[Immune-related genes and their determined immune cell microenvironment to predict the prognosis of gastric adenocarcinoma].

Objective: Through bioinformatics analysis to screen key immune-related genes (IRGs) and cancer-related pathways in gastric adenocarcinoma (GAC) therapy, combining immune cell microenvironment to predict the prognosis of GAC. Methods: RNA sequencing and clinical data were obtained from public databases. Differentially expressed IRGs between GAC and normal tissues were identified by integrated bioinformatics analysis. Univariate and multivariate Cox regression analyses were applied to screen survival-associated IRGs. Then, we established the risk signature model and found another database for external validation. In addition, we explored the relationship with the immune cell microenvironment in each GAC sample using CIBERSORT algorithms. Results: A total of 78 differentially expressed IRGs were screened, including 47 up-regulated and 31 down-regulated genes. Subsequently, a five-IRGs signature (BMP8A、MMP12、NRG4、S100A9 and TUBB3) was significantly associated with the overall survival of GAC patients. Survival analysis indicated that patients in the high-risk group have a poor prognosis. The results of the multivariate analysis revealed that the risk score was an independent prognostic factor. Further analysis showed that the prognostic model had excellent predictive performance in both TCGA and GEO validated cohorts. Besides, the results of tumor-infiltrating immune cell analysis indicated that the risk score could reflect the status of the tumor immune microenvironment. Conclusion: BMP8A, MMP12, NRG4, S100A9 and TUBB3 with the risk signature model are associated with prognosis in patients with GAC, combined with tumor-infiltrating immune cells to provide new markers for immunotherapy in GAC.

[Establishment of a nomogram model for predicting lymph node metastasis in patients with cN0 gastric cancer based on combination of preoperative C-reactive protein/albumin ratio].

Objective: To investigate the relationship between systemic inflammatory markers such as neutrophil/lymphocyte ratio (NLR) and C-reactive protein/albumin ratio (CAR), and lymph node metastasis in patients with cN0 gastric cancer. To establish a nomogram model to predict the risk of lymph node metastasis in patients with cN0 gastric cancer. Methods: The preoperative systemic inflammatory markers and clinical data of 134 patients with cN0 gastric cancer were retrospectively analyzed, and these markers of patients with negative (pN0) or positive (pN+ ) lymph node metastasis in postoperative pathological diagnosis were compared. The receiver operating characteristic (ROC) curve was used to evaluate the predictive effect of preoperative systemic inflammatory markers on lymph node metastasis. The influencing factors for lymph node metastasis were assessed by univariate analysis and multivariate logistic regression analysis. A nomogram subsequently established by R software was validated by Bootstrap resampling as internal validation. Results: Compared with pN0 group, NE (P=0.022), CRP (P<0.001), NLR (P<0.001), PLR (P=0.003) and CAR (P<0.001) were higher, LY (P=0.003) and Alb (P=0.042) were lower in pN+ group. ROC curve analysis showed that the area under the curve (AUC) of postoperative pathological lymph node metastasis in patients with cN0 gastric cancer diagnosed by NLR, PLR and CAR were 0.687, 0.651 and 0.694, respectively, and the best cutoff values were 2.12, 113.59 and 0.02, respectively. The corresponding sensitivity and specificity were 62.9% and 72.2%, 77.4% and 48.6%, 74.2% and 58.3%, respectively. Univariate analysis showed that tumor size, depth of invasion, NLR, PLR and CAR were associated with lymph node metastasis in cN0 gastric cancer patients (all P<0.05). Multivariate analysis showed that depth of invasion, NLR and CAR were independent influencing factors of lymph node metastasis in patients with cN0 gastric cancer. OR were 8.084, 3.540 and 3.092, respectively (all P<0.05). The C-index of the nomogram model was 0.847 (95% CI: 0.782-0.915). The predicting calibration curve was properly fit with the ideal curve in calibration chart. Conclusion: Combination of NLR and CAR to establish a nomogram model has a good consistency and can accurately predict the risk of lymph node metastasis in patients with cN0 gastric cancer.

First Report of Powdery Mildew (Pseudoidium neolycopersici) on Croton (Codiaeum variegatum var. pictum) in China.

