Objective:To develop a real time fluorescent quantitative reverse transcriptase polymerase chain reaction(RT-PCR) system for determining the expression of Dicer mRNA in human hepatoma cell lines and 20 samples of primary hepatocellular carcinoma(PHC)tissues.Methods: The specific primers,designed according to the complete sequence of Dicer mRNA,and the fluorescence dye SYBR Green Ⅰwere used for RT-PCR amplification.The fluorescence was monitored in a real time manner.The expression levels of Dicer mRNA in samples were calculated according to the standard curve and the nonspecific amplifications were excluded by melting curve analysis.The mRNA levels of Dicer were presented as the ratios of Dicer mRNA to 18S rRNA.Results: The linear detection range of this system was from 5?10~(2)to 5?10~(9)copies/?l and the coefficient of variation values for intra-experimental and inter-experimental reproducibility ranged from 4.13% to 19.72% and from 6.25% to(18.76%,) respectively.The expression levels of Dicer mRNA in HBV positive hepatoma cell line HepG2.2.15 or in HBV negative hepatoma cell line HepG2 were significantly lower(P

Objective To observe the preventive effect of early rehabilitation intervention and intermittent pneumatic compression therapy device (IPC) combined with low molecular weight heparin for cerebral hemorrhage deep vein thrombosis (DVT) formation,thus to provide basis for clinical intervention.Methods 124 patients with cerebral hemorrhage were selected,and they were randomly divided into the observation group and control group,62 cases in each group.The control group received low molecular weight heparin and routine intervention,the observation group received early rehabilitation intervention and IPC treatment on the basis of control group.The venous blood hemorheological indicators were detected,and the incidence of DVT was recorded.Results The whole blood viscosity and D-dimer of the observation group after intervention were (4.17 ± 1.12)mPa/s and (214.84 ± 31.40)ng/mL,which were lower than (5.21 ± 1.08) mPa/s and (331.90 ± 35.38) ng/mL of the control group,the differences were statistically significant(t =8.52,8.90,all P ＜ 0.05).The incidence rate of DVT of the observation group was 1.64％ (1/62),which was lower than 17.74％ (11/62) of the control group,the difference was statistically significant(x2 =9.17,P ＜ 0.05).Conclusion Early rehabilitation intervention and IPC combined with low molecular weight heparin for preventing cerebral hemorrhage DVT formation can significantly increase the venous blood flow velocity,reduce blood viscosity,decrease the incidence of DVT.It has better clinical value for the prevention of DVT formation of cerebral hemorrhage.

The quality of sleep has a great relationship with health and working efficiency. The result of sleep stage classification is an important indicator to measure the quality of sleep, and it is also an important way to diagnose and treat sleep disorders. In this paper, the method of detrended cross-correlation analysis (DCCA) was used to analyze sleep stage classification, sleep electroencephalograph signals, which were extracted from the MIT-BIH Polysomno graphic Database randomly. The results showed that the average DCCA exponent of the awake period is smaller than that of the first stage of non-rapid eye movement (NREM) sleeps. It is well concluded that the method of studying the sleep electroencephalograph with this method is of great significance to improve the quality of sleep, to diagnose and to treat sleep disorders.
Electroencephalography ; Humans ; Signal Processing, Computer-Assisted ; Sleep Stages

Metabonomics is the branch of science concerned with the quantitative understandings of the metabolite complement of integrated living systems and its dynamic responses to the changes of both endogenous factors (such as physiology and development)and exogenous factors (such as environmental factors and xenobiotics). As a holistic approach, metabonomics detects, quantifies and catalogues the time related metabolic processes of an integrated biological system, ultimately, relates such processes to the trajectories of the pathophysiological events. Ever since its birth in 1999, metabonomics has already been described in more than 800 scientific papers and half dozen patents, amongst which almost 700 papers were experimental articles. Now, metabonomics has been established as an extremely powerful analytical tool and hence found successful applications in many research areas including molecular pathology and physiology, drug efficacy and toxicity, gene modifications and functional genomics, and environmental sciences. This holistic approach has thus become an important part of systems biology and has now evolved to be a unique part in global systems biology. The essence of metabonomics and some of the present applications were reviewed to illustrate the rapid development of this extremely exciting new frontier.

