Objective To investigate the role of opioid receptors in remifentanil-induced attenuation of renal ischemia/reperfusion (I/R) injury in rats.Methods Seventy-five male Sprague-Dawley rats weighing 250-300 g were randomly divided into 5 groups ( n =15 each):sham operation group (group S),group I/R,remifentanil group (group R),naloxone group (group N),and naloxone + remifentanil group (group NR).Renal ischemia was induced by clamping the bilateral renal arteries for 45 min using an atraumatic clamp followed by reperfusion.In groups R and NR,remifentanil was infused at 1.0 μg· kg-1 · min-1 via the caudal vein starting from 15 min before ischemia until 30 min of reperfusion,while groups S,I/R and N received the equal volume of normal saline instead of remifentanil.In groups N and NR,naloxone 0.3 mg/kg was injected via the caudal vein at 20 min before ischemia and at 35 min after ischemia respectively,while groups S,I/R and R received the equal volume of normal saline instead of naloxone.Blood and urine samples were collected from the femoral vein and urinary bladder respectively at 24 h of reperfusion for determination of the levels of serum creatinine (Cr) and blood urea nitrogen (BUN),urinary N-acetyl-β-D-glucosaminidase (NAG) and γ-glutamyl transpeptidase (γ-GT).The rats were sacrificed at 24 h of reperfusion and the renal tissues were removed for determination of nalondialdehyde (MDA) content and superoxide dismutase (SOD) activity.Pathological changes in renal tissues were observed with light microscope.Results Compared withgroup S,the levels of serum Cr and BUN,urinary NAG and γ-GT,and MDA were significantly increased,while the activity of SOD was significantly decreased in the other 4 groups ( P ＜ 0.05 or 0.01 ) and pathological changes in renal tissues were observed in the other 4 groups.Compared with group I/R,the levels of serum Cr and BUN,urinary NAG and γ-GT levels,and MDA were significantly decreased,while the activity of SOD was significantly increased ( P ＜ 0.01 ),and the pathological changes were reduced in group R,and no significant change was found in the parameters mentioned above in groups N and NR ( P ＞ 0.05).The pathological changes were similar in groups I/R,N and NR.Compured with group R,serum Cr and BUN concentrations,urinary NAG and γ-GT levels and MDA concent were increased,while SOD activity were decreased ( P ＜ 0.05 or 0.01 ).Conclusion Opioid receptors mediate remifentanil-induced attenuation of renal I/R injury in rats.

Objective To investigate the effect of remifentanil on nucleotide-binding oligomerization domain 1 (NOD1) mRNA expression in rats with renal ischemia-reperfusion (I/R) injury.Methods Sixty male Sprague-Dawley rats,weighing 220-250 g,were randomly divided into 3 groups (n =20 each):sham operation group (S group),I/R group and remifentanil group (R group).Renal ischemia was induced by occlusion of bilateral renal arteries for 45 min followed by 24 h reperfusion in groups I/R and R.Remifentanil 1.0 μg· kg-1 · min-1 was infused until 30 min of reperfusion starting from 15 min before ischemia in group R,while the equal volume of normal saline was given instead in S and I/R groups.The animals were sacrificed at 15 min before ischemia and at 3,6,24 h of reperfusion and the kidneys were removed for microscopic examination and polymorphonuclear leukocyte (PMN) count and for measurement of NOD1 mRNA expression (by RT-PCR).The apoptotic rate was determined by flow cytometry double staining method.Results Compared with group S,NOD1 mRNA expression was up-regulated,and the apoptotic rate and PMN count were significantly increased at each time point during reperfusion in group I/R,and the apoptotic rate and PMN count were significantly increased at each time point during reperfusion,and NOD1 mRNA expression was up-regulated at 6 and 24 h of reperfusion in group R (P ＜ 0.01).Compared with I/R group,NOD1 mRNA expression was down-regulated,and the apoptotic rate and PMN count were significantly decreased at each time point during reperfusion (P ＜ 0.05 or 0.01),and the pathological changes were significantly attenuated in group R.Conclusion Remifentanil can reduce the renal I/R injury by down-regulating the expression of NOD1 mRNA and inhibiting inflammatory response and apoptosis.

