OBJRCTIVE:To study the feasibility and the preparation of bone morphogenetic protein-2(BMP 2 )carrying gel microspheres by dextran-glycidyl methacrylate(DEX-GMA)and to make an initial study on the drug-loading and drug releasing function of gel.METHODS:The orthogonal test was conducted with the reaction temperature,the addition of DEX-GMA and Span-80,the stirring speed and so on as investigation factors,the best preparation technics of BMP 2 -DEX-GMA gel microspheres was optimized and the finished products of which were given an initial determination.RE-SULTS:The BMP 2 -DEX-GMA gel microspheres preparation could be made by suspension polymerization,the optimized preparation technics including the reaction temperature was30℃,the addition of DEX-GMA and Span-80were0.6%and0.06%respectively,the stirring speed was300r/min,the BMP 2 -DEX-GMA gel microspheres from the above formulation take good shapes with the particle diameter at20?m～50?m,the envelopment ratio was(78.52?4.34)%,the quantity of drug-loading was(10.68?1.34)%,and the swelling ratio was85.9%,which has a good stability and redispersibility.CON-CLUSION:The preparation technics of gel microspheres drug-loading system is simple and the loading dosage is high,which can be used as a bioactive drugs carrier.

A method for simultaneous determination of 25 kinds of fungicides in cereals, vegetables and fruits using SPE-LC-Q-TOF/MS technique was developed.The samples were extracted with acetonitrile containing 1% (V/V) acetic acid, purified by solid phase extraction (SPE) with a Crabon/NH2 cartridge, eluted with acetonitrile-toluene(3∶1, V/V), separated by a reversed phase C18 column, gradiently eluted with acetonitrile and 0.1% formic acid solution (Containing 5 mmol/L ammonium acetate), determined by LC-Q-TOF/MS, and quantified by external standard method.A data base of the accurate mass numbers and a library which contains 25 kinds of fungicides were established.The automatic retrieval of detection results was carried on according to the characteristics of the compound, such as accurate mass, retention time, isotope peak distribution, isotopic ratios, and so on.Based on the above results, the qualitative identifications of the 25 new fungicides were accomplished without the contrast of standard substances.The results indicated that 25 fungicides showed good linearity in the range of 0.02-200 μg/L, and the limits of detection (LOD) and the limits of quantification (LOQ) were 0.01-5.00 μg/kg and 0.02-20.00 μg/kg, respectively.The linear relative coefficients were greater than 0.995.The recoveries were in the range 71.8%-114.0% and the relative standard deviations (RSD) were ranged from 0.1% to 21.3% (n=3).The method has some advantages such as simplicity, rapidity and high sensitivity, and is suitable for the rapid determination of the common fungicides in cereals, vegetables and fruits.

Objective To investigate effects of xeroderma pigmentosum B(XPB) gene on IL-6 induced proliferation and apoptosis in human vascular smooth muscle cells (VSMC).Methods Recombinant plasmid pcDNA3.1-XPB and vacant vector plasmid pcDNA3.1 were transfected stably into VSMC by liposome,and these cells were incubated with IL-6 at a 100 U/ml for 48 hours.The experiments were divided into six groups:blank control group; pcDNA3.1 group; pcDNA3.1-XPB group;IL-6 group; IL-6 + pcDNA3.1 group; IL-6 + pcDNA3.1-XPB group.The expression levels of XPB,Bcl-2,Bax and wild type p53 (wt-p53) were detected through reverse transcription polymerase chain reaction (RT-PCR) and Western blotting.The cell survival,cell cycle and apoptosis were examined with 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry,respectively.Results The transfection of pcDNA3.1-XPB increased the expression of XPB,Bax and wt-p53 (P＜0.05 or P＜0.01),decreased the expression of Bcl-2 (P＜0.05 or P＜0.01),and reduced the IL-6 induced effects on decreasing the expression of Bax and wt-p53 and increasing the expression of Bcl-2(P＜0.05 or P＜0.01).The over expression of XPB inhibited the cell growth(q=2.95,P＜ 0.05),and reduced the positive effects of IL-6 on VSMC growth ( 102.6 +6.2) ％ vs.(124.5 + 7.9) ％,q=3.49,P＜0.05.The over expression of XPB increased the apoptosis rate of VSMC(P＜0.01 ) and the cell amounts of G0/G1 phase (q=2.99,P＜ 0.05),decreased the cell amounts of S phase(q=3.05,P＜0.05),and reduced the IL-6 induced effects on decreasing the apoptosis rate of VSMC(5.9±2.1)％ vs.(0.3±0.1)％,q=7.53,P＜0.01; the cell amounts of G0/G1 phase(70.9±6.7) ％ vs.(54.8±2.9) ％,q=6.91,P＜0.01 ;and on increasing the cell amounts of Sphase(20.2+3.6)％ vs.(36.4+7.2)％,q=8.54,P＜0.01.Conclusions XPB gene could inhibit VSMC proliferation,promote VSMC apoptosis,and reduce the effects that IL-6 promotes VSMC proliferation and inhibits VSMC apoptosis.Therefore,XPB gene is likely to be potential molecular target for treatment of atherosclerosis.

