Objective To evaluate the role of P2X3 receptors in dorsal root ganglion in development of incisional pain in rats.Methods Twenty-four healthy male SD rats weighing 200-220 g were randomly divided into 3 groups(n = 8 each): control group(group C),incisional pain(group IP)and P2X3 receptor antagonist + IP group(group A).In group IP and A,a 1 cm longitudinal incision was made in the plantar surface of left hindpaw according to the method described by Brennan et al.in isoflurane-anesthetized rats.P2X3 receptor antagonist TNP-ATP 200 nmol was injected into the plantar surface of left hindpaw 30 min after plantar incision was made in group A,while equal volume of normal saline was given instead of TNP-ATP in group C and IP.The behavior of the hindpaw of the rats were assessed using cumulative pain score within 1 h after injection.The animals were sacri ficed 2 h after injection and the dorsal root ganglion was removed for determination of P2X3 receptor expression and intracellular Ca2+ concentrations.ResultsThe cumulative pain scores,P2X3 receptor expression and Ca2 + concentrations were significantly higher in group IP and A than in group C(P ＜ 0.05).The cumulative pain scores,P2X3 receptor expression and Ca2+ concentrations were significantly lower in group A than in group IP(P ＜0.05).Conclusion P2X3 receptors in dorsal root ganglion is involved in the development of incisional pain through increasing intracellular Ca2+ concentrations in rats.

Objective To evaluate the role of the expression of heat shock protein 70 (HSP70) and heme oxygenase-1 (HO-1) in the reduction of renal ischemia-reperfusion (I/R) injury by ischemic postconditioning in tats.Methods One hundred and forty healthy male SD rats weighing 250-280 g were randomized into 4 groups ( n = 35 each) : sham operation group (S group) ; I/R group; ischemic postconditioning group (IPo group); quercetin (an inhibitor of HSP) + ischemic postconditioning group (Q + IPo group). Renal I/R was produced by clamping bilateral renal pedicels for 45 min followed by reperfusion. In group S, bilateral kidneys were only exposed through a midline incision but their- pedicels were not clamped. In IPo and Q + IPo groups, 45 min ischemia was followed by three 10 s episodes of ischemia at 10 s intervals for reperfusion and in addition intraperitoneal quercetin 100 mg/kg was injected at 1 h before ischemia in group Q + IPo. Blood samples from hearts were obtained at 0, 1, 3, 6, 12, 24 and 48 h of reperfusion (T0-6) and the rats were then sacrificed and kidneys removed to detect the expression of HSP70 and HO-1 mRNA and protein in renal tissues. The blood samples obtained at T3 were used to determine serum creatinine (Cr) and urea nitrogen (BUN) concentrations and the expression of caspase-3 mRNA . The apoptosis in the renal tissues was detected using TUNEL and apoptotic index ( AI) was calculated. Microscopic examination was performed with light microscope. Results Compared with group S, the serum Cr and BUN concentrations and AI were significantly increased at T3,the expression of caspase-3 mRNA was up-regulated at T3, and the expression of HSP70 and HO-1 mRNA and protein was up-regulated at T0-6 in the other groups (P ＜ 0.05) . Compared with group I/R, the serum Cr and BUN concentrations and AI were significantly decreased at T3, the expression of caspase-3 mRNA was down-regulated at T3, and the expression of HSP70 and HO-1 mRNA and protein was up-regulated at T1-5 in group IPo ( P ＜ 0.05) . Compared with group IPo, the serum Cr and BUN concentrations and AI were significantly increased at T3, the expression of caspase-3 mRNA was up-regulated at T3, and the expression of HSP70 and HO-1 mRNA and protein was down-regulated at T1-5, in group Q + IPo ( P ＜ 0.05) . The microscopic examination showed that the renal I/R injury was significantly attenuated by ischemic postconditioning and the degree of injury in group IPo was similar to that in group I/R. Conclusion The expression of HSP70 and HO-1 is involved in the reduction of renal I/R injury by ischemic postconditioning in rats.

