Aim To establish an allogenetic mouse skin trans-plant model,in order to provide a research model for immunosup-pressive drugs. Methods Skins from the ears of C57BL/6 mice were transplanted to the back of BALB/c mice and skin isografts ( BALB/c mice to BALB/c mice) were used as control. Cyclos-porin A( CsA) was used as a model compound to test the imm-nosuppresive effect on allogenetic graft rejection. Following the transplation and CsA treatment, the graft rejection score and graft skin survival rate were quantified. Four and nine days after transplantation,serum IL-4,IL-12 and IFN-γ levels were meas-ured using ELISA kits. Twelve days after transplantation, mice were sacrificed. The weight of spleen and thymus was obtained, and CD4 + and CD8 + population of spleenic T cells were ana-lyzed using flow cytometer. Histological features were assessed by hematoxylin-eosin( HE) staining of formalin-fixed, paraffin-em-bedded graft skins. Results After transplantion, the graft rejec-tion score increased and graft skin survival rate decreased gradu-allly. Serum IL-12 and IFN-γ levels of allograft mice increased markedly. Compared with those of isograft mice, mice with skin allograft displayed a significant increase in the percentage of the CD8 + T cell subpopulation. Remarkable inflammation, such as edema, inflammatory cell infiltration were observed in allograft mice. Compared with saline treated mice, CsA significantly re-duced the graft rejection score and improved survival rate of skin grafts. And also, CsA treated mice had smaller spleen and thy-mus. Mice that received high doses of CsA had significantly less CD8 + T cells than those treated with saline. Moreover, allograft skins in mice that received CsA had less inflammation. Conclu-sions Allogenetic mouse skin transplantation exhibits acute graft rejection. CsA can inhibit the rejection in a dose dependent manner.

Alcoholic liver disease ( ALD ) , a chronic progres-sive disease, threatens human health seriously. An increasing number of studies have shown that gut flora dysbiosis plays an important role in the development of ALD. Intestinal microbiota maintains a steady state under normal conditions, regulating gut flora normal physiological function. However, chronic alcohol consumption produces intestinal bacteria overgrowth and dysbio-sis, including the alteration of the composition of intestinal mi-croflora, the increment of gut permeability and bacterial translo-cation. Subsequently, the host immune is activated, promoting the production of inflammatory cytokines in liver, which plays a central role in the development of ALD. Notably, the supple-ment of prebiotics or probiotics reverses the intestinal flora disor-der,ameliorating the clinical symptoms effectively in ALD pa-tients. The evidence impies that the modulation of dysbiosis may be effective in the prevention and treatment of ALD. This review summarizes the research progress on the mechanism of the devel-opment of dysbiosis-mediated ALD, to provide a theoretical basis for the research on intestinal flora and ALD.

Aim To investigate the role of Wnt/β-cate-nin signaling pathway on the baicalin-induced osteo-genic differentiation in rat bone marrow derived mesen-chymal stem cells ( rBMSC ) . Methods rBMSC was isolated and cultured by adherence screening method. Alkaline phosphatase ( ALP) amount, CFU-FALP and mineralized nodules were compared between each ba-icalin group and vehicle control group at different time points. Real time q-PCR was employed to evaluate the mRNA level of Wnt signaling-related marker ( Wnt10a, GSK-3β,β-catenin and LEF1) after baica-lin treatment. Protein expression of β-catenin and Runx2 was measured by Western blot. Results Ba-icalin significantly increased ALP activities from day 3 to day 7 . The formation of CFU-FALP and mineralized nodules remarkably increased after rBMSC was treated with1, 10, 50 μmol · L-1 baicalin. mRNA levels of Wnt10a, β-catenin, GSK-3β, LEF1and osteocalcin were enhanced significantly in baicalin-treated group compared to control group. Protein expression of β-catenin and Runx2 was also elevated. Conclusion Baicalin ( 0. 1 to 50 μmol · L-1 ) promotes the osteo-genic differentiation and maturation of rBMSC, in which Wnt/β-catenin signaling pathway might be in-volved.

To explore novel antifatigue agents targeting with AMPA receptor, 10 compounds were synthesized and their structures were confirmed by 1H NMR, ESI-MS and elemental analysis. 1-BCP was treated as the leading compound. The antifatigue activities were evaluated by weight-loaded forced swimming test, and the AMPA receptor binding affinities were tested with radioligand receptor binding assays. The results unveiled that 5b appeared to possess potent antifatigue activities and high affinity with AMPA receptor, which deserved further studies.

Intestinal microflora is an important part of the organ-ism, promoting digestion and absorption of nutrients, maintaining intestinal normal physiological function, regulating immune sys-tem. Intestinal microflora maintains steady state under normal conditions, but intestinal microbiota dysbiosis occurs when surrounding environment c hanges, such as age, diet, obesity and other metabolic diseases as well as antibiotics. Many recent studies have found intestinal flora could cause a variety of disea-ses, and colon cancer is closely related with intestinal microbiota dysbiosis. Some researches suggest improving the intestinal flora dysbiosis can reduce the incidence of colon cancer and inhibit the growth and the worsening of colon cancer. However, under-lying mechanisms remain unknown. So this article summarizes the research progress on the development of colon cancer and in-testinal microbiota dysbiosis, in order to provide reference for re-search on intestinal flora and colon cancer treatment.

