Objective: To investigate the analgesic mechanism of Tuina (Chinese therapeutic massage) by observing the effect of the N-methyl-D-aspartate receptor subunit 2B (NR2B)/postsynaptic density-95 (PSD-95) pathway on the dendritic structure of spinal cord dorsal horn in rats with lumbar disc herniation. Methods: Fifty Sprague-Dawley rats were randomly divided into a blank group, a model group, a Tuina group, a blocker agent group, and a blocker agent + Tuina group. The sciatic nerve chronic constriction injury (CCI) model was prepared by the sciatic nerve ligation method. From the 4th day after modeling, rats in the Tuina group and the blocker agent + Tuina group were subject to daily Tuina intervention, and those in the blocker agent group and the blocker agent + Tuina group were daily intrathecally injected with NR2B blocker agent (MK-801). The spontaneous pain score was used to observe the pain behavior of all rats. The expression levels of NR2B and downstream PSD-95 were measured by immunohistochemistry, and the dendritic structure changes were observed by Golgi staining for rat spinal cord dorsal horn after 14 d of continuous intervention. Results: Compared with the blank group, the degree of rat spontaneous pain after CCI was elevated in both the model and the Tuina groups (P<0.01) and was reduced in the Tuina group after the Tuina intervention compared with the model group (P<0.05). Compared with the model group, the rat spontaneous pain level after blocking NR2B was reduced in both the blocker agent group and the blocker agent + Tuina group (P<0.05). The NR2B and PSD-95 protein levels were significantly higher in the model group compared with the blank group (P<0.01); the total number of dendritic branches was increased (P<0.01), and the total dendritic length became longer (P<0.01) in the spinal cord dorsal horn. The rat NR2B and PSD-95 protein levels were significantly decreased in the Tuina group compared with the model group (P<0.01); the total dendritic branch number was reduced (P<0.01) and the total length was shortened (P<0.01) in the spinal cord dorsal horn. After blocking NR2B, the expression levels of NR2B and downstream PSD-95 protein were significantly lower in both the blocker agent group and the blocker agent + Tuina group compared to the model group (P<0.01). The total branch number was significantly reduced (P<0.01), and the total length was significantly shortened (P<0.01) of the dendrites in the spinal cord dorsal horn. Conclusion: Tuina may exert an analgesic effect by remodeling the dendritic structure in the spinal cord dorsal horn in rats with lumbar disc herniation, and its mechanism may be related to the inhibition of NR2B/PSD-95 signaling pathway.

OBJECTIVE: To investigate the role of nuclear factor erythroid-2-related factor 2(NRF2) signaling pathway in trichloroethylene(TCE) induced oxidative stress in human liver cancer HepG2 cells. METHODS: HepG2 cells in the exponential growth phase were randomly divided into control group and low-, medium-and high-dose groups, and the cells were stimulated with TCE at the final concentrations of 0, 2, 4 or 8 mmol/L respectively for 24 hours. After TCE exposure, the cells were collected. The activity of superoxide dismutase(SOD) was measured by 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulphenyl)-2 h-tetrazole monosodium salt method.The activities of catalase(CAT) and glutathione peroxidase(GSH-Px) were detected by enzymatic method. The level of malondialdehyde(MDA) was detected by thiobarbituric acid method. The level of 8-hydroxydeoxyguanosine(8-OHDG) was detected by enzyme-linked immunosorbent assay. The mRNA expression of NRF2, heme oxygenase(HO1), glutamate-cysteine ligase catalytic subunit(GCLC), NAD(P) quinone oxidoreductase 1(NQO1) were detected by real-time polymerase chain reaction and the protein expression of NRF2, HO1, GCLC, NQO1 were detected by Western blotting. RESULTS: The activity of CAT in HepG2 cells decreased in the 3 doses drug exposure groups(all P<0.05). The activity of GSH-Px increased(all P<0.05), while the level of MDA in HepG2 cells decreased in low-dose group compared with the control group(P<0.05). There was no statistical significance in SOD activity and 8-OHDG level of HepG2 cells among these 4 groups(all P>0.05). The mRNA and protein relative expression of NRF2 and GCLC in HepG2 cells decreased in the low-and medium-dose groups(all P<0.05) compared with the control group. The mRNA relative expression of HO1 decreased(all P<0.05). The HO1 protein relative expression of HepG2 cells increased in low-dose group compared with the control group(P<0.05). The mRNA and protein relative expression of NQO1 in low-dose group increased(both P<0.05). The protein relative expression of HO1 in HepG2 cells in medium-dose group was lower than that in control group(P<0.05).The mRNA and protein relative expression levels of NRF2 and NQO1 increased in HepG2 cells of high-dose group(all P<0.05) and the mRNA relative expression of HO1 increased in HepG2 cells of high-dose group(all P<0.05) compared with the other 3 groups. CONCLUSION: The low and medium dose TCE stimulation can cause oxidative stress in HepG2 cells and decrease the antioxidant enzyme activity. The high dose TCE stimulation to HepG2 cells can activate NRF2 signaling pathway, thus upregulating the expression of downstream antioxidant enzymes HO1, GCLC and NQO1, and relieving the oxidative damage caused by TCE.

