Objective To investigate the effects of 1,25(OH)_2D_3 on cell proliferation and cell cycle progression in mouse osteoblasts.Methods Sterile bones of skull of mouse were taken from 30 newborn mouse,and the osteoblast were separated by enzyme digestion methods.After 1,25(OH)_2D_3 in different concentrations were added into culture medium,the effects of 1,25(OH)_2D_3 on cell proliferation of mouse osteoblasts and on cell cycle progression were examined by mono-nuclear celldirect cytotoxicity assay(MTT)reduction assay and flow cytometry respectively.Results After 24,48,72 h of 1,25(OH)_2D_3 incubation,the cell number of osteoblast had significant difference among groups of 1,25(OH)_2D_3 of 10~(-8),10~(-9),10~(-11)mol/L.Significant differences were found in the cell cycle progression in response to 1,25(OH)_2D_3 treatment from the Gl(84.30?1.90)to the G2-M (7.70?0.667)and S(8.00?1.42)phases when compared with those in the control group. Conclusions Cell proliferation of mouse osteoblasts can be inhibited by 1,25(OH)_2D_3 in a concentration-dependent manner.

Objective To analyze the value of sentinel lymph node biopsy (SLNB)in predicting axillary lymph node status by injection of fluorescence agent and methylene blue.Methods 156 breast cancer patients receiving surgery from Oct.2013 to Jun.2014 were studied and they were randomly divided into the experimental group(n =78) and the control group(n =78).The fluorescent agent combined with methylene blue and methylene blue were used respectively as tracers for SLNB.Axillary lymph nodes dissection was made during surgery and combined with pathology the status of sentinel lymph node(SLN) metastasis was distinguished between true negative,false negative,true positive,and false positive.Results A total of 164 SLNs were detected by the method of fluorescent agent combined with methylene blue with the detection rate of 97.44％.An average of 2.10 SLNs were detected for each patient.The accuracy rate was 97.44％,the sensitivity was 97.44％,the false negative rate and the false positive rate was 0％ and 0％.A total of 139 SLNs were detected by the method of methylene blue with the detection rate of 89.74％.An average of 1.78 SLNs were detected for each patient.The accuracy rate was 89.74％,the sensitivity was 89.74％,the false negative rate and the false positive rate was 10.26％ and 3.85％.There was statistical difference between the two groups in the average detection number and the false negative rate (P ＜ 0.05)while no statistical difference was found in the detection rate,accuracy,or sensitivity between the two groups (P ＞ 0.05).Conclusion Fluorescent agent combined with methylene blue as tracer for lymph nodes has the advantages of higher detection rate and less trauma,which is worth of clinical application.

BACKGROUND:The microcosmic and submicroscopic organizations of tissue engineering scaffold matedals’superficial structure have all important effect on the eell adhesion and growth.By means of nano.Technique and three-dimensional porous technique,the resultant nano-hydroxyapatite/collagen(n-HAC)call imitate the component and microstructure of natural bone.OBJECTIVE:To observe the biocompatibility of human bone m arrow mesenchymal stem cells(MSCs)cultured in vitro with nHAC.DESIGN,TIME AND SETTING :Single samples observation was performed in the Experimental Center of School ofMedical Technology,Jiangsu University from September 2005 to December 2006. MATERIALD:nHAc was provided by the Material Science and Engineering Department of Tsinghua University.Humanbone marrow mesenchymal stem cells were derived from healthy adult volunteers.All the subiects signed the informedconsents. METHODS:Whole bone marrow culture and successive adherence method was used to culture MSCs in vitro,and the cells were then induced to differentiate into the phenotype of osteoblasts by the revulsants(methylprednisolone,vitamin C,β-glycerophosphate and basic fibroblast growth factor).MSCs at passage 3 were co-cultured with nHACfor 14 days.MAIN OUTCOME MEASURES:The cytological characteristics of the osteoblast were identified throue,alkalinephosphatase immunohistochemistry method and Von Kossa stain.The growth condition with or without nHAC wasevaluated through invert microscope and scanning electron microscope,respectively.RESULTS:The cultured MSCs proliferated into uniform fibroblast-like cells rapidly.MSCs reached confluence and started to form multilayers averaging from 10 to 12 days,passaged stably as well.Then the MSCs passaged from 7 to 9 days.Cytochemistry evaluation showed that MSCs in induced culture were positive for alkaline phosphatase and Von Kossa stain,and deposited calcified matrix.It showed a typical ostcoblast feature in morphology and biology.In coculture model ofMSCs with nHAC,cells would attach to the inner surface of nHAC.At 8 days,the osteoblasts proliferated in the nHAC and the secretion of the matrix was observed.Lots ofcells adheredon the surfaceand pores of nHAC at 14 days.There wereextensive prominent connections among cells. CONCLUSION:THE nHAC is suitable for MSCs to adhere,grow and proliferate,with a good compatibility.