Objective To study the expressions of Toll-like receptor (TLR)2 and TLR4 in colonic mucosa of patients with irritable bowel syndrome (IBS) and in normal subjects. Methods Thirty patients with diarrhea predominant IBS and 12 healthy volunteers were enrolled. The expressions of TLR2 and TLR4 in colonic mucosa were examined by immunohistochemistry (IHC).TLR2 and TLR4 were semi-quantitative analyzed with average absorbence. Results Contrast to healthy controls, the lamina propria of IBS patients showed edema and looseness with lots of inflammatory cells infiltration. There was no difference in expression of TLR2 between healthy controls and IBS patients (P>0.05). Compared with healthy controls, TLR2 in crypt epithelium and TLR4 in luminal surface of IBS patients were significantly up-regulated (TLR2 : 6.7 % vs. 50.0 %,TLR4: 40.0% vs. 0, P<0. 05). The TLR4 expressed in intestine epithelial cell of both the apical surface and the basolateral surface in 86.7% of patients with IBS, and in 50% of healthy controls.The positive cells of TLR4 in lamina propria were higher in patients with IBS than those in healthy controls (70. 084 ± 21. 887 vs. 20. 577 ± 4. 546, P<0.01). The A values of TLR2 and TLR4 in colonic mucosa of the patients with IBS were higher than those in healthy controls (TLR2:0. 3079±0. 0283vs. 0.3886±0. 0510,TLR4:0. 3044±0. 0481 vs. 0. 3971 ±:010996,P<0. 01). Conclusions Inflammatory cells infiltrated into colonic mueosa in patients with IBS suggested that inflammation might participate in the pathogenesis of IBS. Up-regulated expressions of TLR2 and TLR4 in IBS patients supposed that they might contribute to the occurrence of IBS.

Objective To study the expressions of TLR4, CD14, MD-2 and NF-kB in colonic mucosa in patients with diarrhea-irritable bowel syndrome (IBS-D) , and compared with normal subjects. The purpose of this study is to explore the role of TLR4 and TLR4 signal transduction pathway in the pathogenesis of IBS-D. Methods The expressions of TLR4, CD14, MD-2 and NF-kB in colon mucosa were examined by immunohistochemistry (IHC) in 30 IBS-D patients and 12 healthy volunteers separately. The average absorbance (A value) of TLR4 was analyzed. The positive expression rates of CD14, MD-2 and NF-kB of colonic mucosa were studied. Results Compared with healthy controls, significant upregulation of TLR4 expression relative to controls was found in colon mucosa of IBS-D. A value of TLR4 in IBS-D was significantly higher (0.3971 ±0.0996 vs 0. 3044 ±0.0481). The positive rate and intensity of NF-kB in IBS-D were significantly higher than those in healthy. The number of positive cells of MD-2 showed significant increase in lamina propria of IBS-D against controls. The percent of CD14 positive was upregulated in lamina propria in IBS-D. The expressions of MD-2 and CD14 in intestine epithelial cell were low or negative. Conclusions There is the activation of the signal transduction pathway of TLR4/NF-kB in the colonic mucosa of patients with IBS-D. Up-regulated expression of TLR4 in IBS patients suppose that it might contribute to occurrence of IBS-D.

