Objective To discuss the safety of using etomidate combined with remifentanil by target controlled infusion ( TCI) for painless bronchofibroscopy. Methods Sixty patients were divided into two groups: painless bronchoscopy group (treatment group, 24 patients) and the routine bronchoscopy group (control group, 36 patients). Treatment group received TCI of remifentanil and intravenous injection of etomidate fat emulsion. Control group was subjected to surface anesthesia with 2%lidocaine. SpO2 , blood pressure, heart rate and breath changes during examination and complete awakening were continuously monitored. Bronchofiberscopy time, body movement during examination, bucking and satisfaction degree after examination were also recorded. Results Treatment group patients felt senseless and painless during bronchoscopy, without memory of bronchoscopy and pain. Patients in control group had discomfort, body movement and acute bucking, and most of them had painful memory. There were significant differences between the two groups (P<0. 01). In treatment group, after examination, blood pressure, respiratory frequency, heart rate and SpO2 were significantly decreased (P<0. 01). During examination, the blood pressure, respiratory frequency and heart rate were increased, and SpO2 decreased in control group compared to the baseline (P<0. 01). There was no significant difference in SpO2 between treatment group and control group during examination (P>0. 05). Conclusion TCI etomidate combined with remifentanil during bronchoscopy achieved satisfying anesthetic effect.

Objective:To investigate the mechanisms of IL-1β promoted lung cancer cells proliferation. Methods: The“Transwell? Inserts” system was used to coculture lung cancer cells A549,NCI-H520 with macrophages. BrdU ELISA used to measure the effect of macrophages promoted lung cancer cells proliferation. Expression of mRNA of IL-1β in A549 and NCI-H520 cells were analysed by Real-time PCR analysis. IL-1β was responsible for macrophage-promoted lung cancer cells growth, IL-1β neutralizing antibody was added. The autophagy marker Beclin1 protein was detected by Western blot. Results:The BrdU ELISA assay showed that after coincubation with macrophages in the proportion of 1:0. 5,the OD value of A549 increased from(0. 41±0. 06)to(1. 13±0. 10). There was statistical significance(P<0. 05). It also showed that the growth of the A549 cell was dependent on the macrophage number (P<0. 05). The OD value variability of NCI-H520 cells was as same as A549 cell upon cocultured with macrophages. Real-time PCR results showed that the expression of IL-1β mRNA in macrophages was remarkably enhanced in a time dependent manner upon coincubated with lung cancer cell,and the expression level was higher than lung cancer cells. Addition of IL-1β neutralizing antibody markedly inhibited macrophage-promoted lung cancer cells proliferation. The OD value of these two cells were decreased from ( 3. 63 ± 0. 33) to (1. 46±0. 18),from (2. 94±0. 38) to (1. 53±0. 20),respectively (P<0. 05). After treatment with IL-1β,the expression of Beclin1 was significantly inhibited in tumor cells. Conclusion:Over-expression of IL-1βfrom macrophages and lung cancer cells is re-sponsible for proliferation of tumor cells in coculture condition. Inhibition of autophagy in tumor cells may be the important mechanisms of IL-1β promotes lung cancer cells proliferation.

Objective To explore the dynamic changes of serum S100B and glial fibrillary acidic protein (GFAP) levels and their clinical significance in patients with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP).Methods By means of enzyme-linked immunno-sorbent assay (ELISA),the serum S100B and GFAP levels from 33 DEACMP patients were assayed,and the condition changes were analyzed with three types of scale:the activity of daily living scale ( ADL),information-memory-concentration test (IMCT) and Hasegawa' s dementia scale(HDS).The comparison with 32 patients of acute carbon monoxide poisoning without DEACMP was also conducted.Results (1) The serum S100B( (0.60 ±0.21)ng/ml) and GFAP( (226.58 ±90.05 )ng/ml) in DEACMP group at acute stage were significantly higher than those in acute-CO-poisoning group ( (0.50 ± 0.20) ng/ml,( 183.04 ± 73.01 ) ng/ml) and those in DEACMP group at convalescent stage ( (0.51 ±0.16) ng/ml,(183.25 ±81.76)ng/ml) (all P values ＜0.05).(2)In DEACMP group,the serum S100B and GFAP at acute and convalescent stages were significantly correlated (at acute stage:r=0.466,P=0.006; at convalescent stage:r=0.365,P=0.037 ).(3)The S100B and GFAP in ineffective DEACMP patients at acute stage were significantly higher than those in the other groups ( all P values ＜ 0.05 ).(4) In DEACMP group,the ADL,HDS and IMCT scores( (45.21 ± 9.69),(8.26 ± 6.31 ),(9.91 ± 7.52) ) at acute stage were significantly different from those at convalescent stages( (33.67 ± 13.62),( 15.91 ± 10.83),( 19.06 ± 10.37 ) ) ( all P values ＜0.01 ).Conclusion There is secondary brain insult (SBI) in DEACMP; glial activation may play an important role.The S100B and GFAP levels may be associated with the prognosis of DEACMP.

