Objective To explore the role and mechanisms of ω-3 polyunsaturated fatty acids (ω-3PUFA) alone or in combination with dexamethasone ( DEX) in inducing cell apoptosis and anti -proliferation in multiple myeloma(MM).Methods DEX resistance MM cell line MM1R were treated with different concentra-tions of Eicosapentaenoic acid(EPA)or Docosahexaenoic acid(DHA)alone or in combination with DEX for 24hrs or 48hrs.Cell proliferation was detected by MTT .Cell cycle and apoptosis were measured by flow cytometry .The levels of apoptosis related proteins were analyzed by Western blot .Results The proliferation of MM1R was in-hibited by different concentrations (10,20,50,100μM)of EPA or DHA alone or in combination with 10μM DEX in a dose-and time-dependent manner .Inhibition effect was significantly higher in combinative groups than in single agent groups(P<0.05).The percentage of G0/G1 phase and cell apoptosis rate in MM1R treated with dif-ferent concentrations of EPA or DHA alone was increased in a dose -dependent manner ,and being significantly higher in combinative groups than in single agent groups (P<0.05).The expressive levels of cleaved caspase -3 and Bax were up-regulated ,while pro-caspase-3 and BCL-2 were down-regulated in a dose-dependent manner.Conclusion ω-3PUFA can inhibit DEX resistant MM cell proliferation ,arrest cell cycle and induce cell apoptosis ,and has a synergistic anti -resistant effect in combination with DEX ,may serve as a new ,effective MM drugs.

Objective To construct and evaluate a novel PAMAM dendrimer vector-DNA vaccine for schistosomiasis japonica.Methods Lysine was used to modify 4.0G PAMAM.and the modified product PAMAM-Lys was synthesized.Agarose gel electrophoresis was used to confirm the composite ratio of plasmid DNA and dendrimer.Micrestructure of the compound was observed by using transmission electronic microscopy,and the stability was analyzed by using electrophoresis.The viability of the cells transfected with dendrimers was evaluated by using a MTT technique in vitro.Fiftyty mice were immunized with purified plasmid pJW4303,pJW4303-Sj23 dendrimer PAMAM-Lys and compound PAMAM-Lys/pJW4303-Sj23,respectively.The specific antibodies of the mice in each group were detected to access the immunoreactivity.Results The agarese gel electrophoresis showed that when the charge ratio of the dendrimer vector and DNA was between 2 and 4,the positive and negative charges could be counteraeted completely,and the compound was blocked completely by DNA electrophoresis.The obscrvation results with transmission electronic microscopy showed that the composition of dendrimer vector and DNA caused shrink of DNA structure.Dendrimer-DNA compound had a good stability.MTT showed the modified dendrimer vector and DNA compound system produced a lower cell toxicity on 293T cell than the unmodified Ones.Thk levels of specific antibodies of the mice immunized with PAMAM-Lys/pJW4303Sj23 were significantly higher than those of the mice immunized with naked DNA vaccine(P<0.05).Conclusions The lysinemodified PAMAM-lys is an excellent vector,and has an appropriate biocompatibility.Lysine-modification can reduce the cell toxicity of PAMAM dendrimer significantly.PAMAM-Lys can enhance the immunoreactivity of DNA vacmine which merits further application in schistosomiasis DNA vaccine.

Objective To explore the role of spinal cord p300 in neuropathic pain induced by chronic constriction nerve injury (CCI) in rats.Methods Thirty two Sprague-Dawley (SD) rats were randomly divided into sham and CCI groups,14 days after surgery,immuno-fluorescence staining and Western blot were used to detect distribution and expression of p300 protein.After rats were successfully implanted with an intrathecal catheter and accepted CCI surgery,another 24 rats were randomly divided into three groups (n =8):dimethyl sulfoxide (DMSO) group,p300 acetyltransferase inhibitor C646 group,and control C37 group.Each rat were administered through the intrathecal catheter from day 7 to 14 and mechanical withdraw threshold were tested.Results (1) The p300 positive cells were detected mainly in neurons,and p300 protein in spinal cord of CCI group were significantly higher than sham group (P ＜0.05).(2) C646 alleviated significantly neuropathic pain in rats,without significant changes in pain threshold after injection of C37 and DMSO.Conclusions The p300 protein in spinal cord was involved in the development of neuropathic pain in rats,the mechanism may be referred to its acetyltransferase activity.

