Pirh2 (p53-induced protein with a Ring-H2 domain),one of the ubiquitin-protein E3 ligases,can promote p53 degradation via ubiquitin-proteasome pathway,and repress the biological functions of p53.Researchers have found that the Pirh2 is overexpressed in many cancers,which suggests Pirh2 is probably correlated with tumorigenesis and cancer development,and it may become a promising cancer therapeutic target.

With the indepth studies of molecular mechanisms of renal cell carcinoma ( RCC),people have made great progress in the targeted therapy drugs which targeted at vascular endothelial growth factor (VEGF) and their receptors,which dramatically improve the treatment outcome for patients with advanced or metastatic RCC.However,recent clinical trails show that the targeted therapy drugs may cause adverse events,such as hypertension,bone marrow toxicity,pneumonitis and so on.It is important to manage the adverse events promptly not only to improve the therapeutic efficacy but also to improve the quality of life for patients with RCC.

Objective The diolistic assay has been modified to make it simpler and more efficient in labeling neurons and glia. Methods CNS neurons and glial cells were labeled with DiI diolistic assay in fixed tissue and living brain slices of C57/B6J mice. Results The method allowed the visualization of the fine structure of neurons and glia including synaptic structures such as dendritic spines. Conclusion With the method, the labeling efficacy of cell's fine structure is improved, making it preferable for the analysis of dendritic spine. In addition, the ability to label the living neuron and glia will extend its application vastly.

AIM:To study the urinary pharmacokinetics of five anthraquinones after oral administration of Xiexin Decoction(Radix et Rhizoma rhei,Rhizoma coptidis and Radix scutellariae) in rats.METHODS:A high-performance liquid chromatographic method with fluorescence detection(HPLC-FLD) was established and validated the quantification for five anthraquinones(aloe-emodin,rhein,emodin,chrysophanol and physcion) in rat urine.SD rats were given 12 g/kg of Xiexin Decoction.Urine was collected before and after perfusion.Anthraquinones components in urine were measured by HPLC-FLD.Urinary pharmacokinetic parameters were determined according to urinary output-time data.RESULTS:After oral administration of Xiexin Decoction all the five anthraquinones were excreted from the urine.The excretion T_ 1/2 of aloe-emodin,rhein,emodin,chrysophanol and physcion were 3.46?1.18,3.24?0.60,4.69?1.99,4.49?1.63,5.65?1.74 h,respectively.The amounts of aloe-emodin,rhein,emodin,chrysophanol and physcion excreted from urine during 0～48 h were(11.28?4.30)?g,(116.73?17.46)?g,(5.48?2.92)?g,(9.53?2.67)?g,(0.41?0.20)?g,respectively.CONCLUSION:After oral administration of Xiexin Decoction five anthraquinones were excreted from urine and a small quantity of five anthraquinones excreted from urine in rats is less than 10% of oral dose.

Objective To explore the clinical effect of operative treatment for grade Ⅳ pronation-external rotation ankle injury.Methods 60 patients with pronation extorsion type Ⅳ degree of ankle injury were selected as the research subjects.According to the digital table,they were divided into two groups,30 cases in each group.The group A adopted ordinary steel plate,the group B adopted locking plate fixation.The clinical curative effect,postoperative recovery index,AOFAS score and complications were observed.Results The total effective rate of group A was 93.3％,which of group B was 90.0％,the difference was not statistically significant (x2 =0.218,P ＞ 0.05).The swelling subsided time [(24.2 ± 5.0) d vs.(22.9 ± 4.1) d],fracture healing time [(42.4 ± 9.5) d vs.(41.3 ± 9.4) d],postoperative recovery time of walking [(46.6 ± 9.2) d vs.(45.7 ± 9.0) d],AOFAS score [(87.3 ± 2.6) vs.(88.4 ±2.2)] between the two groups had no statistically significant differences(t =1.101,0.451,0.383,1.769,all P ＞0.05).The group A had 1 case in infection,the incidence rate of complications was 3.3％.The group B had no complication.The difference was not statistically significant in the incidence of complications between the two groups (x2 =1.017,P ＞ 0.05).Conclusion In the operative treatment for patients with grade Ⅳ pronation-external rotation ankle injury,whether choose ordinary steel plate fixation,or choose the locking plate fixation,the clinical effects of the two methods are both good,and the postoperative healing is fast.