Objective To evaluate the effect of remifentanil on Toll-like receptor 2 (TLR2) mRNA expression during renal ischemia-reperfusion (Ⅰ/R) in rats.Methods Fifty-four male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 3 groups (n=18 each) using a random number table:sham operation group (group S),Ⅰ/R group and remifentanil group (group R).Renal Ⅰ/R injury was produced by clamping the bilateral renal arteries for 45 min followed by reperfusion in Ⅰ/R and R groups.Bilateral renal arteries were only exposed but not clamped in group S.Remifentanil 1.0 μg · kg-1 · min-1 was infused via the tail vein starting from 15 min before ischemia until 30 min of reperfusion in group R,while the equal volume of normal saline was given instead in S and Ⅰ/R groups.The animals were sacrificed at 15 min before ischemia and 6 and 24 h of reperfusion,and the renal specimens were obtained for examination of the pathological changes (with light microscope) and for determination of the contents of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) (by ELISA) and expression of TLR2 mRNA (by RT-PCR) and cell apoptosis (by double staining and flow eytometry).The apoptotic rate was calculated.Results Compared with group S,TLR2 mRNA expression was significantly up-regulated,and the contents of TNF-α and IL-6 and apoptotic rate were increased at 6 and 24 h of reperfusion in Ⅰ/R and R groups.Compared with group Ⅰ/R,TLR2 mRNA expression was significantly down-regulated,and the contents of TNF-α and IL-6 and apoptotic rate were decreased at 6 and 24 h of reperfusion in group R.The pathological changes were significantly attenuated in group R as compared with group Ⅰ/R.Conclusion The mechanism by which remifentanil reduces renal Ⅰ/R injury is related to down-regulation of TLR2 expression and decrease in TLR2 activity and inhibition of inflammatory responses in renal tissues and cell apoptosis in rats.

Objective Systemic analysis the effect of the application of continuity nursing in community diabetes.Methods Retrieve documents from the PubMed,Cochrane Library,EMBASE,Science Direct,CBM,WANFANG Data and CNKI.Dating from the dates of establishment to May,2014.Randomized controlled trials (RCTs) on continuity of patient care for patients with diabetes were included.Two reviewers independently screened the literature according to the inclusion and exclusion criteria,extracted the data and assessed the quality of the included studies,and then meta-analysis was performed by RevMan 5.1 software.Results 5 RCTs (156 cases with diabetes) were involved into this Meta-analysis.The experimental group was superior to the control group in FBG,2h PBG,HbA1c and the level of treatment compliance,with statistically significant differences,P＜0.05.The descriptive analysis showed that the differences in the score of diabetes knowledge and the quality of life were statistically significant,P＜0.05,because the data could not be converted.Conclusions The continuity nursing can improve the compliance and the quality of life of community diabetes,ameliorate certain biochemical indicators.Study of large sample and RCTs should be take because of the existence of some restraining factors.

Objective To investigate the effect of remifentanil on protein kinase C (PKC) activity during renal ischemia-reperfusion (I/R) in rats.Methods Seventy-five male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 5 groups (n=15 each) using a random number table:sham operation group (group S),I/R group,remifentanil group (group R),naloxone group (group N),and naloxone + remifentanil group (group NR).Renal ischemia was induced by clamping the bilateral renal arteries for 45 min using an atraumatic clamp followed by reperfusion.In R and NR groups,remifentanil 1.0 μg · kg-1 · min-1was infused via the caudal vein starting from 15 min before ischemia until 30 min of reperfusion.In N and NR groups,naloxone 0.3 mg/kg was injected via the caudal vein at 20 min before ischemia and 35 min of ischemia,respectively.The rats were sacrificed at 24 h of reperfusion and the kidneys were removed for determination of the ultrastructure of the renal tubular epithelial cells (using transmission electron microscope),activity of PKC in renal tissues (by ELISA),and expression of the PKC in renal tissues (by immuno-histochemistry).Results Compared with group S,the activity of PKC in renal tissues was significantly increased in the other four groups,and the expression of the PKC in renal tissues was up-regulated in group R.Compared with group I/R,the activity of PKC in renal tissues was significantlyincreased,the expression of PKC in renal tissues was up-regulated,and the pathological changes were attenuated in group R.Compared with group R,the activity of PKC in renal tissues was significantly decreased,the expression of PKC in renal tissues was down-regulated,and the pathological changes were aggravated in N and NR groups.Conclusion The mechanism by which remifentanil attenuates renal I/R injury may be related to up-regulation of PKC expression and increase in PKC activity through activating opioid receptors in rats.