Objective To establish a method of analyzing total uranium and 235U/238U ratio in urine using inductively coupled plasma mass spectrometry (ICP-MS) and uncertainty assessment.Methods Urine sample was digested with HNO3 and H2O2,and total uranium was determined using ICP-MS method directly.The digested urine sample was separated to concentrate uranium with tributyl phosphate (TBP) column,and 235U/238U ratio was analyzed using ICP-MS.The uncertainty was evaluated through sample pre-treatment,measurement and standard curve calculation.Results The recovery of total uranium in urine was 98.4％-102.4％,detection limit was 0.002 μg/L.The relative expanded uncertainty of total uranium concentration in urine was 0.26 (k =2).235 U/238 U ratio was 0.001 1 (k =2).Conclusions This study offers a low detection limit,good recovery and precision method for rapid determination of total uranium and 235U/238 U ratio in urine samples.It is potential used for both in occupational exposure assessment and nuclear emergency situation.The uncertainty evaluation of total uranium and 235U/238U ratio in urine are reliable.

Objective To investigate the damage control surgery(DCS)in the treatment of severe abdominal trauma and the clinical value of learning from experience.Method Forty-severl cases of severe abdominal trauma patients treated with DCS were analyzed retrospectively.Results Forty-one cases (87.23%)were cured,liver abscess after re-operation was 3 cases(6.38%),intestinal fistula,biliary fistula,pancreatic fistula was 1 case(each 2.13%),they were cured by conservative treatment,6 cases(12.77%)were died,the causes of death were nothing to do with the surgery.Conclusion For patients with severe abdominal trauma actively adopt DCS,is safe and effective,with clinical value.

Objective To screen the Chinese drugs with liver channel tropism acting on the characteristic differential expressed protein of human liver cancer cell protein tyrosine kinase (PTKs) system by protein chip technology, and to analyze the difference in different Chinese drugs with liver channel tropism in relation to PTKs regulation. Methods Forty BALB/C nude mice were chosen; a piece of subcutaneous tumor mass was implanted into the left lobe of liver parenchyma to reduplicate the orthotopically implanted tumor models. After modeling for 10 days, the tumorigenicity was confirmed, and all the nude mice models were divided into four groups; different Chinese medicine extracts were injected intra-peritoneally into corresponding treatment groups respectively, and the methods of treatment in the 4 groups were as follows: In liver channel tropism drug pair of Huayu Xiaozhen group, rhizoma sparganii and curcuma zedoary with dosage containing crude drug 4.5 g·kg-1·d-1 was given, in liver channel tropism drug pair of Gongdu Sanjie group, the extract of centipede and scorpion with dosage containing crude drug 0.3 g·kg-1·d-1 was intra-peritoneally injected, in spleen channel tropism drug pair control group, astragalus and rhizoma atractylodis macrocephalae with dosage containing crude drug 6.3 g·kg-1·d-1 was given, and in the model group, equal amount of 0.9% normal saline was intra-peritoneally injected. After the drug treatment for 3 weeks, the mice were sacrificed and the liver cancer tissue was obtained; after its total protein was extracted, protein chip technology was applied to screen the Chinese drug pair with liver channel tropism acting on differential expressed PTKs of human liver cancer cells, and the results were analyzed by cluster analysis.Results After the end of the experiment, there were 6 animals in Huayu Xiaozhen drug pair group, 5 in Gongdu Sanjie drug pair group, 5 in control drug pair group and 7 in model group. The protein chip screening results showed that the adjusting fold greater > 1.50 or < 0.67 was defined as the difference with statistical significance. Compared with model group, there were 42 up-regulated expressions of PTKs in Huayu Xiaozhen drug pair group, including 29 receptor tyrosine kinases (RTKs) and 13 non-receptor protein tyrosine kinases (nrPTKs) of which the erythropoietin having adjusting fold over 5.0 produced liver cell receptor B1 (EphB1), epidermal growth factor receptor (ErbB2, ErbB4) etc. 3 RTKs; there were 7 RTKs with adjusting fold 3.0 - 5.0 including EphA1, EphA3, EphA7, fibroblast growth factor receptor 2-α (FGFR2-α), hepatocyte growth factor receptor (HGFR), macrophage colony-stimulating factor receptor (M-CSFR) and vascular endothelial growth factor receptor 2 (VEGFR2), and 2 nrPTKs with adjusting fold 3.0 - 5.0 were non-receptor tyrosine kinase BMX (BMX) and Janus kinase 1 (JAK1). In the Gongdu sanjie drug pair group, there were 23 up-regulated expressions: 15 RTKs and 8 nrPTKs, and 6 down-regulated expressions; the RTKs with adjusting fold 3.0 - 5.0 were ErbB4, M-CSFR and 1 nrPTKs that was megakaryocyte-associated tyrosine kinase (MATK). In the control drug pair group, there were 28 up-regulated and 8 down-regulated expressions. The results of cluster analysis showed that in Huayu Xiaozhen drug pair group, there were 17 differential expressed PTKs in accord with the screen criteria of which 9 were RTKs [receptor tyrosine kinase-like orphan receptor 2 (ROR2), stem cell factor receptor (SCFR), anaplasia lymphoma kinase (ALK), platelet-derived growth factor receptor β (PDGFR-β), insulin-like growth factor-IR receptor (IGF-IR), ErbB2, ErbB3, EphB1 and EphA2], 1 nrPTKs [fps/fes related tyrosine kinase (FER)] and 7 PTKs 3 RTKs (M-CSFR, FGFR2-α, EphA3) and 4 nrPTKs [acetate kinase 1 (ACK1), bruton tyrosine kinase (Btk), non-receptor tyrosine kinase ABL1 (ABL1) and BMX]. In Gongdu Sanjie drug pair group, there were 7 differential expressed PTKs in accord with the screen criteria 5 RTKs (M-CSFR, FGFR1, ROR2, EphB1, ErbB2) and 2 nrPTKs [src-related kinase lacking C-terminal regulation and N-terminal myristylation sites (SRMS), FER]. Conclusions The drug pair of centipede and scorpion with Gongdu Sanjie action possesses a more effective anti-HCC role than the drug pair of rhizoma sparganii and curcuma zedoary with Huayu Xiaozhen action, the mechanism is possibly via the regulation of PTKs signal pathway. The liver channel tropism drug pair of rhizoma sparganii and curcuma with action of promoting blood circulation and removing blood stasis possibly has an independent anti-HCC effective pathway outside the PTKs signal system.

Objective To assess left ventricular(LV) longitudinal strain in patients with coronary heart disease by two-dimensional speckle tracking imaging (2DSTI),and to explore the clinical value of 2D longitudinal strain in detecting myocardial ischemia. Methods Forty-four patients with coronary heart disease (CHD group) and 28 age-matched subjects (control group) were enrolled into this study. The two-dimensional data were obtained in apical 4-chamble, 2-chamber and long axis view. And the longitudinal strains of every segments, the average longitudinal strain of LV 18 segments (SL18), the average longitudinal strain of 12 segments (SL12,excluded the 6 apical segments) were analyzed. Results In the patients with CHD, the longitudinal strain of ischemia segments and the global LV longitudinal strain were significantly decreased than that of the control subjects. Both in patients with CHD and in control subjects,the longitudinal strains in apical segments were higher than that of middle and basal segments. There was significant difference between SL18 and SL12 ( P=0.027 in CHD group and P =0.003 in control group).Receiver operating curve (ROC) analysis demonstrated that the cutoff point of SL18 to detect myocardial ischemia was - 18.8％ (sensitivity 80.2％ and specificity 74.1％ ) ,and the cutoff point of SL12 to detect myocardial ischemia was - 17.8％ ( sensitivity 81.7％ and specificity 85.6％ ). Conclusions 2D longitudinal strain was sensitive to detect myocardial ischemia, SL12 was better than SL18 in detecting myocardial ischemia. 2DSTI might be useful for identifying patients with severe CHD.

Objective To assess the correlation between time during admission to percutaneous coronary intervention (PCI) and left ventricular function recovery of acute myocardial infarction (AMI) by two-dimensional speckle tracking imaging (STI).The clinical value of STI in assessing therapeutic effect of AMI treated by PCI and estimation of the prognosis were discussed.Methods Sixty-one AMI patients who had AMI for the first time and had been treated by primary PCI were enrolled.Dynamic images were acquired before PCI,at 7 days after PCI and 30 days after PCI and analyzed by STI.The time during admission to PCI of AMI patients was recorded accurately.Dynamic images were analyzed for longitudinal peak systolic strain (LPSS) values (global,infarcted area) by STI.According to the comparison of left ventricular ejection fraction(LVEF) before PCI and 30 days after PCI,patients were divided into left ventricular function improved group (ΔLVEF≥5 ％) and not-improved group,and the values of LPSS and time during admission to PCI were compared between the two group respectively.Results Compared to not-improved group,the time during admission to PCI in improved group was lower ( P ＜0.001),infarcted segmental LPSS at 7 days after PCI ( P ＜0.05) and both global ( P ＜0.001) and infarcted segmental LPSS ( P ＜0.001) at 30 days after PCI in improved group were higher than those in not-improved group.Linear regression analysis showed that both global and infarcted segmental LPSS were significant correlated to LVEF respectively ( P ＜0.001,all).Infarcted segmental LPSS at 7 days after PCI were correlated to the time during admission to PCI ( P ＜0.05).LVEF ( r =0.303,P ＜0.05),global ( r =0.300,P ＜0.05)and infarcted segmental LPSS ( r =0.590,P ＜0.001) at 30 days after PCI were correlated to the time during admission to PCI.Conclusions STI provides reliable and useful clinical information for the assessment of therapeutic effect of AMI treated by PCI and estimation of the prognosis by sensitively presenting the close correlation between time during admission to PCI and left ventricular function recovery of AMI patients.

Objective To assess the left ventricular function of patients with acute myocardial infarction (AMI) and its correlation with cardiac troponin T (cTnT) and specific manifestation of electrocardiogram (ECG) by automated function imaging (AFI) of two-dimensional speckle tracking imaging.Methods Forty-six AMI patients who had AMI for the first time and had been treated by primary percutaneous coronary intervention and 30 healthy controls who were age and sex-related to infarct group were involved.The values of cTnT within 24 hours after admission of AMI patients were recorded and the values of ST segment elevation were measured accurately.All the subjects were analyzed for longitudinal peak systolic strain (LPSS) values and the bull' s eyes by AFI.Results Compared to control group, left ventricular ejection fraction (LVEF), global and infarcted LPSS of infarct group were significantly different and the values of ST elevation of infarct group were higher than those of control group.Both global,infarcted segmental LPSS were significant closely correlated to LVEF and cTnT,respectively (P＜0.001,all).Both global, infarcted segmental LPSS were correlated to ST elevation (P ＜0.05,all).Global LPSS had the closest correlation with LVEF (r = -0.565, P＜0.001) and so did infarcted LPSS with cTnT (r = 0.432, P ＜0.01).Conclusions As a procedural simple and rapid diagnostic tool,AFI provides reliable and useful information of the assessment of AMI.Both global and infarcted segmental LPSS have well described left ventricular function of AMI patients.Compared to LVEF, LPSS was more closely correlated to cTnT and ST segment elevation, which meant that LPSS was more sensitive and more closely related to real infarct size and actual involved range of AMI.

Rhynchosia volubilis Lour has been a major drug in a folk prescription for contraception in China, whereas its mechanism remains unknown. Its antifertility effects on male mice and antimicrobial activities on sexually transmitted infection (STI) pathogens were previously reported. This study was undertaken to develop the n-Butanol extract of Rhynchosia volubilis Lour (BERVL) as a spermicidal agent with STI prevention. The spermicidal activities of BERVL with different doses were assessed using selected high-motile sperms of normal human semen samples, and their inhibitory effects on Lactobacillus acidophilus were determined. The mechanism of the spermicidal activity was explored by aqueous Eosin Y and Hoechst 33342/PI staining. The results showed spermicidal activities and inhibitory effects of BERVL on Lactobacillus acidophilus were dose-dependent. Dose of 90 mg/mL BERVL terminated all progressive sperm motility within 2 min, and had slight inhibitory effect on Lactobacillus acidophilus, suggesting it was an effective and safe dose for contraception use. About 80% sperms exposed to BERVL displayed changes consistent with high permeability of head membrane. It is concluded that BERVL as spermicide has advantages over N-9 with strong ability to instantaneously kill human sperm and possesses light inhibitory effect on Lactobacillus acidophilus.