contrast phase microscope each day. The calcified nodules were stained by alizarin red. The SEMFs were arranged in spiral appearance after 3 to 5 days.

RESULTSThe SEMFs showed characteristic distribution 3 to 5 days after SEMFs treatment. On the 9(th) day after treatment, the activity of alkaline phosphatase (ALP) significantly increased in the 0.5-h group, whereas the ALP histochemical straining results and the area of calcified nodules were consistent with ALP activity. In the 48-h and 96-h groups, the genetic expression levels of osteoprotegerin and collagen-1 were significantly higher than that in the control group; particularly, the mRNA expression increased in the 0.5-h group.

CONCLUSIONThe SEMFs at 50-Hz 3.6-mT could suppress the proliferation of osteoblasts maturation but stimulate the differentiation and maturation of osteoblasts in vitro.

Animals ; Cell Differentiation ; radiation effects ; Cell Proliferation ; radiation effects ; Cells, Cultured ; Electromagnetic Fields ; Male ; Osteoblasts ; cytology ; radiation effects ; Rats ; Rats, Sprague-Dawley

AIMTo observe the antagonistic effect of hydroxysafflor yellow A (HSYA) on the platelet activating factor (PAF).

METHODSWashed rabbit platelet (WRP) aggregation and rabbit polymorphonuclear leukocytes (PMNs) aggregation induced by PAF were observed by turbidimetric assay in vitro. The PAF receptor antagonistic effect of HSYA was investigated by radio ligand binding assay (RLBA).

RESULTSIn RLBA the specific binding inhibition effect of HSYA was found to be concentration-dependent in three different [3H]PAF concentrations. In the experiments, WRP aggregation and rabbit PMNs aggregation induced by PAF (9.55 x 10(-10), 9.55 x 10(-6) mol.L-1) were both inhibited by HSYA in a concentration-dependent manner in vitro. The IC50 of HSYA to inhibit WRP and rabbit PMNs aggregation was 0.99 and 0.70 mmol.L-1, respectively.

CONCLUSIONThe PAF receptor binding can be antagonized by HSYA.

Animals ; Carthamus ; chemistry ; Cell Aggregation ; drug effects ; Chalcone ; analogs & derivatives ; isolation & purification ; pharmacology ; In Vitro Techniques ; Male ; Neutrophils ; drug effects ; Plants, Medicinal ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Membrane Glycoproteins ; antagonists & inhibitors ; Quinones ; isolation & purification ; pharmacology ; Rabbits ; Receptors, G-Protein-Coupled ; antagonists & inhibitors

OBJECTIVETo study the antioxidant constituents from the root of Rubus crataegifolius.

METHODThe constituents isolation and purification from the root of R. crataegifolius was carried by reported column chromatography including silica gel, toyopearl, and their structures were elucidated on the basis of spectral compounds. DPPH method was used to evaluate the free radical scavenging activity of the isolated compounds.

RESULTNine compounds were isolated from the root of R. crataegifolius, and their structures were identified as follow: euscaphic acid (1), kaempferol-3-O-beta-D-galactopyranoside (2), tormentic acid (3), 2alpha, 19alpha, 24-trihydroxyurs-12-ene-3-oxo-28-acid (4) , 2alpha-hydroxy-oleanolic acid (5), ursolic acid (6), daucosterol (7), beta-sitosterol (8) and polydatin (9). By experiment of antioxidant activity, the result showed compounds 2 and 9 revealed DPPH free radical scavenging rates were 95.60% and 75.23% at the concentration of 50 mg x L(-1).

CONCLUSIONCompounds 1-8 were isolated from this plant for the first time, and compounds 2 and 9 showed the significant antioxidant activity.

Antioxidants ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Rosaceae ; chemistry

OBJECTIVETo evaluate the effect of capsaicin on nude mice xenografted with colorectal carcinoma cells, and to explore its mechanism of action.

METHODSA nude mouse model of colorectal cancer was established by subcutaneous inoculation of human colorectal carcinoma HT-29 cells. Terminal deoxynucleotidyl transferase-mediated nicked labeling assay (TUNEL) was undertaken to detect the cell proliferation and apoptosis in the xenograft tissue in nude mice. Immunohistochemical (IHC) staining and Western blot were used to detect the expression of HSP27, Cyt-C and active caspase-3.

