FBLN3 inhibited the invasion and metastasis of colorectal cancer through the AKT/mTOR pathway.

Fibulin-3(FBLN3) levels are different in different types of cancers. We found that fibulin-3 was downregulated in colorectal (CRC) cells, particularly in the SW480 cell line. By comparison, transfecting SW480 cells with a lentivirus overexpressing fibulin-3 RNA could inhibit proliferation, induce G1/S arrest, and promote cell apoptosis. Fibulin-3 overexpression further suppressed the invasion and metastasis of CRC. These effects were regulated through the AKT/mTOR signaling pathway.

Lung microdialysis study of florfenicol in pigs after single intramuscular administration.

For most bacterial lung infections, the concentration of unbound antimicrobial agent in lung interstitial fluid has been considered as the gold standard for estimating the antibacterial efficacy. In this study, the pharmacokinetics of florfenicol (FF) in porcine lung interstitial fluid was investigated after single intramuscular administration at two different doses (20 and 50 mg/kg). Twelve pigs underwent thoracotomy under general anesthesia. Then, the CMA/30 probe was implanted into the lung and perfused at 1 µL/min. The microdialysis (MD) samples were collected on a preset schedule and analyzed by high-performance liquid chromatography (HPLC). Noncompartmental pharmacokinetic analysis was performed. FF exhibited rapid distribution and slow elimination in porcine lung interstitial fluid. The main pharmacokinetic parameters at 20 and 50 mg/kg were 4.88 ± 0.54 and 10.36 ± 2.52 µg/mL for the maximum concentration (Cmax ), 3.25 ± 0.32 and 3.50 ± 0.27 h for the time to Cmax (Tmax ), 9.47 ± 6.84 and 7.75 ± 3.23 h for the half-life (t1/2 ), 0.10 ± 0.06 and 0.10 ± 0.04 1/h for the terminal elimination rate constant (λz ), 13.85 ± 7.97 and 11.42 ± 2.79 h for the mean residence time (MRT), 37.77 ± 8.13 and 71.15 ± 16.99 h·µg/mL for the area under the curve from time 0 to 18.25 h (AUC0-18.25 ), and 51.18 ± 20.11 and 88.78 ± 27.58 h·µg/mL for the area under the curve from time 0 to infinity (AUC0-∞ ), respectively.

Diffusion-limited PBPK model for predicting pulmonary pharmacokinetics of florfenicol in pig.

For most bacterial lung infections, the concentration of unbound antimicrobial agent in lung interstitial fluid has been thought to be responsible for antimicrobial efficacy. In this study, a diffusion-limited physiologically based pharmacokinetic (PBPK) model was developed to predict the pulmonary pharmacokinetics of florfenicol (FF) in pigs. The model included separate compartments corresponding to blood, diffusion-limited lung, flow-limited muscle, liver, and kidney and an extra compartment representing the remaining carcass. The absorption rate constant and renal and hepatic clearance of FF were determined in vivo. Other parameters were taken from the literature or optimized based on existing pharmacokinetic data. All mathematical operations during the development of the model were performed using acslXtreme version 3.0.2.1 (Aegis Technologies Group, Inc., Huntsville, AL, USA). The model accurately predicted the concentration-time courses of FF in lung interstitial fluid, serum, and plasma following different dosing schedules, except at the dose of 15 mg/kg. When compared with the tissue residue data, the model generally underestimated the FF concentration at the injection site, whereas it gave good predictions of FF concentrations in lung, liver, and kidney at early time points. The model predictions provide a scientific basis for the dosage regimen design of FF.

Correlation between hepatitis B virus DNA levels and diagnostic tests for HBsAg, HBeAg, and PreS1-Ag in chronic hepatitis B.

The aim of this study was to investigate the diagnostic value of the serum markers HBsAg and HBeAg and PreS1 protein (PreS1-Ag) in quantifying the levels of hepatitis B virus (HBV) DNA in patients with chronic hepatitis B (CHB). One thousand CHB patients were recruited from Beijing You'an Hospital between June and December 2012. Serum HBsAg and HBeAg levels were detected by electrochemiluminescence immunoassay. Enzyme-linked immunosorbent assay and fluorescence quantitative PCR were used to determine the level of PreS1-Ag and HBV DNA, respectively. We observed a low correlation between HBsAg and HBV DNA (r = 0.172, P < 0.001) expression; however, the correlation coefficient increased gradually with the increase in HBV DNA levels, and was more significant when HBV DNA log10 > 7 (r = 0.597, P < 0.001). Additionally, HBsAg and HBV DNA showed a significant positive correlation in the HBeAg+ group (r = 0.321, P < 0.001), whereas no correlation was observed in the HBeAg- group (r = -0.016, P = 0.825). HBV DNA expression was correlated with HBeAg (χ2 = 83.07, P < 0.001) and PreS1-Ag (χ2 = 36.01, P < 0.001). HBV DNA-positive rate was higher in HBeAg/PreS1-Ag++ patients (72.26%) than that in the single-positive groups (P < 0.001). Therefore, serum HBsAg is not a good marker for the prediction of HBV replication, and co-detection of HBeAg and PreS1-Ag, which can better predict HBV DNA replication, can be used as a reliable method for the clinical diagnosis and treatment of CHB.

