Background:Intestinal metaplasia(IM)and dysplasia in Barrett’s esophagus(BE)can be detected by endoscopic biopsy combined with pathological examination. Aims:To evaluate the efficacy of acetic acid stain-guided biopsies in detection of IM and dysplasia in BE under standard endoscopy. Methods:One hundred patients found to have columnar-lined esophagus by endoscopy from January 2012 to November 2014 at Shanghai Jing’an District Central Hospital were enrolled. Under standard endoscopy,acetic acid was sprayed and multi-point biopsies were performed in visible abnormal regions. This was followed by four-quadrant biopsies at every 1 to 2 cm of the remaining BE segments. Detection rates of columnar epithelium,IM and dysplasia were compared between acetic acid stain-guided biopsies and four-quadrant biopsies. Results:A total of 96 patients were diagnosed as BE. Detection rates of columnar epithelium by acetic acid stain-guided biopsies and four-quadrant biopsies were 96. 9%(93 / 96)and 90. 6% (87 / 96),respectively,no significant difference was found between the two methods(McNemar test,P = 0. 146),however,consistency of the two methods was poor(κ = 0. 340). A total of 32 patients were diagnosed as BE with IM. Detection rates of IM by acetic acid stain-guided biopsies and four-quadrant biopsies were 84. 4%(27 / 32)and 40. 6%(13 / 32),respectively,difference in detection rate between the two methods was significant(McNemar test,P = 0. 007),and consistency of the two methods was poor(κ =0. 266). Five cases of low-grade dysplasia were detected by acetic acid stain-guided biopsies only. Conclusions:Acetic acid stain-guided biopsies can detect BE with IM and dysplasia better than four-quadrant biopsies.

Objective To investigate the relationship between self-efficacy and positive degree in patients with cervical spondylosis.Method A questionnaire survey was conducted among 98 patients with cervical spondylosis.Results The self-efficacy score was 6.30±1.56,the score on the positive degree was 53.76 ±12.53.The self-efficacy and positive degrees were positively correlated (r=0.531,P＜0.001).Conclusions The patient's self-efficacy and positive degree are at medium level.Nursing staff should take corresponding measures to improve self-efficacy of cervical spondylosis patients,in order to improve its positive degree and achieve the purpose of effective self-management and health promotion.

Molecules can enter the nucleus by passive diffusion or active transport mechanisms, depending on their size. Small molecules up to size of 50-60 kDa or less than 10 nm in diameter can diffuse passively through the nuclear pore complex (NPC), while most proteins are transported by energy driven transport mechanisms. Active transport of viral proteins is mediated by nuclear localization signals (NLS), which were first identified in Simian Virus 40 large T antigen and had subsequently been identified in a large number of viral proteins. Usually they contain short stretches of lysine or arginine residues. These signals are recognized by the importin super-family (importin α and β) proteins that mediate the transport across the nuclear envelope through Ran-GTP. In contrast, only one class of the leucine-rich nuclear export signal (NES) on viral proteins is known at present. Chromosome region maintenance 1 (CRM1) protein mediates nuclear export of hundreds of viral proteins through the recognition of the leucine-rich NES.

Objective To establish heterologous expression system of Na+-glucose cotransporter 2 (SGLT2) gene.Methods Human SGLT2 cDNA from normal kidney, generated by RT-PCR,was subclonedintoPEXL-GFP vector andtransfectedinto HEK293cells. After 24hours of incubation, the expression of SGLT2-GFP fusion protein was detected by Western blotting and laser confocal microscopy.Transport activity of SGLT2-GFP fusion proteins in cultured human HEK293 cells was evaluated with the uptake test of glucose analogue.ResultsSGLT2-GFP fusion protein was expressed in cultured human HEK293 cells.Furthermore, confocal microscopy using green fluorescent protein(GFP) revealed a punctate membrane pattern of SGLT2.Glucose analogue uptake increased in HEK293 cells transfected with SGLT2-GFP at least by 3.5 folds compared with HEK293 cells transfected with GFP vector only(P＜0.01).Conclusion Heterologous expression of SGLT2 gene in HEK293 cells is successfully established, which provides valuable approach for the functional and pathological study of SGLT2 gene.

