OBJECTIVETo observe the suppression of the expression of androgen receptor (AR) gene in PC3 cells after AR-specific siRNAs transfection, and to search for the siRNA (s) with the greatest suppressing efficiency.

METHODSFive AR-specific siRNAs were selected, RNAi expression vectors were constructed and transfected into PC3 cells, and the AR expression was detected by real time FQ-PCR and Western blot. A nonsense small RNA was set as negative control.

RESULTSCompared with the control group, the AR expression decreased in various degrees in the 5 experimental groups (P < 0.05), and siRNA1, siRNA4 and siRNA5 showed the greatest suppressing efficiency as compared with the other experimental groups, with statistically significant difference (P < 0.05).

CONCLUSIONThe AR-specific siRNAs could suppress the endogenous expression of target gene. Three siRNAs with great suppressing efficiency were identified and the expression vectors were constructed successfully. It can be applied in the future researches in vivo.

Blotting, Western ; Cell Line, Tumor ; Genetic Vectors ; genetics ; Humans ; Male ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics ; Receptors, Androgen ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Transfection

OBJECTIVETo develop a system for quick screening of efficient siRNA targeted HBx mRNA.

METHODSUsing recombination DNA technique, the fusion expression plasmid of HBx and EGFP was constructed, and siRNA expression cassettes (SECs) containing U6+1, H1 or tRNA(Val )promoter were prepared via one-step overlapping extension PCR. By co-transfection with recombinant plasmid and SECs into AD293 cell, the inhibition effects on the transient expression of HBx-EGFP fusion protein were analyzed by FACS and semi-quantitated RT-PCR analysis.

RESULT(1)HBx-EGFP fusion protein expression plasmid pHBx-EGFP was constructed successfully, which expressed green fluorescence in cell mainly located at plasma or the periphery of nucleus in granules. (2) Co-transfection with recombinant plasmid and SECs into AD293 cells resulted in inhibition of HBx-EGFP expression. SEC-siHBx388 showed significant inhibition effect on HBx-EGFP expression compared with SEC-siHBx271, indicating that siHBx388 is effective siRNA site and could be screened out with our screening system. In addition,the results of that U6+1-, tRNA(Val) and H1-siHBx388 reduced HBx-EGFP expression by 21.7%, 12.9% and 12.4% of control respectively indicated that both tRNAVal and H1 promoter was high efficient in driving effect of siHBx388.

CONCLUSIONCombination of the HBx expression carrying reporter gene and PCR-based multi promoter SECs may develop a useful system to be applied in identification of optimal HBx- siRNA and its matching promoter.

Base Sequence ; Cells, Cultured ; Genetic Therapy ; Green Fluorescent Proteins ; Humans ; Luminescent Proteins ; genetics ; Molecular Sequence Data ; Plasmids ; RNA, Small Interfering ; analysis ; Recombinant Fusion Proteins ; biosynthesis ; Trans-Activators ; antagonists & inhibitors ; genetics ; Transfection

OBJECTIVETo evaluate the clinical significance of AFP-L3 in patients with hepatocellular carcinoma.

METHODSSerum AFP-L3 variants were separated by micro centrifugal column, and detected by chemiluminescence.

RESULTSAFP and AFP-L3 levels were higher in patients with hepatocellular carcinoma than those in patients with chronic hepatitis (P<0.001); as a diagnostic target, the sensitivity and specificity of AFP-L3 were 72.3 percent and 97.2 percent, respectively. Eight patients with hepatitis have higher AFP-L3, but none of them were found with carcinoma by CT three months later.

CONCLUSIONAFP-L3 is very useful in the diagnosis of patients with hepatocellular carcinoma.

Adult ; Carcinoma, Hepatocellular ; blood ; diagnosis ; Centrifugation ; Female ; Humans ; Liver Neoplasms ; blood ; diagnosis ; Male ; Middle Aged ; alpha-Fetoproteins ; analysis ; isolation & purification

OBJECTIVETo investigate the expression of clusterin protein in bladder transitional cell carcinoma (BTCC) and it's association with tumor cell proliferation and apoptosis.