BACKGROUND: It has been demonstrated that electromagnetic field (EMF) can adjust proliferation and differentiation of bone marrow mesenchymal stem cells, but the specific mechanism is not clear. OBJECTIVE: To investigate the effects of EMF-activated ERK1/2 pathway on proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells.METHODS: The 3rd passage of rat bone marrow mesenchymal stem cells were received EMF treatment (15 Hz, 1 mT, sine wave), 20 μmol/L PD98059 + EMF treatment, or only PD98059 treatment. Simultaneously, a normal control group was established. Western blotting was applied to detect the activation of ERK signal pathway after EMF exposure. MTT assay was used to determine the activation of proliferation of cells. And alkaline phosphatase (ALP) activity in cells was detected by an ALP kit. RESULTS AND CONCLUSION: The ERK1/2 phosphorylation, proliferation and ALP activity of rat bone marrow mesenchymal stem cells were remarkably increased after exposure to EMF (P < 0.01). PD98059 could effectively block the increasing of ERK1/2 phosphorylation and cell proliferation (P < 0.01), but elevate ALP activity in a certain level (P < 0.01). EMF stimulation can fast activate ERK1/2 signal pathway and then promote the proliferation of rat bone marrow mesenchymal stem cells, however, ERK1/2 signal pathway activation has a less effect on osteogenic differentiation of bone marrow mesenchymal stem cells.

Objective To observe the therapeutic effect of general combinable external fixator for the treatment of femoral intertrochanteric fracture(FIF).Methods One hundred and seventeen FIF patients were randomized into two groups: the treatment group(N =56) received treatment with general combinable external fixator,and the control group(N =61) received bone traction.A follow-up of 6~13 months was carried out.The therapeutic effect and the incidence of complications were compared in both groups.Results The total effective rate was 96.43% in the treatment group and 50.82% in the control group,and the incidence of complications was 7.1%(4/56) in the treatment group and 45.9%(28/61)in the control group,the difference being significant(P

Objective To study the effects of electromagnetic field (EMF) on the activation of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) pathways in mesenchymal stem cells (MSCs) and their interaction, and to explore the cellular signal transduction mechanism of the biological effects induced by EMF.Methods The 3rd-passage rat bone marrow MSCs were randomly divided into a control group, an EMF group, an EMF + PD98059 group and an EMF + SB202190 group.Cells in the EMF group were cultured in the electromagnetic field, those in the EMF + PD98059 and EMF + SB202190 groups cultured in the electromagnetic field after PD98059 or SB202190 was added, and those in the control group were cultured normally.Different groups of cells were exposed to electromagnetic fields (15 Hz, 1 mT, sine wave form) for different exposure duration.The activated phosphorylated and non-phosphorylated p38MAPK were measured using Western blotting analysis with their specific corresponding antibodies.The alkaline phosphatase (ALP) activity in cells in different groups was detected according to the instructions of ALP kit.MTT assay was applied to investigate the proliferation of cells.Results Electromagnetic fields could rapidly induce the activation of p38 MAPK (P ＜ 0.05) and the phosphorylation of p38 MAPK elevated after 15 min exposure to EMF.The phosphorylation of p38 MAPK was significantly lower in the EMF + SB202190 group than that of the EMF group.After 5 days of EMF exposure, the ALP activity of cells was significantly improved, and the effect could be inhibited by SB202190.The bone marrow mesenchymal stem cells proliferation increased significantly after being exposed to EMF for 3 days, and it could not be blocked by SB202190.Phosphorylation of ERK and MAPK increased significantly when the p38 MAPK pathway was blocked by SB202190 and exposed to EMF for 5 minutes, and it also increased significantly when the ERK MAPK pathway was blocked by PD98059 and received 30 minutes of EMF exposure.Conclusion EMF can quickly activate ERK and p38 MAPK pathways to induce cell proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.Moreover, in EMF there is a mutual interference between ERK and p38 MAPK pathways.