Triptolide, a compound extracted from the traditional Chinese medicine preparation of Tripterygium wilfordii Hook F., has been reported to have anti-inflammatory and anti-cancer activities. However, its effect on ovarian cancer invasion is unknown. We observed that MMP7 and MMP19 expression increased in ovarian cancer tissue. Triptolide treatment inhibited the migration and invasion of ovarian cancer cells SKOV3 and A2780 at the concentration of 15 nM. We also observed that triptolide suppressed MMP7 and MMP19 promoter activity in a dose-dependent manner, down-regulating the expressions of these promoters on mRNA and protein level. Moreover, triptolide enhanced E-cadherin expression in ovarian cancer cells. In vivo, triptolide inhibited tumor formation and metastasis in nude mice, and suppressed MMP7 and MMP19 expression; it also enhanced E-cadherin expression in tumor in a dose-dependent manner. Over expression of MMP7 and MMP19, or suppression of E-cadherin expression partially abolished the inhibitory effect of triptolide on invasion of ovarian cancer cells. To summarize, triptolide significantly inhibited the migration and invasion of ovarian cancer cells by suppression of MMP7 and MMP19 and up-regulation of E-cadherin expression. This study shows that triptolide is a good candidate for the treatment of ovarian cancer and reduction of metastasis.
Animals ; Antineoplastic Agents, Alkylating/*pharmacology ; Cadherins/*genetics/metabolism ; Cell Line, Tumor ; Cell Movement/drug effects ; Cell Proliferation/drug effects ; Cystadenocarcinoma, Serous/*drug therapy/pathology ; Diterpenes/*pharmacology ; Epoxy Compounds/pharmacology ; Female ; Gene Expression Regulation, Enzymologic/drug effects ; Humans ; Matrix Metalloproteinase 7/genetics/*metabolism ; Matrix Metalloproteinases, Secreted/genetics/*metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Ovarian Neoplasms/*drug therapy ; Paclitaxel/pharmacology ; Phenanthrenes/*pharmacology ; Promoter Regions, Genetic ; Up-Regulation/drug effects ; Xenograft Model Antitumor Assays

Triptolide, a diterpenoid triepoxide from the traditional Chinese medicinal herb Tripterygium wilfordii Hook. f., is a potential treatment for autoimmune diseases as well a possible anti-tumor agent. It inhibits proliferation of coloretal cancer cells in vitro and in vivo. In this study, its ability to block progress of colitis to colon cancer, and its molecular mechanism of action are investigated. A mouse model for colitis-induced colorectal cancer was used to test the effect of triptolide on cancer progression. Treatment of mice with triptolide decreased the incidence of colon cancer formation, and increased survival rate. Moreover, triptolide decreased the incidence of tumors in nude mice inoculated with cultured colon cancer cells dose-dependently. In vitro, triptolide inhibited the proliferation, migration and colony formation of colon cancer cells. Secretion of IL6 and levels of JAK1, IL6R and phosphorylated STAT3 were all reduced by triptolide treatment. Triptolide prohibited Rac1 activity and blocked cyclin D1 and CDK4 expression, leading to G1 arrest. Triptolide interrupted the IL6R-JAK/STAT pathway that is crucial for cell proliferation, survival, and inflammation. This suggests that triptolide might be a candidate for prevention of colitis induced colon cancer because it reduces inflammation and prevents tumor formation and development.
Animals ; Cell Transformation, Neoplastic/*drug effects ; Colitis/complications ; Colonic Neoplasms/chemically induced/*drug therapy/metabolism/pathology ; Dextran Sulfate/toxicity ; Dimethylhydrazines/toxicity ; Diterpenes/*administration & dosage ; Epoxy Compounds/administration & dosage ; Humans ; Interleukin-6/biosynthesis ; Janus Kinases/metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred ICR ; Mice, Nude ; Neoplasm Transplantation ; Phenanthrenes/*administration & dosage ; STAT3 Transcription Factor/metabolism ; Signal Transduction/*drug effects ; Tumor Burden/drug effects ; rac1 GTP-Binding Protein/*biosynthesis

To observe the expression of cyclooxygenase (COX)-1 and COX-2 in brain after spared nerve injury (SNI) and compare the analgesic effects of COX inhibitors with different selectivity. Radioimmunoassay, RT-PCR and Western blotting techniques were used to evaluate the change of brain COX expression at different time points( before SNI, 1 h, 12 h, 1 d, 3 d, 7 d, 14 d, 30 d and 60 d after SNI); By exploring hot plate test, we observed the reacting time of animals after injection of saline, NS-398, SC-560 and indomethacin at different time points. The results showed that: ( 1 ) The expression of brain COX-1 didn't increase significantly until 14 d after SNI, while that of COX-2 increased significantly and rapidly after SNI and reached peak at the time point of 1 d ( all P ＜0.05 ); (2) NS-398 showed significant analgesic effect on neuropathic pain after SNI at the early phase ( P ＜ 0.05 ), but didn't persist for over 30 d; ( 3 ) Indomethacin and SC-560 didn't show significant analgesic effects until 14 d. These results suggest that brain COX-1 is involved in the late phase of neuropathic pain and may play a role in the persistence of pain, while brain COX-2 is involved in the early phase of neuropathic pain and may play a role in the pain origination.