MiRNA-411 attenuates inflammatory damage and apoptosis following spinal cord injury.

OBJECTIVE: To investigate the role and regulate the target of miRNA-411 on spinal cord injury. MATERIALS AND METHODS: The microglia cultured in vitro was activated by lipopolysaccharide (LPS) to express the inflammatory phenotype. The inflammatory response through miRNA-411 transfection in microglia was measured to certain whether increased miRNA-411 suppressed interleukin-18 (IL-18) level to attenuate the inflammation amplification via downregulating JNK pathway. Furthermore, we established spinal cord injury (SCI) model in SD rats and further explored the glial inflammatory degree and neurological recovery following miRNA-411 treatment. Lastly, we estimated the hindlimbs function of SCI rats with miRNA-411 administration or not within four weeks at post-SCI. RESULTS: In vitro, miRNA-411 inhibited IL-18 expression and downregulated JNK pathway, along with that inflammatory microglia were declined. In SCI rats, we detected the decreased amounts of inflammatory microglia and reduction of the inflammatory factors after miRNA-411 treatment. IL-18 and JNK pathway was also restrained resulted from increased miRNA-411. In addition, apoptosis degree in injury site reduced and survived axons were relatively multiple in the miRNA-411 group compared with the SCI group. The Basso-Beattie-Bresnahan (BBB) locomotor scores of miRNA-411 treated rats were superior to those in rats with no treatment. CONCLUSIONS: MiRNA-411 increase ameliorates the inflammatory microglia-induced neurological lesion and promotes neural recovery by JNK pathway inhibition via negative targeting IL-18 in SCI.

LncRNA MSTO2P promotes proliferation and autophagy of lung cancer cells by up-regulating EZH2 expression.

OBJECTIVE: This study was designed to investigate the specific mechanism underlying the regulatory effect of long noncoding ribonucleic acids (lncRNAs) MSTO2P on lung cancer (LCa) cell proliferation and autophagy via regulating enhancer of zeste homolog (EZH2) expression. PATIENTS AND METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to analyze the levels of MSTO2P and EZH2 in 40 pairs of LCa tissues and corresponding adjacent tissues, as well as in LCa cell lines (H1299, H23, A549) and human bronchial epithelial cells (BEAS-2B). Besides, the effect of MSTO2P on cell proliferation ability was detected by cell counting kit-8 (CCK-8) and plate cloning experiments. The interaction between MTS02P and EZH2 as well as their effects on cell autophagy ability were examined by qRT-PCR and Western blot. RESULTS: The qRT-PCR results showed that MSTO2P expression in LCa tissues was remarkably higher than that in adjacent tissues. Meanwhile, compared with human bronchial epithelial cells, the level of MSTO2P was remarkably up-regulated in LCa cells. After down-regulating MSTO2P, the cell proliferation ability was weakened, and the protein levels of autophagy-related genes including Agt5, LC-3I, and LC-3II were remarkably down-regulated. At the same time, EZH2 expression in LCa tissues was also remarkably up-regulated relative to adjacent tissues, and it was positively correlated with the expression of MSTO2P. In addition, after down-regulating MSTO2P, the EZH2 level was also remarkably reduced. Further experimental results revealed that EZH2 down-regulation could impair the cell proliferation ability and down-regulate the expressions of autophagy genes such as Agt5, LC-3I, and LC-3II. CONCLUSIONS: LncRNA MSTO2P promotes LCa cell proliferation and autophagy by up-regulating EZH2. Therefore, MSTO2P may be a potential therapeutic target for LCa.

Clinical and experimental studies on stomach carcinoma treated with Yangwei Kangliu granules.

