ObjectiveTo investigate effects of pamidronate on the proliferation and differentiation of osteoblasts of rats in vitro.MethodsOsteoblasts isolated from newborn rat calvaria were treated with various concentrations of pamidronate, the proliferation of osteoblasts was evaluated with the method of methyl thiazole tetrazolium (MTT) and activity of alkaline phosphatase (ALP) in medium was measured with kit of ALP detecting.ResultsThe proliferation of osteoblasts increased under the stimulation of Pamidronate range 10-6-M-10-12 M(P<0.05), but was inhibited at the concentration of high level (10-4 M). The activity of ALP decreased in the experiment.ConclusionPamidronate can act on the osteoblasts directly and increase the proliferation of bone cells, but inhibit the differentiation of the same cells.

To establish the online two-dimensional liquid chromatography by using double gradient liquid chromatography system and UV detector, in order to simultaneously determine the content of paeoniflorin, paenol, amygdaloside and cinnamic acid. A pump of the two-dimensional liquid chromatography was adopted as the one-dimensional separation pump. C18 (3.0 mm x 150 mm, 3 microm) was used as the analytical column, with acetonitrile as the organic phase and 0.08% phosphoric acid + 0.08% triethylamine as the aqueous phase for gradient elution at the flow rate of 0.5 mL x min(-1). Another pump of the two-dimensional liquid chromatography was adopted as the two-dimensional separation pump. PAII C18 was used as the analytical column, with acetonitrile as the organic phase and 20 mmol, pH 3.0 monopotassium phosphate as the aqueous phase for gradient elution at the flow rate of 0.8 mL x min(-1). The detection wavelengths were set at 218, 230, 275 nm by using wavelength time-switching program. The linearity range of paeoniflorin, amygdaloside, paeonol and cinnamic acid were 5.55-222 (r = 0.999 7), 6.6-264 (r = 0.999 8), 3.3-132 (r = 0.999 5) and 0.315-12.6 mg x L(-1) (r = 0.999 7), respectively. The average recoveries of the four components were between 96.12% and 103.9%. The experiment proved that this method was so rapid and accurate in determination results that it could be used for evaluating drug quality.
Capsules ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Online Systems ; Time Factors

To evaluate the immunities of biodegradable microsphere as a release delivery system for DNA vaccine against Infectious Bursal Disease Virus, in our study, silk fibroin/chitosan microsphere adjuvant was prepared with a precipitation/coacervation method. Both glutaraldehyde and Na2SO4 solution were used in cross-linking. No immune chicken were intramuscularly inoculated at 14 day-old and boosted 2 weeks later. The results show that glutaraldehyde destroyed the DNA activity of the vaccine whereas Na2SO4 solution did not. Factors of the chitosan concentration 0.5% (pH 5.0), silk fibroin concentration 0.6%, plasmid DNA (500 microg/mL) dissolved in 2% Na2SO4 solution were optimized to produce microsphere, with a loading capacity of 89.14%. The average particle size of SF-CS/pCI-VP2/4/3 microsphere is 1.98 microm, and it can protect the loading DNA vaccine from DNase I digestion. Data from anti IBDV ELISA antibodies in the serum show that immunization activity of the microsphere groups were generally higher than plasmid vaccine group (P < 0.05), and the SF/CS compound microspheres group was better than that of sole CS microsphere group. The developed SF/CS microspheres are a very promising vaccine delivery system.
Adjuvants, Immunologic ; chemistry ; Animals ; Birnaviridae Infections ; prevention & control ; veterinary ; Chickens ; Chitosan ; chemistry ; Fibroins ; chemistry ; Infectious bursal disease virus ; Microspheres ; Plasmids ; Poultry Diseases ; prevention & control ; Vaccines, DNA ; chemistry ; Viral Vaccines ; chemistry

Objective To assess the value of the urine flow acceleration(UFA)versus maximum urinary flow rate (Qmax) for diagnosis of bladder outlet obstruction (BOO) in benign prostate hyperplasia (BPH).Methods A total of 50 men with BPH and 50 normal men were included in this study.Urodynamic examinations were performed in all patients according to the recommendations of the International Continence Society.Prostate volume,UFA and Qmax of each patient were analyzed and the results were compared between two groups.Results The UFA and Qmax of BPH group were much lower than that of the control group [(2.05±0.85)ml/s2 vs.(4.60±1.25)ml/s2 ; (8.50±1.05)ml/s vs.(13.00±3.35)ml/s,P＜0.05].The prostate volume in BPH group was increased compared with control group [(28.6±9.8) ml vs.(24.2±7.6)ml,P＜0.05].As diagnosis standard of UFA＜2.05 ml/s2 and Qmax＜ 10 ml/s,the sensitivity and specificity of UFA and Qmax in diagnosing BOO were (88％,75 ％)vs.(81％,63％).While compared with the result of P-Q chart,the Kappa values in correspondence analysis were 0.55 vs.0.35.The sensitivity,specificity and Kappa value of UFA in diagnosing BOO in BPHs were slightly higher than that of Qmax in comparison with the gold standard (BOO diagnosed by P-Q figure).Conclusions The UFA is a useful urodynamics parameter in diagnosing BOO of BPH.

