Purpose To study the expression of inhibitor of differentiation 2 (Id2) in Ewing sarcoma of bone as well as the correlation with cell cycle proteins. Methods The expression of Id2, c-Myc, Cyclin D1, p53, p16 and p27 in 45 cases of Ewing sarcoma were detected by immunohistochemical EnVision methods, the relationship between the expression of Id2 and clinicopathologic characteristics and the correlation with cell cycle proteins were analyzed. Results Total expression of Id2 was found in 88. 9% of Ewing sarcoma. Strong expression of Id2 was detected in 33. 3% cases. Higher frequencies of c-Myc and Cyclin D1 immunostaining were observed (66. 7% and 71. 1%, respectively), but lower frequencies of p53, p16 and p27 expression were present (20. 0%, 35. 6% and 28. 9%, respectively). Id2 overexpression displayed a negative relationship with p27 expression (P<0. 05). Among the follow-up of 24 cases, overexpression of Id2 was found more frequency in dead and metastases cases (41. 2%) when compared to that of progres-sion-free survival cases (14. 3%). Conclusions Id2 was extensively expressed in Ewing sarcoma of bone. Overexpression of Id2 and its correlation with p27 expression suggest that this is an important pathway involved in development of Ewing sarcoma. Moreover, Id2 overexpression may predict poor prognosis.

Mononuclear cells(MNCs) isolatedfrommammal'sbonemarrowandperipheral bloodbymeansof Ficoll density grandient centrifugation,which are composed of monocytes and lymphocytes,can differentiate into different progenitor cells with different functions in the development of atherosclerosis under different inducing conditions. This article described the induction conditions of MNCs,the identification methods of different progenitor cells,and the relationship between these progenitor cells and the development of atherosclerosis,thus provide a new idea for the prevention of the atherosclerosis.

Objective The expression of TGF-β1 and IL-10 in the grafts of inbred mice with rejection of heart transplantation were studied and the interaction of them in the rejection of heart transplantation in inbred mice investigated.Methods Allografts were divided into 2 groups:control group (n =70),cyclosporin A-treated group (CsA group,n =70).Hearts from inbred BABL/c mice were transplanted into a cervical location in the other recipients and the survival time of the allografts was observed.The local expression of TGF-β1 and IL-10 was detected at day 1,3,7,11,14,21 by reverse transcription polymerase chain reaction(RT-PCT) respectively.Results The survival time of the allografts was (20.3 ± 1.7) days in control group,and(32.2 t 3.4) days in CsA group (P ＜ 0.01).The levels of the two cytokines expression were up-regulated in CsA group.The up-regulation of TGF-β1 was closely correlated with the survival of the grafts.Conclusion The expression and production of the two cytokines is up-regulated probably cause of administ ration of cyclosporin A,and favorite to the heart graft survival,and action of these two cytokines are probably interrelated.

Objective To investigate the inhibitory effect of blocking PI3K/AKT pathway by wortmannin on hypoxia-inducible factor 1α (HIF-1α) and the effect on the expression of glycolysis associated genes in human esophageal carcinoma cell lines TE1 and TE13,and to analyze the relation between PI3K/AKT-HIF-1α pathway and glycolysis in esophageal carcinoma cells. Methods Esophageal carcinoma cell lines TE1 and TE13 pretreated with wortmannin (2 μmol/L) were incubated under normoxic and hypoxic conditions.And each cell line was divided into four groups.The expression of HIF-1α and glycolysis associated genes GLUT-1,LDHA and HK-Ⅱ at protein level were measured by.Western blot.The expression of HIF-1α,GLUT-1,LDHA and HK-Ⅱ at mRNA level was determined by real-time PCR. The activities of LDH and HK-Ⅱ and lactic acid (LA)concentration in the culture supernatant were tested with spectrophotometer method.Results Under normoxic condition,HIF-1α was expressed in TE1 cells and the expression of HIF-1α was inhibited by wortmannin (2 μmol/L),the most significant inhibitory effect was at 12 hours,therefore 12 hours was selected for the subsequent hypoxia experiment.Compared with untreated cells,the expression of HIF-1α、HK-Ⅱ 、GLUT-1、LDH-A at protein level significantly decreased in TE1 and TE13 cells after pretreated with wortmannin (P ＜ 0.05),and the expression of HIF-1α、HK-Ⅱ at mRNA level significantly decreased (P＜ 0.05).Under normoxic and hypoxic conditions,the HK-Ⅱ and LDH activities in TE1 and Te13 esophageal carcinoma cells significantly decreased after treated with wortmannin compared with untreated cells (P＜0.05).Under hypoxia condition,the enzyme activity increased in untreated cells (P＜ 0.05). Under normoxic and hyp0xic conditions,the lactic acid concentration in the culture supernatant obviously decreased in cells treated with wortmannin compared with untreated cells (P＜ 0.05). Under hypoxia condition,lactic acid concentration increased in wortmannin treated cells (P ＜ 0.05). Conclusions Under normoxic and hypoxic conditions,wortmannin decrease lactic acid concentration through inhibiting the expression of HIF-1α and glycolysis associated genes, which indicate PI3K/AKT-HIF-1α pathway was closely related to glycolysis in esophageal carcinoma cells.

