Objective To study the effects of CD2-associated protein (CD2AP) on podocyte adhesion and extension ability and to explore its possible mechanism. Methods Conditionally immortalized murine podocyte cell line was cultured in RPMI 1640 medium at 33℃permissive conditions. The podocytes were transfected with CD2AP small interfering RNA (siRNA) and serambing sequences labeled with fluorescein were taken as control. The transfected podocytes were trypsinized and seed into collagen IV coated plates. The relative cell adhesion and cell area were examined 90 min later. Apoptotic rates of CD2AP siRNA transfected podoeytes and different PAN concentrations incubated podoeytes were detected by flow cytometer. The distribution of F-actin was observed under laser scanning confoeal microscope. Nephrin protein expression and its phosphorylation level were examined by immunofluorescence and Western blot. Results The relative ceil adhesion of CD2AP siRNA transfected podocytes was apparently lower than that of control group[(41.72±6.07)% vs (64.46±8.53)%, P<0.05]. The cell area analysis had the similar result. The apoptotic rate of CD2AP siRNA transfected podocytes was significantly higher than that of the controls [(5.73±0.61)% vs (3.26±0.45)%, P<0.05]. 100 mg/L PAN could markedly induce podocytes to apoptosis and impair cell adhesion ability (P<0.05). Nevertheless, no significant difference was found in cell body spreading (P>0.05). The distribution of F-actin in CD2AP depletion podocytes was apparently altered. The expression of nephrin protein and its phosphorylation level was conspicuously descended to some degree (P<0.05). Conclusions CD2AP depletion facilitates podocyte apoptosis and impairs cell adhesion function. Cytoskeleton confusion and nephrin signaling weakness caused by CD2AP depletion may he partly responsible for the decline of cell adhesion and spreading.

Objective To investigate the efficacy of escitalopram combined with psychoanalysis in the treatment of refractory depression. Methods A total of 63 patients were randomly divided into escitalopram group ( n = 32) and escitalopram combined with psychoanalysis group( n = 31 ). All patients were evaluated with Hamilton depression Rating Scale(HAMD). Results After treatment,the scores of HAMD in two groups were both significantly lower than those before treatment. In the 8th ( ( HAMD ( 17.35 ± 2.98 ) ), 12th ( ( HAMD (9. 26 ±3.46) )weekend of treatment, the scores of HAMD in study group were significanlly lower than those in control group(8 th:21.97 ± 3.26; 12 th: 15.28 ± 3. 18 ). There were no significant differences in side effects between study group and control group. Conclusion Escitalopram angumented with psychoanalysis takes effects better than escitalopram single and doesn't increase side effects in the treatment of refractory depression.

Objective To construct a new recombinant immunotoxin expression vector fused with a murine interleukin18(IL18) gene and a truncated pseudomonas exotoxin (PE38) gene, and examine the expression of IL-18-PE38 fusion protein in Escherichia coli (E. coli). Method Murine IL-18 (mIL-18) cDNA was cloned from murine liver tissue through reverse transcription-polymerase chain reaction (RT-PCR). The mIL-18 cDNA was ligased with a PE38 gene carried by PRKL expression vector through T4 DNA ligase and constructed into fusion protein expression plasmid PRKL-IL18-PE38. The recombinant vector was identified by restriction endonucleases digestion, PCR and DNA sequencing. After transformed into E.coli BL21 and induced by IPTG, the expressed product was obtained and the molecular weight and specificity were determined by SDS-PAGE and Western-blotting. Result The new recombinant immunotoxin expression vector was constructed successfully. DNA sequencing revealed that the mIL-18 and PE38 gene were consistent with NCBI Gene Bank. The IL-18-PE38 fusion protein was expressed in E.coli BL21, and Western-blotting analysis indicated that the molecular weight of the expression product is about 56 kDa, and could react with the specific antibody against mIL-18. Conclusion IL-18-PE38 recombinant immunotoxin expression vector will provide the basis for study on the targeted cytotoxic activity to Th1 cells and may have some potential value in the treatment of Th1 cell-mediated autoimmune diseases.

