Objective: To investigate the signal transduction mechanisms of time-dependent effects of 50 Hz 1.8 mT sinusoidal electromagnetic fields(SEMFs)on osteoblastic activity. Methods: Newborn rat calvarial osteoblasts(ROBs)were treated with 50 Hz 1.8mT SEMFs for 30, 60, 90, 120, and 150 min, respectively.Intracellular alkaline phosphatase(ALP)activity was assayed, and protein expression of Runt-related transcription factor 2(Runx2), Smad1/5/8 and mitogen-activated protein kinases(MAPKs)were examined by Western blot.The Dual-Luciferase Reporter Assay System was used to measure the activity of the Wnt pathway.Immunofluorescence staining and laser confocal microscopy were applied to examine the nuclear translocation of Smad1/5/8 and β-catenin.Changes in ALP activity were determined after inhibiting p38 MAPK using a specific inhibitor. Results: ALP activity of ROBs increased after 30, 60, 90, 120 and 150 min of SEMFs treatment, compared with the control group(51.41±5.21, 59.47±4.02, 67.56±4.68, 63.69±3.92, and 58.16±3.61 vs.45.53±3.24), and reached the highest value at 90 min and then started to decline.Protein expression of Runx-2, p-Smad1/5/8, p-p38 and β-catenin increased after SEMFs treatment, and reached the highest value at 90 min and then gradually returned to baseline levels.The values for Wnt pathway activity for the control group and with SEMFs treatment at 30, 60, 90, 120 and 150 min were 0.49±0.06, 0.52±0.09, 0.75±0.05, 0.77±0.42, 0.58±0.08 and 0.42±0.09, respectively.Wnt pathway activity, the nuclear translocation of β-catenin and Smad1/5/8 reached the highest level at 90 min.Values of ALP activity in the control group and the SEMFs group were 44.60±3.84 and 71.54±7.34, respectively, before specifically inhibiting p38 MAPK, and were 52.08±0.83 and 52.15±10.77, respectively, after p38 MAPK inhibition, indicating that ALP activity could not be increased with inhibition. Conclusions 50 Hz 1.8 mT SEMFs increase osteoblastic activity by activating the BMP-2/Smad1/5/8, Wnt/β-catenin and p38 MAPK signal pathways.The optimal duration of treatment is 90 min per day.

OBJECTIVE:To investigate the effect of total saponins of Man medicine Thladiantha dubia root(TSTR)on the ex-pressions of CD3+,CD4+and CD8+ in spleen tissue of rats with type Ⅱ collagen-induced arthritis (CIA),and explore its mecha-nism in the treatment of rheumatoid arthritis(RA). METHODS:8 rats were taken as normal control group(NC group),the others 72 rats were injected mixture of bovine type Ⅱ collagen and Freund's complete adjuvant in tail and back to induce CIA model. The 50 modeled rats were randomly divided into model group(MC group),tripterygium polyglycoside(TG)group(12 mg/kg,posi-tive control),TSTR low-dose,medium-dose,high-dose groups (20,40,80 mg/kg),10 in each group. Rats in medicine groups were intragastrically administrated for 35 d,once a day;rats in NC group and MC group were intragastrically administrated equal volume of distilled water. Ankle swelling degree of rats was determined,arthritis indexes were calculated,HE staining was used to observe the lesions in synovial tissue,and immunohistochemistry was used to detect the expressions of CD3+,CD4+,CD8+ in spleen tissue of rats. RESULTS:After 35 d of administration,ankle swelling degree,arthritis indexes,CD4+ expression in spleen tissue,and CD4+/CD8+ ratio in MC group were significantly higher than NC group (P<0.05 or P<0.01),expressions of CD3+, CD8+were significantly lower than NC group(P<0.05);and there was congestion and massive inflammatory cell infiltration in sy-novial tissue. The ankle swelling degree,arthritis indexes,expression of CD4+ in spleen tissue,and CD4+/CD8+ ratio in administra-tion groups were significantly lower than MC group(P<0.05);the expressions of CD3+,CD8+ in spleen tissue in administration groups were significantly higher than MC group (P<0.05);TG group,TSTR medium-dose and high-dose groups showed mild congestion and a small amount of inflammatory cell infiltration in synovial tissue of rats,and TSTR low-dose group showed no ob-vious congestion or inflammatory cell infiltration. CONCLUSIONS:Up-regulating the expressions of CD3+,CD8+ and down-regu-lating the expression of CD4+ may be one of the mechanisms of TSTR in the treatment of RA;and with best efficacy when the TSTR dose of 20 mg/kg.