AIM:To observe the expression of nuclear factor-kappa B ( NF-κB) , N-methyl-D-aspartic acid re-ceptor 2B (NR2B) and inducible nitric oxide synthase (iNOS) in the spinal cord in a rat model of chronic constriction in-jury (CCI) of the sciatic nerve.METHODS:Fifty-six adult male Sprague-Dawley rats weighing 180~220 g were random-ly divided into sham group (n=8) and CCI group (n=48).The mechanical withdrawal threshold (MWT) and paw with-drawal latency (PWL) of the hind paws were measured 1 d before CCI and 1 d, 4 d, 7 d, 14 d and 21 d after surgery.The L4~L6 segment of the spinal cord was taken for determining the expression of NF-κB, NR2B and iNOS by RT-PCR and Western blotting.RESULTS:At 1 d, 4 d, 7 d, 14 d and 21 d after surgery, the MWT and PWL in CCI group were obviously lower than those in sham group .The expression of NF-κB, NR2B and iNOS at mRNA and protein levels in-creased significantly.Positive correlations were found between the mRNA expression of NF-κB and iNOS (r=0.842, P<0.05), and between the mRNA expression of NR2B and iNOS (r=0.833, P<0.05).CONCLUSION:The generation and maintenance of hyperalgesia in sciatic nerve injury rats may attribute to the activation of NF -κB and NR2B and concom-itant increase in iNOS .

BACKGROUND:Activation of Notch signaling plays a critical role in stem cel differentiation, and this effect seems to be cel-type dependent. Little is reported on the role of activation of Notch1 signaling in the differentiation of c-Kit+ bone marrow mesenchymal stem cels. OBJECTIVE:To analyze the influence of activation of Notch1 signaling on the differentiation of c-Kit+ bone marrow mesenchymal stem cels. METHODS:The Notch1 intracelular domain (N1-ICD) was obtained from the cDNA library by PCR and cloned intoBamHI/AgeI digested adenoviral GV314 plasmid to construct N1-ICD overexpressing shuttle plasmid, and the positive clones were verified by sequencing. N1-ICD shuttle plasmid and helper plasmids pBHGloxΔE1,3 Cre were used to co-transfect HEK293T cels to obtain N1-ICD overexpressing adenoviral particles (N1-ICD-Ad). The c-Kit+ subpopulation were isolated from bone marrow mesenchymal stem cels of the Sprague-Dawley rat femurviamagnetic activated cel sorting. After transfection of the c-Kit+ BMSCs with N1-ICD-Ad adenovirus, we assessed the activation of Notch1 signaling and differentiation in c-Kit+ bone marrow mesenchymal stem cels by quantitative RT-PCR and immunofluorescent staining. RESULTS AND CONCLUSION:N1-ICD coding sequence was successfuly generated from the cDNA library, and then was cloned into the linearized adenoviral vectors GV314. The resistant clones were verified by sequencing. With the assistance of packaging plasmids, recombinant N1-ICD-Ad adenovirus plasmids were successful packaged in HEK293T cels, and its title was 2×1012 PFU/L. c-Kit+ bone marrow mesenchymal stem cels with the purity of 91.6% were successfuly isolated from the bone marrow mesenchymal stem cels of the Sprague-Dawley rat femur. Compared with the blank and negative controls, N1-ICD-Ad infection in the c-Kit+ bone marrow mesenchymal stem cels led to substantial accumulation of N1-ICD in the cytoplasm and nuclei, significantly unregulated expressions of Hes1 (a downstream gene of Notch) and cardiomyocyte differentiation genes Nkx2.5 and cTnT, significantly increased the expression of von Wilebrand factor, an endothelial cel differentiation gene, and mildly increased the expression of smooth muscle22α, a smooth muscle cel differentiation gene. These experimental results indicate that the activation of Notch1 signaling contributes to multi-lineages differentiation of c-Kit+ bone marrow mesenchymal stem cels, and the construction of N1-ICD overexpressing adenoviral vector makes the foundation for further research on the role of Notch1 signaling in stem cel biology.

