OBJECTIVE:To establish a HPLC method for determining the concentration of dexketoprofen in rabbit’s pla_ sma.METHODS:Dexketoprofen was extracted from samples with ether.HPLC was performed on C 18 column with methyl al?cohol-50mmol/L sodium dihydrogen phosphate as mobile phase.Detection wavelength was260nm.RESULTS:The linear range was0.05～25?g/ml.Recovery was99.92%～100.54%.Within-day and between-day RSDs were0.56%～2.94%and0.6%～2.89%respectively.CONCLUSION:This method is simple,accurate and good in repetitiveness.Satisfactory results have been obtained in determining plasma concentrations of dexketoprofen in6rabbits after taking dexketoprofen-?-cy?clodextrin inclusion microspheres.

Objective To prepare microspheres of dextro ketoprofen ? cyclodextrin (S KP ? CD) for prolonging the drug releasing time in vitro . Methods Microspheres containing S KP ? CD were prepared by complex coacervation method with gelatin and acacia. Trap efficiency, drug loading, and drug content were determined. The dissolubility of S KP in the intestinal liquid was compared with that in the gastric liquid. Results In comparison with S KP, microspheres of S KP ? CD possessed slow releasing property. Conclusion Drug microspheres prepared by this simple, easy, and accurate method are of slow releasing property.

Objective To investigate the relationship between serum protein S100b in intracerebral hemorrhage (ICH) patients and brain damage. Methods A total of 37 hypertensive cerebral hemorrhage patients, 24 male and 13 female, were enrolled as intracerebral hemorrhage (ICH) group, aged from 45 to 85 years with an average of (69.2?8.9) years. The control group were matched with ICH group, including 30 healthy subjects, 19 male and 11 female, aged 52 to 70 (61.2?5.0) years. The concentration of serum protein S100 was detected using double antibody sandwich ELISA. Evaluation of blood volume was calculated with the fomula based on cranial CT data: V=S?L?Slice??/6. The nerve function of the patients was evaluated by the CSS during acute phase and Barthel index score at 3 month after stroke. Results Serum protein S100 concentration was significantly elevated in patients with ICH (0.54?0.41 ?g/L), as compared to controls (0.17?0.04 ?g/L)(P

Objective To explore the mechanism of Xiaoying decoction on experimental autoimmune thyroiditis (EAT) rats in view of regulatory T cells and Th17 cells.Methods SD rats were divided into five groups,normal control group,model control group,tripterygium glycoside group,Xiaoying decoction low and high dose groups.Except for normal control group,the other groups were established the model of EAT.Rats in the Xiaoying decoction low,and high dose groups were given Xiaoying decoction of 17.24 and 68.95 g·kg-1;rats in the tripterygium glycoside group were given tripterygium glycoside 6.25 mg·kg-1.The serum free triiodothyronine(FT3),free thyrocyte(FT4) and thyroglobulin antibody were detected by RIA method.Thyrocyte morphology was observed under optical microscope.The expression levels of Foxp3 mRNA and IL-17 mRNA were detected by real-time PCR.The changes of Treg cells and Th17 cells were analyzed by flow cytometry.Results Compared with the normal control group,FT3,FT4 and TgAb were increased in the model control group (P ＜0.01,P ＜0.01,P ＜0.05).Compared with the model control group,FT3,FT4 and TgAb were decreased in tripterygium glycoside group and Xiaoying decoction high dose group (P ＜ 0.05).The infiltration score in the normal control group,model control group,tripterygium glycoside group,Xiaoying decoction low and high dose groups were 0,4,4,3.5,2.Compared with model control group,the infiltration was improved (P ＜0.01).Foxp3 mRNA expression was decreased while IL-17 mRNA increased in the model control group as compared with the normal control group(P ＜ 0.01).In contrast,the expression of Foxp3 mRNA was increased and IL-17 mRNA expression was decreased in Xiaoying decoction low and high dose groups,tripterygium glycoside group as compared with the model control group (P ＜0.05).Compared with the normal control group,rats in the model control group had fewer Treg cells and more Th17 cells (P ＜0.01).Compared with the model control group,the percentage of Treg cells was elevated and Th17 cells was reduced in tripterygium glycoside group and Xiaoying decoction high dose groups (P ＜ 0.01).Conclusion The therapeutic mechanism of Xiaoying decoction on EAT rats may be related to changing the percentage of regulatory T cells and Th17 cells with up-regulating the expression of Foxp3 mRNA and down-regulating the expression of IL17mRNA.