Purpose To explore the correlation between quantitative parameters from intravoxel incoherent motion (IVIM) with prognostic factors of rectal adenocarcinoma.Materials and Methods Eighty-six patients with rectal adenocarcinoma who were underwent surgery without neoadjuvant therapy in our hospital between September 2015 and July 2016 were selected in this retrospective study.The image data included multiple b-values diffusion weighted imaging (DWI) examination and the corresponding D,D* and f values of the lesions.Relationships between the quantitative parameters and tumor pathology indexes including histological differentiation grade,tumor T/N stage,lymphangiovascular invasion state,the expression level of epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor-2 (Her-2) were assessed.Results The average D values of different differentiation degree (high,middle and low) of rectal adenocarcinoma were (0.541±0.093)×10 3mm2/s,(0.490±0.156)×10-3mm2/s and (0.342 ± 0.147)× 10-3 mm2/s,and the difference was statistically significant (P＜0.05).TheD values were significantly different between the lymphangiovascular invasion and non invasion state [(0.511 ±0.154)× 10-3 mm2/s vs (0.387±0.130)×10-3 mm2/s,P＜0.05)].However,there were no significant differences in the mean D,D* and f values among different tumor T/N stage (P＞0.05).The average f value of EGFR or Her-2 high expression group was higher than that of low expression group (0.379±0.076 vs 0.298±0.099,P＜0.01;0.356±0.097 vs 0.298±0.098,P＜0.05,respectively).Conclusion Quantitative parameters of IVIM in rectal adenocarci-noma can be used as noninvasive imaging biomarkers to predict the biologic behavior of tumor and the prognosis of the patients.

Objective To construct, express and purify human scFv antibody (S1) against the recombinant SAG1 of Toxoplasma fused to magainin, and observe its protective effect against Toxoplasma in infected mice. Methods The S1 scFv antibody gene amplified from phagmid S1/pIT-2 fused to magainin was cloned into procaryotic expression vector pET-32c. The recombinant plasmid S1M/pET-32c proved by DNA sequencing was transformed into E.coli BL21, and induced for fusion expression of S1M with IPTG. The expressed S1M was purified with Ni 2+ chelating HiTrap HP column and detected with SDS-PAGE. The effect of reduction of infection of Toxoplasma was observed through in vivo and in vitro experiments in mice. Results The fused gene of S1 and magainin was successfully cloned into procaryotic expression vector pET-32c proved by DNA sequencing. The recombinant S1M protein about 43 kDa was expressed in E.coli as inclusion body, and prepared with Ni 2+ column purification. Tachyzoite of Toxoplasma preincubated with S1M showed decreased infectivity in mice, the result of in vivo experiments showed that mice treated with S1M hadlonger survival time than the mice untreated. Conclusion The purified targeting antimicrobial peptide S1M could reduce the infectivity of tachyzoites of Toxoplasma in a certain extent and has a potential value for biological therapy of toxoplasmosis; otherwise, the constructed targeting antimic robial peptide S1M also provides a new model for biological therapy of toxoplasmosis.

Objective To enhance the protective immunity effects against Schistosoma japonicum infection by priming with cocktail DNA vaccines and boosting with cocktail protein vaccines in infected BALB/c mice.Methods Plasmids and proteins for immunization were prepared and diluted in no bacterial saline solution to final concentration of 1.5 mg/ml,and mixed with pcDNA3.1-SjC23,pcDNA3.1-SjCTPI,pcDNA3.1-(CDR3)6 plasmid DNAs by equal volume to form the cocktail DNA vaccine,and also mixed with recombinant proteins SjC23-HD,SjCTPI,and NP30 by equal volume to form the cocktail protein vaccine.Seventy female BALB/c mice of 4-5 weeks old were randomly divided into 5 groups(A,B,C,D,E).In Group A(control group),each mouse was immunized with 100 ?l saline solution by intramuscular(i.m.);in Group B(pcDNA3.1 control group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 for three times at week 0,3,6;in Group C(pcDNA3.1 and cocktail protein group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 for three times at week 0,3,6 and immunized with 100 ?l mixed protein vaccines plus 100 ?l FCA by subcutaneous at week 9;in Group D(cocktail DNA vaccines group),each mouse was immunized(i.m.)with 100 ?l mixed DNA vaccines for three times at week 0,3,6;in Group E(cocktail DNA vaccines plus cocktail proteins),each mouse was immunized(i.m.)with 100 ?l mixed DNA vaccines for three times at week 0,3,6 and immunized with 100 ?l mixed protein vaccines plus 100 ?l FCA by subcutaneous at week 9.Four weeks after the last DNA immunization or two weeks after protein boosting,all the mice were challenged with(40?1)cercariae of Schistosoma japonicum by abdominal skin penetration at the same time.Forty-two days post-challenge,the mice were sacrificed and perfused,and the numbers of recovered worms and eggs in liver were counted.The blood was collected from the tail veins of all the mice two days before the first immunization and challenge,respectively,the serum was prepared for detection of IgG,IgG1 and IgG2a.Two days before the challenge,the spleen cells of two mice from each group were cultured and stimulated with ConA and soluble egg antigen(SEA),and the supernatant was collected for detection of IL-2,IL-4 and IFN-?.Results The worm reduction rates in Group C,D and E were 17.70%,32.88% and 45.35%,respectively,compared with the control group.The worm reduction rates in Group D and E were significantly higher than that in Group C(P