Objective To explore neurobiological mechanisms of the withdrawal-induced aversion,the changes of protein kinase A(PKA) were measured in shell of accumbens nucleus (AcbSH) of CPA model rats.Methods 1.All 36 male SD rats were divided into three groups,model group ( MN group),and control group (MS group and SN group).MN group was injected with morphine,6.5 days,10mg/kg,intraperitoneally (IP),twice per day,naloxone injection,0.3 mg/kg,ip,along with conditioned place aversion training,to develop the CPA model.The MS group was administrated equivalent volume of morphine and saline.Also the SN group was injected with equivalent volume of saline and naloxone.2.During the development of CPA,the expression of protein kinase A was assayed with immunohistochemistry in the AcbSH.Results Before the development of CPA,PKA expressions were no significant differences among the three groups in the AcbSH (F=2.306,P=0.130).However,after development of CPA,PKA expressions showed significant differences among the three groups(F =36.516,P =0.000).The average gray intensity of MN group (109.50 ± 4.661 ) was apparently higher than the MS group (126.50 ±3.697,P＜0.01),than the SN group (133.50 ±6.364,P＜0.01).Conclusions 1.Protein kinase A expression,leading to the aversion in the AcbSH probably is a key pathway contributing to the development of CPA.2.The neuroadaptation mediated by PKA may be one of important molecular underpinnings of CPA.

objective To establish a new method for isolation and identification of mixed erythrocytes with different blood groups from the patients following ABO-incompatible allogeneic haematopoietic stem cell transplantation(allo-HSCT),and investigate its application in post-transplantation survival erythrocytes.Methods The erythrocyte blood group antigens from the patiehts following ABO-incompatible allo-HSCT were agglutinated by the antibodies known.centrifuged at 800×g and 50×g to isolate agglutinated and unagglutinated erythrocytes respectively step by step.These erythrocytes collected were counted and identification respectively.Results The sensitivity,accuracy,recovery and reproducibility of the new method were 1%,100%,92.5%and 100%,respectively.The post-transplantation survival erythrocytes mixed with patient's erythrocytes from 18 patients following ABO-incompatible allo-HSCT were successfully isolated and identified by provided methods.It was the first time to identify the survival erythrocytes 11 th to 72 th day after allo-HSCT.The amount of the survival erythrocytes had been changing following post-transplantation periods.Conclusions The method can be widely employed in the isolation and identification of mixed erythrocytes with different blood groups.It provides reliable method to study the clinic significance of the changes of the survival erythroeytes from the patients following ABO-incompatible allo-HSCT.

Objective To study the correlation of HLA-G levels with acute rejection and CMV active infection post-kidney transplantation.Methods A total of 132 initial kidney transplantation recipients were divided into kidney function stable group (F),acute rejection group (AR),CMV group according to whether they had active CMV infection and acute rejection.Forty-one healthy donors served as control group (H).HLA-G levels and mRNA expression were analyzed by using flow cytometry,ELISA,RT-PCR and Western blotting.Immunohistochemical staining was used to detect the HLA-G expression in kidney biopsies.Results The expression levels of mHLA-G1 were low in all 4 groups pre-transplantation.Only CMV group had significantly more CD14+ mHLA-G1+ cells post-transplantation (P＜0.05).sHLA-G5 levels were higher in F group than in H group (P＜0.05),but there was no significant difference among other groups pre-transplantation (P＞0.05).sHLA-G5 levels were increased significantly in CMV group as compared with F group (P＜0.05),and those in F group were higher than in H and AR groups (P＜0.05).Renal tissue biopsies from 21 renal transplantation recipients with AR indicated that HLA-G5 was expressed negatively in 17 patients,positively in 3 patients and 1 weakly positively.HLA-G was positive in the kidney tissue of 9 patients out of 9 patients with active CMV infection.In total 132 recipients,AR incidence was significantly lower in CMV ( + ) group (7.1 ％,2/28) than that in CMV ( - ) group (24.0 ％,25/104).Conclusion The sHLA-G5 may contribute to predict AR and CMV active infection; AR and CMV active infection may be correlation with immune balance in kidney transplantation recipients.

