Prostaglandin (PG) E2 is an active substance in pathological and physiological mechanisms, such as inflammation and pain. The in vitro high-throughput assay for screening the inhibitors of reducing PEG2 production is a useful method for finding out antiphlogistic and analgesic candidates. The assay was based on LPS-induced PGE2 production model using a homogeneous time-resolved fluorescence(HTRF) PGE2 testing kit combined with liquid handling automation and detection instruments. The critical steps, including the cell density optimization and IC50 values determination of a positive compound, were taken to verify the stability and sensibility of the assay. Low intra-plate, inter-plate and day-to-day variability were observed in this 384-well, high-throughput format assay. Totally 5 121 samples were selected from the company's traditional Chinese medicine(TCM) material base library and used to screen PGE2 inhibitors. In this model, the cell plating density was 2 000 cells for each well; the average IC₅₀ value for positive compounds was (7.3±0.1) μmol; the Z' factor for test plates was more than 0.5 and averaged at 0.7. Among the 5 121 samples, 228 components exhibited a PGE2 production prohibition rate of more than 50%, and 23 components exhibited more than 80%. This model reached the expected standards in data stability and accuracy, indicating the reliability and authenticity of the screening results. The automated screening system was introduced to make the model fast and efficient, with a average daily screening amount exceeding 14 000 data points and provide a new model for discovering new anti-inflammatory and analgesic drug and quickly screening effective constituents of TCM in the early stage.

Brucellosis is one of the most widespread zoonotic diseases, with the most frequent complication being osteoarticular changes. The aim of this study was to assess the changes of C-terminal telopeptide of type II collagen (CTX-II) in patients infected with brucellosis. A total of 84 brucellosis patients and 43 volunteers were selected and divided into brucellosis vs. control groups. Serum samples were subjected to serological tests for brucellosis, and CTX-II levels in all samples were measured simultaneously with ELISA. The results showed that serum CTX-II levels in human brucellosis were higher than those of healthy controls, without a statistically significant difference, but serum CTX-II levels in male patients were significantly higher than those of female patients (P<0.05). This finding could indicate the biological changes in the cartilage and bone in human brucellosis.
Adult ; Brucellosis ; blood ; epidemiology ; China ; epidemiology ; Collagen Type II ; blood ; genetics ; Female ; Gene Expression Regulation ; immunology ; Humans ; Male ; Middle Aged ; Peptide Fragments ; blood ; genetics ; Sex Factors

OBJECTIVETo compare therapeutic effects of clavicular hook-plate fixation and modified Weaver-Dunn surgery combined with clavicular hook-plate fixation in treating Tossy type III acromioclavicular joint dislocation.

METHODSForty-one patients with Tossy type III acromioclavicular dislocation treated by operation were retrospectively analysis from January 2012 to January 2014. The patients were divided into clavicular hook-plate fixation group (group A) and modified Weaver-Dunn surgery combined with clavicular hook-plate fixation (group B) according to surgical procedures. In group A, there were 15 males and 6 females aged from 17 to 51 years old with an average of (31.60 ± 12.58) years old, preoperative Constant-Murley score was 40.25 ± 9.80, and treated with clavicular hook-plate fixation. In group B, there were 13 males and 7 females aged from 18 to 48 years old with an average of (29.40 ± 11.27) years old, preoperative Constant-Murley score was 41.45 ± 8.81, and treated with modified Weaver-Dunn surgery combined with clavicular hook-plate fixation. Operative time, blood loss, imaging changes before and after operation, postoperative complications were compared; Constant-Murley score at 3, 6 and 12 months after operation were evaluated.