METHODSA tissue microarray (TMA) containing 87 informative cases of BTCCs was constructed firstly. The methods of immunohistochemistry and terminal deoxynucleotidyl transferase-mediated nick end-labeling were then used to examine the expression of clusterin and Ki-67 protein and the status of cell apoptosis in BTCC, respectively, and the correlations between different markers and the clusterin expression associated with patients' clinico-pathological features were evaluated.

RESULTSIn TMA of 87 BTCCs, 37 (43%) cases were observed overexpression of clusterin. A significant association of clusterin expression with BTCC's pathological grade, as well as with tumors clinical stage was observed (P < 0.01), where the frequency of overexpression of clusterin in poor differentiated BTCCs (G(3), 71%) and tumors in more advanced stage (T(2-4), 62%) was significantly higher than that in well differentiated BTCCs (G(1-2), 29%) and tumors in early stage (T(a-1), 28%). In addition, a significant correlation between clusterin expression and tumors apoptotic index (AI) was evaluated (P < 0.01), in which 57% of BTCCs with overexpression of clusterin were observed a lower AI, while 72% of tumors with normal expression of this protein showed a higher AI, but no correlation between clusterin and Ki-67 expression.

CONCLUSIONSThe overexpression of clusterin is associated positively with BTCC's malignant clinical phenotypes including tumor's differentiation and invasive depth, and it is correlated inversely with AI of tumor cells.

Apoptosis ; Biomarkers, Tumor ; metabolism ; Carcinoma, Transitional Cell ; metabolism ; pathology ; Clusterin ; metabolism ; Female ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Ki-67 Antigen ; metabolism ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Urinary Bladder Neoplasms ; metabolism ; pathology

OBJECTIVETo study the changes of serum interleukin-2 (IL-2), interleukin-8 (IL-8) and immunoglobulin (IgG, IgA, IgM) in patients with esophageal cancer, and to probe the relationship between the levels of IL-2, IL-8, IgG, IgA and IgM and the progress of cancer.

METHODSThe serum levels of IL-2 and IL-8 were detected by enzyme-linked immunosorbent assay for 72 case of primary esophageal cancer, 68 advanced esophageal cancer and 120 healthy controls, and the level of immunoglobulin (IgG, IgA, IgM) in patients with esophageal cancer was dynamically observed.

RESULTSThe IL-2 level in patients with early esophageal cancer [(1.69 +/- 0.53) ng/ml] or late esophageal cancer [(1.11 +/- 0.60) ng/ml] was lower than the control group [(2.78 +/- 0.51) ng/ml] (P < 0.01), the late esophageal cancer group was lower than early esophageal cancer group (P < 0.05). The level of IL-8 in patients with early esophageal cancer [(85.48 +/- 6.14) ng/L] or late esophageal cancer [(121.41 +/- 6.22) ng/L] was much higher than the control group [(54.48 +/- 12.20) ng/L] (P < 0.01), the late esophageal cancer group was much higher than early esophageal cancer group (P < 0.01); There was correlation between the levels of IL-2 and IL-8 and the worsen-extent of the tumour in patients with early esophageal cancer or late esophageal cancer. But the level of IgG [(12.23 +/- 2.50) g/L], IgM [(1.60 +/- 0.80) g/L] in the patients with esophageal cancer compared with the level of IgG [(11.65 +/- 3.70) g/L], IgM [(1.46 +/- 0.71) g/L] in the health control group have no significant difference (P > 0.05), the level of IgA [(3.50 +/- 1.10) g/L] in patients with esophageal cancer Compared with the control group [(1.88 +/- 1.08) g/L] has significant difference (P < 0.01), and along with the worsen-extent of the tumor in patients the level of IgA has the increased tendency.

CONCLUSIONThe IL-8 might accelerate the pathogenesis of esophageal cancer, and the IL-2 might restrain. The positive correlation between the level of IgA and the patients with esophageal cancer is observed in this study; the immune maladjustment of IL-2, IL-8 and IgA might be correlative to esophageal cancer, and the IL-2, IL-8 and IgA levels might be an available index for the severity of esophageal cancer, Which may be of some help for clinic practitioners to judge the progress, curative effect and prognosis of the cancer.