Objective　To analyze the correlation between the length,angle,thickness,arc length,opening direction,plaque and thrombus of carotid web and the incidence of ischemic stroke.Methods　Forty six patients with carotid web diagnosed by DSA were selected,of which 21 were ischemic stroke patients as the study group and 25 were non ischemic stroke patients as the control group,Color Doppler Ultrasound was used to measure the length,angle,thickness and arc length of the carotid web in the two groups respectively,and to observe the opening direction,whether there were plaques and thrombosis.Results　The length,angle and arc length of the base of the web in the study group were longer than those in the control group (P<0.05).The incidence of plaques and thrombus in the study group was higher than that in the control group (P<0.05),but there was no significant difference in the thickness and opening direction of the web between the two groups (P>0.05).Conclusion　The length,angle,arc length of the base,plaque or thrombus are closely related to the occurrence of ischemic stroke,which can be used as an effective index to evaluate the prognosis of patients with carotid web.

OBJECTIVE: To study the value of plasma miRNA23-a and miRNA-451 as potential biomarkers for early diagnosis of non-small cell lung cancer (NSCLC). METHODS: Fifty patients with NSCLC and 50 healthy control subjects were recruited for testing the plasma levels of miRNA23-a and miRNA-451 and their expression levels in the tumor tissues using qRT-PCR. The correlations of the plasma levels of miRNA23-a and miRNA-451 with their expressions in the tumor tissues were analyzed. The diagnostic power of CEA, miRNA23-a and miRNA-451 for NSCLC was evaluated using the receiver-operating characteristics (ROC) curves and the area under the ROC curves (AUC). In the NSCLC cell line A549, we tested the effect of inhibition of miRNA-23a and miRNA-451 on the expression levels of SPRY2 and MIF mRNA using qRT-PCR. RESULTS: The expression levels of miRNA-23a and miRNA-451 in NSCLC tissues was significantly associated with smoking, tumor size, lymph node metastasis and TNM stage ( < 0.05). Compared with those in the control group, miRNA-23a level was significantly increased while miRNA-451 was significantly down-regulated in the tumor tissues and plasma of NSCLC patients. The plasma levels of miRNA-23a and miRNA-45 were strongly correlated with their expression levels in the tumor tissues. ROC analysis showed that for the diagnosis of NSCLC, the AUC, sensitivity and specificity of either miRNA-23a or miRNA-451 were significantly higher than those of CEA ( < 0.05). The combination of miRNA23-a and miRNA-451 markedly improved the AUC (0.900), sensitivity (78%) and specificity (86%) for the diagnosis. In A549 cells, inhibition of miRNA23-a and miRNA-451 resulted in significantly increased expression levels of SPRY2 mRNA and MIF mRNA, respectively. CONCLUSIONS miRNA-23a and miRNA-451 can be used as potential biomarkers for early diagnosis of NSCLC, and their combined detection can be more effective for the diagnosis.
Biomarkers, Tumor ; Carcinoma, Non-Small-Cell Lung ; genetics ; Case-Control Studies ; Early Detection of Cancer ; Humans ; Intracellular Signaling Peptides and Proteins ; Lung Neoplasms ; genetics ; Membrane Proteins ; MicroRNAs ; ROC Curve