RESULTSThe tumor growth of the groups C10 and C20 was significantly slower than that of the group NS. The integrated optical density (IOD) of both the group C5 (2532.14 ± 578.11) and group C10 (6364.03 ± 1137.98) was significantly higher than that of the group NS (760.12 ± 238.05), (P < 0.05). The integrated optical density (IOD) of the group C20 was (15743.96 ± 1855.95), significantly higher than that of the groups C10, C5 and NS (all were P < 0.01). Immunohistochemistry showed that the cytoplasmic expression of HSP27 was strongly positive in the group NS, and significantly reduced with the increasing dose of capsaicin in the treated groups. The expression of active caspase-3 and Cyt-C in the group NS was weakly positive, and was significantly increased with the increasing dose of capsaicin in the groups C5 and C10 (P < 0.05), and the expression of active caspase-3 and Cyt-C of the group C20 was significantly higher than that of the groups C5, C10 and NS (P < 0.01). Western blot analysis showed that both the expressions of HSP27 of the group C5 (0.73 ± 0.05) and the group C10 (0.41 ± 0.03) were significantly lower than that of the group NS (P < 0.05). The expression of HSP27 of the group C20 (0.22 ± 0.06) was significantly lower than that of the groups C5, C10 and NS (P < 0.01). The expressions of active-caspase-3 and Cyt-C in the group C5 were (2.57 ± 0.34) and (2.03 ± 0.38), significantly higher than those of the group NS (P < 0.05). The expressions of active-caspase-3 and Cyt-C in the group C10 were (4.23 ± 0.45) and (3.13 ± 0.44), also significantly higher than those of the group NS (P < 0.05). The expressions of active-caspase-3 and Cyt-C in the group C20 were (5.78 ± 0.48) and (4.92 ± 0.52), significantly higher than those of the group C5, C10 and NS (P < 0.01). TUNEL analysis showed that there was a significant difference of cell apoptosis in comparison of each two groups. The higher dose of capsaicin was used, the more apoptosis was observed.

CONCLUSIONSCapsaicin can significantly inhibit the tumor growth and induce cell apoptosis in the colorectal carcinoma xenograft in nude mice. Its mechanism of action is possibly related with the down-regulation of HSP27 expression and up-regulation of expression of active caspase-3 and Cyt-C in the colorectal carcinoma xenograft in nude mice.

Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Capsaicin ; administration & dosage ; pharmacology ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Cytochrome c Group ; metabolism ; Dose-Response Relationship, Drug ; Female ; HSP27 Heat-Shock Proteins ; metabolism ; HT29 Cells ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Random Allocation ; Tumor Burden ; Xenograft Model Antitumor Assays

Aim To observe the effect of epidurally application of osthole on the model of nucleus pulposusinduced inflammatory radicular pain and the expression of p38 MAPK signaling related pathway in the spinal dorsal horn of rats.Methods The model of radicular pain was generated by putting nucleus pulposus to the L5 dorsal root ganglion (DRG).50％ MWT was measured using Von Frey filaments to calculate mechanical pain threshold before and after operation.50 μL of 20 g · L-1 osthole was administered epidurally in group Ost and 50 μL of 100 mL · L-1 DMSO in group DMSO at postoperative day (POD).The expression of phosphorylated p38 (p-p38),IL-18 and IL-18R in the lumbar spinal dorsal horn was detected by Western blot.IL-18 mRNA was assessed by real-time PCR.Results The mechanical pain threshold significantly decreased after operation (P ＜ 0.05),while the expression of protein p-p38 MAPK,IL-18,IL-18R and IL-18 mRNA was significantly different.Compared with DMSO group,50％ MWT was significantly increased and accompanied with the decrease of protein p-p38,IL-18,IL-lgR and IL-18 mRNA in Ost group after drug administration (P ＜ 0.05).The correlation analysis between protein concentration of p38 MAPK and IL-18 mRNA showed that the Spearman correlation coefficient was 0.9 (P ＜ 0.05).Conclusion p-p38 and IL-18 of spinal dorsal horn participate in the rat model with inflammatory radicular pain induced by nucleus pulposus,and IL-18R plays a role in maintenance of the pain.Osthole administered epidurally in the early stage of pain could alleviate the pain for a long time,which may be related with inhibiting p38 MAPK signaling related pathways.