Carcinoma, Hepatocellular ; complications ; Hemangioma ; complications ; diagnosis ; immunology ; pathology ; Hepatitis B, Chronic ; complications ; Hepatocytes ; cytology ; pathology ; Humans ; Liver Cirrhosis ; complications ; Liver Neoplasms ; complications ; Male ; Middle Aged ; Platelet Endothelial Cell Adhesion Molecule-1 ; immunology ; Spleen ; immunology ; pathology ; Tomography, X-Ray Computed

Objective To study the effect of Jade -Screen Powder (JSP)on regulating expression of 5 microRNAs associated with helper T cells in asthmatic mouse model.Methods Forty Balb /c mice were randomly di-vided into 4 groups,1 0 mice for each group,namely normal control,asthma model,JSP treatment and Dexamethasone treatment.The mouse models of allergic inflammation on both upper and lower airways were established by ovalbumin sensitization and challenge.Interleukin(IL)-1 3 and IL -1 7 expressions were detected from lung homogenates by ELISA.Hematoxylin and eosin staining was also performed to observe the pathological changes in the lung tissue.The expressions of miR -1 46a,miR -1 46b,miR -21 0,miR -1 26 and miR -21 a were detected by quantitative real time PCR from splenocytes.Results The lower levels of IL -1 3 [(6.382 ±1 .690)μg/L]and IL -1 7 [(24.21 2 ± 1 .250)μg/L]were found in JSP treatment group compared with those in the asthma model group [(20.1 54 ±7.960)μg/L;(50.31 2 ±5.770)μg/L,rseparately],there was significant difference in IL -1 3 between JSP group and the asthma model group,as well as IL -1 7 (t =3.785,P =0.005;t =9.891 ,P =0.000).Same findings were found in Dexamethasone treated group as well [IL -1 3:(9.366 ±3.460)μg/L,IL -1 7:(29.1 32 ±4.960)μg/L;t =2.779, P =0.024;t =6.225,P =0.000].However,upregulation of miR -21 0 was observed in JSP treatment group (2.052 ± 0.871 )compared with that in the asthma model group (4.034 ±1 .379)(3.95 folds,t =2.71 8,P =0.026).Mean-time,the expression of miR -1 26 in JSP group (4.920 ±0.924)and Dexamethasone group (3.862 ±1 .51 0)in-creased compared with asthma model group (6.024 ±0.447)(2.1 5 folds,t =2.405,P =0.043,and 4.48 folds,t =-3.069,P =0.01 5).Conclusions Th2 and Th1 7 T cells participate in the pathogenesis of asthma and the asthmatic process can be inhibited by JSP.JSP may affect the helper T cells by regulating miR -21 0 and miR -1 26.