Objective To measure the cytokines levels in peripheral blood from kidney transplantation recipients by using cytometric bead array and to analyze their change and the clinical significance in pre- and post- kidney transplantation, inducting with basiliximab and graft rejection. Methods A total of 72 renal transplantation recipients were divided into two groups, kidney function stable group(n =53) and acute rejection group (n = 19). And they were also grouped by induction with basiliximab or not,32 in basiliximab group and 40 in without basilixmab group. The levels of IFN-γ, TNF-α, IL-10, IL-5,IL-4, IL-2 were measured by cytometric bead array in peripheral blood of 72 kidney transplantation recipients and 30 healthy donors at differential time. The data was analyzed according to the following grouping:donors and recipients, kidney function stable group and acute rejection group post transplantation and with or without basiliximab group. Results The levels of TNF-α, IL-10, IL-5, IL-4, IL-2 in recipients before transplantation were ( 1.65 ±0. 10) ,(2. 55 ±0. 19) ,( 1.88 ±0. 14) ,(1.85 ±0. 12) ,(2. 12 ±0. 09) ng/L,respectively. While they were (3.04 ±0. 17), (3.33 ±0. 26), (4.03 ±0.25), (2.73 ±0. 16), (4.03 ±0. 26) ng/L respectively in healthy donors. There was statistical significance between the two groups ( t =6. 890, 2. 375, 7. 851,3.955,7.153, P<0. 01, <0. 05, <0.01, <0.01, <0.01). While the level of IFN-γ in recipients before transplantation was (2. 50 ±0. 18) ng/L,compared with (3. 00 ±0. 24) ng/L in healthy donors. There was no statistical significance between the two groups( t = 1. 625, P > 0. 05 ). The levels of IFN-γ and IL-10 in kidney function stable group were (2. 71 ± 0. 11 ) ng/L and (3.91 ± 0. 52) ng/L,while they were ( 3.30 ± 0. 36 ) ng/L and ( 12. 01 ± 5.35 ) ng/L in acute rejection group. There were statistical dirrerences between the two groups ( t = 5. 061, 11. 465, P < 0. 01, < 0. 05 ). Before induction with basiliximab, the levels of IFN-γ, TNF-α, IL-10 in recipients were (2.90 ±0. 21 ), ( 1.67 ±0. 12),(2. 45 ± 0. 16) ng/L respectively. But they were ( 2. 78 ± 0. 17 ), ( 1.58 ± 0. 07 ), ( 2. 77 ± 0. 24 ) ng/L respectively after induction with basiliximab, which showed significantly different ( t = 5. 605, 6.011,4. 126, P <0. 01, <0. 01, <0. 05). Four weeks after kidney transplantation in recipients with basiliximab,the levels of IFN-γ, IL-10, IL-4 were (2. 90 ± 0. 31 ), (9. 08 ± 0. 16), (2. 73 ± 0. 11 ) ng/L. While they were (3.28 ±0. 11 ), (4. 17 ±0. 21 ), (2. 11 ±0. 20) ng/L respectively in recipients without basiliximab induction, which were significantly different from those with basiliximab induction (t = 4. 268,4. 263,3.762, P <0. 01, <0. 01, < 0. 05 ). Conclusions Six kinds of cytokines can be measured by cytometric bead array simultaneously and accurately. The data suggests that the detection of multiple cytokines in kidney transplantation recipients by cytometric bead array can provide more guidance for clinical diagnosis and therapy.

ObjectiveTo determine the correlation of human leukocyte antigen-G (HLA-G)expression with CMV active infection after kidney transplantation. MethodsA total of 215 first-time kidney transplantation recipients in one transplantation center were divided into CMV ( + ) group and CMV ( - ) group according to whether they had active CMV infection. mhla-g1 expression on leukocytes was analyzed by flow cytometry. The concentrations of soluble HLA-G5 were detected by using ELISA. The sHLA-G5 cutoff levels by ROC curve was employed to predict the active CMV infection. The expression of sHLA-G5 mRNA and protein in leukocytes was analyzed by using RTPCR and Western blotting respectively. Immunohistochemical staining was used to detect the HLA-G expression in kidney biopsies of 12 cases. ResultsThe expression of mHLA-G1 in peripheral blood was low in both CMV ( + ) group and CMV ( - ) group. Also when CMV-PP65 was positive, there was no significant change in mHLA-G1. In CMV ( + ) group, the proportion of CD14+ mHLA-G1 +cells[(45. 53 ± 17.32)％]in peripheral blood was increased as compared with that in CMV (-)group[(10. 22 ± 5.78)％]. The expression of sHLA-G5 was increased significantly in CMV ( + )group. The optimal cutoff value of sHLA-G5 predicting the active CMV infection was 202. 9 μg/L,with high diagnostic accuracy. HLA-G was positive in the kidney tissue of 10 patients out of 12 patients with active CMV infection. Both RT-PCR and Western blot analysis showed that sHLA-G5 was significantly higher in CMV ( + ) group than that in CMV ( - ) group. ConclusionROC curve analysis of sHLA-G5 with the cutoff value of 202. 9 μg/L can be used to predict the active CMV infection. The HLA-G levels in peripheral blood were significantly increased and HLA-G expression in the tubular epithelial cells of the graft could be a protection mechanism of the kidney function.