Objective: To compare the effects of 50 Hz 1.8 mT sinusoidal magnetic field (SEMF) and 50 Hz 0.6 mT pulsed electromagnetic field(PEMF) on the maturation and mineralization of rat calvaria osteoblasts. Methods: Primary cultured rat calvarial osteoblasts were divided into 3 groups:blank control group, SEMF group and PEMF group. The rats in SEMT and PEMT groups were treated with 50 Hz 1.8 mT SEMF or 50 Hz 0.6 mT PEMF for 90 min/d, respectively. Western blotting and Real-time RT-PCR were used to detect the protein and mRNA expressions of Collagen-1, bone morphogenetic protein 2 (BMP-2), osterix (OSX) and Runt-associated transcription factor 2(Runx-2). The alkaline phosphatase(ALP) activity was detected by ALP test kits at d6 and d9 after treatment, and by ALP staining using azo coupling at d10 after treatment. The formation of calcium nodules was observed by alizarin red staining. Results: Compared with blank control group, the protein and mRNA expressions of Collagen-1, BMP-2, OSX and Runx-2 in SEMT and PEMT groups were significantly increased (P <0.01 or P <0.05); while the mRNA expressions of Collagen-1 and BMP-2 in PEMF group were significantly higher than those in SEMF group. After 6 days treatment, the activity of ALP in PEMF group was significantly higher than that in blank control group (P<0.05), while such difference was not observed in SEMF group (P0.05); after 9 days treatment, the activities of ALP in both PEMF and SEMP groups were significantly higher than that in blank control group (all P<0.05), but the difference between PEMF and SEMF groups was not significant (P0.05). After 10 days treatment, ALP staining was increased in both PEMF and SEMF groups compared with that in blank control group (all P<0.01), and the stained area was bigger in PEMF group than that in SEMF group (P<0.05). After 12 days treatment, calcium nodules were increased in PEMF and SEMF groups compared with that in blank control group (all P<0.01), and more calcium nodules were observed in PEMF group than SEMF group (P<0.05). Conclusion: Both 50 Hz 1.8 mT that in SEMF and 50 Hz 0.6 mT PEMF can promote the maturation and mineralization of osteoblasts, and the effect of PEMF is more marked.
Animals ; Calcification, Physiologic ; drug effects ; Cell Differentiation ; Cells, Cultured ; Electromagnetic Fields ; Gene Expression Regulation, Developmental ; radiation effects ; Magnetic Fields ; Osteoblasts ; cytology ; radiation effects ; Rats ; Skull ; drug effects

Objective To explore differently expressed proteins in serum samples obtained from Deficit Schizophrenia as well as Non‐Deficit Schizophrenia patients with proteomic techniques .Methods Make the comparative study of serum proteins between DS and NDS patients with two‐dimensional electrophoresis .The screened proteins were identified by mass spectrometer .Results Dur‐ing 18 differently expressed protein points screened in this study ,15 proteins were identified by mass spectrometer .Among of them , 12 proteins are up‐regulated while 3 proteins are down‐regulated in DS patients group .Conclusion It′s the first time to make the comparative study of serum protein profiles between DS and NDS patients by proteomic analysis .We expect that this study will contribute to better understanding of the pathophysiology of DS and explore the potential disease‐associated biomarker ,improve the future study for clinical medication of schizophrenia .