AIM: To observe the change of subunit of NADPH oxidation enzyme complex nox - 1 protein in cardiocyte hypoxia - reoxygenation injury and the role of cardiotrophin -1.METHODS: Cardiomyocytes from the hearts of 1 -3 d old neonatal rats were prepared by a modified method. Five groups were included in the study: control; hypoxia/ reoxygenation; hypoxia/reoxygenation + CT - 1; CT - 1 + hypoxia/reoxygenation + LY294002 (PIK3/Akt inhibitor) ; CT -1 + hypoxia/reoxygenation + PD98059 (ERK inhibitor) ; CT - 1 + hypoxia/reoxygenation + DMSO. The concentration of CT -1 was 10 μg/L. The survival rate of myocytes was evaluated by MTS method. Apoptosis, mitochondrial permeability transition pore ( △ψm) and reactive oxygen species ( ROS) were detected by flow cytometry. Nox - 1 protein was determined by Western blotting. RESULTS: Apoptosis of cardiomyocytes and the level of ROS (19.7% ±1.4% vs 2.1% ± 0.5% , 14.07% ± 1.25% vs 3.54% ± 0.86% , P < 0.05 ) increased markedly after hypoxia/reoxygenation, but cardio-myocyte survival rate and the level of△ψm (40.55% ±4.25% vs 86.28% ±7.15% , P <0.01) decreased significantly. The expression of nox - 1 protein was upregulated markedly. With CT - 1 intervention, cardiomyocyte survival rate increased markedly, apoptosis, both ROS and expression of nox - 1 protein reduced significantly. The level of△ψm increased obviously. The effect of CT - 1 was inhibited by LY294002.No significant effect was observed on cells survival in DMSO group, which confirmed that LY294002 was specifically involved in blocking the protective effect of CT - 1.CONCLUSION : The expression of subunit of NADPH oxidation enzyme complex nox - 1 protein is upregulated markedly in cardiocyte hypoxia - reoxygenation injury.CT - 1 protects cardiac cells against hypoxia - reoxygenation injury by downregulating the expression of nox -1 protein to decrease the level of ROS.

OBJECTIVETo explore the role of early growth response factor-1 (Egr-1) signal pathway in acute intrahepatic cholestatic liver injury in rats.

METHODSA single dose (50 mg/kg) of alpha-naphthylisothiocyanate (ANIT) was administered by gavage to each experimental rat to induce intrahepatic cholestasis. Liver tissue and serum specimens were collected from rats at 24, 48 and 72 h after the intoxication. The values of Egr-1, cytokine induced neutrophil chemoattractant 1(CINC-1), macrophage inflammatory protein-2 (MIP-2) mRNA, the protein expression of inducible nitricoxide synthase (iNOS) and the values of tumor necrosis factor alpha (TNF alpha) and interleukin 6 (IL-6) were assayed by real-time PCR, immunohistochemistry, and ELISA, respectively. The levels of MDA, SOD, NO and MPO were assayed by thiobarbituric acid method, xanthine oxidase method, nitric acid deoxidizing assay, and colorimetric method, respectively.

RESULTSIn the model group at 24, 48, 72 h after intoxication, the values of CINC-1 and MIP-2 mRNA were higher than those of the controls (P < 0.05). In the model group at 24, 48 h after intoxication, the value of Egr-1 mRNA was higher than that of the controls (P < 0.05), but there was no significant difference at 72 h (P < 0.05). Of the model group, the absorbance value of iNOS was lower than that of the controls at 24, 48 and 72 h (P < 0.05). Of the model group at 24, 48, 72 h after intoxication, the values of TNF alpha, IL-6, MPO and MDA were higher than those of the controls (P < 0.05), but the values of NO and SOD were lower than those of the controls (P < 0.05).

CONCLUSIONSEgr-1 signal pathway is involved in acute intrahepatic cholestatic liver injury induced by ANIT. After Egr-1 was activated, CINC-1 and MIP-2 are activated consequently and attract neutrophils into the liver. TNF alpha and IL-6 are activated at the same time, so neutrophils are activated and the resulting lipid peroxidation and MDA increased, injuring the liver. iNOS and NO may play a protective role in acute intrahepatic cholestatic liver injury induced by ANIT.

1-Naphthylisothiocyanate ; Acute Disease ; Animals ; Chemokine CXCL1 ; metabolism ; Chemokine CXCL2 ; metabolism ; Cholestasis, Intrahepatic ; chemically induced ; metabolism ; pathology ; Early Growth Response Protein 1 ; metabolism ; Interleukin-6 ; metabolism ; Liver ; chemistry ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism