Objective Using apomorphine, a potent dopamine receptor agonist and rotating T-maze, the effect of apomorphine on the visual discrimination learning and reversal learning in rats was investigated. Methods All rats were trained in a visual discrimination task (food reward and light stimulus) in rotating T-maze. After reaching the acquisition criterion, rats were trained in a reversal task (food reward and without light stimulus) in the same maze. During the period of visual discrimination task, apomorphine was administrated either 30 minutes prior to learning or after learning immediately. Results The results showed that apomorphine, which was given either 30 minutes prior to visual discrimination learning or after learning, could impair the acquisition of discrimination learning( 259.20±26.29 and 264.00±16.97, compared to 168.00±16.97 and 163.20±20.08) and apomorphine, which was given only after visual discrimination learning, could impair the acquisition of reversal learning (451.20±39.44 compared to 360.00±29.39). Conclusion The results showed that apomorphine, which was given either 30 minutes prior to visual discrimination learning or after learning, could impair the acquisition of discrimination learning and apomorphine, which was given only after visual discrimination learning, could impair the acquisition of reversal learning.

To observe the changes in hemorheology, endotoxin and TNF ? in blood after hemoperfusion(HP) with adsorbent immobilized polymyxin B (PMB) on sepsis in rats. Wistar rats were randomly divided into three groups:LPS+HP+PMB,LPS+HP and LPS. All the rats received intravenously injection of lipopolysaccharide (LPS, Escherichia coil O111:B4,1mg/kg). Plasma of the rats in the group of LPS+HP+PMB was passed through a column containing sepharose coated activated charcoal immobilized polymyxin B at the 4th hour after LPS injection. The treated plasma was transfused bak after being mixed with blood cells. In LPS+HP group, the column did not contain immobilized polymyxin B. The animals of LPS group received LPS only. Quantitative endotoxin determination in blood was done with limulus amebocyte lysate test,TNF ? of the plasma assayed with ELISA, and hemorheology parameters were also observed after hemoperfusion. In LPS+HP+PMB group, the concentration of plasma was significantly decreased after hemoperfusion, but it was still significantly higher than the baseline value, and there was a decrease of blood cell ratio after hemoperfusion. The results suggest that specific adsorbent could remove endotoxin in the circulation and improve hemorheology.

Objective To investigate the synthesis, in vivo biodistribution of 99Tcm-HYNIC-PEG4-E[PEG4-c (RGDfk)] 2 (99 Tcm-Galacto-RGD2), and its potential usage for targeted imaging of mice orthotopic glioma.Methods 99Tcm-Galacto-RGD2 was synthesized straightforward and its radiochemical purity and stability and distribution in mice were analyzed.MicroSPECT-CT imaging was done in a mice orthotopic glioma model, which had been set up with U87MG cells, after administration of 99Tcm-Galacto-RGD2.Region of interest (ROI) of glioma was drawn on SPECT-CT section images to quantify tumor uptake (％ ID/cm3).Glioma was harvested for pathological examination.Linear-regression was used to analyze the relationship between integrin αvβ3 and tumor uptake (％ID/cm3).Results The radiochemical purity of 99Tcm-Galacto-RGD2 was (97.7 ±0.8)％ and stable in vitro.Hynic-Galacto-RGD2 could specifically bind to integrin αv β3 of tumor cells with a IC50 of (18 ± 3) nmol/L.After tail vein injection, 99Tcm-Galacto-RGD2 was rapidly discharged from the blood, liver, kidneys and had a relative low concentration in normal brain tissue.MicroSPECT-CT imaging demonstrated that, after 60 min of injection, this drug was well uptaken by glioma tumor than that after 30 min (t =7.13 ,P ＜0.05), and the tumor to normal brain tissue (T/B) uptake ratio of 99Tcm-Galacto-RGD2 was 13.92± 3.43.Injection of HYNICGalacto-RGD2 2 min prior to 99Tcm-Galacto-RGD2 injection extensively reduced the uptake of radioactive drug in tumor tissue (t =11.36, P ＜ 0.05).Bland-Altman analysis showed that tumor volume based on SPECT-CT imaging measurement had almost same value with the tumor reference volume (95％ CI =-11.94％-11.92％).In addition, the tumor uptake of 99Tcm-Galacto-RGD2 and cellular integrin αvβ3 expression level had a linear relationship (R2 =0.896).Conclusions Stable 99Tcm-Galacto-RGD2 can be synthesized easily and is applicable for microSPECT-CT imaging analysis of orthotopic glioma in mice together with the evaluation of integrin αvβ3 level in tumor.