AIM: To study the anti-cancer mechanism of Yangwei Kangliu (YWKL) granules from the view point of red blood cell (RBC) immunity and to investigate the relationship between RBC immunity and T lymphocyte immunity. METHODS: Fifty patients with advanced gastric carcinoma were treated with a combination of YWKL granules and chemotherapy. Venous blood samples were obtained before treatment and after one course of treatment. The rosette rate of c-3b-receptor (RBC-C-3bRR), tumor and red cell (RRTR) and RBC immune complex (RBC-ICR) were measured under microscopy by counting the rosettes formed by sensitized or unsensitized yeast adherence. The T lymphocyte subset was observed by the method of APAAP. Control patients were treated with chemotherapy alone (n = 20). In addition, mouse tumor studies were performed to investigate the dynamic changes of RBC-C-3bRR, RRTR and RBC-ICR in response to treatment with YWKL granules (n = 30). Mice treated with chemotherapy alone (n = 30) or water alone (n = 30) were used as controls. RESULTS: The clinical therapeutic effect of combination treatment with YWKL granules and chemotherapy (i.e. the treatment group) was markedly superior to that of chemotherapy alone (i.e. the control group) (P < 0.01). In the treatment group, the rosette rates of RBC-C-3bRR and of RRTR were significantly increased (P < 0.01) after treatment, the rate of RBC-ICR was markedly decreased (P < 0.01), and the ratio of CD4 to CD8 was obviously elevated (P < 0.01). Moreover, CD8 was much lower (P < 0.01) and the ratio of CD4 to CD8 was much higher (P < 0.01) than that in the control group. The RRTR rate was positively correlated with the ratio of CD4 to CD8. In mice, on day 9 of bearing cancer, the tumor weight in the group treated with YWKL granules alone was much lower than that of the tumors in the control mice groups; in addition, the YWKL treated mice showed higher RBC immune function than the mice of the two control groups. On day 13 of bearing cancer, however, the differences in both tumor weight and RBC immune function had disappeared. CONCLUSION: The anti-cancer mechanism of YWKL granules may involve enhancement of RBC immunity and of T lymphocyte immune function, which is supported by the finding of RBC immune function being correlated with T lymphocyte immune function.

Constructive learning of recurrent neural networks: limitations of recurrent cascade correlation and a simple solution.

It is often difficult to predict the optimal neural network size for a particular application. Constructive or destructive methods that add or subtract neurons, layers, connections, etc. might offer a solution to this problem. We prove that one method, recurrent cascade correlation, due to its topology, has fundamental limitations in representation and thus in its learning capabilities. It cannot represent with monotone (i.e., sigmoid) and hard-threshold activation functions certain finite state automata. We give a "preliminary" approach on how to get around these limitations by devising a simple constructive training method that adds neurons during training while still preserving the powerful fully-recurrent structure. We illustrate this approach by simulations which learn many examples of regular grammars that the recurrent cascade correlation method is unable to learn.

[Phagosonogram (PSG) in cancer of the gastric cardia--a preliminary study of 100 patients and 100 normal subjects].

Phagosonography of 100 normal subjects and 100 patients with cardiac cancer (CC) is described. The apparatus was self-designed and self-manufactured. The pre-ejection period and other 8 quantitative indexes in PSG were compared and abnormal level existed in patients with CC (P less than 0.01). The phagosonograms drawn in 54 patients with esophageal, cardiac or gastric benign diseases served as control. The diagnostic accuracy of PSG was 97% positive, 3% suspicious in CC and 100% negative in the normal subjects. It was 3.7% suspicious in the esophageal, cardiac or gastric diseases. PSG is very important in the early diagnosis of CC and is also of value in the prediction of a successful operation. The authors believe that PSG can reflect the changes in cardiac and esophageal motor function. In CC, the PSG changes are mainly due to the malfunction in the dilation and contraction of the lower esophageal sphincter. This malfunction shows itself earlier than the pathomorphology and provides a basis for the early diagnosis of CC. Phagosonography is a new method in the research on the motor function of the sound producing viscera, the diagnosis of their diseases and the observation of the treatment result. Being able to be performed with the patients in physiological and non-invasive state, it is suitable for screening.

[Clinical and experimental study in treating gastric cancer with replenishing qi and invigorating spleen oral liquid combined with chemotherapy].

