Purpose　To study the effects of α-tocopherol on activator protein-1(AP-1)binding and TGF-β1 expression induced by high glucose in rat mesangial cells and further to clarify the molecular mechanism of antioxidant in treating diabetic nephropathy.　Methods　AP-1 binding of the rat mesangial cells exposed to high glucose was detected by gel shift assay.The Jun,Fos compositions of AP-1 dimer were determined by supershift assay.Protein expression of TGF-β1 was detected by Western blot.Additionally,the effects of α-tocopherol on AP-1 binding and TGF-β1 expression induced by glucose in rat mesangial cells were also studied.　Results　High glucose stimulated AP-1 binding of mesangial cells in time-and-dose-dependent manners .This AP-1 binding increase involved JunD and Fos as shown by gel supershift.Glucose also increased protein expression of TGF-β1 at same time.The increased AP-1 binding and TGF-β1 were inhibited with pretreatment with α-tocopherol in glucose-treated mesangial cells.　Conclusions　This study suggests that α-tocopherol can significantly inhibit AP-1 activity and TGF-β1 expression by glucose in rat mesangial cells,which may be one of its antioxidation mechanisms to retard diabetic nephropathy.

Objective To investigate the possible role of protein kinase(PK) activity in the induction of AP-1 by ox-LDL in mesangial cells and to elucidate the upstream signal pathways involved in the ox-LDL-induced AP-1 binding. Methods Rat mesangial cells were randomly divided into the normal cells, ox-LDL-treated cells and PK inhibitor + ox-LDL-treated cells. Treatments with the PKC inhibitor bisindolylmaleimide I, the PKA inhibitor H89, the PTK inhibitor genistein (GEN), the MEKi inhibitor PD98059, or the p38 MAPK inhibitor SB203580 were applied prior to a 24-hour incubation of ox-LDL in mesangial cells. The phosphorylation of MAPK families was detected by Weatern blot analysis. AP-1 binding was determined by gel shift assay. Results Ox-LDL stimulated the phosphorylation of JNKi/SAPK and p38 MAPK( P 0. 05) . Bisindolylmaleimide I at 50, 100, 200 nmol/L appreciably reduced the ox-LDL-induced AP-1 binding. H89 at 0. 5, 5 umol/L significantly inhibited AP-1 binding by ox-LDL. GEN at 25, 50 umol/L did not reduce the AP-1 binding by ox-LDL, but when GEN rose up to 100 umol/L, the ox-LDL-induced AP-1 binding significantly decreased in mesangial cells. However, SB203580 and PD98059 did not reduce the ox-LDL-induced AP-1 binding in the present study. Conclusion Multiple protein kinases may involve in the stimulation of AP-1 by ox-LDL in mesangial cells.

AIM: To establish the quality standard for Jinqiaomai Tablets ( Fagopyri Dibotryis Rhizoma). METHODS: Fagopyri Dibotryis Rhizoma was identified by TLC. The contents of (-)-epicatechin and procyanidin B2 were determined simultaneously by HPLC,the Symmetry C18 column (5 ?m,3. 9 mm id ? 150 mm) was used. The water (adjust pH value to 3. 00 ? 0. 02 with phosphoric acid)-acetonitrile (92 ∶ 8) was used as a mobile phase,the detecting wavelength was at 280 nm. The column temperature was at 35 ℃. RESULTS: Fagopyri Dibotryis Rhizoma could be identified. The calibration curve of (-)-epicatechin was linear in the range of 0. 113 6-2. 272 ?g with the correlation of 0. 999 968 (n = 8). The average recovery of (-)-epicatechin was 97. 12% ,RSD = 0. 92% (n = 6). The calibration curve of procyanidin B2 was linear in the range of 0. 166 4-1. 664 ?g with the correlation of 0. 999 702(n = 8). The average recovery of procyanidin B2 was 98. 26% ,RSD = 0. 61% (n = 6). CONCLUSION: The method is specific,reliable and accurate,so it can be used for the quality control of Jinqiaomai Tablets.