Objective To investigate the effect of maternal deprivation (MD) on neurobehavior and PP1Cγgene expression in hippocampus. Methods Male pups were randomly divided into MD group(thirty-five)and control group(twenty-four). From PND 1 to PND 21 ,pups in the MD groups underwent daily maternal deprivation for 3 h ( Postnatal day). Neurobehavior was observed to investigate neurodevelopment, Morris water maze was used to measure spatial learning and memory,and Real-Time quantitative PCR was employed to analyze PP1Cγ gene expression. Results Several significant deficiencies were observed in bodyweight and grasping reflex while a great enhancement in hot-plate test in rat pups suffering from MD( (26.23 ± 2.81 )g vs. (30. 38 ± 3.85 )g;( 19.37 ± 11.89) s vs. (22.39 ± 17.62 ) s; (4.36 ± 1.76 ) s vs. ( 5.26 ± 2.55 ) s; P ＜ 0. 05 ), but deficiencies in neurological reflexes were subtle ( ( 0.83 ±- 0.30 ) s vs. ( 0. 83 ± 0. 34 ) s; ( 3.68 ± 1.63 ) s vs. ( 5.61 ± 3. 01 ) s;( 3.00 ± 0.00 ) vs. ( 3.00 ± 0. 00); P ＞ 0. 05 ). MD had a subtle influence on spatial learning and memory (P ＞0.05). Meanwhile,MD could lead to PP1Cγ expression down-regulation on PND 22 ( (2.19 ±0.62) vs. (3.52 ±0.86), P＜0. 05)which was in line with early neurobehavior results. No difference was found compared with MD group and control group on PND60 ( ( 1.73 ± 0. 78 ) vs. ( 1.33 ± 0. 34); P ＞ 0.05 ). However, there was the up-regulation of PP1Cγexpression on PND 90 ( (2.85 ± 0. 34) vs. ( 1.34 ± 0.93 ); P ＜ 0.05 ). Conclusion MD alters early neurobehavior and hippocampal PP1Cγgene expression in the Wistar rats,but has a subtle effect on learning and memory. At the same time,MD can make PP1Cγexpression in the hippocampus varying with the age.

Objective To study the effects of tacrolimus(Tac) concentrations on the number of NK cells and receptor expression in peripheral blood of renal transplantation receptors.Methods A total of 60 first-time kidney transplantation recipients in our institute from Dec.2007 to July 2009 were followed up.Tac maintenance immunosuppressive therapy was given to all recipients.The recipients were divided into low-concentration Tac group (6.84 + 1.72μg/L,n =30) and highconcentration Tac group ( 11.88 + 2.59 μg/L,n =30) according to concentrations of Tac.Twenty healthy volunteers served as controls.Before and 6 months after operation,concentrations of Tac were analyzed by using micro particle immunoassay chemiluminescent method.NK cells and their receptors (CD85j,CD158d,CD94 and NKG2D) were detected by using flow cytometry.The concentrations of soluble HLA-G5 were detected by using ELISA.Results The number of NK cells in lowconcentration Tac group and high-concentration of Tac group preoperatively was significantly reduced as compared with control group (P ＜ 0.05 ). The percentage and number of NK cells in low concentration Tac group and high-concentration Tac group at 6th month after operation were significantly reduced as compared with control group (P＜0.05).The number of NK cells in lowconcentration Tac group was significantly greater than in high-concentration Tac group (P＜ 0.05).There was no significant differende in the expression of CD85j,CD158,CD94 and NKG2D before operation between two groups(P＞0.05).The expression of CD85j and CD158d in two groups was increased,but that of CD94 and NKG2D was decreased at 6th month post-transplantation as comapred with that preoperation.In low-concentration Tac group,the expression of CD85j and CD158d was increased as compared with that in high-concentration Tac group (P＜0.05 ).Spearman correlation analysis revealed that the CD85j and CD158d expression had a positive correlation with sHLA-G5(P＜0.01 ),but the NKG2D had a negative correlation with sHLA-G5(P＜0.01 ).Conclusion There was correlation between the concentrations of Tac and NK cells count and NK receptors. Low concentrations of Tac can safely and effectively protect kidney function.The number of NK cells andtheir inhibitor receptors are increased in the recipients with low concentration of Tac.