AIM: To study compatibility rationality of combination of Paeonia lacliflora and Glycyrrhiza uralensis. METHODS: The effective combination of paeoniflorin(44% purity),glycyrrhizic acid(50% purity) and liquorice flavones(52% purity),glycyrrhizic acid(50% purity) and liquorice flavones(52% purity) were respectively administered to rats.Pharmacokinetic change of these constituents in rat blood was studied. RESULTS: The pharmacokinetic parameters of these constituents in rat blood showed that the increases in AUC and C_(max) of effective combination group were more than that of glycyrrhizic acid group or that of liquorice flavones group.T_(max) of the former was extended with respect to the latters.Clearance of effective combination markedly slowed down. CONCLUSION: The effective combination of paeonia lacliflora and Glycyrrhiza uralensis have the advantage of either Paeonia lacliflora or Glycyrrhiza uralensis.

Objective To investigate the relationships and clinicopathologic features of the expressions of cytochrome P450 2E1 (CYP2E1) and PC cell-derived growth factor (PCDGF) in esophageal squamous cell carcinoma (ESCC),and to determine the role of CYP2E1 and PCDGF in angiogenesis.Methods The expression levels of CYP2E1 and PCDGF in 42 surgical cancer specimens and 20 adjacent normal esophageal mucosa specimens from patients with ESCC were detected by immunohistochemistry.Results The positive expression rates of CYP2E1 and PCDGF in ESCC were 83.3 ％ (35/42) and 88.1％ (37/42),respectively,which were obviously higher than those of normal mucosa (P ＜ 0.05).Expression of PCDGF was correlated with the degree of histological differentiation (r =0.444,P ＜ 0.05),depth of tumor invasion (r =0.332,P ＜ 0.05),lymph node metastasis (r =0.476,P ＜ 0.05),and TNM classification (r =0.450,P ＜ 0.05).The expression of CYP2E1 was negatively correlated with tumor tissue differentiation (r =-0.518,P ＜ 0.05),and positively correlated with depth of invasion in ESCC (r =0.388,P ＜ 0.05).The expression of CYP2E1 was related to that of PCDGF in the tumor (r =0.483,P ＜ 0.05).Conclusion The expressions of CYP2E1 and PCDGF are synergistically involved in tumor growth,infiltration andmetastasis.Overexpression of CYP2E1 and PCDGF can be used as the important predictors for evaluating the biological behavior of ESCC and predicting prognosis of patients.

Objective To make a molecular genetic analysis in a Chinese family with piebaldism,in order to find the causative mutation of this disease.Methods DNA and RNA were extracted from blood samples of the proband and other 13 members in this family.Ploymerase chain reaction (PCR),reverse transcription PCR and DNA sequencing were performed to detect the mutation of kit gene.Results A novel heterozygous mutation c.2472+1G>A in kit gene.which leads to the loss of 3' splicing site in exon 17 followed by the absence of exon 17,was found in all affected members,but not in an unaffected member in the family.Conclusion The novel mutation c.2472+1G>A may be associated with piebaldism initiation in this family.

Objective To determine whether the adjustment of the pusher of GTF was useful to decrease the degree of tilting of the femoral Ginther Tulip filter (GTF) in an in vitro caval model. Methods The caval model was constructed by placement of a 25 mm × 100 mm and two 10 mm ×200 mm Dacron graft inside a transparent bifurcate glass tube. The study consisted of two groups: left straight group (GLS) (n =100) and left curved group (GLC) (n = 100). In the GLC, a 10° to 20° angle was curved on the introducer.The distance (DCH) between the caval right wall and the hook was measured. The degree of tilting (DT) was classified into 5 grades and recorded. Before and after the GTF being released, the angle (ACM1, 2)between the axis of IVC and the metal mount, the distance (DCM1) between the caval right wall and the metal mount, the angle (ACF) between the axis of IVC and the axis of the filter and the diameter of IVC (DIVC) were measured. The data were analyzed with Chi-Square test, t test, rank sum test and Pearson correlation test. Results The degree of GTF tilting in each group revealed a divergent tendency. In group LC, the apex of the filter tended to be grade Ⅲ compared in group LS (x2 value 37. 491 ,P ＜0.01).The differences of most variables between GLS and GLC were considered as statistical significance(16. 60° vs.3.05°, 20.60° vs. 3.50°, -3.90°vs. -0.40°, 2.98 mm vs. 10.40 mm, - 10.95° vs. -0.485°,13. 17 mm vs. 10.06 mm, - 1.70° vs. 0.70°, t or Z values - 12. 187, - 12. 188, -8.545, -51.834,-11. 395,9. 562, -3. 596, P ＜ 0. 01). There exist significant positive correlation between ACM1 and ACF,ACM1 - ACM2 and DCH1 - DCH2 in each group, respectively(r values 0. 978,0. 344,0. 879,0. 627 ,P ＜0. 01),while significant negative associations are detected between DCH1 and ACF in each group , ACP and ACF in group LC(r values -0.974, -0.322, -0.702,P＜0.01). Conclusion The technique of adjusting the orientation of filter pusher had minimized the incidence and extent of GTF filter tilting in vitro.

