[ ABSTRACT] AIM:To observe the effect of endogenous nitric oxide synthase ( NOS) inhibitor asymmetric dimeth-ylarginine ( ADMA ) and its signaling pathways on NO levels and skeletal muscle contractility in 4-week running rats. METHODS:The 4 weeks running rat model was established.The twitch tension, tetanic tension and the fatigue test of sole-us muscle induced by electrical stimulation ex vivo were detected.The ATP content, mitochondrial DNA levels and the mR-NA expression of peroxisome proliferator-activated receptor γcoactivator-1α(PGC-1α), nuclear respiratory factor (NRF) were measured to reflect the mitochondrial function and biosynthesis in the skeletal muscle.Serum ADMA concentration was detected by high performance liquid chromatography.The endogenous ADMA enzymes PRMT1 and 2 subtypes of ADMA me-tabolism enzyme DDAH, 3 subtypes of NOS protein expression in the skeletal muscle were determined by Western blot.NOS activity and nitric oxide ( NO) content were analyzed by colorimetric method.RESULTS: Compared with normal control group, the twitch tension, tetanic tension and the anti-fatigue capability of soleus muscle in running group were significantly enhanced, ATP content, mitochondrial DNA content and the mRNA expression of PGC-1αand NRF were significantly in-creased (P<0.01).In addition, the protein expression of constitute type NOS (cNOS) and NOS activity were significantly increased (P<0.01), but the increase in NO content was relatively smaller in soleus muscle in exercise group (P<0.05). Moreover, serum ADMA concentration in running group was increased, while the DDAH2 expression in skeletal muscle was decreased.CONCLUSION:Short-term endurance exercise enhances the twitch tension, tetanic tension and fatigue resist-
ance of soleus muscle.The mechanism may be that increased cNOS expression feedbacks to increase ADMA concentration, thus maintaining the increase in NO synthesis at a relatively low level, and resulting in promoting skeletal muscle mitochon-dria biosynthesis and mitochondrial function.

Objective To investigate the effects of cold exposure with different time courses on cardiovascular function and skeletal muscle contraction in Sprague-Dawley (SD) rats.Methods The male SD rats were randomly divided into control,12-h cold,24-h cold,and 48-h cold groups.The rats in all cold group were maintained in 4 ℃ environment for indicated time and allowed free access to standard rat chow and water ad libitum.After the animal model was established,the heart,thoracic aorta,soleus,and extensor digitorum longus (EDL) were rapidly isolated from rats under anesthesia and then fixed into the respective organ perfusion system that was connected to physiological recorders and data acquisition system.Lanendorff heart persusion was used to detect some parameters of cardiac function,such as coronary flow (CF),mean left ventricular systolic pressure (mLVSP),mean left ventricular diastolic pressure (mLVDP) and maximum ascending and declining rate of left ventricular pressure (± dp/dt max) ; and then the heart was suffered from 30 min ischemia followed by 60 min reperfusion,the changes of above cardiac function indexes were recorded again.Organ bath method was employed to detect contraction response to potassium chloride (KCl) or phenylephrine (PE) and endothelium-dependent relation responses to acetylcholine (ACh) of aortic rings.The twitch tension,titanic tension,and fatigue test of soleus and EDL were examined by electrical stimulation to reflect contractive function of skeletal muscle.Results The coronary flow (CF) were significantly reduced in 24-h and 48-h cold groups compared to control group [(9.0 ± 1.7) ml/g and (9.8 ± 1.3) ml/g vs (11.6 ± 1.2) ml/g,P ＜ 0.05] ; 48-h cold exposure also showed significantly increased mLVSP (P ＜0.01) and mLVP (P ＜ 0.05) of rats.After ischemia-reperfusion,the heart re-beat time was shorter in all cold exposure groups (all P ＜ 0.01),mLVP and mLVDP were higher in 24-h and 48-h cold group than control group (all P ＜ 0.01).The contraction of aortic rings responses to 60 mmol/L KCl [(0.63 ±0.13) g and (0.79 ±0.11)g vs (1.28±0.17)g,P ＜0.01] and 1 μmol/L PE [(0.62 ±0.18)g and (0.68 ±0.13)g vs (1.25 ± 0.18)g,P ＜ 0.01] were decreased while the endothelium-dependent relaxation responses to low concentration ACh (3 × 10-8 ～ 3 × 10-7) were increased in 24-h and 48-h cold groups relative to control group (all P ＜ 0.05).The 48-h cold also reduced titanic tension of SOL and EDL in comparison with control [(4.14 ± 0.62) N/cm2 vs (5.50 ± 0.75) N/cm2,P ＜ 0.05 and (3.00 ± 0.57) N/ cm2 vs (4.70 ± 0.85) N/cm2,P ＜ 0.01].Conclusions 24 ～ 48 h cold exposure could reduce CF but enhance the tolerance of heart from rats to ischemia reperfusion injury and also decrease the contraction of aortic rings response to KCl or PE but increase the sensitive responses to ACh of aortic rings from rats.The 48-h cold could decrease the contraction of skeletal muscle elicited by electrical stimulation.