RESULTSIn group A, operative time was 40.50 ± 24.36) min, blood loss was (75.30 ± 30.36) ml; In group B, operative time was (60.10 ± 23.55) min, blood loss was (100.70 ± 40.12) ml. Twenty-one patients in group A were followed-up from 12 to 18 months with an average of (14.8 ± 3.1) months; 20 patients in group B were followed-up from 12 to 14 months with an average of (13.6 ± 1.5) months. There were no significant differences in operative time, blood loss and follow-up time between two groups. Complications were in six patients of group A and 3 patients of group B, and there were no significant meaning between two groups. At 6 months after operation, Constant-Murley score in group A was 88.85 ± 4.23, 92.15 ± 3.82 in group B; and had significant meaning between two groups (t = -2.56, P = 0.022 < 0.05). While there were no differences in Constant-Murley score in other times.

CONCLUSIONBoth of clavicular hook-plate fixation and modified Weaver-Dunn surgery combined with clavicular hook-plate fixation are effective operative methods for the treatment of Tossy type III acromioclavicular dislocation. Clavicular hook-plate fixation has advantage of less trauma, while modified Weaver-Dunn surgery combined with clavicular hook-plate fixation could reconstruct coracoclavicular ligament more stronger, clavicular hook plate could take out earlier, also improve shoulder joint function earlier.

Acromioclavicular Joint ; injuries ; Adolescent ; Adult ; Bone Plates ; Case-Control Studies ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Shoulder Dislocation ; surgery

Learning/memory impairment is one of the most serious problems induced by stress, and the underlying mechanisms remain unclear. Opiates and opioid receptors are implicated in multiple physiological functions including learning and memory. However, there is no clear evidence whether the endogenous opioid system is involved in the formation of the stress-induced spatial reference memory impairment. The aim of the present study was to evaluate the role of μ opioid receptor in the stress-induced spatial reference memory impairment by means of Morris water maze (MWM) test in a mouse elevated platform stress model. The mice were trained in the MWM for four trials a session for 4 consecutive days after receiving the elevated platform stress, and intracerebroventricular injection of μ opioid receptor agonist DAMGO, antagonist CTAP or saline. Retention of the spatial training was assessed 24 h after the last training session with a 60-s free-swim probe trial using a new starting position. The results showed that intracerebroventricular injection of μ opioid receptor agonist DAMGO but not antagonist CTAP before MWM training impaired the memory retrieval of mice. Elevated platform stress before MWM training also impaired memory retrieval, which could be reversed by pre-injection of CTAP, and aggravated by DAMGO. These results suggest that endogenous opioid system may play a crucial role in the formation of the stress-induced memory impairment.
Animals ; Enkephalin, Ala(2)-MePhe(4)-Gly(5)- ; pharmacology ; Maze Learning ; Memory Disorders ; Mice ; Receptors, Opioid, mu ; physiology ; Spatial Memory ; Stress, Physiological

OBJECTIVEThis study was to investigate the expression of miR-10a in the different FAB subtype of acute myeloid leukemia (AML) and its relationship with drug resistance.

METHODSForty de novo patients with AML, 16 patients with non-malignant hematologic disease and three AML cell lines HL-60, U937 and HL-60/ADR were enrolled in this study, the MiR-10a expression in bone marrow mononuclear cells of above-mentioned patients and 3 AML cell lines was detected by TaqMan RT-PCR. The correlation of miR-10a with clinicopathological factors of AML patients was analyzed.

RESULTSThe miR-10a expression level in HL-60 cell line was higher than that in U937 cell line (P = 0.039). And its expression level in de novo AML patients was higher than that in patients with non-malignant hematologic disease (P < 0.01). FAB-AML-M3 patients exhibited higher expression of miR-10a than that in M1, M2 and M4 (P < 0.05); HL-60/ADR cell line showed higher miR-10a expression than that in HL-60 cell line (P < 0.01) . Except M3, the patients without CR (non-CR) after the first cycle of chemotherapy showed a higher level of miR-10a as compared with CR patients (P < 0.01).

CONCLUSIONThe high expression of miR-10a may be closely related to over-proliferation of promyelocyte and drug resistance of acute myeloid leukemia cells, except M3.

