Background and purpose:Hsp90 is cell chaperone protein that interacts with many proteins,but itself can not degrade its client proteins.Hsp90 inhibitor can inhibit tumor cell proliferation,induce cell apoptosis,cell growth arrest and increase the degradation of Hsp90 client proteins like Survivin.In order to explore the co-effects of Hsp90 inhibitor 17-AAG and Survivin on LoVo cells and the possible mechanisms,we observed the effects of 17-AAG on proliferation and cycles of LoVo cells and the protein level of Survivin.Methods:LoVo cells were treated with 17-AAG.The cell proliferation inhibition rate was evaluated by MTT assay.The cell cycle was detected by ? ow cytometry.The expression of Hsp90 client protein Survivin was detected by Western blot.Results:17-AAG time-dose-dependently inhibit the proliferation of LoVo cells,after 100 ng/ml,500 ng/ml and 800 ng/ml 17-AAG exposure for 24 hrs,the cell proliferation inhibition rate was 21.00%,40.81%,60.34% respectively,after exposure for 48 hrs,the cell proliferation inhibition rate was increased to 27.29%,48.17%,80.97% respectively,after exposure for 72 hrs,the cell proliferation inhibition rate was to 34.45%,67.81%,88.42%;17-AAG arrested cell cycle,when LoVo cells were exposed to 100 ng/ml 17-AAG for 72 hrs,the cell ratio of G0/G1 phase was(61?3)%,when to 500 ng/ml for 72 hrs,cell ratio of G0/G1 phase was increased to(74?3)%,compared to(48.2?0.8)% LoVo cells without 17-AAG(P

Female ; Follow-Up Studies ; Hearing Loss ; etiology ; Hearing Tests ; Humans ; Infant, Newborn ; Male ; Risk Factors

Objective To investigate the effects of montelukast(MK),a selective cysteinyl leukotriene receptor 1 antagonist on interleukin(IL)-5 and eotaxin expression as well as serum total immunoglobulin E(IgE) production during MK treatment of mouse acute asthma airway inflammation.Methods Sensitized Balb/c mice were challenged by ovabumin(OVA)to establish the acute asthmatic mo-(del).Twenty-five mg/kg of MK(MK group) or Saline(Saline group)were given by intravenously for 3 days.Cellular infiltration of bronchoalveolar larvage fluid(BALF) were assessed by Wright staining.Production of IL-5 and eotaxin in the lung or BALF and serum IgE were determined by enzyme linked immunosorbent assay(ELISA).The expression of IL-5 and eotaxin mRNA were measured by semi-quantified reverse transcriptase-polymerase chain reaction(RT-PCR).Plasma MK level was determined by liquid chromatography.Results After 24 h OVA challenge,the numbers of total white blood cells,neutrophils and eosinophils(EOS)of BALF were(26.0?18.9)?10~7/L,(5.92?8.09)?10~7/L and(0.74?0.88)?10~7/L in MK treatment group.They were significantly reduced compared with those in Saline group,respectively(P80%.The level of IL-5 in BALF and lung tissue were(48.52?14.45) ng/L and(27.40?9.62) ng/g protein in MK group,which significantly declined compared with that in saline group;BALF eotaxin level declined too.Serum IL-5 and total IgE level were also significantly reduced;IL-5 mRNA expression in lung significantly decreased.Eotaxin and its mRNA expression in lung were not decreased significantly.Conclusion MK(exerts) its anti-inflammatory effect mainly through the suppression of IL-5 and IgE production.