AIM:To investigate the role of reactive oxygen species (ROS) in the regulation of intracellular Ca2+induced by angiotensin II ( Ang II) in the primarily cultured medullary neurons .METHODS:Primarily cultured me-dullary neurons were prepared from 14-day-old embryos of Sprague-Dawley rats in the study .The identification of medullary neurons was assessed by double-labeling immunofluorescence .To explore the role of ROS , mainly the superoxide ( O2 ·-) , the O2 ·-generation was measured using the fluorogenic probe dihydroethidium ( DHE) .To determine intracellular free cal-cium concentration ( [ Ca2+] i ) , the neurons were loaded with the Ca 2+-specific dye Fura-2/AM.The cell viability after adding Ang II was also examined using CCK-8 assay.RESULTS:Most of the cultured cells were medullary neurons , more than 80%of which were glutamate positive neurons .Ang II (5 μmol/L) increased the level of ROS within 10 min in the medullary neurons .Ang II at 5μmol/L induced a significant [ Ca2+] i increase in the medullary neurons , and the effect of Ang II occurred rapidly and reached a peak within 20 min after administration.The level of [Ca2+]i started to decline after washout .The Ca2+elevation induced by Ang II was significantly decreased by apocynin or TEMPOL .No significant differ-ence in the cell viability between control group and 5μmol/L Ang II treatment group was observed .CONCLUSION:ROS is involved in the regulation of [Ca2+]i induced by Ang II in the primarily cultured medullary neurons , suggesting a poten-tial intracellular signaling mechanism involved in the Ang II-mediated oxidant regulation of central neural control of blood pressure.

Objective To evaluate the efficacy and safety of covered metallic stents in the management of proximal esophageal strictures.Methods Retrievable covered self-expanding metallic stents were inserted individually in 43 patients with malignant strictures of the proximal esophagus.Main outcome measures included improvement of dysphagia,sealing of fistula and complications were evaluated.Results A total of 45 stents were placed in 43 patients,with a success rate of 95.6％ (43/45) at the first stent placement attempt.The dysphagia score improved significantly from 3.3 ± 0.6 to 1.8 ± 0.9 (P ＜ 0.001),with 18 having an excellent response and 25 having a good response.Improvement rate of dysphagia was 95.6％ (43/45).Fistula sealing was achieved in 7 of 9 patients (77.8％).Early complications included severe pain (n =5),mediastinal emphysema (n =1),moderate fever (n =9) and collapse of the proximal end of the stent (n =1),while late complications included migration (n =3) and tissue overgrowth (n =6).Conclusion Retrievable covered metallic stent placement is effective and safe for the palliation of obstructive dysphagia and sealing of fistulas in malignant proximal esophageal strictures.