Objective To evaluate the effects of anisodamine on apoptosis in cardiomyocytes and inflammatory response in overtrained rats. Methods Twenty-four male Wistar rats, weighing 200-220 g, were randomly divided into 3 groups ( n = 8 each) : control group (group C) , overtraining group (group O) , anisodamine group (group A) . The model of overtraining-induced acute heart injury was established by exhausting swimming. Anisodamine 10 mg/kg was given intraperitoneally 20 min before overtraining in group A. Blood samples were taken at 6 h after overtraining for measurement of serum CK-MB activity. The rats were then sacrificed and myocardial tissues taken for determination of TNF-α content and NF-κB activity (by immunohistochemistry) . The apoptosis rate was detected by flow cytometry. Results The CK-MB activity, apoptosis rate, TNF-α content and NF-κB activity were significantly higher at 6 h after overtraining in groups O and A than in group C, while lower at 6 h after overtraining in group A than in group O ( P < 0.05) . Conclusion Anisodamine can inhibit apoptosis in cardiomyocytes by reducing inflammatory response in overtrained rats.

Objective To explore the regulation of Bub1 mRNA expression in endometrial carcinoma cells by estrogen and paclitaxel. Methods The high differentiated endometrial adenocarcinoma cells ( Ishikawa cell line) were cultured in DMED/F12 supplemented with 10% fetal bovine serum(FBS) or phenol red-free DMED/F12 supplemented with 5% dextran-charcoal FBS (dcFBS). Firstly, the cells were stimulated by 10 nmol/L estradiol (17β-E2 ) or no hormonal stimulation as control group, and the cell proliferation was quantified at 24, 48 and 72 hours using cell counting kit-8 (CCK-8) method. Then the cells were stimulated with different concentrations of 17β-E2 (0.1, 10, 1000 nmol/L) at different periods (5,15,30 minites and 2,4,8,12,16,24,30 hours), the expression of Bub1 mRNA was detected by real-time quantitative PCR. Ishikawa cells were cultured with non-serum DMEM/F12 to be synchronized, and then were treated with different concentrations of paclitaxel( 10,100 nmol/L) for 8 and 24 hours. While, nonsynchronized Ishikawa cells were exposed to 100 nmoL/L paclitaxel for different periods(4, 8, 16, 24,48 hours), and real-time quantitative PCR was also used to detect the expression levels of Bub1 mRNA.Data were presented as folds change relative to control group, in which values ＜ 1 were down-regulated, and those ＞ 1 were up regulated. Results The proliferation rate of cells in the presence of 17β-E2 was significantly highter than that of the control group after treated 24 hours (A value: 0. 70 ±0. 08 vs. 0. 86 ±0.10, P = 0.049). Time-dependent experiments revealed that addition of 17β-E2 could increase cell numbers during 72 hours period, while the expression level of Bub1 mRNA was decreased in Ishikawa cell.Dose-dependent experiments revealed maximal estradiol stimulation effects at 10 nmol/L( P = 0. 020). After being treated with serum-free culture, Ishikawa cells were exposed to 10 nmol/L paclitaxel for 8 and 24 hours, and the expression of Bubl mRNA decreased (0. 403 ± 0. 008 vs. 0. 775 ± 0. 144, P = 0. 251 ).Compared to the control cells, the mRNA expression levels of Bub1 in cells treated by paclitaxel for 8 hours was significantly decreased ( P = 0. 009 ), while there was not significantly decreased at 24 hours ( P =0. 396). When exposed to 100 nmol/L paclitaxel for 8 and 24 hours, the expression of Bubl mRNA was also decreased(0. 697 ±0. 017 vs. 0. 850 ±0. 004, P =0. 061 ). Compared to the control cells, Bub1 mRNA expression was also significantly decreased (P = 0. 038 and P= 0. 019, respectively). While with serum freetreatment culture, when Ishikawa cells exposed to 100 nmoL/L paclitxel for 4, 8, 16, 24 or 48 hours, the expression of Bub1 mRNA significantly increased ( 1. 127 ± 0. 105 vs. 1. 614 ± 0. 154 vs. 2. 092 ± 0. 179vs. 1. 381 ± 0. 061 vs. 1. 519 ± 0. 182, P = 0. 002 ), of which was signicantaly increased at 16 hours treatment. Conclusion Bub1 exrpession could be regulated by estradiol and paclitaxel, in which deregulated Bubl expression may contribute to chemotherapeutic efficacy of paclitaxel.