Objective To investigate the relationship between the serum concentrations of brain derived neurotrophicfactor (BDNF) and its G196A polymorphism in the amphetamine induced-psychosis inpatients.Methods The cross-sectional study included 233 amphetamine abuses and 110 healthy participants who served as controls.The serum concentration of BDNF was measured by sandwich ELISA,and the genotype of BDNF G196A polymorphism was determined used polymerase chain reaction (PCR) techniques.The data were analyzed using SPSS 12.0 statistics software.Results The serum concentration of BDNF in case group((205.81±75.36) pg/ml) were significantly higher than that in control group((95.04±31.63) pg/ml;t=15.02,P＜0.01).There was no significant difference about the BDNF serum concentrations between the inpatients with the amphetamine induced psychosis and the inpatients with the amphetamine abuse (P＞0.05).The BDNF serum concentration showed no significant difference in the genotype distributions and allele frequencies (P＞0.05).The genotype distributions and allele frequencies of BDNF G196A showed no significant difference among three groups(P＞0.05).Conclusion The BDNF serum concentration is correlated with methamphetamine abuse,while the BDNF G196A gene polymorphism may not be associated.

Objective To investigate the effects of tanshinol on the proliferation,apoptosis and NF-?B activation in rat hepatic stellate cells(HSCs) after IL-1? inducement,and to elucidate the anti-fibrotic molecular mechanisms of tanshinol.Methods The rat HSCs was isolated with collagenase in situ liver recirculation perfusion and cultured in vitro.The cells were divided into 5 groups: normal control,IL-1? treatment group(10 ng/ml),and tanshinol group 1,2 and 3.The later 3 groups were pretreated with tanshinol at the concentrations of 0.062 5,0.125 and 0.25 mmol/L respectively followed by 10 ng/ml IL-1? treatment 1 h later.MTT colorimetric assay was used to detect the proliferation of HSCs.AO/EB immunoflurorescence microscopy and combination Annexin-V-FITC/PI double-labelimmunofluorescence with flow cytometer were employed to examine the apoptosis of HSCs.Synthesis and secretion of collagen Ⅲ were detected by the quantitative immunocytochemical assay and ELISA respectively.The amounts of cytoplasm p-I?B? and NF-?B p65,and nuclear NF-?B p65 in HSCs were determined by Western blotting.Immunocytochemical staining and Western blotting were used to observe nuclear translocation of NF-?B p65.Results IL-1? increased the proliferation of HSCs(P

Objective To study the safety and feasibility of endobiliary intraductal radiofrequency ablation (RFA) to reopen occluded self-expandable metal stents in patients with malignant biliary obstruction.Methods 11 patients with malignant biliary obstruction and blocked metal stents were prospectively studied.During ERCP, after biliary cannulation, the blocked metal stents underwent RFA using a bipolar radiofrequency probe which was introduced into the stenotic bile duct via a guide wire.This was followed by a balloon to repeatedly remove debris and then endoscopic nasobiliary drainage.The patients were closely observed and followed up.Results RFA was successfully carried out in all the patients and patencies were achieved when compared with pre-RFA.The median post-RFA luminal diameter of the strictures showed significant improvement: 6 (4 ～ 10) mm versus 2 (0 ～ 5) mm, and the mean post-RFA total bilirubin level decreased sharply : (39.4 ± 8.7) μ mol/L versus (130.1 ± 38.2) μmol/L.Following this intervention, 3 patients developed fever, which were controlled with conservative therapy.There was no mortality, haemorrhage, bile duct perforation or bile leak.Of the 11 patients, 3 were dead and 6 were alive at a median follow-up of 187 (75 ～ 304) days.The median stent patency was 135 (75 ～ 203) days and the median survival was 278 (75 ～ 304) days.Four patients had their stents patent at the time of the last follow-up or death.Seven patients had their stents blocked on 113, 124, 154, 203, 96, 135 and 112 days post-procedure.Condusions Endobiliary intraductal RFA is technically feasible and safe to reopen occluded metal stents in malignant biliary obstruction.This efficacy needs to be confirmed by future randomized studies.