Cell Line, Tumor ; Drug Resistance, Neoplasm ; Humans ; Leukemia, Myeloid, Acute ; MicroRNAs

OBJECTIVETo investigate the effect of n-3 polyunsaturated fatty acids (n-3PUFAs) on gut microbiota and endotoxin levels in portal vein of rats fed with a high-fat diet (HFD).

METHODSThirty-six male Sprague-Dawley rats were randomly divided into four groups and fed with normal control diet (CD), HFD, CD supplemented with n-3PUFAs, and HFD supplemented with n-3PUFAs, respectively. Fresh fecal samples were collected to analyze the gut microbiota 10 weeks after feeding. DNA was exacted from the fresh fecal samples. Quantitative PCR was used to detect the composition of the gut microbiota. The endotoxin levels were detected through modified azo chromogenic substrate limulus amebocyte lysate assay.

RESULTSThe differences in body weight before breeding in each group were not statistically significant among these four groups (P=0.613). The increase in the body weight was significantly larger in the HFD group than in the CD group (P=0.0002), CD+n-3PUFAs group (P=0.0001), and HFD+n-3PUFAs group (P=0.022). There were significantly more firmicutes (P=0.002) and enterobacteriales (P=0.022) and significantly less bacteroidetes (P=0.026) and bifidobactera (P=0.034) in the gut of rats from HFD group than those from the CD group. There were significantly more bacteroidetes in the fecal samples of the rats from the CD+n-3PUFAs group compared to those from the CD group (P=0.043). There were significantly more firmicutes (P=0.044)and enterobacteriales (P=0.012) and less bacteroidetes (P=0.042) in the fecal samples of the rats from HFD group compared to those from the HFD+n-3PUFAs group. The endotoxin in plasma form portal vein of rats in HFD group were significantly higher than in CD group (P=0.007) and HFD+n-3PUFAs group (P=0.042) but showed no significant difference between CD+n-3PUFAs and CD group (P=0.210).

CONCLUSIONSHFD can increase body weight and change gut microbiota. Supplementation of n-3PUFAs can partially counteract such gut dysbiosis, lower endotoxin level in portal vein blood, and improve the body weight.

Animals ; Body Weight ; Diet, High-Fat ; adverse effects ; Endotoxins ; blood ; Fatty Acids, Omega-3 ; pharmacology ; Intestines ; microbiology ; Male ; Microbiota ; drug effects ; Portal Vein ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effects of eukaryotic translation initiation factor 5A2 (EIF5A2) down-regulation by small interfering RNA (siRNA) on aggressiveness of human gastric cancer cell and its potential mechanisms.

METHODSThe expressions of EIF5A2 in human gastric cancer cell lines (MKN28 and HGC27) and immortalized gastric mucosal epithelial cells (GES-1) were measured by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blotting. EIF5A2 gene in MKN28 cells was silenced by RNA interference and the inhibitory effect was evaluated by both qRT-PCR and Western blotting. Cell proliferation was assessed by CCK-8 assay. Cell migration and invasion were assessed by Transwell assay. The possible downstream targets of EIF5A2, such as CyclinD1, CyclinD3, matrix metallopeptidase-9 (MMP-9), E-cadherin, vimintin, C-myc, and metastasis-associated protein 1 (MTA1) expression levels, were examined by Western blotting.

RESULTSHigh expressions of EIF5A2 were found in MKN28 cells and human gastric adenocarcinoma tissues. Both EIF5A2 mRNA and protein expression in MKN28 cells were significantly down-regulated by siRNA#1 and siRNA#2, especially siRNA#1. Knockdown of EIF5A2 caused an apparent suppression of MKN28 cell proliferation (all P<0.01), migration (P<0.001), and invasion (P<0.001). After the knockdown of EIF5A2 in MKN28 cells, E-cadherin levels were upregulated, whereas vimentin, Cyclin D1, Cyclin D3, C-myc and MTA1 levels were downregulated.