AIM To investigate the effect of fenofibrate and simvastatin on the serum free fatty acids of alcoholic fatty liver in rats. METHODS The rat model of alcoholic fatty liver was reproduced by chronic ethanol ingestion plus olive oil diet. The model rats were divided into three groups as follows: finofibrate treatment group(finofibrate 80 mg?kg -1 po, once a day),simvastatin treatment group (simvastatin 4 mg?kg -1 po, once a day)and control group without either above-mentioned treatment. Experimental rats were treated for four weeks and then sacrificed for blood sampling. Serum free fatty acids were analyzed by gas chromatography. RESULTS Fenofibrate significantly ameliorated the decrease in polyunsaturated fatty acids induced by ethanol [oleic acid:(38.212?7.788) ?g?L -1 vs (31.620?6.142) ?g?L -1,linoleic acid:(37.269?8.065) ?g?L -1 vs (30.254?9.063) ?g?L -1,arachidonic acid:(11.646?2.601) ?g?L -1 vs (9.012?1.236) ?g?L -1] accompanied by the improvement of the fat infiltration of the liver, but demonstrated no effect on the increase in serum saturated fatty acids by ethanol. In the contrast, simvastatin can aggravate the decrease in polyunsatrurated fatty acids and significantly increase the levels of satrurated fatty acids in serum induced by ethanol along with the pathological aggravation of alcoholic fatty liver. CONCLUSION The results of present study revealed that fenofibrate and simvastatin exerted different effect on the serum free fatty acids of alcoholic fatty liver. Polyunsatrurated fatty acids in the serum play an important role in the pathogenesis and treatment response of alcoholic fatty liver.

Objective The aim of this study is to summarize preverbal communication skills in children with cochlear implants,and to provide a clinical baseline of assessment in early stage after cochlear implantation.Methods 23 prelingually deaf children with homebred cochlear implants participated in this study.The age at cochlear im-plantation ranged from 12 to 66 months with a mean of 35 months.The evaluation was performed at switch-on ses-sion and 1,3,6,9 and 12 months after the first programming session.Tait video analysis was used to assess the preverbal communication skills including turn-taking,autonomy,eye contact and auditory awareness.CAP and SIR were used to evaluate the auditory and speech abilities.ResuIts The skills of turn-taking and auditory aware-ness increased significantly,especially in the first six months after switch-on session (P<0.05).The scores of autonomy and eye contact were relative low and there were no significant differences during the first year after surgery (P>0. 05).There was a correlation between the score of CAP/SIR and turn-taking or auditory awareness.ConcIusion Children with cochlear implants established vocal-auditory mode of early communicative behavior over time.Pre-verbal communication skills CAP and SIR have relativity and can comprehensively evaluate the auditory,speech and communication skills of children with cochlear implants.

Objective: To analyze the clinical features in patients with hypertrophic cardiomyopathy (HCM) combining left ventricular apical aneurysm (LVAA) . Methods: A total of 1194 HCM patients treated in our hospital from 2007-01 to 2013-01 were studied. There were 23 (1.93%) patients combining with LVAA including 19 male and 4 female; coronary artery disease (CAD) was excluded and the patients received echocardiography and coronary angiography (CAG) examinations. Results: There were 21/23 LVAA patients having left mid-ventricular obstruction and 7 of them combining simultaneous left ventricular outflow obstruction.The average pressure gradient in those 21 patients was (56.8 ± 12.9) mmHg and the rest 2 patients suffered from apical hypertrophic cardiomyopathy.The mean maximum thickness of left ventricular wall was (21.8 ± 6.3) mm and the dimension of left ventricle was (39.4 ± 5.2) mm. Electrocardiography showed that 3 patients had paroxysmal ventricular tachycardia;CAG indicated that 6 patients combined with coronary artery muscular bridge at left anterior descending (LAD) artery. The patients were followed-up for (2.7 ± 1.3) years and adverse cardiovascular events occurred in 5 patients during that period. Conclusion: HCM combining LVAA was most frequently happened in patients with left mid-ventricular hypertrophic cardiomyopathy, some of them combining simultaneous left ventricular outflow obstruction and had the higher occurrence rate of adverse cardiovascular events. Early and accurate diagnosis is very important for guiding clinical treatment.