The nephrotoxicity of gentamicin (GM) has been widely recognized. Heat shock protein 72 (HSP72) has been reported to be a cytoprotectant. However, its cytoprotective effect against GM induced kidney injury has not yet been studied. In this study, we investigated the cytoprotective effect of HSP72 on GM-induced nephrotoxicity in vitro. Human Kidney tubular cell line, HK-2 cells were divided into four groups: control group, GM group (cells incubated with GM only), heat shock (HS) group (cells incubated at 43 degrees C for 30 min), and GM plus HS group, respectively. Lactate dehydrogenanse (LDH) release increased time-dependently from 24 hr to 96 hr compared to the data of cells treated with GM only. Results of NAG activities, superoxide dismutase (SOD) activities and malondialdehyde (MDA) content were similar to that of the LDH release. The amount of HSP72 positive cells increased significartly at 72 hr after cells were treated with GM only. Both HSP72 protein and gene expression increased significantly at 72 hr when cells were treated with GM. On the other hand, HS induced HSP72 expression markedly. Pretreatment of HS inhibited HK-2 cells from GM-induced injury. It could reduce LDH release and NAG activity. HS also increased SOD activity, and decreased MDA content when cells were damaged by GM. These findings suggested that HS may protect kidney cells from GM-induced injury. Pre-induction of HSP72 may provide therapeutic strategies for nephrotoxicity induced by GM.
Reactive Oxygen Species/metabolism ; RNA, Messenger/analysis ; Oxidation-Reduction ; L-Lactate Dehydrogenase/secretion ; Kidney Tubules, Proximal/chemistry/*drug effects ; Humans ; Heat ; HSP72 Heat-Shock Proteins/analysis/genetics/*physiology ; Gentamicins/*toxicity ; Cytoprotection ; Cells, Cultured

BACKGROUND: How to choose traditional Chinese remedy to treat cerebrovascular disease, not only improving the brain blood supply but also not affecting the blood pressure and heart rate, has been a promising research.OBJECTIVE: To study the role of yangyintongnao granule on average blood pressure and heart rate in anaesthetized dog.DESIGN: Complete grouping design and randomized controlled study based on hybrid dog.SETTING: Department of Anesthesiology, Xijing Hospital, Fourth Military Medical University of Chinese PLA and Department of Pharmacology, Faculty of Preclinical Medicine, Fourth Military Medical University of Chinese PLA MATERIALS: The experiment had finished by Cardiovascular Laboratory of Physiology Department in the Fourth Military Medical University of Chinese PLA from March to June 2003. Totally 23 healthy hybrid dogs in either sex were selected. These dogs were divided randomly into 4 groups: high dose group of yangyintongnao( n = 8), moderate dose group of yangyintongnao ( n = 6), low dose group of yangyintongnao( n = 5) and saline group( n = 4).METHODS: Anesthetic dogs in high, moderate and low dose group of yangyintongnao granule were given different doses of yangyintongnao granule: 2 g/kg, 1 g/kg and 0. 5 g/kg respectively. All doses of drug were calculated based on the body mass of dogs, and the drugs were dissolved in 100 mL saline and given through the gastric canal. Dogs in saline group were perfused with equal saline. Aortic average blood pressure was measured by femoral artery intubation via a piezometric transducer. Heart rate was obtained from R-R intervals of a standard Ⅱ lead of electrocardiograph(ECG). The mean blood pressure and heart rate were recorded 0. 5, 1, 1.5, 2.0,3.0, 4. 0, 5.0, 6. 0 hours after treatment.MEAN OUTCOME MEASURES: The changes of blood pressure and heart rate at different time before and after medication.RESULTS: Totally 29 dogs were brought into the final analysis. Blood pressure: The mean blood pressure was reduced -5.4% to -6. 2% respectively in high dose group and moderate dose group after treatment. But in low dose group the average blood pressure sometimes increased sometimes decreased, mainly decreased. It increased by 6.6% ( P ＞ 0. 05) and decreased by -4. 1% ( P ＞ 0.05) at maximum. The average blood pressure in saline group changed by - 9.6% ( P ＞ 0.05). Heart rate: The heart rate in high and medium dose group gradually reduced as time went on. It reduced by - 4. 4%, - 12.2% and - 9.5% respectively in high, moderate and low doses group. The change in each group was not statistically significant as the same in saline group( P ＞ 0.05).CONCLUSION: Yangyintongnao granule has no significant influence on average blood pressure and heart rate.