Objective To provide a simple, convenient, and safe anesthesia method for the establishment of a M1 segment of middle cerebral artery occlusion model in rhesus monkey or other large laboratory animals.Method Twenty male rhesus monkeys weighing 7-11 kg (ages 7-9 years) from Academy of Military Medical Sciences were used in this study.Sumianxin injection combined with 0.1 mg/kg ketamine was given before endotracheal intubation (ID:4.5-5.5#).The animals were then transported to an interventional operation room, where the intravenous access was established and a urinary catheter was inserted into the urinary bladder.Mechanical ventilation was used during the surgery, propofol was continuously injected in a speed of 2-4 mg/kg/h, and Sumianxin-ketamine could be given if necessary to maintain adequate anesthesia depth.The dose was adjusted according to vital signs of the rhesus such as body movements, physiological parameters, and demand of surgery.Brain MRI examination was performed before and after thrombolysis.Anesthetic injection was suspended and the animals were allowed to have a spontaneous breathing every time before the MRI text.Heart rates, temperature, non-invasive blood pressure, and SpO2 were monitored during the whole surgery.Blood samples were taken from the radial artery for blood gas analysis after anesthesia induction and during operation.Results All the 20 animals underwent the operation successfully, no animal had restlessness, respiratory depression, arrhythmia and other serious complications.At the end of the study, animals awake soon.Fifteen of them survived longer than 24 hours, only 5 died from serious cerebral hemorrhage and larger cerebral infarction.Conclusions General endotracheal anesthesia is safe for rhesus monkeys during such interventional operation and MRI examination.

AIM: To investigate the role of N-cadherin in the delamination of neural crest cells. METHODS: The normal expression of N-cadherin in neural tube was identified using in situ hybridization. The cells with N-cadherin over expression were obtained by transfection of wild-type N-cadherin (wt-N-cadherin) ,and the cells with N-cadherin silencing expression were obtained by transfection of dominant-negative N-cadherin (dn-N-cadherin). The migration of cranial neural crest cells was determined by the technique of immunohistochemistry. RESULTS: Either overexpression or down-regulation of N-cadherin significantly affected the migration of cranial neural crest cells. CONCLUSION: Delamination and migration of the cranial neural crest cells rely on the relative N-cadherin expression in the neural tube during neurulation.

Objective To provide coronary sectional anatomical basis for imaging diagnosis of the injure and disease of the knees.Methods 5 right knees of male adults cadavers were used . After the MR imaging examination,all specimens were frozen and cut into 6 coronary sections . Results The morphological charactenstics relation and the law of variation of the articular facets,articular capsule,articular cavity,cruciate ligaments,menisci,synovial plicae and its surrounding structures on all coronary sections of knees were observed, and compared with the corresponding MR images .Conclusion The variation of morphology and structures of all coronary sectional specimens of knee joint provided foundation of dependable anatomy and clinical value for medical imaging diagnosis .

The protective effects of Da Chai Hu Granules (DCHKL) on islet cells which were incubated with 4 mmol x L(-1) alloxan (AXN) were studied. The viability of islet cells were measured with MTT. Insulin released into medium and in islets was detected by radioimmunoassay. Cell apoptosis rate was determined by flow cytometry. The expression of anti-apoptotic gene Bcl-2 and pro-apoptotic gene Bax in islet cells were measured with RT-PCR (reverse transcription polymerase chain reaction). Serum containing DCHKL can promote the activity of islet cells significantly (P < 0.01). Basal insulin secretion and high glucose-stimulated insulin secretion increased significantly (P < 0.01). Serum containing DCHKL can inhibit apoptosis of islet cells, the ratio of apoptosis was decreased. Serum containing DCHKL increased expression of Bcl-2 mRNA and decreased expression of Bax mRNA. DCHKL can significantly promote proliferation of islet cells and increase the amount of basal secretion of pancreatic islet cells and high glucose-stimulated insulin secretion. The expression of Bcl-2 increased significantly. The expression of Bax decreased significantly. DCHKL have a protective effect on the islet cells.