[ABSTRACT]AIM:ToinvestigatetheeffectofD4F,anapolipoproteinA-Imimeticpeptide,onoxidizedlow-density lipoprotein ( ox-LDL)-induced macrophage apoptosis and activation of caspase-12, a key molecule in endoplasmic reticulum stress ( ERS )-associated apoptotic pathway, and to elucidate the underlying molecular mechanisms. METHODS:RAW264.7 macrophages were pretreated with D4F (12.5, 25 and 50 mg/L), 4-phenylbutyric acid (PBA, 5 mmol/L) or diphenyleneiodonium ( DPI, 5 μmol/L) for 1 h and then treated with ox-LDL (100 mg/L) or tunicamycin ( TM, 4 mg/L) for 24 h.The cell viability and apoptosis were determined by MTT assay and TUNEL detection, respective-ly.The levels of malondialdehyde ( MDA) and reactive oxygen species ( ROS) in the cells and the activities of superoxide dismutase ( SOD) and nicotinamide adenine dinucleotide phosphate ( NADPH) oxidase were determined.The protein level of caspase-12 was examined by Western blot analysis.RESULTS: Similar to the ERS inhibitor PBA, D4F protected RAW264.7 macrophages from ox-LDL or TM ( an ERS inducer)-induced decrease in the viability and increase in apoptotic rate in a dose-dependent manner.Like DPI (an oxidative stress inhibitor), D4F significantly inhibited ox-LDL-induced ox-idative stress, as expressed by the decreased generation of ROS and MDA ( P<0.01) , the increased activity of SOD and the decreased activity of NADPH oxidase (P<0.05).Moreover, similar to PBA and DPI, D4F significantly suppressed ox-LDL-induced activation of caspase-12 in a concentration-dependent manner ( P<0.05) .Furthermore, D4F also inhibi-ted the caspase-12 activation induced by TM (P<0.05).CONCLUSION: D4F inhibits macrophage apoptosis induced by ox-LDL, and the mechanism is at least partially by reducing oxidative stress and inhibiting the activation of caspase-12.

Objective To investigate the effects of sulforaphane(SFN)in regulating the macrophage glycolysis via the arachidonate 5-lipoxygenase(ALOX5)/nuclear factor kappa B(NF-κB)signaling pathway on the progression of diabetic nephropathy(DN).Methods Bioinformatics analysis was used to identify the target genes of SFN in the treatment of DN.Human proximal tubular epithelial cell line(HK-2 cells)was induced with 30 mmol/L high glucose(HG)to create an in vitro model of DN.HK-2 cells were divided into the following groups:normal glucose(NG)group,HG group,HG+SFN(3 mmol/L)group,HG+ALOX5 group,HG+SFN(3 mmol/L)+ALOX5 group,HG-treated macrophages+HK-2 group,HG+SFN(3 mmol/L)treated macrophages s+HK-2 group,HG+ALOX5 transfection treated macrophages+HK-2 group,HG+SFN(3 mmol/L)+ALOX5 transfection treated macrophages+HK-2 group.CCK-8 assay was used to detect cell viability,Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)method was used to detect cell apoptosis;glucose and lactate levels in the cells were measured using assay kits;Western blot was performed to detect the expression of ALOX5,NF-κB,and glycolysis-related proteins hexokinase-2(HK2),pyruvate kinase M2(PKM2),glucose transporter 1(GLUT1)in each group.Diabetic nephropathy(DN)mouse models were established using streptozotocin(STZ)and treated with SFN(0.5 mg/kg).Various biochemical parameters were measured in the mice,and kidney tissue pathology was examined using H&E staining.Western blot was used to detect the expression of glycolysis-related proteins(HK2,PKM2,GLUT1)in kidney macrophages.Results Bioinformatics analysis revealed ALOX5 as the target gene of SFN in treating DN.Compared to the HG group,SFN treatment enhanced HK-2 cell viability and in-hibited apoptosis(P<0.05);concurrently,SFN treatment suppressed HG-induced macrophage glycolysis-related protein and attenuated macrophage-mediated HK-2 cellular injury(P<0.05).Western blot results showed that SFN inhibited the expression of ALOX5 and NF-κB(P<0.05).The mouse experiment results showed that SFN treatment improved kidney function and pathological changes in the kidney of DN mice,and inhibited the related protein expression of acrophage glycolysis in kidney tissue(P<0.05).Conclusion SFN improves the progression of DN by inhibiting the expression of macrophage glycolysis-related protein through the ALOX5/NF-κB signaling pathway.