Objective To investigate the role of mitochondrial KATP (mito-KATP) channel in reduction of renal ischemia-repeerfusion (I/R) injury by ischemic postconditioning (IPo) in rats. Methods Thirty-five adult male SD rats weighing 250-280 g were randomly divided into 5 groups ( n = 7 each): group Ⅰ sham operation (group S); group Ⅱ I/R; group Ⅲ IPo; group Ⅳ 5-HD + I/R and group V 5-HD + IPo. The rats were anesthetized with intraperitoneal (IP) chloral hydrate 300 mg/kg. Bilateral kidneys were exposed and their pedicles were occluded for 45 min with atraumatic mini-clamp followed by 6 h reperfusion in group Ⅱ - Ⅴ . In group Ⅲ and Ⅴ 3 cycles of 10 s reperfusion followed by 10 s ischemia were applied immediately after 45 min kidney ischemia. In group Ⅳ and Ⅴ 5-HD (a specific blocker of the mito-KATP channel) 10 mg/kg was given IP at 30 min before ischemia. Blood samples were obtained at 6 h of reperfusion for determination of serum creatinine (Cr) and urea nitrogen (BUN) concentrations. The animals were then killed. Bilateral kidneys were removed for determination of mitochondrial membrane potential in the renal tubular epidural cell and intracellular reactive oxygen species (ROS)content and free Ca2+ concentrations. Results Renal I/R significantly increased serum Cr and BUN concentrations and intracellular free Ca2+ concentration and ROC content and decreased mitochondrial membrane potential as compared with sham operation group. IPo significantly attenuated the I/R-induced changes mentioned above. The protective effects of IPo against renal I/R injury was reversed by 5-HD. Conclusion Mito-KATP channel is involved in reduction of I/R-induced renal injury by ischemic postconditioning.

Objective To compare the effect of TNF-α preconditioning and ischemic preconditioning on hepatic ischemia-reperfusion injury ( IRI) and investigate the underlying mechanisms of TNF-αpreconditioning. Methods Fourty healthy male Wistar rats were random-ly divided into four groups which were Sham-operated group ( SO) ,ischemia-reperfusion group ( IR group:produced by total inflow occlusion for 30 min) ,ischemic preconditioning group ( IPC group:induce with 10 min hepatic ischemic and open 10 min before IR) and TNF-αpre-conditioning group ( TPC group:intraperitoneal injection with 1 μg/kg TNF-a 30 min prior to IR) . The sample of blood and hepatic tissue of all groups were taken after experiment. The protein levels of NF-κBp65 and Bcl-2 in the hepatic tissue were detected by immunohistochemis-try. Results There was significant difference (P<0. 05) between IR group and IPC group,TPC group on the level of ALT,AST and the expression of NF-κBp65 and Bcl-2,apoptosis index in hepatic tissue. There was no significance difference (P>0. 05) between IPC group and TPC group. Conclusion TNF-α preconditioning decreased the intensity of hepatic IRI,just as ischemic preconditioning,by induces an de-crease in the NF-κBp65 expression and an increase in the Bcl-2 expression.

Objective To evaluate the effects of TAT-heme oxygenase-1 (TAT-HO-1) fusion protein on liver injury in rats undergoing orthotopic liver transplantation (OLT).Methods Adult male Lewis (inbred) rats (aged 8-10 weeks,weighing 180-230 g) were used as donors and Brown Norway rats (aged 8-10 weeks,weighing 180-230 g) as recipients.The recipient rats were randomly divided into 2 groups (n=6 each) using a random number table:OLT group and TAT-HO-1 group.The livers were harvested according to the method described by Kamada.In OLT group,the donor livers were flushed and preserved with 4 ℃ HTK solution,while the livers were flushed and preserved for 6 h with 4 ℃ HTK solution containing TAT-HO-1 50 μg/ml in group P.Blood samples were obtained at 7 days after transplantation for measurement of activities of alanine aminotransferase and aspartate aminotransferase in serum.Hepatic specimens were obtained at 7 days after transplantation and stained with haematoxylin and eosin for examination under light microscope.Rejection activity index was calculated according to Banff criteria.The contents of transforming growth factor-beta 1 and interleukin-6 in liver tissues were determined using ELISA.Kupffer cells were isolated and cultured for 48 h to determine the levels of transforming growth factor-beta 1 and interleukin-6 in culture medium.Results Activities of alanine aminotransferase and aspartate aminotransferase in serum,rejection activity index and levels of transforming growth factor-beta 1 and interleukin-6 in liver tissues and culture medium of Kupffer cells were significantly decreased,and the pathological changes of livers were mitigated in group TAT-HO-1 as compared to group OLT.Conclusion TAT-HO-1 fusion protein applied during cold storage of donor livers can attenuate liver injury in rats undergoing OLT.