BACKGROUND: In addition to physical disability, stroke may also result in psychological impairments usually manifested by depression and anxiety.Regardless of the primary or secondary onset of anxiety, anti-depressants should be given for treatment of the anxiety and depression besides routine treatment of the primary condition underlying the symptoms.OBJECTIVE: To compare the effect of different treatment protocols with or without anti-depressants and different anti-depressants on poststroke anxiety and depression as well as the neurological functions.DESIGN: Randomized controlled double-blind clinical trial.SETTING: Departments of Psychiatry and Neurology of the Second Hospital Affiliated to Lanzhou University.PARTICIPANTS: Ninety patients aged 41-72 years with post-stroke anxiety and depression, who were admitted in the Departments of Psychiatry and Neurology of the Second Hospital Affiliated to Lanzhou University between July 1999 and December 2002, were enrolled in this study and randomized equally into Paxil group, imipramine group and control group.METHODS: After emergency management for 1-2 weeks in the acute stage, the stroke patients showed clear consciousness and stable life signs without any understanding problems. Patients in the control group received conventional treatment combined with rehabilitative training, while patients in the other two groups were given additional Paxil (20 mg/day) or imipramine (50-150 mg/day) for totally 12 weeks. The neurological deficits and capacity for independent living of the patients were assessed with Hamilton Depression Scale (24 items) and Hamilton Anxiety Scale (14items) at 2, 4, 8, and 12 weeks during the treatment. A reduction of the score for Hamilton anxiety and depression scale by over 75% suggested a cure of depression and anxiety, 50% but ＜ 75% obvious improvement,25% but ＜ 50% improvement, and ＜ 25% non-response. Basically recovered neurological function was indicated by a reduction of neurological deficit score by 90%-100%, remarkable improvement by 46%-89%, improvement by 18%-45%, and non-response or exacerbation by a reduction less than 17%.MAIN OUTCOME MEASURS: ① Neurological function recovery of recovery of the patients after treatment; ② Poststroke anxiety and depression status before and after treatment; ③ Therapeutic effects on depression,neurological functions, severity of neurological deficit, and capacity of independent living. ④ Adverse events and side effects.RESULTS: One patient in Paxil group and 3 in the control group failed to be available for follow-up study, and 3 patients in imipramine group withdrew from the study due to adverse events, so that 83 cases were analyzed.At 2 and 4 weeks in the treatment, the scores for neurological deficits and capacity for independent living exhibited obvious changes (P ＜ 0.01),which gradually stabilized at 8 and 12 weeks (P ＜ 0.05), and significantly greater improvement in the neurological function and capacity for independent living was observed in Paxil group than in the control group (P＜ 0.01), but the differences between imipramine group and the control group and between Paxil group and imipramine group were not statistically significant (P ＞ 0.05). The scores for Hamilton Depression Scale and Hamilton Anxiety Scale were obviously lower in Paxil group and imipramine group at 2, 4, 8 and 12 week than those in the control group (P＜ 0.01-0.001), but similar between the former two groups at 12 weeks (P＞ 0.05). Paxil and imipramine on resulted in curative rates of anxiety and depression of 86.6% and 85.1%, respectively, which were obviously higher than that of the control group (46.6%); the improvement rate ofneurological function in Paxil group, imipramine group and control groupwas 89.6%, 70.3% and 56.6%, respectively, with that of Paxil groupsignificantly higher than that of the control group (P ＜ 0.01), but the difference between imipramine group and control group was not signifi cant (P ＞ 0.05).CONCLUSION: Patients with poststroke anxiety and depression shouldreceive appropriate interventions with anti-depressants in addition to treat ment of neural function impairment. Paxil and imipramine haye similar effect in treating anxiety and depression, but the former can be for its less side effects, better compliance on the part of patients and good effect inpromoting neurological function recovery.