Objective To synthesize 99Tcm- (hydrazinonictinamide- [Lys3] -bombesin) (tricine)(trisodium triphenylphosphine-3,3',3"-trisulfonate) ((HYNIC-[Lys3]-BBS) (tricine) (TPPTS)) and evaluate its biodistribution and binding capability with tumor tissue in Balb/c nude mice bearing human pancreatic cancer xenografts. Methods HYNIC was conjugated to the [Lys3] -BBS at pH = 9.0 with SnCl2 as reducing agent and both tricine and TPPTS as coligands for 99Tcm-labeling. 99Tcm-HYNIC-[Lys3]-BBS)(tricine) (TPPTS) was purified by Sep-Pak C18 cartridge and was analysed by HPLC. The radiochemical purity and radiolabeling yield were measured. The stability of 99Tcm-(HYNIC-[Lys3]-BBS) (tricine)(TPPTS) in serum, biodistribution (% ID/g) in the normal mice and imaging of the Balb/c nude mice bearing human pancreatic cancer xenografts in vivo were studied. Results The radiolabeling yield was (90 ±2)% and the radiochemical purity was over 95%. The radiochemical purity after 4 h in serum was over 85%. The distribution in normal mice showed rapid clearance from blood (the uptake was (0.07 ±0.01) %ID/g at 2 h postinjection). 99Tcm-(HYNIC-[Lys3]-BBS) (tricine) (TPPTS) was excreted mainly via the kidney with little radioactivity accumulation in the liver and gastrointestinal tract (the uptake of liver, stomach, intestine was (0.27 ±0.03), (0.06 ±0.03), (0.04 ±0.00) %ID/g at 2 h postinjection). Marked uptake of radioactivity was found in tumor tissue of the Balb/c nude mice bearing human pancreatic cancer with maximum T/NT ratio of 3.71 ± 0.57 at 2 h postinjection. Conclusions 99Tcm-(HYNIC-[Lys3]-BBS)(tricine) (TPPTS) can be easily prepared with high radiolabeling yield and radiochemical purity. The stability in serum and good biodistribution charateristics make it useful for the diagnosis of human pancreatic cancer with over-expression of the gastric-releasing peptide(GRP) receptor.

Objective To investigate the inhibitive effect of shRNA (short hairpin RNA) targeting APRIL gene on the pancreatic cancer cells in vitro and in vivo, in order to explore the feasibility of gene therapy for pancreatic cancer. Methods The LV-shAPRIL targeting APRIL gene had been constructed before, and was used to infect the CFPAC-1 cells. Cell proliferation and apoptosis were examined by MTT and flow cytometry. Then CFPAC-1 cells were used to construct the model of transplantation tumor into the nude mice, the tumor growth was assessed after LV-shAPRIL treatment. Results 96 hours after the LV-shAPRIL infection into CFPAC-1 cells, the cell proliferation was significantly inhibited when compared with control group and lentivirus infection group (P＜0.05 ). Flow cytometry showed the apoptosis ratio of the CFPAC-1 cells was (17.35±0.96)% in LV-shAPRIL group, which was higher than that in control group and lentivirus infection group (P＜0.05 ). After LV-shAPRIL injection into the model of nude mice, the tumor growth was slower than that in the two control groups. The tumor's volume of the LV-shAPRIL group was(821.8±123.3) mm3 and the mass was (2.16±0.18)g at 27 day, and were obviously depressed, when compared with two control groups (P±0.05). Conclusions LV-shAPRIL targeting APRIL gene can inhibit the growth of the CFPAC-1 cells in vitro and vivo. This may provide a new gene therapy approach for pancreatic cancer.