Female ; Foreign Bodies ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Pharynx ; Tomography, Spiral Computed

Objective To investigate the infection following the lymphocytes deleted agent (ATG) and IL-2 receptor antagonists (Basilixinab and Daclizumab)-based induction therapy after renal trausplantation.Methods A retrospective analysis was carried out on 701 kidney transplant recipients between Jan. 1,2005 to Dec.31,2010.According to exclusive and inclusive criteria,finally 549 patients were evaluated,including 429 patients treated with ATG (ATG group) and 120 patients with anti-CD25 monoclonal antibodies (monoclonal antibodies group; 86 patients with Basiliximab,and 34 patients with Daclizumab).The incidence of acute rejection,infection rate,infection time,hospital stay,severe infection rate and mortality were analyzed.After operation,the patients received an immunosuppression therapy including Tacrolimus (cyclosporine A),Mycophenolate-Mofetil and prednisone to present rejection. Part of the patients were treated with ganciclovir and sulfamethoxazole sulfadiazine and trimethoprim for infection prevention.Results The acute rejection rate in ATG group and monoclonal antibodies group was 15.9％ (68/429) and 10.0％ (12/120),and there was no statistically significant difference (P＞0.05).The infection rate in ATG group was 11.9％ (51/429),including 13.7％ (7/51) with severe infection,and mortality was 7.8％(4/51).The infection rate was 15.0％ (18/120) in monoclonal antibodies group,including 11.1％ (2/18) with severe infection,and mortality was 5.6％ (1/18).There was no statistically significnat difference in infection rate,severe infection rate and mortality between two groups (P＞0.05).The hospital stay in ATG group and monoclonal antibodies group was 25.8 days and 19.1 days respectively (P＜0.05).Dead cases had not received regular anti-infection treatment,and the patients age was over 50 years.Conclusion The infection risk and mortality between these two induction therapies are identical,but hn comparison to the patients using ATG,the infection of patients using anti-CD25 monoclonal antibodies is easier to control.Anti-infection prophylaxis is important to reduce infection rate and decrease infectious mortality.

Objective To construct and express the recombinant plasmid pET28α-Sj26GST-Sj32 of Schistosoma japonicum(Sj) in Escherichia coli BL21 (DE3). Methods Total RNA was extracted from Sj adult worms by ultrasound-breaking, Sj26GST and Sj32 antigen gene was respectively amplified by RT-PCR from the total RNA; Sj26GST-Sj32 fusion gene obtained with gene splicing by overlap extension(SOEing) was cloned into prokaryotic expression plasmid pET28α and transformed into Escherichia coli BL2 (DE3) to construct pET28α-Sj26GST-Sj32;BL21 (pET28α-Sj26GST-Sj32) was induced with isopropyl-β-D-thiogalactopyranosid (IPTG), and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE)and Western blotting. Results The 1991 bp Sj26GST-Sj32 fusion gene was successfully amplified by gene SOEing and cloned into pET28α by restriction analysis and PCR identification, the recombinant plasmid pET28α-Sj26GST-Sj32 was successfully constructed; the relative molecular mass of the expressed recombinant protein was approximately 69 × 103 by SDS-PAGE, and the amount of the expressed protein was 25% of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pET28α-Sj26GST-Sj32 is successfully constructed and highly expressed in Escherichia coli in fused form with His-tag, and the expressed fusion protein shows specific antigenicity.

Objective To construct and express the recombinant plasmid pET32α-Sj26GST of Schistosoma japonicum(sj)in Escherichia coli(E.coli)B121(DE3).Methods The total RNA was extracted from sj adult worms by ultrasound-breaking,Sj26GST antigen gene was amplified by RT-PCR from the total RNA,then cloned into prokaryotic expression plasmid pET32α(+) and transformed into E.coli B12(DE3)to construct pET32α-Sj26GST;BL21(pET32α-Sj26GST)WaS induced with isopropyl-β-D-thiogalactopyranosid(IPTG),and the expressed products were analyzed and identified by SDS-PAGE and Western blot.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and cloned into pET32α(+)by restriction analysis and PCR identification,the recombinant plasmid pET32α-Sj26GST was successfully constructed;the relative molecular mass of the expressed recombinant protein was approximately 49×103 by SDS-PAGE,and the amount of the expressed protein was 24%of the total bacterial proteins;the fusion protein could be recognized by sera from rabbits infected with sj by Western blot.Conclusions The recombinant plasmid pET32α-Sj26GST is successfully constructed and highly expressed in E.coli in fused form with Trx-tag and His-tag,and the expressed fusion protein shows specific antigenicity.