61 cases of the gastric cancers have been treated with Replenishing Qi and Invigorating Spleen (RQIS) combined with chemotherapy. The patients were divided into treated group and control group randomly, The results showed that: rate of the adverse effects, e.g. nausea, vomiting, anorexia, diarrhea etc. were higher in the chemotherapy used patients, combined treated groups were lower significantly (P < 0.01-0.001). Rate of chemotherapy completion was 95% in the treated group, 78% in control group, the difference was significant (P < 0.01). WBC and blood platelet were reduced in the treatment group and chemotherapy group, they were 30% and 45% respectively (P < 0.01). Superoxide dismutase was increased obviously (P < 0.01). It revealed that RQIS could scavenge the free radical potently. 60 cases of gastric cancers have been treated over 1 year, the survival rates of 1 year, 3 years, 5 years were 98.36%, 82.69%, 60.00% respectively after following up. Among them, survival rate of III stage patients was 57.14%. Animal experimental study showed that the inhibition rates of lung metastasis were 92.2% and 83.7% in mice B16 of RQIS groups. The difference was significant than that of control group. They could elevate the tolerance to chemotherapy in mice.

Evaluation of the rpoB macroarray assay to detect rifampin resistance in Mycobacterium tuberculosis in Beijing, China.

The aim of this study was to evaluate the modified rifampin oligonucleotide (RIFO) macroarray method to detect mutations in the hot-spot region of the rpoB gene, associated with rifampin (RIF) resistance in Mycobacterium tuberculosis. The study sample included 123 strains of M. tuberculosis isolated in the Beijing, China, area in 2002-2005, including 73 RIF-resistant and 40 pansusceptible strains. The genotypic assay successfully identified 91.8% of the RIF-resistant strains, whereas no mutations were found in RIF-susceptible strains. The most frequently detected rpoB mutations were in the codons 516, 526, and 531, together accounting for 74% of RIF-resistant strains. Spoligotyping subdivided all strains into 11 unique profiles and 3 profiles shared by 3, 4, and 103 strains, respectively. The 113 strains belonged to the Beijing family genotypes, defined by the specific spoligotype signature (absence of signals 1-34) and deletion of the RD105 region. The rpoB S531L (TCG-->TTG) mutation was found in 57.4% of the RIF-resistant strains of the Beijing genotype. A mutation in the rpoB hot-spot region was found in 51 of the 55 (92.7%) multidrug-resistant strains (i.e., resistant to at least RIF and isoniazid), thus demonstrating the added utility of the modified RIFO method to predict multidrug resistance. The RIFO method is relatively simple to perform and allows straightforward interpretation of results; consequently, it can be used in clinical diagnostic laboratories as a fast complement to phenotypic methods.

Plasma glucose levels and diabetes are independent predictors for mortality and morbidity in patients with SARS.

AIMS: To investigate the relationships between a known history of diabetes and ambient fasting plasma glucose (FPG) levels with death and morbidity rates in patients with severe acute respiratory syndrome (SARS). METHODS: In this retrospective analysis, the clinical and biochemical characteristics of 135 patients who had died from SARS, 385 survivors of SARS and 19 patients with non-SARS pneumonia were compared. RESULTS: All patients were treated according to a predefined protocol. Before steroid treatment, the mean FPG level was significantly higher in the SARS group (deceased vs. survivors vs. non-SARS pneumonia group: 9.7 +/- 5.2 vs. 6.5 +/- 3.0 vs. 5.1 +/- 1.0 mmol/l, P < 0.01). In the SARS group, the percentage of patients with a known history of diabetes was significantly higher in the deceased patients than in the survivors (21.5% vs. 3.9%, P < 0.01). Among patients with no known history of diabetes and before commencement of steroid therapy, those who had hypoxaemia (SaO(2) < 93%) had higher FPG levels than those who did not have hypoxia in both the survivor (8.7 +/- 4.9 vs. 6.3 +/- 2.1 mmol/l, P < 0.001) and deceased (9.8 +/- 4.8 vs. 7.2 +/- 1.5 mmol/l, P < 0.001) groups. A known history of diabetes [odds ratio (OR) 3.0, 95% confidence interval (CI) 1.4, 6.3; P = 0.005] and FPG > or = 7.0 mmol/l before steroid treatment (OR 3.3, 95% CI 1.4, 7.7, P = 0.006) were independent predictors of death. During the course of the illness, FPG levels were negatively associated with SaO(2) (beta =-0.682 +/- 0.305, P = 0.025, general estimation equation model) in SARS patients. Survival analysis showed that FPG was independently associated with an increased hazard ratio (HR) of mortality (HR = 1.1, 95% CI 1.0, 1.1, P = 0.001) and hypoxia (HR = 1.1, 95% CI 1.0, 1.1, P = 0.002) after controlling for age and gender. CONCLUSIONS: A known history of diabetes and ambient hyperglycaemia were independent predictors for death and morbidity in SARS patients. Metabolic control may improve the prognosis of SARS patients.