Objective:To explore the influence of continuous quality improvement of nursing care on the compliance of senile patient with gastroscopy.Methods: 100 senile patients with gastroscopy were selected, and they were divided into control and improvement groups ( each group included 50 cases ) depended on the different time of hospitalization. The control group was treated with routine nursing quality method, while the improvement group was treated with continuous quality improvement method. A series of indicators in the endoscopic examination, such as the success rate of one time intubation, the used time, yes or no cough and other adverse reactions and the patient's satisfaction for nursing services were compared between the two groups.Results:After 5 months of continuous quality improvement nursing, the success rate of one time intubation in the improvement group was significantly higher than that in the control group, and the checking time of improvement group was significantly less than that of control group, the differences were statistically significant(t=7.162,t=6.61; P<0.01). Besides, on check process, the incidence of adverse reactions in the improvement group was significantly lower than that in the control group and the satisfaction of nursing service was significantly higher than that of the control group, the differences were statistically significant(x2=23.077,x2=10.699,P<0.01).Conclusion:The application of continuous quality improvement nursing for senile patients during gastroscopy can improve the success rate of one time intubation, shorten the gastroscopy time, reduce the incidence of adverse reactions, enhance compliance of gastroscopy and satisfaction for nursing service, and its clinical effect is obvious.

Objective To investigate the influence of hepatitis B virus X (HBx) protein on the activity of NLRP3 inflammasome as well as the mechanism of accelerating HBV-related hepatocellular carcinoma (HCC).Methods The HepG2 cell strains were divided into the 5 groups:blank control group (without plasmid transfection),empty vector group [transfected with pE green fluorescent protein (GFP)-N1 vector plasmid],fulllength HBx protein group (transfected with pEGFP-N1-X plasmid),HBx1-127 group (transfected with pEGFP-N1-X1-127 plasmid),HBx1-101 group (transfected with pEGFP-N1-X1-101 plasmid).(1) The expressions of HBx protein and NLRP3 inflammasome were detected by Western blot [Lipopolysaccharide (LPS) + ATP intervention was performed in the blank control group].(2) The HepG2 cells in the full-length HBx protein group were respectively intervened by glibenclamide and ammonium pyrrolidinedithiocarbamate (APDC),and the expressions of IL-1β and IL-18 were detected by enzyme-linked immunosorbent assay (ELISA).(3) The expressions of reactive oxygen were detected by flow cytometry.The measurement data with normal distribution were presented by (x) ± s.The one-way ANOVA was adopted in the comparison among groups while the t test was used in the pairwise comparison.Results (1) The results of Western blot showed:① the relative expressions of HBx recombinant plasmid fusion protein inside the HepG2 cells in the blank control group,empty vector group,full-length HBx protein group,HBx1-127 group and HBx1-101 group were 0.07 ±0.03,0.92 ±0.13,0.84 ±0.11,0.30 ±0.06 and 0.29 ± 0.05,respectively.The expressions in the HBx1-127 group and the HBx1-101 group represented the expressions of HBx1-127 protein and HBx1-101 protein.There were statistically significant differences among the 5 groups (F =61.790,P ＜ 0.05).The relative expression of full-length HBx protein group was significantly different from that of blank control group,HBx1-127 group and HBx1-101 group (t =12.070,7.465,7.801,P ＜0.05).There was no statistically significant difference between full-length HBx protein group and empty vector group (t =0.867,P ＞0.05) and between the HBx1-127group and HBx1-101 group (t =0.146,P＞0.05).② The relative expression of NLRP3 inflammasome protein inside the HepG2 cells in the blank control group,full-length HBx protein group,HBx1-127 group,HBx1-101 group and LPS + ATP group were 0.29 ±0.06,0.83 ±0.14,0.27 ±0.06,0.27 ± 0.05 and 0.90 ± 0.16,respectively,with a statistically significant difference among the 5 groups (F =29.550,P ＜ 0.05).The relative expression of NLRP3 inflammasome protein of LPS + ATP group was significantly different from that of blank control group,HBx1-127 group and HBx1-101 group,respectively (t =6.310,6.565,6.741,P ＜0.05).There were statistically significant differences between the full-length HBx group and the HBx1-127 group or HBx1-101 group (t =6.381,6.584,P ＜ 0.05) and no statistically significant difference between LPS + ATP group and full-length HBx protein group (t =0.580,P ＞ 0.05).