OBJECTIVETo detect the estrogenic activity of genistein and apigenin with ER-positive cell line MCF-7 human breast cancer cells.

METHODMTT method was adopted to study the impact of genistein and apigenin on MCF-7 proliferation in vitro. Real-time RT-PCR method was used to detect their impact on ERalpha, ERbeta, PR and PS2 mRNA expression levels.

RESULTGenistein and apigenin promoted the proliferation of MCF-7. Genistein 1 x 10(-10) mol x L(-1) group showed a significant increase in the expression of ERa mRNA levels or a 17. 76 times more than the control group and a 1.75 times more than the E2 group. Apigenin notably promoted the PR mRNA expression or a 4. 57 times more than the control group and a 1.11 times more than the E2 group. Both of them had different effect in promoting ERalpha, ERbeta, PR or PS2 mRNA.

CONCLUSIONBoth genistein and apigenin have a strong estrogen-like effect. Although they have different effect in promoting estrogenic response genes (such as ERa, ERbeta, PR and PS2 mRNA), genistein shows a stronger activity than apigenin. It also suggests that the signaling pathways of phytoestrogens showing estrogen-like effect are not completely identical with estrogen pathways. The B-cycle position of flavonoids is one of the key sites to estrogen-like activity, and isoflavones (cycle B on site 3) show stronger estrogen-like activity than flavones (B-cycle lies in site 2).

Apigenin ; pharmacology ; Cell Proliferation ; drug effects ; Estrogen Receptor alpha ; genetics ; metabolism ; Estrogen Receptor beta ; genetics ; metabolism ; Female ; Gene Expression ; drug effects ; Genistein ; pharmacology ; Humans ; MCF-7 Cells ; Phytoestrogens ; pharmacology ; Presenilin-2 ; genetics ; metabolism

This study is to investigate the effect of osthol on osteoclasts' activity, bone resorption as well as apoptosis in vitro, and explore the mechanism of osthol in preventing osteoporosis. Osteoclasts were separated from long-limb bones of new born rabbits, cultured in 24-well plate with glass slices and bone slices, and treated by 1 x 10(-5) mol x L(-1) osthol. Osteoclasts were identified by observing live cells with phase contrast microscope, HE staining, TRAP staining and toluidine blue staining of bone resorption pits. The numbers of bone resorption pits were counted as well as the surface area of bone resorption on bone slice. Osteoclasts were stained with acridine orange to detect the cell apoptosis. The ratio of apoptotic osteoclasts was observed under fluorescence microscope. The gene expression of RANKL, OPG, TRAP and p-JNK1/2 protein expression were examined using real time PCR and Western blotting, respectively. Comparing with the control group without osthol, the rates of apoptotic osteoclasts increased obviously and the number and area of bone resorption pits decreased evidently with 1 x 10(-5) mol x L(-1) osthol. There is significant difference between control group and experiment group treated by 1 x 10(-5) mol x L(-1) osthol. Therefore, the osthol through RANK+RANKL/TRAF6/Mkk/JNK signal pathway inhibits the osteoclasts activity, enhances osteoclasts apoptotic and inhibits the bone resorption.

ObjectiveTo investigate current status and demand of continous general practice (GP) education in Shanghai.Methods Two hundred and twenty general practitioners from 10 community health service centers of Shanghai underwent a questionnaire survey.Chi-square test was used for data analysis.ResultsThe effective response rate was 96.57％(212/220).About 87 percent of the participants considered continous GP education was helpful.Nearly 55 percent of the participants were satisfied withcurrentcontinousGP education.Unsatisfaction withcontinuing GP education,wishto reparticipate continous GP education,expected method and content of training were partly related to professional title and age.Ninety-seven percent of the participants thought the assessment of continuous GP education should include clinical practical capability evaluation.Conclusion The present continous GP education in Shanghai is not matched with GP's demand.Continous GP education should meet different needs of general practioners in different ways.