Objective To construct of shRNA lentiviral expression vector targeting APRIL (aproliferation-inducing ligand) gene in CFPAC-1 cell of human pancreatic cancer. Methods We used gene engineering to screen RNA interference targeting sequence of APRIL gene. The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed, synthesized and cloned into the pGCL-GFP vector. The resulting lentiviral vector containing shAPRIL were named LV-shAPRIL. Then it was conformed by PCR and DNA sequencing identification. 293T cells were eotransfected with LV-shAPRIL,pHelper 1.0 and pHelper 2.0 to product ientivirus. The titer of virus was tested according to the expression level of GFP in the 293T cells. After recombinant lentivirus infection into CFPAC-1 cells, we used real-time RT-PCR and Western blotting to examine APRIL mRNA and protein expression at different cell culture period.Results PCR analysis and DNA sequencing conformed that shAPRIL DNA was successfully inserted into the lentiviral vector. The titer of concentrated virus were 5 × 107 TU/ml. APRIL expression in CFPAC-1 cells were inhibited significantly at both mRNA and protein level. APRIL mRNA expression were decreased 73%, 70%and 71% , respectively, after the infection of 4 days, 4 weeks and 8 weeks by LV-shAPRIL. APRIL protein expression were decreased 66%, 63% and 62%, respectively , after the infection of 4 days , 4 weeks and 8weeks by LV-shAPRIL. Conclusions ShRNA lentiviral expression vector targeting APRIL gene has been successully constructed, and it can effectively inhibit the expression of APRIL gene in CFPAC-1 ceils. This study lays a foundatin for in vivo research APRIL gene scilence in pancreatic cancer cell using the model of nude mice.

Objective To explore rational surgical treatment for rectal villous adenoma with canceration. Methods Sixty-two cases of rectal villous adenoma with canceration undergoing different surgical procedures were analyzed retrospectively. Among them 28 underwent local excision including 20 with transanal resection and 8 with Kraske resection; Thirty-four underwent extended excision consisted of 11 Miles operation, 21 with Dixon operation and 2 with other procedures. Results Among 62 cases, 82%(51/62)were in early stage. 68%(42/62) and 26%(16/62) of the patients were of high and middle differentiation in pathology respectively. Six cases suffered from recurrence or metastasis in both local and extended excision groups. The 5-year survival rate after local and extended excision were 79% and 76% respectively. Conclusions Most rectal villous adenomas with malignant change were less invasive; Prudent selection of surgical modality is both effective and less dangerous for the treatment of this entity.

AIM:To establish an HPLC method for determining the contents of caffeic acid,quercetin and campherenol in Hedyotis diffusa Willd. METHODS:The samples were separated on an Alltima C 18 (250 mm? 4.6 mm,5 ?m) column with the mobile phase of MeOH(A)-0.5% glacial acetic acid solution;gradient elution(0～15 min,30%～60% A;15～30 min,60%～60% A).Flow rate was 1.0 mL/min. The detection wavelength was set at 350 nm.Column temperature was at 30 ℃. RESULTS:The contents of caffeic acid,quercetin and campherenol were 14.218～23.695 ?g/g,9.919～25.564 ?g/g and 6.229～18.160 ?g/g in Hedyotis diffusa Willd from different sources. The linear range of caffeic acid was 0.005 0～0.200 0 ?g(r=0.999 9),the average recovery was 102.35%,RSD was 2.31%(n=6);The linear range of quercetin was 0.006 2～0.244 0 ?g(r=0.999 9),the average recovery was 101.84%,RSD was 1.79%(n=6);The linear range of campherenol was 0.007 8～ 0.310 6 ?g(r=0.999 9),the average recovery was 99.04%,RSD was 2.90%(n=6). CONCLUSION:The method for quantifying of caffeic acid,quercetin and campherenol in Hedyotis diffusa Willd is accurate and reliable.