0.05).Compared with control group,the GK activity of liver cell,the expression of PEPCK and the expression of GLUT4 in model group decreased signifi cantly(P

Objective To investigate the incidence of infection and the effect of anti-infection prophylaxis in renal transplanted patients after Basiliximab induction therapy. Methods A total of 204patients who have received renal transplantation and Basiliximab induction therapy from January 1,2001 to December 31, 2010 in our hospital have been retrospective analysed in this study. These patients were divided into a prophylaxis group (118 cases) with Ganciclovir + Sulfadiazine +Trimethoprim therapy and a control group (86 cases) without any anti-infection prophylaxis.Furthermore, 440 transplanted patients in the same peroid without any induction therapy were also analysed. They were also devided into two groups: an anti-infection prophylaxis group (206 cases)and a control group (234 cases) without any anti-infection prophylaxis. Results In the prophylaxis group with Basiliximab induction therapy, there were 23 patients (19. 5 %, 23/118)experienced hospitalization due to infection, 3 cases (13. 0 %,3/23) among them were severe infection, and 3patients (13.0 %, 3/23) died from vital infection. In the non-prophylaxis control group with Basiliximab induction therapy, 27 patients (31.4 %, 27/86) had infection complication, 7 patients (25.9 % ,7/27) among them were severe infection, and 4 patients(14. 8 % ,4/27)died. The incidence of infection between the above two groups is significantly different (P＜0. 05). In the prophylaxis group without induction therapy, the incidence of infection was 15.0 % (31/206), there were no severe infection cases but 7 patients (22. 6 %, 7/31) died from infection. In the non-prophylaxis control group without induction therapy, the incidence of infection was 12. 8 % (30/234), 3 cases among them were severe infection(10. 0 %,3/30)and 5 patients died from infection (16. 7 %, 5/30).The incidence of infection in Basiliximab induced patients without anti-infection prophylaxis is significantly higher than that in patients without induction therapy and anti-infection prophylaxis (31.4 % vs. 12.8 %,P＜0.01). Conclusion Basiliximab induction therapy increased the risk of infection, but not the rate of mortality. It is necessary to give anti-infection prophylaxis in renal transplanted patients with Basiliximab induction therapy.

Objective To assess the clinical outcome of varicose veins with incompetent great saphenous vein(GSV) treated with ultrasound guided foam sclerotherapy. Methods Forty limbs with moderate to severe symptomatic varicose veins with incompetent GSV in 38 patients were injected with foam sclerosing agent (Fibro-Vein) under ultrasound guidance. There were 36 patients with unilateral varicose veins and 2 with bilateral varicose veins. No of them suffered from deep vein incompetence or perforating vein incompetence. Second injection was performed one month after the initial injection in 7 limbs. Thirty-eight of 40 limbs were followed up with clinical examination and duplex ultrasound scan 30-47 months (mean 40 months) after the treatment. Results Among 38 limbs with follow-up mild debilitation was found in two limbs(5. 3%). There were no other symptoms or complications. Duplex ultrasound demonstrated four type of results: type I, sclerosed GSV trunk with no detectable venous flow in 32 of 38 limbs (84. 2%) ;type II,patent GSV trunk in 3 limbs (7. 9%) ,two of them had mild reflux in the GSV trunk;type III,sclerosed GSV trunk and mild reflux in the GSV tributaries, 1/38(2. 6%) ; type IV,sclerosed proximal GSV trunk,patent distal GSV communicated with a superficial vein and mild reflux in the veins, 2/38 (5. 3% ). Conclusions Clinical examination and duplex ultrasound scan demonstrated excellent results of varicose veins with incompetent GSV treated with ultrasound guided foam sclerotherapy 40 months after the treatment. Sclerotherapy is less invasive treatment option for varicose veins with incompetent GSV with satisfactory clinical and cosmetic outcome.