Objective To investigate the current status about nursing undergraduate students' knowledge,attitude and practice (KAP)regarding nosocomial infection.Methods The self-administered questionnaires were employed to survey 108 undergraduate nursing students on the basis of a simple random sampling method.Results In the knowledge dimension,the nursing students earned 78.3％ of accuracy rate when responding to the questionnaires.The students also demonstrated positive attitudes towards nosocomial infection and occupational safety,particularly the female students.In terms of practice,the students performed relatively poor as 36.1％ of the students were unclear about the classification of medical garbage and 22.2％ of the students used their non-clean hands to touch their eye-glasses.Conclusions The undergraduate nursing students have demonstrated adequate knowledge and proper attitude towards nosocomial infection and occupational safety,however,some behaviors need to be changed.Nursing schools and hospitals should be aware of these findings and provide more training programs regarding nosocomial infection and occupational safety so that they can help students formulate good habits to prevent and control nosocomial infection.

Objective To investigate the changes of CA125 between primary cytoreductive surgery and interval debulking surgery for prediction the rate of optimal interval cytoreductive surgery and prediction the recurrence and the prognosis in patients with epithelial ovarian cancer.Methods A total of 39 cases with suboptimal primary cytoreductive surgery admitted from Jan.1996 to Jan.2009 were retrospectively analyzed.The median age of patients was 56 years( range:41 -68 years).Based on the changes in CA125level between primary cytoreductive surgery and interval debulking surgery,all cases were divided into four groups,group A (CA125 reduced to normal after primary cytoreductive surgery,n=6),group B (CA125reduced to normal after 1 - 2 cycles of chemotherapy,n =11 ),group C ( CA125 reduced to normal after 3 -4 cycles of chemotherapy,n =14),and group D ( CA125 did not reduced to normal after the chemotherapy,n =8 ), and all received platinum-based chemotherapy.The response to chemotherapy evaluated by pathological examination versus CA125 level,and recurrence and prognoses were also analyzed.Results ( 1 )The rate of optimal interval cytoreductive surgery in group A,B,C and D were 6/6,8/11,9/14 and 2/8respectively,in which there were statistically different between group A or B and group D (P ＜0.05).(2)The clinical benefit rates evaluated by the pathological examination in group A,B,C and D were 4/6,4/11,5/14 and 0,respectively and there were statistically different between group A and group D (P =0.030).( 3 ) There was significant difference in the recurrence rate between group A and group D (3/6 vs.8/8,P =0.024),while there were not significant differences between group B or C and group D ( all P ＞ 0.05 ).The rate of drug-resistant recurrence in group A,B,C and D were 1/6,3/11,5/14 and 7/8,respectively,in which there were significant differences between group A,B or C and group D ( all P ＜ 0.05 ). ( 4 ) The median progression-free survival (PFS) for patients in group A,B,C and D were 32,10,18 and 3 months,respectively,in which there were significant differences in the PFS between group A,B or C and group D (P =0.012,P =0.003,P =0.032 ).The median overall survival (OS) were 44,45,44 and 16 months,respectively.There were significant differences in the OS between group A,B or C and group D ( P =0.022,P =0.004,P =0.000 ).Conclusion The change of CA125 between primary cytoreductive surgery and interval debulking surgery may be predict the recurrence type and the prognosis in patients with epithelial ovarian cancer.

Objective To explore the clinical manifestation, diagnosis and prognosis of Leigh syndrome in children. Method Clinical data from 4 cases of Leigh syndrome conifrmed by genetic testing were retrospectively analyzed. The related literature were reviewed. Results In 4 cases, 3 were boys and one was a girl, 3 cases were onset in infant and one case was in school age. The main manifestations were mental retardation, low muscle tone, convulsions, feeding dififculties, drooping eyelids, extraocular muscle paralysis and nystagmus, irritation, activity intolerance etc. The brain magnetic resonance imaging (MRI) revealed symmetry long T1, T2 abnormal signal in brainstem, bilateral globus pallidus, thalamus, cerebellar dentate nuclei, and periaqueductal, 3 cases involved midbrain, one case involved thalamus, and one case involved cerebellar dentate nuclei;2 cases had encephalatrophy. Electromyography was normal in all cases. The levels of lactate in blood and cerebrospinal lfuid were increased. Mitochondrial DNA (mtDNA) detection found the mutation of mtDNA 8993 T>G in one case, and the mutation of mtDNA 9176 T>C in another 3 cases. The case onset in school age died of respiratory failure one month later, and another 3 cases were still in follow up, there were mental retardation, but no signiifcant setback. Conclusion The clinical manifestations of Leigh syndrome in children are diverse. The diagnosis is based on the typical clinical manifestations and MRI, blood and/or cerebrospinal lfuid lactate levels. The genetic testing is the golden standard for diagnosis.