Objective To investigate the proliferation, immune phenotype and cytotoxicity on different cell lines of cytokine-induced killer (CIK) cells collected from healthy donors. Methods Peripheral blood mononuclear cells (PBMC) from healthy donors were induced to become CIK cells by adding cytokines including rhIL-2, rhIFN-γand CD3 McAb. The proliferation of CIK cells was tested by blood cell recording board. The CIK cells were analyzed on different time points by FACS. The cytotoxicity of CIK cells against different tumor cell lines, such as K562, BJAB, A549, MCF-7 and HepG2, was detected by MTT assays on day 13. Results CIK cells quickly proliferated from day 5, and expanded by 182-fold after 20-day culture. The immunophenotypes of CD3+, CD3+CD8+and CD3+CD56+were (97.83±1.03)%, (77.12±1.60)%and (27.58± 2.02)%. The percentages of CD3+, CD3+CD8+and CD3+CD56+increased noticeably (P＜0.01). According to the effector-tar-get ratio of 40∶1, the activity of CIK cells against tumor cells K562, BJAB, A549, MCF-7 and HepG2 were (88.89±7.22)%, (75.42±9.52)%, (63.19±5.67)%, (43.53±5.67)%and (42.63±7.69)%. The experiments showed that CIK cells possessed high-er antitumor cytotoxic activity. Conclusion CIK cells can be largely capacity cultured by adding cytokines in vitro. CIK cells were a highly efficient cytotoxic cell against tumors, and had clinical application potentials.

Objective To evaluate the anesthetic efficacy of etomidate target-controlled infusion (TCI) in combination with remifentanil in patients undergoing gynecological laparoscopy.Methods Sixty ASA physical status Ⅰ or Ⅱ patients,aged 25-56 yr,with body mass index 18-27 kg/m2,undergoing elective gynecological lapa-roscopy,were equally and randomly divided into 2 groups:propofol TCI combined with remifentanil group (group PR) and etomidate TCI combined with remifentanil group (group ER).Anesthesia was induced with iv injection ofmidazolam 0.1 mg/kg,fentanyl 4 μg/kg and cisatracurium 0.15 mg/kg in both groups,and with TCI of propofolwith the target effect-site concentration (Ce) of 2.5 μg/ml in group PR or with TCI of etomidate (Ce 0.8 μg/ml) ingroup ER.The patients were mechanically ventilated after endotracheal intubation.Anesthesia was maintained withTCI of propofol (Ce 2.0-2.5 μg/ml) in group PR or with etomidate (Ce 0.5-0.7 μg/ml) in group ER,and with ivinfusion of remifentanil 0.1-0.2 μg· kg-1 · min-1 and intermittent iv boluses of cisatracurium 5 mg.BIS value was maintained at 40-60.Before anesthesia (baseline,T0),at the end of operation (T1),and at 24 and 48 h after operation (T2-3),venous blood samples were collected for determination of serum cortisol and aldosterone concentrations by radioimmunoassay.The emergence time,extubation time and requirement for vasoactive agents during operation were recorded.The development of injection pain and muscle twitch during induction of anesthesia,intraoperative awareness,and post-operative agitation,nausea and vomiting were also recorded.Results Compared with the baseline value at T0,the serum cortisol concentration was significantly decreased at T1 in group ER (P ＜0.05),while no significant change was found in serum aldosterone concentrations at each time point in the two groups (P ＞ 0.05).Compared with group PR,the requirement for vasoactive agents and incidence of injection pain were significantly decreased,and the incidence of muscle twitch was increased (P ＜ 0.05),and no significant change was found in the emergence time,extubation time,and incidences of post-operative agitation,nausea and vomiting in group ER (P ＞ 0.05).Conclusion Compared with propofol TCI in combination with remifentanil,etomidate TCI combined with remifentanil is helpful in maintaining the hemodynamics stable and exerts transient inhibition of adrenocortical function with less injection pain in patients undergoing gynecological laparoscopy.