OBJECTIVETo explore the incidence, pathogenesis, risk factors, prophylaxis and treatment of acute kidney injury (AKI) after myeloablative allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSClinical data of 120 patients received myeloablative allo-HSCT were retrospectively analyzed.

RESULTSSerum creatinine level in the patients showed significantly higher than baseline value at 28-60 days after transplantation (P<0.05). 73 patients (60.8%) developed AKI at a median of 33 days after allo-HSCT, including grade 2 in 32 patients (26.7%). Patients with grade 1 AKI showed significant higher serum cyclosporine A (CsA) levels (P<0.05). Hepatic veno-occlusive disease( HVOD), acute graft-versus-host disease (aGVHD) and total bilirubin > 40 micromol/L were high risk factors of occurring AKI (P<0.05). 19 patients died within 100 days after allo-HSCT, grade 2 AKI was a high risk factor of mortality (P< 0.05). 180-day survival rate was significantly lower in patients with grade 2 AKI after allo-HSCT (P<0.05).

CONCLUSIONAKI is one of the major complications after myeloablative allo-HSCT. Prophylaxis and treatment of AKI might reduce mortality in early stage of transplantation.

Acute Kidney Injury ; etiology ; prevention & control ; Adolescent ; Adult ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; Retrospective Studies ; Risk Factors ; Transplantation Conditioning ; Transplantation, Homologous ; Young Adult

This study was purposed to investigate the effect of prostaglandin E2 (PGE2) on proliferation of peripheral blood T lymphocytes, and to evaluate the regulatory role of PGE2 on immunological balance between Th1/Th2 and Tc1/Tc2 lymphocytes. The peripheral blood mononuclear cells (PBMNC) were stimulated by anti-human CD3 monoclonal antibody (mAb) and anti-human CD28 mAb, and were cultured in the presence of different concentration of PGE2 for 120 hours. The proliferation of peripheral blood T lymphocytes was assayed according to the manufacture protocol of BrdU Kit; the IFN-gamma and IL-4 levels in supernatants cultured for 24, 48, 72 and 120 hours were detected by ELISA; the ratios of CD4+IL-4+ T cells/CD4+ IFN-gamma+ T cells and CD8+IL-4+ T cell/CD8+IFN-gamma+ T cells were determined by flow cytometry. The cells cultured without PGE2 were used as control. The results indicated that (1) with the raising of concentration of PGE2, the inhibitory rate of T cell proliferation in vitro significantly increased (p=0.001). There was significant positive correlation between inhibitory rate of T cells and PGE2 concentration (correlation coefficient=0.889, p=0.000). (2) the difference between the IFN-gamma concentrations in supernatant cultured for 120 and 72 hours in test groups had no statistical significance (p=0.917). The IFN-gamma concentration increased continually with prolonging of culture time in control group (p=0.046). The IFN-gamma concentrations produced at different times in test group were significantly lower compared with those in control group (p<0.05). The IL-4 concentrations produced at different time had no significant change in test groups (p=0.400). The IL-4 concentration in 24 hours in control group was significantly higher than that at 48, 72 and 120 hours in control group (p=0.007, 0.003 and 0.002). After cultured for 24 hours the IL-4 concentration in test group was significantly lower than that in control group (p=0.037), but after cultured for 48, 72 and 120 hours, the IL-4 concentration in test group did not show statistical difference in comparison with control group (p>0.05). (3) the proportions of CD4+IFN-gamma+T cells in test group and in control group had no significant difference (p=0.767). The proportion of CD4+IL-4+T cells in test group was slightly higher than that in control group (p=0.051). The ratio of CD4+IL-4+T cells to CD4+IFN-gamma+ T cells in test group was significantly higher than that in control group (p=0.011). The proportions of CD8+IFN-gamma+ T cells in test group and in control group had no statistical difference (p=0.441). The proportion of CD8+IL-4+T cells in test group was significantly higher than that in control group (p=0.015). The ratio of CD8+IL-4+ T cells to CD8+IFN-gamma+ T cells in test group were obviously higher than that in control group(p=0.038). It is concluded that the PGE2 inhibits the proliferation of T lymphocytes in vitro. PGE2 influences the production of IFN-gamma and IL-4, and significantly influences peak appearance of IFN-gamma produced by T lymphocyte. PGE2 can continuously inhibit the production of IFN-gamma, but its continuous effect on IL-4 is no significant. PGE2 enhances the ratio of CD4+IL-4+T lymphocytes to CD4+IFN-gamma+T lymphocytes and the ratio of CD8+IL-4+T lymphocytes to CD8+IFN-gamma+T lymphocytes, and regulates development of T cells toward Th2/Tc2 cells.
Cell Proliferation ; drug effects ; Dinoprostone ; pharmacology ; Flow Cytometry ; Humans ; Lymphocyte Activation ; drug effects ; Lymphocyte Count ; T-Lymphocytes, Cytotoxic ; drug effects ; immunology ; Th1 Cells ; drug effects ; immunology ; Th2 Cells ; drug effects ; immunology