Objective To investigate the mechanisms of erythropoietin-producing hepatocellular A3 (EphA3) in the invasion of hepatocellular carcinoma (HCC) cells.Methods Hepatic cell HL-7702 and HCC cell and HCC cell lines HepG2 and MHCC97H were cultured.The expression of EphA3 in the HepG2 and MHCC97H cells was suppressed by siRNA interference,and then were divided into the untreated group,the control group and the siRNA intervention group.The expression of EphA3 was detected by RT-PCR and Western blot.The invasion ability of HepG2 and MHCC97H was detected by Transwell chamber.The protein expression of VEGF and activity of vascular endothelial growth factor (VEGF) were detected by western blot and ELISA.All data were analyzed using the analysis of variance or LSD-t test.Results The relative mRNA expressions of EphA3 in HL-7702,HepG2,and MHCC97H cells were 0.94 ±0.13,1.76 ±0.16 and 3.62 ±0.14,respectively,and the protein expressions of EphA3 in the 3 cells were 0.96 ±0.12,1.59 ±0.11 and 3.82 ±0.11.There was significant difference in the EphA3 expression between HL-7702 cells and HepG2,MHCC97H cells (t =2.511,6.437 ; 2.321,6.895,P ＜ 0.05).The relative mRNA expressions of EphA3 in the HepG2 cells in the untreated group,the control group and the siRNA intervention group were 0.95 ±0.11,0.96 ±0.12 and 0.31 ±0.15,respectively.There was significant difference in the mRNA expression of EphA3 in the HepG2 cells between the siRNA intervention group and the control group (t =4.051,P ＜ 0.05).The relative mRNA expressions of EphA3 in the MHCC97H cells in the untreated group,the control group and the siRNA intervention group were 0.97 ± 0.16,0.95 ± 0.14 and 0.40 ± 0.11,respectively.There was significant difference in the mRNA expression of EphA3 in the MHCC97H cells between the siRNA interference group and the control group (t =5.237,P ＜0.05).The relative protein expressions of EphA3 in the HepG2 cells in the untreated group,the control group and the siRNA intervention group were 0.97 ± 0.16,0.95 ± 0.15 and 0.32 ± 0.17,respectively.There was significant difference in the protein expression of EphA3 in the HepG2 cells between the siRNA interference group and the control group (t =4.145,P ＜ 0.05).The relative protein expressions of EphA3 in the MHCC97H cells in the untreated group,the control group and the siRNA intervention group were 0.95 ± 0.11,0.96 ± 0.12 and 0.38 ±0.17,respectively.There was significant difference in the protein expressions of EphA3 in the MHCC97H cells between the siRNA interference group and the control group (t =4.327,P ＜ 0.05).The numbers of HepG2 cells penetrated the Watrigel in the untreated group,the control group and the siRNA intervention group were (111 ±4)/10HPF,(109 ±5)/10HPF and (51 ±3)/10HPF,respectively.There was significant difference in the number of HepG2 cells between the siRNA interference group and the control group (t =7.582,P ＜ 0.05).The numbers of MHCC97H cells penetrated the Watrigel in the untreated group,the control group and the siRNA intervention group were (402 ± 6)/10HPF,(397 ± 7)/10HPF and (152 ± 7)/10HPF,respectively.There was significant difference in the number of MHCC97H cells between the siRNA interference group and the control group (t =9.479,P ＜ 0.05).The relative protein expressions of VEGF in the HepG2 cells in the untreated group,the control group and the siRNA intervention group were 0.98 ± 0.11,0.96 ± 0.13 and 0.57 ± 0.11,respectively.There was significant difference in the protein expression of VEGF of the HepG2 cells between the siRNA interference group and the control group (t =3.167,P ＜ 0.05).The relative protein expression of VEGF in the MHCC97H cells in the untreated group,the control group and the siRNA intervention group were 0.97 ±0.14,0.98 ±0.12 and 0.34 ± 0.15,respectively.There was significant difference in the protein expression of VEGF of the MHCC97H cells between the siRNA interference group and the control group (t =4.278,P ＜ 0.05).The relative activities of VEGF proteins of HepG2 cells in the untreated group,the control group and the siRNA intervention group were 0.96 ±0.15,0.94 ±0.11 and 0.47 ±0.13,respectively.There was significant difference in the activity of VEGF protein in the HepG2 cells between the siRNA interference group and the control group (t =3.981,P ＜ 0.05).The relative activities of VEGF proteins in MHCC97H cells in the untreated group,the control group and the siRNA intervention group were 0.98 ±0.12,0.97 ±0.12 and 0.38 ±0.14,respectively.There was significant difference in the activity of VEGF protein in the MHCC97H cells between the siRNA interference group and the control group (t =4.059,P ＜ 0.05).Conclusions EphA3 plays an important role in the invasion of HCC cells via regulating the protein expression and activity of VEGF.EphA3 might be a new target for the treatment of HCC.