Adult ; Aged ; Case-Control Studies ; Esophageal Neoplasms ; blood ; pathology ; Female ; Humans ; Immunoglobulin A ; blood ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Interleukin-2 ; blood ; Interleukin-8 ; blood ; Male ; Middle Aged ; Neoplasm Staging

With the advancement of manned space science and technology,the scope of human exploration into outer space has been gradually expanded.The time and distances that astronauts flight on-board are becoming longer and longer.However,long-term microgravity can negatively affect astronauts' physical and psychological aspects,thus influencing their health and leading to the failure of work tasks,and even threatening their lives.This paper reviews USA,Russia and Europe research results in the field of astronauts on-board physical training methods and health protection measures,with the purpose of learning from their successful experience.Overall,it is suggested that for cross complementation in sport science,we should make the introduction of advanced and successful training systems to solve problems existing in astronauts on-board training,ensuring the maintenance of health.

Objective To explore the effect of high altitude exposure and short-term acclimation on the platelet-associated parameters by studying the changes of platelet-associated parameters in healthy young man.Methods Four hundred and sixtytwo young men were recruited from Chengdu (the elevation of 500m) during Jun.2012 to Aug.2013 according to the inclusion and exclusion criteria,of which 193 had been living in Chengdu (plain group),and 269 flew from Chengdu to Lhasa (3700m) in 2 hours,and then 147 of them were exposed in the high altitude for 1 day (acute high altitude exposure group),and another 122 for 7 days (short-term acclimation group).The demographic data were collected and the blood routine and platelet-associated parameters were measured of all the participants,and then the information collected were compared between the 3 groups.Results Compared to the plain group,the platelet count (PLT),platelet distribution width (PDW) and plateletcrit (PCT) reduced and the mean platelet volume (MPV) increased significantly (P＜0.05) in the acute high altitude exposure group;while all the indexes in short-term acclimation group returned to approach the levels in the plain group,but statistical differences still existed in PLT,PDW and PCT (P＜0.05).The platelet activating factor (PAF) and epinephrine (Epi) decreased markedly in acute high altitude exposure group than in plain group (P＜0.05);while in short-term acclimation group,the PAF returned to approach the level in plain group,and the Epi was further down (P＜0.05).In addition,no marked difference of 5-HT level was observed in the 3 groups (P＞0.05).Pearson correlation analysis indicated that high altitude-induced reduction of oxygen saturation (SpO2) was positively related to the changes of PLT.Conclusions Acute high altitude exposure may reduce PLT,PDW and PCT levels,but elevate MPV and enhance the platelet activity.The reduction of SpO2 might be associated with the changes of PLT,PDW and PCT.Platelet-associated parameters may be recovered to normal with a compensatory effect after short-term acclimation.

OBJECTIVETo investigate the chemical constituents of the herbs of Taraxacum mongolicum.

METHODThe chemical constituents were isolated by various column chromatographic methods and their structures elucidated mainly by NMR and MS evidences.

RESULTForty-four components were obtained and identified were as artemetin (1), quercetin (2), quercetin-3', 4', 7-trime-thyl ether (3), luteolin (4), luteolin-7-O-beta-D-glucopyranoside (5), luteolin-7-O-beta-D-galactopyranoside (6), genkwanin (7), isoetin (8), hesperetin (9), genkwanin-4'-O-beta-D-lutinoside (10), hesperidin (11), quercetin-7-O-[beta-D-glucopyranosyl (1-->6) -beta-D-glucopyranoside (12), quercetin-3, 7-O-beta-D-diglucopyranoside (13), isoetin-7-O-beta-D-glucopyranosyl- 2'-O-alpha-L-arabinopyranoside (14), isoetin-7-O-beta-D-glucopyranosyl-2'-O-alpha-D-glucopyranoside (15), isoetin-7- O-beta-D-glucopyranosyl-2'-O-beta-D-xyloypyranoside (16), caffeic acid (17), furulic acid (18), 3-O-caffeoylquinic acid (19), 3, 5-di-O-caffeoylquinic acid (20), 3, 4-di-O-caffeoylquinic acid (21), 4, 5-di-O-caffeoylquinic acid (22), 1-hydroxymethyl-5-hydroxy-phenyl-2-O-beta-D-glucopyranoside (23), p-hydroxybenzoic acid (24), p-coumaric acid (25), 3, 5-dihydroxylbenzoic acid (26), gallic acid (27), gallicin (28), syringic acid (29), 3, 4-dihydroxybenzoic acid (30), caffeic acid ethyl ester (31), esculetin (32), rufescidride (33), mongolicumin A [6, 9, 10-trihydroxy-benzoxanthene-1, 2-dicarboxylic acid] (34), mongolicumin B [1 l-hydroxy-2-oxo-guaia-1 (10), 3, 5-trien-8, 12-lactone] (35), isodonsesquitin A (36), taraxacin (37), sesquiterpene ketolactone (38), taraxasteryl acetate (39), phi-taraxasteryl acetate (40) and lupenol acetate (41), palmitic acid (42), beta-sitosterol (43), and stigmasterol (44).