Objective To investigate the effects of fasting with different time courses on the contraction of skeletal muscle and cardiovascular function in Sprague-Dawley (SD) rats.Methods The healthy and male SD rats were randomly divided into (1) control,(2) 12 h fasting,(3) 24 h fasting,and (4) 48 h fasting groups.After animal models of fasting with different time courses were established,the heart,thoracic aorta,soleus,and extensor digitorum longus (EDL) were rapidly isolated from rats under anesthesia and then fixed into the respective organ perfusion system that was connected to physiological recorders and data acquisition system.Langendorff heart perfusion was used to detect some parameters of cardiac function,including left ventricular systolic pressure (LVSP),left ventricular end-diastolic pressure (LVEDP),and maximum ascending and declining rate of left ventricular pressure (± dP/dt max).Then,the heart was suffered from 30 min ischemia followed by 60 min reperfusion,the changes in above cardiac function parameters were recorded again.The contraction response to potassium chloride or phenylephrine and endothelium-dependent relation responses to acetylcholine (ACh) of aortic rings were measured with organ bath.The changes of twitch tension and titanic tension of soleus and EDL were examined to reflect contractive function of skeletal muscle.Results The + dP/dt max was significantly reduced in 24 h and 48 h fasting groups compared to the control (P ＜ 0.01).After ischemia-reperfusion,the heart re-beat time was significantly longer [(24.3 ± 7.0)min vs (14.0 ± 2.9)min,P ＜0.05],both mLVDP and mLVP were significantly higher in 48 h and 24 h fasting groups than control group (P ＜ 0.05),and those changes did not normalized until re-perfusion 60 min.The 48 h fasting increased significantly endothelium-dependent relaxation of aortic rings response to low concentration acetylcholine (P ＜ 0.05),but did not affect the maximum relaxation of aortic rings response to high concentration acetylcholine and contraction of aortic rings responses to potassium chloride and phenylephrine.The 48 h fasting also enhanced significantly titanic tension of soleus compared to the control [(7.01 ± 1.22) N/cm2 vs (5.5 ± 0.75) N/cm2 for48 h-Fasting vs Control,P ＜0.05].Conclusions The 24 ～48 h fasting may induce inhibition of cardiac function,while fasting 48 h can increase the sensitivity of aortic rings responses to ACh and contraction function of soleus in healthy and male rats.

Objective To study the effects of using the dropping retention-enema in the clinical practices.Methods Divided 60 cases who need retention-enema into the experimental group and the control group,there were 30 cases in the each group.The traditional retention-enema method was used in the control group,while the dropping retention-enema method was used in the experimental group.Compared the related factors between the two groups.Results All the factors which can indicated the clinical effects in the experimental group were better than those of in the control group,P

ObjectiveTo explore the factors associated with activity of daily living (ADL) in patients injured from earthquake and to observe the rehabilitation nursing of fractures for them. Methods367 patients were reported and analyzed with Logistic regression. Results and ConclusionThe age (OR=1.016), renal failure (OR=4.648), lower limb fractures (OR=2.404), spine fracture (OR=3.558), right hand fractures (OR=1.190), right foot fractures (OR=4.389) and the complex fractures (OR=2.600) were associated with the poor ADL (<60). The rehabilitation nursing for fracture were reported and discussed.

Objective To observe the anti-tumor effect on human multiple myeloma cell lines U266 and KM3 with a combination of varinostat and melphalan in vitro.Methods The cell proliferation of U266 and KM3 was datected with MTT assay when treated them with vorinostat alone and melphalan alone,then calculate their IC50 values respectively.Fixed the concentrations of vorinostator melphalan,the cell proliferation was datected in combination with melphalan/vorinostat in seriesly concentrations by MTr assay.Then to calculate drug combination index(CI).Results The IC50 value of U266 was 5.0-7.5 μmol/L and that of KM3 was 2.5-5.0 μmol/L when treated by vorinostat alone,the IC50 value of U266 was 40-60 μmol/L and that of KM3 was 60-80 μmol/L treated by melphalan alone.When fixed the concentration of vorinostat(in U266 the concentration was 1.25 μmol/L,in KM3 the concentration was 1.0 μ mol/L),Synergism(CI＜0.9)was observed when the concentrations of melphalan were 20 μmol/L,40 μmol/L,60 μ mol/L and 80 μ mol/L in U266,40 μmol/L,60 μmol/L,80 μmol/L and 100 μmol/L in KM3;When fixed the concertration of melphalan (in U266,was 10 μmol/L,in KM3 was 20 μmol/L),synergism(CI＜0.9)was observed when the concentrations of vorinostat were 1.0 μmol/L,2.5 μmol/L,5.0 μmol/L and 7.5 μmol/L in U266,and 1.0 μmol/L,2.5μmol/L,5.0 μmol/L in KM3.An additive effect was observed with the czombination of vorinostat 7.5 μmol/L plus melphalan 20 μmol/L in KM3(CI=0.93).Conclusion Vorinostat had potential anti-myeloma effect alone,and had synergistic anti-tumor effect with melphalan in vitro.