Objective To study the value of serum human epididymal protein 4 (HE4) ,cytokeratin 19 frag-ment (CYFRA21-1) ,neuron-specific enolase (NES) and gastrin release precursors (Pro-GRP) in the diagnosis of female lung cancer .Methods A total of 100 cases of female lung cancer patients in the hospital were select-ed as the research object ,100 cases of benign lung diseases and 100 female health examiners as the control ,the serum levels of HE4 ,CYFRA21-1 ,NES and Pro-GRP were measured ,and the related statistical analysis was carried out .Results The serum levels of HE4 ,CYFRA21-1 ,NES and Pro-GRP in patients with lung cancer were significantly higher than those of benign lung disease and healthy control group (P<0 .05) ,and there was no significant difference between the benign lung disease group and the healthy control group (P>0 .05) . There was no significant difference in serum HE4 expression in different stages and pathological types of lung cancer (P>0 .05) .The ROC curve analysis showed that the area (AUC) of HE4 ,CYFRA21-1 and NES/Pro-GRP w ere 0 .927 ,0 .758 ,0 .652 and 0 .799 respectively ,and the best critical values w ere 63 .38 ,2 .05 ,14 .05 and 58 .50 respectively ,and the sensitivity was 88 .0％ ,80 .0％ ,60 .0％ ,71 .0％ respectively ,and the speci-ficity was 96 .0％ ,73 .0％ ,87 .0％ and 89 .0％ respectively .HE4 was obviously better than the other 3 items . Combined detection of HE4 ,CYFRA21-1 ,NES and Pro-GRP could also improve the diagnostic sensitivity of lung cancer ,which was 89 .0％ ,but the specificity had decreased by 88 .0％ .Conclusion The level of serum HE4 in female patients with lung cancer is significantly higher ,which can be used as a candidate marker for differential diagnosis of pulmonary benign and malignant diseases .The combined detection of these 4 markers has a high sensitivity for the diagnosis of female lung cancer ,which is suitable for the survey of female lung cancer in clinical .

Objective To explore the association of synovial fluid vasoactive intestinal peptide levels with cartilage damage,radiological changes and symptomatic severity in patients with ankle post-traumatic osteoarthritis. Methods 74 patients with ankle traumatic osteoarthritis undergoing ankle anthroscopic debridement or joint replacement and 69 healthy controls receiving body check were enrolled in the this study. Serum and synovial fluid VIP concentrations were measured by a special radioimmunoassay method. Cartilage degradation biomarker colla-gen type Ⅱ(CTX-II)and inflammatory marker interleukin-6 were determined by enzyme-linked immunosorbent assay (ELISA). The symptomatic and functional severity was evaluated using Teeny & Wiss and AOFAS ankle-hindfoot rating scale. The radiographic progression of PTAOA was identified according to the modified ankle osteoar-thritis Kellgren-Lawrence (K-L) grading system. The mankin score was used for assessing the histopathological severity for cartilage lesions. Receiver operating characteristic (ROC) curve was conducted and the area under curve(AUC)was used to evaluate the diagnostic value of VIP,IL-6 and CTX-II levels for the prediction of the modified K-L grading by comparing with other biomarkers. Results There were no significant differences in serum VIP levels between PTAOA patients and controls. VIP levels in synovial fluid showed a negative correlation with modified ankle K-L grading,Mankin scores,CTX-Ⅱand IL-6. In addition,VIP levels were also positively associated with Teeny&Wiss and AOFAS ankle-hindfoot scores. The AUC area of VIP was similar to CTX-Ⅱat early stage of the disease. Conclusions Synovial fluid VIP levels show an independent and negative correlation with disease severity in patients with PTAOA. Low level of VIP in SF can be used as a potential biomarker for reflecting disease progression.