We investigated the effects of IFN-γ on Chlamydia psittaci (Cps) infection.HeLa cells were treated with different concentrations of recombinant human IFN-γ (5 ng/mL,25 ng/mL,50 ng/mL) after infecting with C.psittaci 6BC,then the number and morphology of C.psittaci inclusion bodies were examined after 48 hours.C57BL/6J mice were intranasally infected with 2 × 106 IFUs C.psittaci 6BC,and intraperitoneally administrated with 10 μg recombinant murine interferon-γ 24 hours prior or post infection,then body weight,activity and survival rate were recorded.The histopathology of mice livers and lungs was analyzed by HE staining on day 5 or day10 post infection.And the chlamydial inclusion bodies were titrated in the lung homogenates of mice sacrificed on day 5 after infection.The inclusion body numbers of recombinant human IFN-γ treated groups (by 5ng/mL,25ng/mL,50ng/mL) were significantly less than that in the control group (23.8±5.1)× 106,(10± 3.58) × 106,(8.0±2.22) × 106,(43.3±11.05)× 106,respectively).And the morphology of inclusion bodies in IFN-γ treated HeLa cells was irregular and much smaller.We also found that IFN-γ could significantly improve the survival rate,reduce acute clinical manifestations and pathological injurery of lung and liver in C.psittaci respiratory tract infected mice model.So we summarized that IFN-γ can mediate strong immunological protection during acute C.psittaci early infection.

Objective To observe the clinical efficacy and toxicity of intraperitoneal chemotherapy combined with intravenous chemotherapy in treatment of cervical cancer.Methods The experimental group of 40 patients received systemic chemotherapy of docetaxel and cisplatin combined abdominal intraperitoneal high-frequency hyperthermia in the control group,40 patients received docetaxel plus cisplatin chemotherapy,The efficacy and toxicity were observed.Results The complete remission rate of observotion group was 82.5％,significantly higher than 62.5％ in control group.Effective rate in observation group was 92.5％ and also significantly higher than 75.0％ in control gronp.After 6 cycles of chemotherapy,the improvement of the experimental group was 85.0％ higher than 57.5 ％ in control gronp.The differemles between the two groups was statistically significant(P ＜ 0.05); the main side effects during chemotherapy are gastrointestinal reactions,since the cells decreased,hair loss,the difference was not statistically significant(P ＞0.05).Conclusion The chemotherapy of docetaxel and cisplatin intraperitoneal chemotherapy can significantly improve the treatment of cervical cancer,without increasing side effects.

Objective To express and purify Chlamydial protease-like activity factor(CPAF)from Chlamydophila pneumoniae,for investigating the effect of its recombinant protein GST-CPAF in inducing human monocytic cells to secrete proinflammatory cytokines and cell apoptosis.Methods The recom-bination expression plasmid pGEX6p-2/CPAF from Chlamydophila pneumoniae was transformed into E.coli.The recombination GST-CPAF was expressed after induction by IPTG,and purified by a agarose gel FF.Human monocytic cells were stimulated by the GST-CPAF to test the production of tumor necrosis factor a(TNF-α)and interleukin-6(IL- 6)by ELISA.Inhibition of cells proliferation with GST-CPAF was assessed by MTT.The THP-1 cell apoptosis stimulated by GST-CPAF was detected by Hoechst33258 fluorescence staining,DNA fragmentation analysis and cell apeptosis was detested bv Annexin V-FITC-propidiuum iodide (PI)staining.Results The recombination protein GST-CPAF was successfully expressed with high level in E.coli,and stimulated human monocytic cells to produce proinflammatory cytokines including TNF-α and IL-6 in a dose-and time-dependent manner.Otherwise,the GST-CPAF inhibited the growth of human monocytic cell in a dose-dependent manner.Apoptosis with nuclear chromatin fragmentation as well as cell shrinkage was observed by fluorescent staining and microscopy,DNA ladders in apoptosis cells were detected after 24 h with the GST-CPAF.Conclusion The GST-CPAF from Chlamydophila pneumoniae can induce the secretion of proinflammatory cytokines TNF-α and IL-6 by human monocytic cells,and inhibited the proliferation of THP-1 cell and apoptosis in vitro.

0.05)between DA and DE when the diameter of gel microspheres was20?m～40?m.CONCLUSI_ ON:The drug-loading pattern of BMP 2 -DEX-GMA gel microspheres can be influenced by its particle size,but its drug release character can not be changed,the controlled release of drugs can be achieved through changing the nature of its drug-loading material.

Objective To evaluate the safety of Renaissance spine robot assisted system in spinal injury.Methods From March 2014 to May 2016,38 patients with spinal disease received spinal surgery assisted by spine robot system.They were 20 males and 18 females,with an average age of 42 years (range,from 12 to 69 years).There were 10 lumbar fractures,8 thoracic fractures and 20 spinal deformities.Pedicle screw implantation was conducted in 30 patients (PS group) and percutaneous vertebroplasty in 8 (PV group).One side was chosen randomly to use Mazor spine robot assisted system (assisted group) and the opposite side the conventional method (non-assisted group).The anteroposterior and lateral X-rays and CT scan of the lumbar and/or thoracic spine were performed in all patients after surgery.The precision of pedicle screws implantation in PS group was evaluated by the Abul-Kasimhierarchy grading system;location of the puncture trajectory,time used for puncture and radiation exposure time in PV group were evaluated.Results 208 pedicle screws were implanted in PS group,including 120 lumbar ones and 88 thoracic ones.For lumbar pedicle screw implantation,the excellent to good rate was 95.0％ (57/60) in the assisted group,significantly higher than that in the non-assisted group (80.0％,48/60) (P ＜ 0.05).For thoracic pedicle screw implantation,the excellent to good rate was 95.5％ (42/44) in the assisted group,significantly higher than that in the non-assisted group (77.3％,34/44) (P ＜ 0.05).There were 24 puncture trajectories in 8 patients in PV group,showing no pedicle penetration or cement leaking in any case.The mean time used for puncture was 5.5 ± 1.4 min in the assisted group,significantly shorter than that in the non-assisted group (17.8 ± 7.5 min) (P ＜ 0.05);the X-ray exposure time was 14.0 ± 4.0 s in the assisted group,significantly shorter than that in the non-assisted group (22.4 ± 6.0 s) (P ＜ 0.05).Conclusions Renaissance spine robot-assisted system deserves more clinical application,because in spinal surgery it can make pedicle screw implantation more precise and safer,and can reduce operation time and X-ray exposure time in percutaneous vertebroplasty.

Acute pancreatitis (AP) is a common abdominal acute inflammatory disorder.Clinical manifestations of AP vary from self-limiting local inflammation to multiple organ failure causing significant mortality.At present,AP treatment methods mainly include non-surgical treatment such as fluid resuscitation and somatostatin,and minimally invasive or open surgical debridement treatment.Either treatment programs,nutritional support treatment is an essential part of them.According to the pathophysiological characteristics of AP onset,many scholars have emphasized that strategic nutritional support therapy is the key to limiting local inflammation,preventing and controlling AP-related complications.This article will provide an overview of the latest advances in nutritional support treatment of AP,including enteral and parenteral nutrition strategies in clinical treatment,and nutritional supplements such as glutamine,omega-3 fatty acids,vitamins and probiotics.

Objective:To explore combined detection of mad2 with anti-nuclear mitotic spindle apparatus antibody(MSA)and anti-centromere antibody(ACA)and their clinical value for the diagnosis of small cell lung cancer(SCLC).Methods:One hundred and twen-ty SCLC patients,110 non-small cell lung cancer(NSCLC)patients,and 115 pulmonary nodule(PN)patients were enrolled in this study. The expression of mad2 was analyzed by qt-PCR.MSA and ACA were detected by indirect immunofluorescence(IIF)staining.Results:mad2 was overexpressed in SCLC and NSCLC samples(P<0.05).There were significant differences between the results obtained for SCLC and NSCLC samples by qt-PCR(P<0.05).AUC in ROC curve for mad2 expression was 0.799 with an intermediate diagnostic value. In the correlative analysis,the odds ratio of MSA and ACA was 6.94 and 5.60,respectively.In the correlation analysis,Kappa value of mad2 with MSA was 0.49,and Kappa value of mad2 with ACA was 0.42.In the parallel analysis,the sensitivity and specificity was 83.31% and 79.34%,respectively,while the Youden Index was 0.62.Moreover,in the serial analysis,the sensitivity and specificity was 65.32% and 93.35%,respectively,and the Youden Index was 0.59.Conclusions:In comparison with the NSCLC and PN samples,mad2 was overexpressed in SCLC samples.Therefore,mad2 ought to play a critical role in the pathology of SCLC.The combined expression of mad2 with MSA and ACA may contribute to enhancing the sensitivity and specificity of detection;this expression may allow early diag-nosis and clinical diagnosis of SLCC and may be a promising treatment for SCLC.