Objective To construct a cardiovascular surgery nursing service action plan system based on patient's demand so as to provide a reference for nurse specialisms of cardiovascular surgery to provide good service for patients.Methods From January 2017 to June 2017, patients of cardiovascular surgery of a ClassⅢ Grade A hospital were investigated with the self-designed patients' demand for nursing service questionnaire according to literature analysis and interview results of patients. Weights of the primary index and secondary index were calculated by analytic hierarchy process so as to construct a cardiovascular surgery nursing service action plan system based on patient's demand.Results The cardiovascular surgery nursing service action plan system based on patient's demand included 5 primary indexes and 78 secondary indexes. The Cronbach's α of patients' demand for nursing service questionnaire was 0.974 with a good internal consistency. Kaiser-Meyer-Olkin (KMO) was 0.843, andP value of Bartlett sphericity test was less than 0.001 with a good construct validity. The normalized weight coefficients of 5 primary indexes (physiological needs, safety needs, belongingness and love needs, self-esteem needs and self-actualization needs) were 0.2205, 0.4798, 0.0771, 0.1339 and 0.0887 respectively with 0.0488 for the consistency ratio (CR). TheCR of 21 secondary indexes of physiological needs, 19 secondary indexes of safety needs, 11 secondary indexes of belongingness and love needs, 20 secondary indexes of self-esteem needs and 7 secondary indexes of self-actualization needs was 0.0488, 0.0508, 0.0823, 0.0877 and 0.0746 respectively. TheCR was all less than 0.1 showed a satisfactory consistency in the results of hierarchy sequencing.Conclusions The construction of cardiovascular surgerynursing service action plan system based on patient's demand with the strict scientific method improves patient satisfaction. It is worth to be a reference for nursing peers and managers of cardiovascular surgery.

Objective: To understand smoking behavior and related factors among college and middle school students in Inner Mongolia Autonomous Region, so as to provide a technical basis for targeted tobacco control interventions. Methods: In September 2023, a total of 93 647 students in Inner Mongolia Autonomous Region were randomly selected by stratified cluster sampling from 12 league cities and 103 banner counties (cities districts) to conduct a questionnaire survey. The χ2 test and binary Logistic regression analyses were used to explore the current situation of adolescent smoking and its related factors. Results: The prevalence of college and middle school students who tried smoking was 10.2%, and the prevalence of current smokers was 4.4%. The binary Logistic regression model showed that students in high school, vocational high school, university, males, those from unstable family, boarders, individuals who sew someone smoking at home, at school, in public places, drinking alcohol, inhaling volatile solvents for pleasure, using cough suppressant except for coughing, using sedative without a doctors permission, being scolded by parents, experiencing school bullying, being addicted to the Internet, having depressive symptoms, anxiety symptoms, and low selfesteem were more likely to attempt smoking and currently smoke (attempt smoking:OR=1.38, 2.59, 1.75, 3.24, 1.18, 1.43, 1.25, 1.43, 1.43, 0.79, 9.18, 1.42, 1.17, 1.65, 1.52, 1.31, 1.58, 1.41, 1.11, 1.17； current smoking:OR=1.61, 4.76, 3.76, 4.45, 1.17, 1.44, 1.27, 1.58, 1.69, 0.68, 7.99, 1.64, 1.12, 2.31, 1.48, 1.37, 1.26, 1.31, 1.35, P<0.05). Teenagers with parents who had graduated from high school and eat breakfast every day had a relatively low risk of trying and current smoking (OR=0.78, 0.83, 0.79, 0.84, 0.78, 0.68, P<0.05). Conclusions Adolescent smoking behaviours in Inner Mongolia Autonomous Region exhibit obvious clustering pattern in the school setting. Associated factors includes demographic characteristics, secondhand smoke exposure, daily behaviours and substance abuse, and psychological behaviours. Schools, families and society should work together to create a smokefree environment to protect adolescents from the harms of tobacco.