BACKGROUND: It has been proved that electromagnetic field can adjust and control proliferation and differentiation of bone marrow mesenchymal stem cells v/a cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) signal transduction system. However, there are few relevant reports about Ca2+ as the second messenger in application. OBJECTIVE: To study the effects of verapamil on the proliferation and differentiation of bone marrow rnesenchymal stem cells stimulated by electromagnetic fields and to conclude influx changes of Ca2+.DESIGN, TIME AND SETTING: Electrostimulative cytological observation in vitro, which was performed in Laboratory of Orthopedic Surgery, Tongji Hospital between April and June 2005.MATERIALS: Six 4-5-week SD rats of clean grade were selected in this study. Verapami| was provided by Sigma Company, USA, and Helmholtz coil-magnetic field producer was made in Department of Electric Machine, Navy Engineering University.METHODS: The bone marrow mesenchymal stem cells were isolated and cultured in vitro with adherence method and digested with trypsin. The fourth-passage cells were harvested, adjusted to 1 × 107 L-1 in density, and divided into A, B, C and D groups in 96-well plate with 200 μ I/well. Cells in the normal control group were not performed with any agent. On the second day of inoculation, cells in the magnetic field (EMF) group were cultured in Helmholtz-coil magnetic field (0.8 mT, 50 Hz) in 0.05% CO2 saturated humidity incubator at 37 ℃, 30 minutes for each, 12 hours for interval, six time in total. Cells in the verapamil group were cultured with 20 μ mol/L verapamil, and cells in the combination group were cultured with 20 μ mol/L verapamil and magnetic stimulation.MAIN OUTCOME MEASURES: Proliferative activity was tested with MTT method, content of alkaline phosphate differentiated to osteoblasts was measured, and cells were stained with modified Gomori Ca-Co staining. RESULTS: Proliferative activity was significantly increased in the EMF group as compared with that in the normal control group after 3-day magnetic stimulation (P < 0.01), but verapamil could inhibit promotive effect on proliferation. Content of alkaline phosphate in the normal control group was similar to that in the EMF group, while those two contents were significantly higher than those in the verapamil group and the combination group (P < 0.01); furthermore, content of alkaline phosphate in the combination group was significant higher than that in the EMF group (P < 0.01). Qualitative analysis of alkaline phosphate showed a coincident result as mentioned above.CONCLUSION: EMF of 50 Hz frequency and 0.8 mT intensity can change intracellular free calcium ion concentration of bone marrow mesenchymal stem cells, and the change play a key role in the cellular proliferation and play a partial role in the differentiation of bone marrow mesenchymal stem cells into osteoblasta.

Objective To study the expression level and clinical significance of Galectin-3 and miRNA-21 in non-small-cell lung carcinoma(NSCLC).Methods One hundred and fifty patients with NSCLC were chosen as cancer group,and 1 50 patients with benign pulmonary diseases were chosen as control group.The expression level of Galectin-3 and that of miRNA-21 between two groups were compared,and the relevance between expression level of Galectin-3 and that of miRNA-21 and clinical feature were analysed.Results In cancer group,the expression level of Galectin-3 was 6.75±2.38,and that of control group was 1.12 ±0.29;the expression level of miRNA-21 was 5.91 ± 1.59,and that of control group was 0.97 ± 0.1 7,and the difference between two groups had statistical significance(P <0.05 ).The relevance between expression level of Galectin-3 and stage,differentiation,lym-phatic metastasis,diameter of carcinoma and PFS,OS of patients had statistical significance(P <0.05).The relevance between ex-pression level of miRNA-21 and stage,differentiation,diameter of carcinoma and PFS,OS of patients had statistical significance(P <0.05).In the diagnosis of NSCLC,the sensitivity of the expression level of Galectin-3 was 90.20%,and its specificity was 70.69%, while the sensitivity of expression level of miRNA-21 was 88.24% and its specificity was 69.97%.The difference between the di-agnostic value of Galectin-3 and that of miRNA-21 had no statistical significance(P >0.05 ).Conclusion The expression level of Galectin-3 and that of miRNA-21 can be applied in the diagnosis and prognosis of non-small-cell lung carcinoma.