Objective To investigate the mRNA expression of a proliferation inducing ligand (APRIL) and its receptors including B cell maturation antigen (BCMA),transmembrane activator.calcium modulator and cyclophilin ligand interactor (TACI) in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SEE).Methods APRIL mRNA、BCMA mRNA and TACI mRNA in PBMCs were detected by real-time quantitative PCR in 66 SLE patients and 25 normal controls.Gene expression level was measured as 2-AACT.Results The expression levels of APRIL mRNA、BCMA mRNA and TACI-mRNA were significantly increased in both active SLE group and stable SLE group compared with those in the normal controls(P<0.01 for all).The expression levels of APRIL mRNA and TACI mRNA in active SLE group were significantly higher than those in stable SLE group(P<0.01,P<0.05,respectively).But there was no significant difierence in the expression levels of BCMA mRNA between the SLE stable and active groups-Beside,the expression levels of APRIL mRNA and TACI mRNA were significantly increased in patients with lupus nephritis (LN) compared to patients with non-LN (P<0.01 for all).Conclusion The expression levels of APRIL and its receptors are significantly elevated in SLE patients.It may suggest that APRIL and its receptors play an important role in the pathogenesis of SLE.

Objective To investigate the mRNA expression level of neurogranin on peripheral blood mononuclear cells(PBMCs)from patients with systemic lupus erythematosus(SLE).Methods Top 20 tags of SLE PBMCs SAGE library were searched from normal lymphocytes SAGE library including navie-T,Th1,Th2, CD8+T, NK and B cells,and their abundance was compared.The mRNA expression level of neuro-granin,a differential over-expressed tag,was detected in 35 cases of SLE and 15 normal controls by reversetranscription-polymerase chain reaction (RT-PCR).Results Neurogranin tag could only be detected in SLE PBMCs SAGE library,but was hardly found in normal lymphocyte SAGE library.However,either SLE pa-tients or normal controls showed a detectable mRNA level of neurogranin on PBMCs by RT-PCR.The mRNA level of neurogranin in active SLE patients was significantly increased than those in controls(P＜0.001).but only slightly increased in inactive SLE patients (P＞0.05).Conclusion Neurogranin,as a novel proapototic factor,is overexpressed on PBMCs of SLE patients.It may be involved in the regulation of abnormal immune responses in lupus.

OBJECTIVE: Discussion of expression and its significance of CD44 in SP cells of nasopnaryngeal carcinoma. METHOD: Flow cytometry was used to sort cultured CNE-2 cells of nasopharyngeal carcinoma for obtaining CD44-SP and CD44+SP cells. Biological differences of CNE-2, CNE-2 SP, CNE-2 NSP, CNE-2 CD44+SP and CNE-2 CD44-SP cells were statistically analyzed by experiments such as cell migration experiments, plate clone formation assay, cell cycle analysis and sensitivity tests to chemotherapeutics. RESULT: Two point 3 perent of SP cells were extracted from CNE-2 cells of nasopharyngeal carcinoma, among which 36.5% was CD44+SP cells. Abilities of proliferation, cell migration and plate clone of CD44+SP cells were significantly higher than other cells (P < 0.01), and its tolerance to chemotherapeutics was significantly higher too (P < 0.01). CONCLUSION The proportion of SP cells in nasopharyngeal carcinoma cells was small, but SP cells had strong activeness in the aspect of cell proliferation with a "seed" characteristic of tumor cells. As CD44+SP cells played an important role in proliferation and chemotherapy resistance of nasopharyngeal carcinoma, it indicated that CD44 can be one of the surface markers of SP cells of nasopharyngeal carcinoma.
Biomarkers, Tumor ; metabolism ; Carcinoma ; Cell Cycle ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Flow Cytometry ; Humans ; Hyaluronan Receptors ; metabolism ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism

[ABSTRACT]OBJECTIVEThis study aims to explore the molecular mechanism and expression of Wnt/β-catenin signaling pathway and tumor marker CD44 in nasopharyngeal carcinoma cells after transfection withβ-Catenin when the Wnt/β-catenin signaling pathway was blocked.METHODSSP cells and CD44+SP cells isolated from the nasopharyngeal carcinoma cell line CNE-2 by flow cytometry were identified. Changes in the number and biological characteristics of CNE-2 and CD44+SP cells in vitro were investigated after the Wnt/β-catenin signaling pathway was blocked through siRNA.RESULTSSP cells accounted for 2.3% of nasopharyngeal carcinoma CNE-2 cells, andCD44+SP cells accounted for 36.5% of the SP cells. CD44+SP cells showed significantly higher in vitro proliferation and resistance to chemotherapy (P<0.05). After transfection withβ-Catenin siRNA, the proliferation, cloning efficiency, and tolerance to chemotherapeutic drugs of the cells were found statistical differences compared with those before transfection ofβ-Catenin siRNA. CONCLUSIONWnt/β-catenin signaling pathway abnormalities are closely related to the biological behavior of nasopharyngeal carcinomaCD44+SP cells. Interference of this pathway can change the characteristics of nasopharyngeal carcinoma stem cells.