Objective To study the effect of overexpression of TRPC6 on Ang Ⅱ-induced apoptosis of mouse podocytes in vitro and to explore the possible mechanisms. Methods Mouse TRPC6 cDNA eukaryotie expression vector pEGFP-NI-mTRPC6 was transfected to conditionally immortalized routine podocyte cell line by liposome. The fluorescent microscopy was used to examine the expression of EGFP after 24 hours. The change of TRPC6 protein expression was observed by Western-blot. Podocytes were treated by different concentrations of Ang Ⅱ. The podocyte intracellular calcium concentration was measured with laser-scanning con_focal microscope. The expression of Bax and Bcl-2 mRNA was assessed by RT-PCR and the expression of Bax and Bcl-2 protein was measured by Western-blot. The apoptotic ratio of podocytes was monitored by flow cytometry and Hoechst staining. Results About 35% of the cells expressed EGFP. An up-regulation of protein expression of TRPC6 was detected in podocytes when transfected with pEGFP-N1-mTRPC6 (P<0.01). The overexpression of TRPC6 promoted the Ang Ⅱ-induced influx of extracellular calcium and elevated the expression of Bax but decreased the expression of Bcl-2 (P<0.01, P<0.05). The apoptotic ratio of podocyte was (2.50±0.72)% when treated by low-dose Ang Ⅱ (10-10 mol/L), and it was increased to (4.33±0.45)% when transfected with pEGFP-N1-mTRPC6 (P <0.05 ). Transfection with pEGFP-NI-mTRPC6 increased apoptosis rate from (15.46± 1.40)% to (18.33±0.87)%(P<0.01) by high-dose Ang Ⅱ (10-6 mol/L). Conclusion TRPC6 plays an important role in the Ang Ⅱ-induced apoptosis of podocytes by promoting the influx of extraeellular calcium, which leads to the apoptosis cascade initiation.

Objective To evaluate the effect of epidural block on postoperative cognitive dysfunction in elderly patients undergoing laparoscopic radical operation for gastric cancer under general anesthesia.Methods Sixty ASA physical status Ⅰ or Ⅱ patients,aged 65-75 yr,with preoperative Mini-Mental State Examination (MMSE) score ＞ 23,scheduled for elective laparoscopic radical operation for gastric cancer under general anesthesia,were randomized into epidural block group (group E,n =30) and control group (group C,n =30) using a random number table.Epidural block was performed at T8,9 interspace before induction of anesthesia.Anesthesia was induced with iv injection of midazolam 0.03 mg/kg,propofol 1.0-1.5 mg/kg,fentanyl 3-4 μg/kg and rocuronium 0.9 mg/kg.The patients were endotracheally intubated and mechanically ventilated.Anesthesia was maintained with iv infusion of remifentanil 0.15-0.25 μg· kg-1· min 1 and propofol target-controlled infusion (target plasma concentration 2.5-3.0μg/ml).In group E,10 ml of mixture of 0.25％ ropivacaine and 0.04 mg/ml butorphanol was injected via the epidural catheter at 30 min before skin incision.In the two groups,the mixture 10 ml mentioned above was administered via the epidural catheter at 10 min before the end of operation followed by patientcontrolled epidural analgesia.The development of postoperative cognitive dysfunction (MMSE score ＜ 21) was recorded within 72 h after operation.Blood samples were collected from the internal jugular vein to detect the concentration of serum protein S-100β.Results The concentration of serum protein S-100β was significantly lower at 6,12 and 24 h after operation,and the incidence of postoperative cognitive dysfunction within 72 h after operation was lower in group E than in group C (P ＜ 0.05).Conclusion Epidural block can decrease the development of postoperative cognitive dysfunction in elderly patients undergoing laparoscopic radical operation for gastric cancer under general anesthesia.