Objective:To investigate the effect of rapamycin on cell growth and migration of gallbladder cancer GBC-SD cells, and to discuss its potential in clinical therapy of gallbladder cancer. Methods: Proliferation of GBC-SD cells treated with different concentrations of rapamycin (12.5, 25, and 50 mmol/L) was examined by MTT assay. Cell cycle distribu-tion and apoptosis of GBC-SD cells treated with different concentrations of rapamycin were determined by flow cytometry. Migration ability of GBC-SD cells was assessed by Transwell assay. The expression of mTOR (mammalian target of rapam-ycin) and its phosphorylation in GBC-SD cells were examined by Western blotting assay. Results: Rapamycin significant-ly inhibited the phosphorylation of roTOR, but had no influence on the expression of roTOR in GBC-SD cells. Rapamycin significantly inhibited the growth of GBC-SD cells in a dose-dependent manner (P < 0.01). Raparnycin induced apoptosis of GBC-SD cells and arrested them at the G_1/S phase. Furthermore, rapamycin also significantly suppressed migration of GBC-SD cells as showed by Transwell assay (P < 0.01). Conclusion: Rapamycin can remarkably inhibit the growth and migration of gallbladder cancer cells, probably by inhibition of p-roTOR pathway, induction of apoptosis and cell cycle ar-rest of gallbladder cancer cells.

Objective:To observe the killing effect of irradiated peripheral blood mononuclear cells (PBMCs) at low dose combined with apogossypolone (ApoG2) on cultured human prostate cancer PC-3 cells.Methods:Human PBMCs were irradated by gamma ray at 1 gray,the irradiation dose rate was 17 Gy/min.The experiment were divided into PC-3 tumor cell control group,PC-3 cells with irradiated and non-irradiated PBMCs co-culture groups,ApoG2 treatment group,irradiated PBMCs and ApoG2 co-treatment group.Acridine orange/ethidium bromide (AO/EB) staining and MTT method were used to observe the killing effect of PBMCs and/or ApoG2.Results:The killing activity of irradiated PBMCs group and ApoG2 treatment group were obviously increased and were higlaer than that of non-irradiated group (P＜0.05).The killing activity of combined group were much higher than that of irradiated group and ApoG2 treatment group (P ＜0.01 ).Conclusion:Irradiated PBMCs at low dose combined with ApoG2 can enhances the anti-tumor effects markedly.

OBJECTIVE: To observe the expression changes of metadherin (MTDH) and matrix metalloproteinase-9 (MMP-9) in the laryngeal squamous cell carcinoma tissues and to investigate the significance. METHOD: The expression of MTDH and MMP-9 in 54 cases of laryngeal squamous cell carcinoma tissues(observation group) and 30 cases of para-carcinoma tissues (control group) was examined by immunohistochemical method, the correlation between them and their correlations with the clinicopathological parameters were analyzed. RESULT: The positive expression rates of MTDH in the observation group and control group were 64.8% (35/54) and 6.7% (2/30), respectively; the positive expression rates of MMP-9 in the observation group and control group were 70.4% (38/ 54) and 13.3% (4/30), respectively; and there was a statistically significant difference between two groups (all P < 0.01). In the laryngeal squamous cell carcinoma tissues, the expression of MTDH protein was related with degree of differentiation, lymph-node metastasis and TNM stage (all P < 0.05); and the expression of MMP-9 protein was related lymph-node metastasis and TNM stage (all P < 0.05). The expression of MTDH was positively correlated with MMP-9 in the laryngeal squamous cell carcinoma tissues (r = 0.371, P < 0.01). CONCLUSION The high expression of MTDH and MMP-9 was closely related to the occurrence, development and metastasis of laryngeal squamous cell carcinoma, joint detection of the two proteins was valuable for early diagnosis and prognosis of laryngeal squamous cell carcinoma.
Carcinoma, Squamous Cell ; diagnosis ; metabolism ; Case-Control Studies ; Cell Adhesion Molecules ; metabolism ; Cell Differentiation ; Head and Neck Neoplasms ; diagnosis ; metabolism ; Humans ; Laryngeal Neoplasms ; diagnosis ; metabolism ; Lymphatic Metastasis ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Staging ; Prognosis ; Squamous Cell Carcinoma of Head and Neck