OBJECTIVETo explore the expression of CD66c (CEACM6) in adult acute leukemia and its significance.

METHODSAcute leukemia cell lines HL-60, K562, LCL721.221 and Jurkat were cultured in vitro. RT-PCR and multi-parameter flow cytometry were applied to analysis of CD66c mRNA and protein expression respectively in the cell lines and patient' s bone marrow leukemic cells. Cytogenetic analysis for 199 bone marrow samples from leukemia patients and Minimal Residual Disease (MRD) detection for 25 CD66c positive B lineage ALL were performed.

RESULTS(1) CD66c expression both on cell surface and in plasma were negative in all the cell lines. (2) Four of 127 AML (3.15%) (mainly of M2 and M4), and 28 of 79 ALL (35.44%) (all of B linage ALL) were CD66c positive the subtypes of the ALL being common B-ALL (20/54) and pre B-ALL (8/11) including 8 Ph + B-linage ALL. (3) Six-month relapse rate was significantly different between the MRD positive and negative patients. (4) CD66c mRNA was strongly expressed in B-linage ALL. For the cell lines, only the HL60 cells weakly expressed CD66c mRNA.

CONCLUSIONCD66c expression could be a useful bio-marker for the MRD analysis in ALL, and is closely associated with its transcription level.

Adolescent ; Adult ; Aged ; Antigens, CD ; biosynthesis ; Carcinoembryonic Antigen ; biosynthesis ; genetics ; Cell Adhesion Molecules ; biosynthesis ; GPI-Linked Proteins ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia, Myeloid, Acute ; metabolism ; Male ; Middle Aged ; Neoplasm, Residual ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; RNA, Messenger ; biosynthesis

The study was purposed to investigate the differentiation ability of mesenchymal stem cells (MSCs) into myocardial cells in vitro. Rat bone marrow-derived MSCs were labeled and co-cultured with neonatal rat cardiomyocytes (CM) for 5 - 7 days. The expression of cell surface antigens was detected by flow cytometry, and the expression of muscle-specific marker myosin and troponin T in labeled cells was detected by immunofluorescence. The results showed that in vitro cultured MSCs expressed CD90, CD44, CD105, CD54, not expressed CD34, CD45, CD31. After co-cultured with neonatal rat CM, labeled MSCs differentiated into cardiomyocyte-like cells expressing myosin and troponin T. It is concluded that MSCs can differentiate into cardiomyocyte-like cells when co-cultured with neonatal myocardial cells in vitro. In co-culture of two kind of cells in ratio of four to one showed obvious efficacy differentiating MSCs into CMs.
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Coculture Techniques ; Mesenchymal Stromal Cells ; cytology ; Myocytes, Cardiac ; cytology ; Rats ; Rats, Wistar