CONCLUSIONFour compounds (14, 15, 34 and 35) were new compounds, compounds 1, 3, 6-13, 20-22, 30 and 31 were isolated from this genus for the first time, while compounds 18, 23-29, 32 and 37-42 were obtained from this species for the first time.

Drugs, Chinese Herbal ; chemistry ; Flavonols ; chemistry ; Mass Spectrometry ; Taraxacum ; chemistry

The reporting of complications following transperitoneal and retroperitoneal open radical nephrectomy (RN) is nonstandardized. This study aimed to compare early complications between the two approaches using a standardized reporting methodology in a large contemporary cohort. Between 1996 and 2009, 558 patients underwent open RN for renal cell carcinoma (RCC) in our two centers (424 from Sun Yat-sen University Cancer Center and 134 from the First Affiliated Hospital of Sun Yat-sen University). Records were reviewed for clinicopathologic features and complications. Complications were graded using the Clavien system based on the severity of impact. One hundred and five patients (18.8%) had one or more early complications (168 complications overall). The overall rates of grades I to V complications were 5.6%, 10.8%, 2.2%, 0.4%, and 0.2%, respectively. Patients who underwent transperitoneal RN did not experience more overall or procedure-related complications than those who underwent retroperitoneal RN (P = 0.911 and P = 0.851, respectively). On subgroup analysis, neither grade I/II nor grades III-V complications were significantly different between the transperitonal RN and retroperitoneal RN groups. Multivariate analysis showed that for any grade of complication, age (P = 0.016) and estimated blood loss (P = 0.001) were significant predictors. We concluded that open RN is a safe procedure associated with low rates of serious morbidity and mortality. Compared with retroperitoneal RN, transperitoneal RN was not associated with more complications. Older patient and more blood loss at surgery were independent predictors for higher early postoperative complication rates.
Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Blood Loss, Surgical ; Carcinoma, Renal Cell ; pathology ; surgery ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Staging ; Nephrectomy ; methods ; Postoperative Complications ; Retrospective Studies ; Young Adult

Aim To observe the inhibitory effect of Alpha-momorcharin (α-MMC) on the inflammatory cytokine storm of Ml-type inflammatory macrophages induced by LPS and explore its possible targeting mechanism. Methods Western blot was used to detect the expression of WIL2-S B lymphocytes, H9 T lymphocytes, THP-1 monocytes and M0 macrophages LRP1 receptor protein. CCK-8 method was used to detect the survival rate of the four cells. ELISA was used to detect the expression level of inflammatory cytokines in Ml macrophages. Western blot was used to detect the expression of TLR4 signaling pathway-related protein in Ml macrophages. Results Macrophages had a high density of LRP1 receptors consistent with monocytes; the survival rate of α-MMC on the four cells was positively correlated with the density of this receptor; α-MMC inhibited the expression of inflammatory cytokinesTNF-α, IL-lβ, IL-6, IL-8, MlP-lα and MCP-1 in Ml macrophages in a dose-and time-dependent manner; α-MMC showed significant inhibition to TAKl/pTAK1, p-JNK, p-APl and p-p65 signaling proteins of the TLR4 signaling pathway, and this inhibition could be blocked by the LRP1 receptor blocker RAP. Conclusions α-MMC selectively inhibits macrophage inflammatory cytokine synthesis by inhibiting TAK1 of the TLR4 signaling pathway, which in turn inhibits the downstream NF-ΚB and MAPK pathways, mediated by the LRP1 receptor. The selective immunosuppressive effect of α-MMC on macrophages may make it a very promising agent for the treatment of acute infectious macrophage activation syndrome (MAS).