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 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.

With the advancement research on target therapy,more and more target drugs have been applied in the treatment of endometrial cancer,including anti-TKI or monoclone antibody and inhibitors in molecular signal pathways like mTOR,all of which have shown good therapeutic effects.As there were only on some phaseⅡclinical trials,so randomized stageⅢclinical trials are required to confirm these effects.In this review,the author discusses the advancement of target therapy in endometrial cancer.

Object To study the chemical constituents from Epimeredi indica and search for the natu-ral compounds with bioactivities. Methods Three compounds were isolated by a combination of RP-18and silica gel column chromatographies, and their structures were identified by spectral methods. ResultsA novel 2- (3-methoxy-4-hydroxy) phenyl-ethanol 1-O-a-L- [(1→3 )-rhamnopyranosyl- 6-feruloyl] gluco-side ( I ) along with known 2-(3, 4-dihydroxy) phenyl ethanol 1-O-?-L-[(1→3)-rhamnopyranoside-4-O-caffeoyl-] glucoside (verbascoside) ( Ⅱ ), and 2-(3, 4-dihydroxy) phenyl ethanol (1→1) (2→2)-[(1→3)-rhamnopyranoside-4-O-caffeoyl glucoside (oraposide) (Ⅲ) were isolated from E. indica. ConclusionCompound I is a new compound named as epimeredinoside A and compounds Ⅱ and Ⅲ are isolated fromthis plant for the first time.

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.