Objective To study the protective effect of codon optimized TPI DNA vaccine against Schistosoma japonicum infection.Methods Sixty female BALB/c mice were randomly divided into 5 groups.The mice were injected through musculus quadriceps fexoris with 100 ?g pcDNA 3.1 control(Group A), pcDNA3.1-TPI(Group B), pcDNA 3.1-TPI-mHSP70(Group C), pcDNA3.1-TPI.opt(Group D), and pcDNA3.1-TPI.opt-mHSP70(Group E) respectively.All mice were immunized for three times with an interval of two weeks.The mice were challenged with(40?1) cercariae of S.japonicum per mouse by abdominal skin penetration 4 weeks after the last immunization, and sacrificed at 42 days post-challenge, the number of worms or hepatic eggs was counted.Blood was taken for the detection of IgG, IgG1, and IgG2a 2 days before immunization and before challenge, respectively.Spleen cells of 2 mice from each group were cultured and stimulated with ConA and rSjCTPI peptide, and the supernatant was collected for detection of IL-2, IL-4, IL-5, IFN-?, and TNF by flow cytometry.Results ELISA showed that the mice in groups B, C, D, and E produced specific IgG and IgG1, IgG2a antibody isotypes, and the ratio of IgG2a/IgG1 was 1.73, 2.06, 2.44, and 3.09, respectively.The levels of IL-2, IFN-? and TNF in groups D and E were higher than that of groups B and C.The worm reduction rate and hepatic egg reduction rate in groups D(36.03%, 41.7%) and E(39.03%, 46.85%) were higher than those of groups B(26.28%, 28.35%) and C(28.38%, 31.39%)(P

Objective To evaluate the effect of continuous aspiration of subglottic secretions (CASS) on the prevention of ventilator-associated pneumonia (VAP) in mechanically ventilated patients. Methods Patients ventilated mechanically at the ICU from October, 2004 to April,2006 were randomly divided into 2 groups: one group received CASS and the other did not (NASS group). CASS was performed immediately after admission for patients in the CASS group. The diagnosis d VAP was made based on clinical presentations, and the evaluation of YAP was done using simplified version of the clinical pulmonary infection score (CPIS). The general status of the patients, days of ventilated treatment, the volume of daily aspirated aubglottic secretions, the morbidity and timing of VAP, days of stay in ICU and mortality within 28 days of hospitalization were recorded. Results One hundred and one patients were included in the study. There were 48 patients in the CASS group who were treated with mechanical ventilation more than 48 hours,and 43 patients in the NASS group. There was no significant difference in the general status of the patients and days of ventilation between 2 groups with the averaged score of APACHE Ⅱ being 20.8± 6.1. The average of CPIS was of 5.6±1.0 when VAP was diagnosed. The mean volume of aspirated subglottic secretions within the first 24 hours in the CASS group (n=48) was (27.2±21.2)ml. The morbidity of VAP in the CASS and the NASS groups was 25.0% and 46. 5% respectively (P=0.032), and the length of time before the onset of VAP in these 2 groups was (7.3±4.2) days and (5.1±3.0) days respectively (P=0.100). There was a significant increase in the percentage of gram-positive cocci from the lower respiratory tracts in the NASS group compared with that in the CASS group (P=0.004). In the CASS group, the volume of the first daily aspirated subglottic secretions in patients with VAP was significantly less than that in patients without VAP(P =0.006). The morbidity of VAP in patients with failed early aspiration (the volume of first daily aspirated secretions≤20 ml) was significantly higher than that in patients in whom the aspiration was effective (P<0.01). The length of mechanical ventilation in patients with VAP was significantly longer than that in patients without VAP(P=0.000). The in-hospital mortality in patients with VAP was significantly higher than that in patients without VAP(P=0.009), and the mortality in 28 days after admission in patients with VAP was significantly higher than that in patients without VAP(P=0.035).Conclusion Effective continuous aspiration of subglottic secretions could significantly reduce the morbidity of early-onset VAP.

Objective To investigate the in vitro effects of quercitrin on the proliferation and the cytotoxicity of human γδT cells.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from healthy subjects and cultured with isopentenyl pyrophosphate and IL -2 to induce human γδT cells.The hu-manγδT cells were cultured with quercitrin at various concentrations for 48 hours.CCK-8 kits were used to analyze the in vitro proliferation and cytotoxic activities of γδT cells.Flow cytometry was performed to meas-ure the expression of granzyme B and perforin in γδT cells.The expression of p-ERK, p-Akt and Bcl-2 at protein level were detected by Western blot .Results The percentage of human γδT cells in PBMCs was in-creased from (2.96±1.83)%to (88.94±2.36)%after 10 days of culture.The quercitrin at concentrations of 10 to 80 μg/ml could promote the growth of γδT cells and up-regulate the expression of granzyme B , per-forin, p-ERK, p-Akt and Bcl-2 in a dose dependent manner .The cytolytic activities of γδT cells against co-lonic carcinoma cells ( HCT116 ) were enhanced by quercitrin .Conclusion Quercitrin could promote the proliferation of γδT cells and enhance the expression of granzyme B and perforin at certain concentrations in vitro.ERK1/2 and Akt signal transduction systems might be involved in the process .