Leukoencephalopathy with vanishing white matter(VWM) is one of the most prevalent inherited white matter disorders in childhood,and it′s the only known hereditary human disease due to the direct defects in protein synthesis process,with the gene defects in EIF2B1-5,encoding the five subunits of eukaryotic translation initiation factor(eIF2B ?,?,?,? and ?) respectively.eIF2B is essential for the protein translation initiation process,and its action is realized via eukaryotic translation initiation factor2(eIF2).Phosphorylation of eIF2? and eIF2B? is an important way to regulate eIF2B function,and thus play a key role in control of the protein translation level under physiological condition.Mutant eIF2B results in functional defects and decrease of the overall protein translation in cells,but in increase the translation of proteins with multiple upstream open reading frames,such as activating transcription factor 4(AFT4),which leads to the susceptibility to un-folded protein response under stress,and the following apoptosis.The exact pathogenic mechanisms of VWM are far from well understood.It′s suggested that level of AFT4 in cells with eIF2B mutations is higher than in wild type cells under physiological condition,which makes the mutant cells more susceptible to endoplasmic reticulum(ER) stress and unfolded protein response(UPR).Under stress,the defect eIF2B leads to a vicious cycle of UPR activation,which may underlie the neurological aggravation in VWM patients after minor stress,a specific cli-nical feature of VWM.Elucidating the pathogenesis of VWM will be helpful to further understand the protein translation process in eukaryotic cells,and provide a clue for possible therapeutic targets and treatment strategies in the future.Abstract:SUMM ARY Leukoencephalopathy with vanishing white matter(VWM) is one of the most prevalent in-herited white matter d isorders in childhood,and i′ts the only known hered itary human d isease due to the d irect defects in protein synthesis process,with the gene defects inEIF2B1-5,encod ing the five sub-units of eukaryotic translation initiation factor(eIF2B?,?,?,?and?) respectively.eIF2B is essential for the protein translation initiation process,and its action is realized via eukaryotic translation initiation factor2(eIF2).Phosphorylation of eIF2?and eIF2B?is an important way to regulate eIF2B function,and thus play a key role in control of the protein translation level under physiological cond ition.Mutant eIF2B results in functional defects and decrease of the overall protein translation in cells,but in increase the translation of proteins with multiple upstream open read ing frames,such as activating transcription factor 4(AFT4),which leads to the susceptibility to un-folded protein response under stress,and the following apoptosis.The exact pathogenic mechanisms ofVWM are far from well understood.I′ts sugges-ted that level ofAFT4 in cells with eIF2B mutations is higher than in wild type cells under physiological cond ition,which makes the mutant cellsmore susceptible to endoplasm ic reticulum(ER) stress and un-folded protein response(UPR).Under stress,the defect eIF2B leads to a vicious cycle ofUPR activa-tion,which may underlie the neurological aggravation in VWM patients afterm inor stress,a specific cli-nical feature ofVWM.E lucidating the pathogenesis ofVWM will be helpful to further understand the pro-tein translation process in eukaryotic cells,and provide a clue for possible therapeutic targets and treat-ment strategies in the future.

Objective To evaluate the effect of remifentanil on cell apoptosis during renal ischemia-reperfusion (I/R) in rats.Methods Seventy-five male Sprague-Dawley rats,weighing 220-250 g,were randomly divided into 3 groups (n =25 each):sham operation group (group S),I/R group,and remifentanil group (group R).Renal ischemia was induced by occlusion of the bilateral renal arteries for 45 min followed by reperfusion in groups I/R and R.Remifentanil was infused at 1.0 μg· kg-1 · min-1 via the caudal vein starting from 15 min before ischemia until 30 min of reperfusion in group R,while the equal volume of normal saline was given instead of remifentanil in groups S and I/R.At 15 min before ischemia (T0) and 3,6,12,24 h of reperfusion (T1-4),5rats were anesthetized and sacrificed,and renal specimens were obtained to detect the apoptotic rate and expression of Bax and Bcl-2 protein (by flow cytometry) and mRNA (by RT-PCR).The ratios between Bcl-2/Bax protein and mRNA expression were calculated.The pathological changes of renal tubules were scored.Results Compared with group S,the pathological scores and apoptotic rate were significantly increased at T1-4,and ratios between Bcl-2/Bax protein and mRNA expression were increased at T1,2,while decreased at T3,4 in groups R and I/R (P ＜0.01).Compared with group I/R,the pathological scores and apoptotic rate were significantly decreased at T1-4,while the ratios between Bcl-2/Bax protein and mRNA expression were increased in group R (P ＜ 0.05 or 0.01).Compared with the baseline value at T0,the pathological scores and apoptotic rates were significantly increased at T1 4,and the ratios of Bcl-2/Bax protein and mRNA expression were increased at T1,2,while decreased at T3,4 in groups R and I/R (P ＜ 0.01).Conclusion Regulation of Bcl-2/Bax expression and inhibition of cell apoptosis in renal tissues are involved in the mechanism by which remifentanil reduces renal I/R injury in rats.