(2) The results of ELISA showed:①) the expression of IL-1β inside the HepG2 cells in the blank control group,full-length HBx protein group,HBx1-127 group,HBx1-101 group and LPS + ATP group was (87 ± 9)pg/mL,(587 ±56)pg/mL,(125 ±12) pg/mL,(113 ± 13) pg/mL and (677 ± 74) pg/mL,respectively,with a statistically significant difference among the 5 groups (F =139.010,P ＜ 0.05).The expression of IL-1 β of LPS + ATP group was significantly different from that of blank control group,HBx1-127 group and HBx1-101 group (t =13.691,12.752,13.001,P ＜0.05).The expression of IL-1β of full-length HBx group was significantly different from that of the HBx1-127 group and the HBx1-101 group (t =14.051,14.283,P ＜ 0.05).There was no statistically significant difference between the LPS + ATP group and the full-length HBx protein group (t =1.691,P ＞0.05).The expression of IL-18 in the blank control group,full-length HBx protein group,HBx1-127 group,HBx1-101 group and LPS + ATP group was (43 ±8)pg/mL,(252 ±38)pg/mL,(70 ± 13)pg/mL,(63 ± 10)pg/mL and (263 ±48)pg/mL,respectively,with a statistically significant difference among the 5 groups (F =44.010,P ＜0.05).The expression of IL-18 of LPS + ATP group was significantly different from that of blank control group,HBx1-127 group and HBx1-101 group,respectively (t =7.848,6.722,7.065,P ＜ 0.05).The expression of IL-18 of full-length HBx group was significantly different from that of HBx1-127 group and HBx1-101 group (t =7.882,8.331,P ＜ 0.05).There was no statistically significant difference between LPS + ATP group and full-length HBx group (t =0.326,P ＞ 0.05).②The expressions of IL-1β and IL-18 in the HepG2 cells of the full-length HBx protein were (587 ± 91)pg/mL and (243 ± 22) pg/mL before the addition of glibenclamide,(115 ± 17) pg/mL and (90 ± 12) pg/mL after the addition of glibenclamide,respectively,with statistically significant differences before and after the addition of glibenclamide (t =8.800,10.566,P ＜ 0.05).The expressions of IL-1β and IL-18 in the HepG2 cells of the full-length HBx protein were (573 ± 89) pg/mL and (252 ± 24) pg/mL before the addition of APDC,(124 ±21)pg/mL and (116 ± 15)pg/mL after the addition of APDC,respectively,with statistically significant differences before and after the addition of APDC (t =8.516,8.269,P ＜ 0.05).(3) The results of flow cytometry showed that the relative expression of reactive oxygen in the HepG2 cells in blank control group,fulllength HBx protein group and LPS + ATP group were 66 ± 14,275 ± 54 and 388 ± 88,with statistically significant differences among the 3 groups (F =22.130,P ＜ 0.05) and between the full-length HBx protein group or LPS +ATP group and blank control group (t =6.489,6.256,P ＜ 0.05).There was no statistically significant difference between full-length HBx protein group and LPS + ATP group (t =1.887,P ＞ 0.05).Conclusion HBx protein may play an important role in the occurrence and development of HBV-related HCC by activating NLRP3 inflammasome through inducing reactive oxygen generation in the HepG2 cells.

Objective To study the clinical effect of low-molecular-weight heparin combined with edaravone in the treatment of progressive cerebral infarction.Methods 60 patients with acute progressive cerebral infarction in 72 hours were randomly divided into treatment group received edaravone combined low-molecular-weight heparin(n=30)and control group received low-molecular-weight heparin(n=30).The primary efficacy was evaluated by NIHSS(the National Institutes of Health Stroke Scale).ADL(Activities of daily living)and clinical effect.Results Our study showed that the score of NIHSS on 7th day post-therapy and 14th day post-therapy in treatment group was lower than that in control group(t=15.987 and 11.756,both P＜0.01).However,the score of ADL in the theatment group on 14th day post-therapy was higher than that of control group(t=62.56,P＜0.01).The overall clinical responserate of treatment group was higher than that of control group(x2=13.695,5.769,all P＜0.01).Conclusion Edaravone combined with low-molecular-weight heparin has remakable effect on the treatment of progressive cerebral infarcti.