ObjectiveTo identify the correlation factors for urosepsis shock after endourology surgery and analyze its clinical characteristics.MethodsFifty-eight patients with urosepsis without shock and 15 patients with shock were studied.Factors which may be correlated with the development of sepsis were analyzed including age,sex,ASA physical status,history of urinary tract infection,preoperative leukocyturia,urine bacteria culture,prophylactic antimicrobial treatment,pyonephrosis,type of surgery,duration of operation,blood leukocyte count and concentration of creactive protein at the end of surgery,the time when the signs of infection (shivering,agitation,nausea or vomiting) first appeared.Changes in BP in patients with urosepsis shock,scores for multiple organ dysfunction syndrome as well as treatment and prognosis were also recorded.ResultsFemale sex and early emergence of signs of infection after surgery were correlated with urosepsis shock.Abrupt hypotension was usually the first manifestation of urosepsis shook.The lowest systolic BP was positively correlated with the time when diagnosis of shock was made and negatively correlated with the length of ICU stay and the highest multiple organ dysfunction syndrome scores but not correlated with shock correction time.All patients with urosepsis shock recovered completely when discharged from hospital.ConclusionFemale sex and patients with early emergence of signs of infection are prone to develop urosepsis shock after endourology operation.The prognosis of urosepsis shock is good if diagnosed and treated in time.

Objective To investigate the effect of laparoscope combined with ureteroscope for the treatment of hepatolithus. Methods 98 patients were randomly divided into two groups:the study group(laparoscope combined with ureteroscope group) and the control group (the open surgery group). The mean blood loss, the recovery of the stomach and intestines function, hospital stay and the rate of complications were observed. Results Compared with the open surgery group,the mean blood loss,the recovery of the stomach and intestines function,and hospital stay in the study group had statistically significant difference (all P＜0. 05). Compared with the open surgery group, the time of surgery and the patients carryingT tubes in the study group had statistically significant difference (all P＜0. 05). Compared with the open surgery group, the rate of gallstone removal、the rate of gallstone residual and the rate of injures in the study group had no statistically significant difference(all P＞0. 05). Conclusion Laparoscope combined with ureteroscope in patients with hepatolithus had the advantage of less trauma, faster postoperative recovery and fewer complications. It was safe and effective,so it was worth to recommend its clinical use.

ObjectiveTo investigate the role of platelet in mouse pulmonary microvascular endothelial cell (PMVEC) injury caused by lipopolysaccharide( LPS)-activated neutrophil.MethodsPMVECs were obtained from pathogen-free C3H/HeN mice of both sexes aged 6-8 weeks weighing 18-25 g according to the method described by Lim YC et al.Platelets and neutrophils were isolated from mouse blood by twice centrifugation and denaity gradient centrifugation respectively.PMVECs were seeded into twelve- or six-well plates ( 1 or 2 ml/well) after 2-5 passages and were randomly divided into 4 groups (n =31 each): group LPS; group platelets (group P);group neutrophils (group N) and group platelets + neutrophils (group PN).Each well contained about 5 × 107/ml platelets and/or 5 × 105/ml neutrophils respectively.PMVECs were incubated with LPS1 μg/ml at 37 ℃ in a 5％ CO2 humidified atmosphere for 1,6,12,18 and 24 h respectively in all 4 groups.The cells were examined with phase contrast microscope for morphological changes and survival condition.Viability rate,apoptotic rate and activation rate of PMVECs were detected by flow cytometry at each time points.ResultsThere was no significant difference in morphology and number of endothelial cells (ECs) among the 4 groups,while the number of activated ECs was significantly increased but the number of living cells decreased in group PN compared with group LPS.The activation rate of ECs was significantly higher after being incubated with LPS for 6-12 h in groups P and N than in group LPS.The viability rate was significantly lower,while the apoptotic rate and activation rate were significantly higher after ECs were incubated with LPS in group PN than in groups LPS,P and N.ConclusionPlatelets play a decisive role in mouse PMVEC injury induced by LPS activated neutrophils.