AIM: To investigate the effect of dihydroartemisinin (DHA) on the proliferation of murine T lymphocytes stimulated by Con A in vitro and its related immunosuppressive mechanism. METHODS: Murine T lymphocytes were stimulated by Con A and treated with different concentrations of DHA. Cell proliferation was measured by carboxyl fluoresce in diacetate succinmidyl ester (CFDA-SE) staining. The expression of CD69, CD25 and CD71,which was the marker of early, middle, later activation of CD3~+ T lymphocytes, was measured by flow cytometry (FCM) combined with two-color immunofluorescent staining of cell surface antigen. Fluorescence calcium indicator fluo-4/AM was used to measure the change of the intracellular calcium concentration ([Ca~(2+)]_i) of murine T lymphocytes. The distribution of the cell cycle was analyzed by PI staining. The expression of CD69, the early activation antigen on CD4~+CD25~(high) Treg was also measured by FCM combined with three-color immunofluorescent staining. RESULTS: The result of CFDA-SE staining showed that DHA efficiently inhibited the Con A-induced proliferation of T-lymphocytes in a time-and dose-dependent manners. DHA showed modestly increased proportions of CD69 and CD25 on Con A-stimulated CD3~+T cells, but inhibited the expression of CD25 in a dose dependent manner. DHA with Con A, but not DHA alone, caused an increase in intracellular calcium concentration of T cells. The results of FCM analysis with PI staining showed that DHA imposed a total cell cycle arrest in G_0/G_1 and prevented cells entering S phase and G_2/M phase. Furthermore, DHA reduced the expression of CD69 on CD4~+CD25~(high) Treg. CONCLUSION: DHA, which exhibits immunosuppressive effect on the proliferation of murine T-lymphocytes, is promising to be developed as an immunosuppressive reagent.

BACKGROUND:In vitro experiments have demonstrated that the biodegradable mesh-like microporous baloon made of macromolecular materials has obvious advantage of anti-leakage, which is capable of maintaining calcium homeostasis, has no inhibitory effects on cel growth and on microscopic interdigitation formation between new bone and bone cement. OBJECTIVE:To evaluate the therapeutic effects of biodegradable mesh-like microporous baloon with calcium bone cement on vertebral fractures based on animal experiments. METHODS:The fracture model was established in 48 New Zealand rabbits, in which a bone dril was introduced after successful puncture at sites near left low extremity of the femur. These rabbit models were randomized into two groups: experimental group with calcium phosphate bone cement and biodegradable mesh-like microporous baloon and control group only with calcium phosphate bone cement. Clinical parameters such as blood cel count, biochemistry, and CT/X ray were examined at 1, 3 and 6 months after implantation of the baloon and bone cement. After that, the specimens were fixed for pathological analysis. RESULTS AND CONCLUSION:The operation was performed under general anesthesia with no eventful infusion of bone cement. The expansion of baloon was satisfactory without definite extravasation of bone cement in the experimental group. In the control group, cement diffusion was found with pulmonary embolism occurring in three New Zealand rabbits. No statistical significance for blood cel counts and biochemistry was found between pre- and postoperation or between two groups. The materials in the two groups had favorable biocompatibility with injured bones without obvious immunological response. In the experimental group, the baloon wal was thinned and partial bone tissues grew into the cement at 1 month; at 3 months, a large amount of bone tissues grew into the cement and cement volume diminished; at 6 months, the baloon disappeared and only a smal amount of cement left in the bone tissues. In the control group, it was difficult to determine when the cement degraded. The biodegradable mesh-like microporous baloon combined with calcium bone cement is superior to bone cement alone in the management of vertebral fractures.

Objective To establish a scientific and sensitive evaluation index system of nursing quality for cancer hospital. Methods The evaluation index system of nursing quality for cancer hospital was formulated based on literature,semi- structured interviews and expert group discussion. Then, developing the evaluation index system of nursing quality for cancer hospital by two rounds of Delphi consultation. Results The experts′ authority coefficient was 0.862. The nursing quality indicators included 5 first-level indicators,9 second-level indicators and 73 third-level indicators. The Kendall coordination coefficients of the importance of the three level indicators were 0.354,0.217,and 0.243, The Kendall coordination coefficients of the feasibility of the three level indicators were 0.234,0.313,and 0.339. Conclusions Scientific nature and concentration indicator system of nursing quality for cancer hospital was developed. It will be used to provide quantitative basis for the control of nursing quality in cancer hospital.