Objective To investigate the effects of positive end-expiratory pressure (PEEP) after recruitment maneuvers (RM) on respiratory mechanics and gas exchange during laparoscopy in obese patients.Methods Sixty-three ASA physical status Ⅱ or Ⅲ patients,aged 42-64 yr,with body mass index 30-40kg/m2,were randomly allocated into 3 groups (n=21 each):PEEP0 group,PEEP5 group and PEEP10 group.PEEP was not given after RM in PEEP0 group.In PEEP5 and PEEP10 groups,a recruiting maneuver was followed by PEEP 5 and 10 cm H2 O,respectively,until the end of pneumoperitoneum.The intraabdominal pressure was set at 12mmHg in the three groups.Parameters of respiratory mechanics including peak airway pressure (Ppeak),airway plateau pressure (Peat),chest wall plateau pressure (PplatCW),airway resistance (Raw),elastance of respiratory system (ERS),elastanc of chest wall (ECW) and elastance of lung (EL) and parameters of gas exchange including oxygenation index (PaO2/FiO2),arterial to end-tidal difference in carbon dioxide (Da-ETCO2),alveolar-arterial oxygen tension difference (DA-aO2),and dead space/tidal volume ratio (VD/VT) were measured before pneumoperitoneum (T0),at 20 min of pneumoperitoneum (T1),at 10 min after the end of recruitment (T2),and at the end of pneumoperitoneum (T3).Results Da-ETCO2,ERS and Raw were decreased at T2,ECW and EL were decreased at T3 in PEEP5 group,and Da-ETCO2,VD/VT,DA-aO2,Pplatcw Raw and EL were decreased at T2.3,and PaO2/FiO2 was increased at T2,ECW was decreased at T3 in group PEEP10 as compared with that in group PEEP0 (P ＜ 0.05).Da-ETCO2 and VD/VT were decreased and PaO2/FiO2 was increased at T2,3,Raw was increased and EL was decreased at T2 in group PEEP10 as compared with that in group PEEP5 (P ＜ 0.01).Conclusion PEEP after RM can improve respiratory mechanics and gas exchange during laparoscopy in obese patients and PEEP maintained at 10 cm H2O after RM provides better efficacy than PEEP at 5 cm H2 O.

Objective To evaluate the incidence of postanesthetic shivering in patients undergoing radical operation for lung cancer under dexmedetomidine-assisted anesthesia.Methods Eighty ASA Ⅰ or Ⅱ patients,aged 50-75 yr,height 158-180 cm,weighing 48-79 kg,scheduled for radical operation for lung cancer under general anesthesia combined with epidural anesthesia,were randomly divided into 2 groups ( n =40 each): normal saline control group (group C) and dexmedetomidine group(group D).Anesthesia was induced with midazolam,fentanyl,propofol and rocuronium.The patients were tracheal intubated and mechanically ventilated.Anesthesia was maintained with sevoflurane inhalation and intermittent epidural ropivacaine.BIS was maintained at 40-55.Group D received dexmedetomidine intravenous infusion at 0.5 μg · kg- 1· h- 1 from immediately after tracheal intubation until 1 h before the end of operation,and group C received the equal volume of normal saline.The shivering within 1 h after operation was scored and recorded.Results The incidence rate of postanesthetic shivering was 50％ in group C and 10％ in group D.Compared with group C,the incidence rate and degree of shivering were decreased in group D( P ＜ 0.05).Conclusion Dexmedetomidine-assisted anesthesia can reduce the incidence of postanesthetic shivering in patients undergoing radical operation for lung cancer.