Objective To observe recombinant plasmids were constructed with the macrophage colony-stimulating factor ( GM-SCF) , interleukin -21 (IL-21) and retinoic acid early transcription factor-1 (Rae-1), and observe the inhibitory effects in subcutaneous liver cancer model in mice with the recombinant plasmids.Methods The recombinant plasmids of GM-SCF, IL-21 and Rae-1 were constructed with RT-PCR method, mouse model was constructed, the model mice were randomly divided into six groups including control, IRES/GFP, IRES/IL21, IRES/GM-SCF, IRES/GM-SCF-IL21 and IRES/combination with 10 mice included in each group, each groups (15 mice) were treated with the corresponding gene therapy.The survival rate were observed after 60 days.The blood levels of interferon -γ(IFN-γ) and interleukin -2 (IL-2) were detected in each group.Results The pGM-CSF-GFP-IRES-Rae-1-IL-21 has been successfully constructed.All mice had demised 14 and 16 days after treatment in the control and IRES/GFP groups, respectively.There were 2, 1, 11 mice remaining after 60 days of treatment in the IRES/GM-SCF, IRES/IL21 and IRES/GM-SCF-IL21 groups respectively.The survival rate of mice at 60 days of treatment was 73.33%, 13.33%, and 6.67% for groups IRES/GM-SCF-IL21, IRES/GM-SCF and IRES/IL21, respectively.The survival rate of the mice was significantly higher in IRES/GM-SCF-IL21 than the other groups.The levels of IL-2 and INF-γof mice 1-6 days after treatment gradually increased in the IRES/combination groups, including IRES/GM-SCF-IL21, IRES/GM-SCF and IRES/IL21.They were highest in the IRES/combination group and lowest (P<0.01) in the IRES/GM-SCF and IRES/IL21 groups, with the IRES/GM-SCF-IL21 group showing intermediate levels.By 6-10 days after treatment, IL-2 and INF-γlevels had stably increased in the IRES/combination groups, but had gradually decreased in the IRES/GM-SCF-IL21, IRES/GM-SCF and IRES/IL21 groups.At the end of treatment, IL-2 and INF-γlevels were significantly (P<0.01) higher in the IRES/GM-SCF-IL21 than were found in either the IRES/GM-SCF group or IRES/IL21 group, which were also significantly (P<0.01) higher than either the IRES/GFP or control groups.The levels of IL-2 and INF-γwere highest in the IRES/combination group ( P<0.01) and not significantly different among the IRES/GM-SCF, IRES/IL21, IRES/GFP, and control groups.Conclusion The inhibitory effects in subcutaneous liver cancer model in mice were obvious significantly, and its mechanism maybe be related to the activation of the body's immune.

Acupuncture Points ; Acupuncture Therapy ; Fasciitis, Plantar ; therapy ; Female ; Humans ; Middle Aged

Objective To understand the main pathogenic bacteria distribution and drug susceptibility in the common bile duct calculi with infection ,so as to provide the basis for rational use of antimicrobial agents .Methods Data′s of 296 cases of bile specimens treated from January 2011 to December 2013 were retrospectively surveyed and drug resistance of bacteria culture results were analyzed .Results Among all the 296 cases ,there were 199 cases of pathogenic bacteria in bile specimens ,the positive rate was 67 .23% ;among them ,there were 21 cases of two kinds of bacteria ,a total of 220 strains of pathogenic bacteria were detected ,and among them there were 158 strains of gram negative bacteria ,accounting for 71 .82% ;there were 46 strains of gram positive bacte‐ria ,accounting for 20 .91% ;there were 16 strains of fungi ,accounting for 7 .27% ;the lowest resistance of gram negative bacilli lies in amikacin ,followed by tazobactam ,aztreonam ,three to four generations of cephalosporins and quinolone .Gram negative bacilli has the highest drug resistance to pseudomonas aeruginosa and e .coli ,and it has a drug resistant rate higher than 50 .00% to most anti‐microbial drugs .Gram positive cocci has 0 drug resistance rate to add ring element ,has a drug resistance rate lower than 30 .00% to vancomycin and rina thiazole amine ,and has highest drug resistance rate (70 .00% -100 .00% ) to erythromycin ,clindamycin and chlorine lincomycin .Conclusion Among all the 296 cases of common bile duct calculi bile with biliary tract infection ,the main path‐ogenic bacteria were e .coli ,klebsiella pneumoniae ,eosinophilic malt narrow and excrement enterococcus bacterium ;and the drug re‐sistance to penicillin is highest .

Objective:To evaluate the in vitro release behavior of doxorubicin(Dox)-loaded microspheres and the stability of Dox during encapsulation process and in vitro release.Methods: Dox-loaded microspheres were prepared by double emulsion(W/O/W) method with poly(lactic-co-glycolic acid)(PLGA) as the carrier material.The physical and chemical characteristics of microspheres,including the mean diameter,morphology,drug entrapment efficiency and loading rate,were evaluated.The in vitro release behavior and its influencing factors were determined by ultraviolet spectrophotometry.Dox stability was evaluated by HPLC method during the encapsulation process and in vitro release.Results: The prepared microspheres had a complete spheric shape and dispersive quality.The mean diameter of the microspheres was 85 ?m;the drug entrapment efficiency was 95.1%;and the loading rate was 14.8%.Releasing rate of the microspheres slowed down with the increase of PLGA concentration and the decrease of W/O value.The encapsulation process had no obvious effect on the stability of Dox,while Dox degraded during in vitro release as the prolongation of time.On day 10,the peak area of degraded material accounted for 2.46%.Conclusion: Dox can be encapsulated in the microspheres by double emulsion method and different release rates of Dox can be achieved by adjusting PLGA concentration and W/O volume ratio.

To develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of epirubicin hydrochloride (EPI) in rat plasma, daunorubicin hydrochloride was used as internal standard. The plasma samples were deproteinated with methanol, and separation was performed on a reversed-phase CAPCELL PAK C18 column (3.0 mm x 50 mm, 3 microm). The mobile phase contained methanol-0.1% formic acid (80:20). Detection was carried out by multiple reaction monitoring on a HP1200-6410 QQQ LC/MS system. Different preparations of EPI solution, EPI-LIP (EPI-liposome) and EPI-LTSL (EPI-thermosensitive liposome) was administered in rats by i.v with the same dosage (12 mg kg(-1)). The pharmacokinetic model and parameters were fitted and calculated by the DAS ver2.0 software. The calibration curve was linear in the range of 0.01-50 microg mL(-1). The limit of quantification was 0.01 microg mL(-1). RSDs of intra- and interbatch precisions were all less than 11.9%. The average extract recovery was 89.3% and 92.1%, respectively. The pharmacokinetics of EPI in rats with all preparations were fitted to three compartments, which all fast distributed and slowly eliminated. The t1/2 alpha, t1/2 beta, t1/2 gamma, AUC(0-infinity), and MRT(0-infinity) of EPI-LTSL group were 7.5, 1.3, 12.6, 12.9, 3.7 times those of EPI solution group; and 1.6, 1.4, 12.3, 2.9, 2.6 times those of EPI-LIP group. Moreover, the CL of the latter two groups was about 13.4 times of the former EPI-LTSL group. EPI-LTSL can significantly improve AUC and prolong the circulation time of EPI in rat plasma.

Objective To assess the efficacy and safety of intraoperative wake-up test during surgery for ossification of the ligamentum flavum(OLF). Methods Between June 2004 and June 2010,35 patients(23 males and 12 females,at age range of 32-71 years,mean 50.5 years) with OLF underwent posterior decompressive laminectomy and excision of OLF.The operation wag performed on the cervical vertebrae in four patients,on the thoracic vertebrae in 24 and on the lumbar spine in seven,with monosegmental OLF in 14 patients and multisegmental OLF in 21.The outcomes were evaluated using Japanese Orthopaedic Association (JOA) scale. Results The operation lasted for mean 190 minutes (120-260 minutes),with the blood loss for mean 350 ml (220-520 ml)and the mean 2.4(1-5) lamin receiving decompression.The follow-up period ranged from 6 to 72 months (mean 29.5 months).The JOA score was improved from 4.302±1.023 preoperatively to 8.327±1.032 at the final follow-up (P<0.01).The neurological symptoms was aggravated during operation in three patients,for whom the muscles force recovered gradually in two patients but one showed no change after special measures.No serious complication was found in the other patients postoperatively. Conclusions The wake-up test can serve as an effective monitoring during the procedure ofthe decompressive laninectomy in the patients with severe OLF and decrease the occurrence of neurologic deficit complications.

Objective To evaluate the effects of compositive cognitive training on reasoning ability for healthy community elderly.Method All the participants were selected from one district in Shanghai by every 50 samples.151 community healthy elderly who accorded with the standard were collected at last.They were divided into cognitive intervention group ( n =90) and control group ( n =61 ) by sequence.The interventions ( includes reasoning,memory training,et al) were conducted in 24 sessions over 12 weeks.All individuals were assessed by Neuropsychological Test Battery for Elderly (NTBE) at baseline,follow-up,half a year follow-up and one year follow-up phase,and Raven' s Standard Progressive Matrices on reasoning classes.Results The scores of Raven' s Standard Progressive Matrices were significantly higher after reasoning training in intervention group( (15.54 ±5.70),(10.35 ± 5.10),t =3.595,P ＜ 0.01 ).Compared with baseline,the reasoning test scores of NTBE were significantly higher after cognitive intervention in treatment group at follow-up phase,and were also significantly higher than control group (P ＜ 0.05 or P ＜ 0.01 ).Conclusion Results support the effectiveness of compositive cognitive training in improving reasoning ability are positive,and can last for one year.

Objective To investigate the changes of serum ferritin and homocysteine of diabetic patients,to explore its correlation with diabetic gastroparesis.Methods Pure 50 patients with diabetes,50 patients with diabetic gastroparesis,and 50 cases of normal control group were chosen.The serum ferritin levels were measured with chemiluminescence method,the serum homocysteine levels were measured with enzyme linked immunosorbent assay (ELISA) method.The changes of serum ferritin and serum homocysteine in each groups were observed.Its relationship with diabetic gastroparesis was studied.Results For all diabetic patients,the serum ferritin and homocysteine levels were significantly higher than those in normal control group (P ＜ 0.01).Patients with diabetic gastroparesis was higher than those of pure diabetes (P ＜ 0.01).Serum ferritin levels was positively correlated with glycosylated hemoglobin and homocysteine (r =0.62,0.78,P ＜ 0.01).Multi-factor analysis showed that serum ferritin,and homocysteine level was positively associated with diabetic gastroparesis (P ＜ 0.05).Conclusions The increases of serum ferritin and homocysteine levels were closely related to the occurrence and development of diabetic gastroparesis.They can be used as one of parameters of diabetic gastroparesis,and provide theoretical basis for clinical intervention therapy.