Objective To reduce the incidence rate of hospital-wide indwelling needle catheter blockage through a nursing project.Methods We set up a nursing project team,conducted a cross-sectional survey to know the using status of hospital-wide catheter,and analyzed the reasons of catheter blockage by root cause analysis method. "Reducing the incidence rate of hospital-wide indwelling needle catheter blockage" was established as the theme;improvement activities of nursing project were planned and developed for six months;and improvement methods were developed and implemented.Results The incidence rate of hospital-wide indwelling needle catheter blockage reduced to 3.6% from 8.1% (χ2=752.177,P<0.01),which reaches the goal of controlling the incidence rate of catheter blockage below 4%,and continuously decreases month by month. After implementation,the score of knowledge questionnaire about indwelling needle catheter blockage of nurses in the wards increased from (78.23±7.14) to (96.4±1.48) (t=54.735,P<0.01).Conclusions The nursing project obviously improves the degree of mastering the knowledge about preventing blockage,and reduces the incidence rate of hospital-wide indwelling needle catheter blockage.

Objective To study the closure time of neonatal ductus venosus and the Z score regression equation, and to explore the variation of closure time of neonatal ductus venosus with gestationalage.Methods Color doppler ultrasound was applied to detect the closure time of neonatal ductus venosus in normal newborns, Z score regression equation ( lnY =a +bX +cX2 ) for the closure time of neonatal ductus venosus ( Y ) was developed by regression analysis which used gestational age ( X) as an independent variable. The Z scores of the closure time of neonatal ductus venosus in different gestational age were calculated by the formula [ Z = ( M - Y )/S x , M for observation value, Y for predictivevalue].Results There were 432 cases in our study.The closure time of neonatal ductus venosus was negatively related to gestational age ( r = -0. 938 , P <0. 001 ) . The nonlinear regression equation was lnY= -5. 228+0. 089X-0. 000228X2, R2 =0. 854, Sx =0. 214(P<0. 001). Based on the predicted mean of the closure time and Sx related to different gestational age, Z scores for specific closure time of neonatal ductus venosus could be calculated by the formula [ Z =( M -Y)/Sx ] . The Z scores were normally distributed,and did not change with gestational age.Conclusions Theclosure & nbsp;time of neonatal ductus venosus is negatively related to gestational age. The Z scores obtained by the predicted nonlinear regression equation are normally distributed.

Objective To study the value of intensive care nursing scoring system(ICNSS) on care resource allocation in intensive care unit(ICU). Methods 108 ICU patients were divided into the observation group (55 cases) and the control group (53 cases) randomly. In the observation group,the ICNSS scale was applied to evaluate nursing workload, and care resource was allocated according to intensive care nursing scoring. In the control group, care resource was allocated according to dynamic monitoring of acute physiology and chronic health evaluation (APACHE Ⅱ ) scoring. The ICU monitoring time,medical cost,incidence of complications during the hospitalization,satisfaction of the nurses and the patients or their relatives were compared between two groups. Results The ICU monitoring time,medical cost and incidenceof complications during the hospitalization in the observation group were significantly less than those in the control group, while the satisfaction of the nurses and the patients or their relatives in the observation group was markedly better than that in the control group. Conclusions Care resource allocation according to ICNSS is worthy of promotion and application in ICU because it can more significantly improve nursing quality,satisfaction of nurses and patients or their relatives than care resource allocation according to APACHE Ⅱ scoring.

Objective To construct, express and identify the anti-idiotypic antibody single chain variable fragment (scFv) against Edwardsiella tarda. Methods By using RT-PCR method, the variable regions of the heavy and light chain of the anti-idiotypic monoclonal antibody (mAb) 1E11 against Edwardsiella tarda were cloned and joined with a (Gly_4ser)_3 linker, and the scFv in the orientation of V_L-linker-V_H was constructed. It was then cloned into vector plasmid pET-28a, expressed in Escherichia coli BL21(DE3), and confirmed by SDS-PAGE, Western blot and ELISA. Results The recombinant scFv could be expressed in E.coli BL21 (DE3) in a fusion protein pattern. The expression product was in the form of an inclusion body and the purified fusion protein was obtained after being purified and refolded. The SDS-PAGE and Western blot analysis showed that the molecular had the binding activity to the antigen. Conclusion The recombinant anti-idiotype scFv has been successfully constructed and expressed in E.coli BL21 (DE3), providing the basis and potential for preparation of genetically engineered vaccine against Edwardsiella tarda.