To develop polyclonal antibodies against predicted B cell epitopes in HIV-1 accessory protein Vpr, the prepared consensus Vpr amino acid sequence was used to predict potential B cell epitopes by online softwares (ABCpred and Bcepred), the synthesized polypeptides of B-cell epitopes were subsequently conjugated with keyhole limpet hemocyanin (KLH) and then used to immunize rabbits. The antibody titers were determined by ELISA, and antibody specifity was analyzed by Western-Blotting and immunoprecipitation, respectively. Amino acid residues 3-19 (N) and 82-95 (C) of Vpr were predicted as the potential B cell epitopes. After inoculation of the conjugation of synthesized peptide to KLH, the antibody titers in rabbit sera against N and C peptides reached more than 1:100000 by ELISA. Western-Blotting analysis showed that the polyclonal antibodies reacted with both wild Vpr and fusion protein of GFP with Vpr, no matter Vpr was derived from HIV-1 B subtype or CRF07_BC recombinant form; Immunoprecipitation analysis showed similar reactions to Western-Blotting results. Two B cell epitopes of Vpr were successfully predicted by Bio-informatics methods and polyclonal antibodies against those peptides could be successfully prepared.
Animals ; Antibodies, Viral ; blood ; immunology ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; HIV Infections ; blood ; immunology ; virology ; HIV-1 ; genetics ; immunology ; Humans ; Peptides ; genetics ; immunology ; Rabbits ; vpr Gene Products, Human Immunodeficiency Virus ; genetics ; immunology

Objective To evaluate the therapeutic effect of Shuxuetong injection on acute ischemic stroke. Methods One hundred and ninety-six patients with acute ischemic stroke were randomly divided into Shuxuetong injection treatment group (n=98) and control group (n=98). The neurological deficiency scores, scores of cognitive dysfunction and clinical outcomes were evaluated; and the cerebral blood flow were evaluated by Doppler before treatment and 2 weeks after the therapy.Results The improvement of neurological function and clinical outcomes in the Shuxuetong injection treatment group was more obvious as compared with that in the control group and the improvement rate of patients in the treatment group was higher than that in the controls; Patients in the Shuxuetong injection treatment group enjoyed less worsening than the controls. The cognitive dysfunction 2 weeks after the therapy were significantly improved as compared with those before treatment (P＜0.05); and the cerebral blood flow after the treatment was significantly increased as compared with that before the treatment (P＜0.05); these improvements in the Shuxuetong injection treatment group were much obvious as compared with those in the controls. Conclusion Shuxuetong is effective in treating cerebral ischemic patients.

OBJECTIVETo investigate the effect of anti-EGFR monoclonal antibody on the chemosensitivity of human colon cancer cells and explore the possible molecular mechanism.

METHODSThe inhibitory effect of irinotecan (CPT-11), oxaliplatin (L-OHP) and fluorouracil (5-Fu), used alone or in combination with anti-EGFR monoclonal antibody, on the proliferation of LoVo cells in vitro was assessed by MTT assay. The expressions of PI3K and Akt protein in the treated cells were examined by Western blotting, and their mRNA expressions were detected by RT-PCR.

RESULTSBoth h-R3 and C-225 treatments significantly increased the chemosensitivity of LoVo cells to irinotecan and oxaliplatin. 5-Fu and h-R3 coadministered showed a synergistic effect on the cells, but 5-Fu and C-225 had an antagonistic action. Treatment with C-225 or h-R3 resulted in lowered expressions of PI3K and Akt in LoVo cells.

CONCLUSIONAnti-EGFR monoclonal antibody can increase the chemosensitivity of human colon cancer cells to most chemotherapeutic drugs, and such effect might be attributed to the blocking of PI3K/Akt signaling pathway by these antibodies.

Antibodies, Monoclonal, Humanized ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cell Line, Tumor ; Cetuximab ; Colonic Neoplasms ; drug therapy ; metabolism ; Humans ; Oncogene Protein v-akt ; metabolism ; Receptor, Epidermal Growth Factor ; immunology ; Signal Transduction