CONCLUSIONKnockdown of EIF5A2 may inhibit MKN28 cell proliferation by downregulating the CyclinD1 and CyclinD3 and suppressing the cell migration and invasion by inhibiting MTA1, C-myc and epithelial-mesenchymal transition.

Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cyclin D1 ; metabolism ; Cyclin D3 ; metabolism ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Genes, myc ; Histone Deacetylases ; metabolism ; Humans ; Peptide Initiation Factors ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; RNA-Binding Proteins ; genetics ; Repressor Proteins ; metabolism ; Stomach Neoplasms ; pathology

Gastric cancer is caused by the interaction of genetic and environmental factors. MicroRNA (miRNA) is involved in many cellular processes including proliferation, differentiation, and apoptosis and plays an important role in pathogenesis of gastric cancer, as demonstrated in many recent studies from perspectives including miRNA profiling, reciprocal modulation between epigenetic and miRNA, and Helicobacter pylori infection. MiRNA is highly stabe in blood, and therefore non-invasive diagnosis of gastric cancer using circulating miRNA may be promising.
Helicobacter pylori ; Humans ; MicroRNAs ; metabolism ; Stomach Neoplasms ; genetics

OBJECTIVETo investigate the expression of doublecortin-like kinase 1(DCLK1)in gastric cancer and its prognostic significance.

METHODSThe expression of DCLK1 was examined by immunohistochemical staining of paraffin-embedded tumor specimens from 122 patients who underwent curative gastrectomy for gastric cancer at Peking Union Medical College Hospital between July 2002 and December 2006. Survival curves were described by the Kaplan-Meier method and compared by the Log-rank test. Univariate and multivariate analysis was performed with the Cox proportional hazard model.

RESULTSThe expression of DCLK1 in tumor cells was significantly upregulated in 51 of 122 patients. High expression of DCLK1 in tumor cells was strongly correlated with pN stage(P=0.029)and lymphovascular invasion(P=0.029). Kaplan-Meier analysis revealed that patients with DCLK1 high expression had a significantly lower 5-year overall survival(OS)rate than that of patients with DCLK1 low expression(39. 0% vs. 65. 8%, P=0.001), as well as a significantly lower 5-year disease-free survival(DFS)rate(37. 0% vs. 64. 5%, P=0.001). Univariate and multivariate analyses showed that DCLK1 expression(both P=0.036)was an independent factor for predicting OS and DFS rate.

CONCLUSIONSHigh expression of DCLK1 in gastric cancer cells is associated with pN stage and lymphovascular invasion. It may be a predictor for poor survival in patients undergoing surgery for gastric cancer.

Disease-Free Survival ; Gastrectomy ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Kaplan-Meier Estimate ; Multivariate Analysis ; Prognosis ; Proportional Hazards Models ; Protein-Serine-Threonine Kinases ; metabolism ; Stomach Neoplasms ; diagnosis ; epidemiology ; metabolism

Human gut microbiota plays a key role in the development of obesity. Intestinal flora can regulate energy absorption and nutrition metabolism, increasing the energy harvesting from diet. Alteration of gut flora produces excessive lipopolysaccharide, which, when absorbed into the blood, can induce inflammatory reactions and promote the high-fat diet-associated obesity and metabolic syndrome. Intestinal flora increase visceral fat deposition by lowering the expression of Fiaf in intestinal mucosa. Different immune status also affects the intestinal flora.The gut microbiota is hypothesized to be an environmental factor that contributes to obesity; by interacting with factors such as host and diet, it adjusts the energy metabolism. Antibiotics or probiotics may alter the composition of intestinal microflora and improve the metabolic syndrome, and thus provides new treatment options.
Animals ; Diet, High-Fat ; Gastrointestinal Tract ; microbiology ; Humans ; Inflammation ; etiology ; Mice ; Obesity ; microbiology ; therapy ; Probiotics ; therapeutic use