Croton (Codiaeum variegatum (Linn.) var. pictum (Lodd.)) is an ornamental plant commonly grown in southern China. In March 2014, severe powdery mildew infections were observed on crotons in gardens of Hainan University (20.1°N and 110.3°E), Haikou, Hainan province. Disease incidence was estimated in a random batch of 100 plants in three replicates, with the average value approaching 80%. Symptoms first appeared as white circular patches on the adaxial surface and expanded to the abaxial surface, petioles, and stems. The top leaves were the most affected. Upper surfaces of the infected leaves were covered by white, dense mycelia. As the disease progressed, infected leaves turned purple on the lower surfaces and curly before becoming necrotic and abscising from the plant. Powdery mildew was more severe in shaded environments, especially during rainy or foggy weather in early spring. Two hundred conidiophores and conidia were observed microscopically. The conidiophores were straight or occasionally flexuous, 62.3 to 127.6 × 6.2 to 10.2 µm, consisting of two to three straight cells. Conidia were born in solitary on the top of conidiophores. Conidia were hyaline, ellipsoidal, 26.4 to 42.2 × 11.7 to 23.4 µm (average 32.5 × 16.5 µm), contained no distinct fibrosin bodies, and produced a subterminal germ tube. The wrinkling pattern of the outer walls of older conidia was angular or reticulated. Appressoria were single and multilobed. Cleistothecia were not observed. Based on morphological characteristics, the fungus was identified as Oidium neolycopersici (2), which was recently renamed Pseudoidium neolycopersici (L. Kiss) (3). The identity was confirmed by sequence analysis. Genomic DNA was extracted from the foliar powdery mildew colonies using Chelex-100 (Bio-Rad, Shanghai, China). The rDNA internal transcribed spacer (ITS) region was amplified with primers ITS1 and ITS4 (5). The ITS sequence of the representative isolates C01 (GenBank Accession No. KJ890378.1) and four other powdery mildew samples collected from crotons in Hainan University was 100% identical to that of P. neolycopersici isolates from tomato plants such as JQ972700 and AB163927. Inoculations were made by gently pressing diseased leaves onto leaves of five healthy plants of croton and tomato ('Money maker'). Five non-inoculated croton and tomato plants served as controls. Inoculated and non-inoculated plants were maintained in an incubator at 25°C with a 12-h photoperiod. After eight days, typical powdery mildew symptoms developed on 93% of the inoculated plants, while no symptom developed on the non-inoculated plants. The pathogenicity tests were repeated three times. The same fungus was always re-isolated from the diseased tissue according to Koch's postulates. The pathogenicity tests further confirmed that the pathogen from crotons is P. neolycopersici (Basionym. Oidium neolycopersici (KJ890378.1)), which is commonly known as the tomato powdery mildew. P. neolycopersici is also a pathogen of Normania triphylla (1) and papaya (4). To our knowledge, this is the first report of P. neolycopersici infecting croton. The avenue of this pathogen entering gardens of Hainan University remains unknown. The gardens are located far away from tomato farms. Also no symptom of powdery mildew on croton was observed during surveys in other locations in Haikou. The origin of the pathogen warrants additional research. References: (1) D. Delmail et al. Mycotaxon 113:269, 2010. (2) L. Kiss et al. Mycol. Res. 105:684, 2001. (3) L. Kiss et al. Mycol. Res. 115:612, 2011. (4) J. G. Tsay et al. Plant Dis. 95:1188, 2011. (5) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Cytotoxicity and radiosensitization effect of TRA-8 on radioresistant human larynx squamous carcinoma cells.

TRAIL induces apoptosis in a variety of tumorigenic and transformed cell lines, but not in many normal cells. Recent studies have demonstrated that death receptor 5 (DR5), one of the two death receptors bound by TRAIL, showed expression in most malignantly transformed cells. This study evaluated effects of a monoclonal antibody (TRA-8) to human death receptor 5, combined with ionizing radiation, on radioresistant human larynx squamous carcinoma cell line (Hep-2R). Cells were treated with TRA-8 alone or in combination with radiation, cell viability inhibition was measured by MTT assay, and the induction of apoptosis was determined by Annexin V staining. Radionsensitivity of Hep-2R cells treated with TRA-8 were investigated with long-term clonogenic assays. Regulation of DR5 expression in cells after radiation was analyzed by indirect immunofluorescence using murine TRA-8 in combination with flow cytometry. The results suggested that TRA-8 enhanced radionsensitivity of Hep-2R cells, and that TRA-8 regulated Hep-2R cell cycle arrest at G2/M phase. Irradiation up-regulated the expression of DR5, and when combined with TRA-8 yielded optimal survival benefit. Therefore, TRA-8 can be used in combination with irradiation in radioresistant human larynx squamous carcinoma cells. Monoclonal antibodies such as TRA-8 may play an important role in the development of an effective treatment strategy for patients with radioresistant cancers.

Enhanced radiosensitivity of SW480 cells via TRAIL up-regulation mediated by Egr-1 promoter.

Radiosensitization of cancer cells to irradiation could improve the efficacy of radiotherapy. The early transcriptional factor (Egr-1) promoter induced expression of downstream genes after irradiation. TNF-related apoptosis-inducing ligand (TRAIL) is known to induce apoptosis in malignant cells, but displayed little or no toxicity on normal cells. In this study, we constructed pcDNA3.1-Egr-1-TRAIL (pEgr.1-TRAIL) recombinant plasmid and evaluated its effect on human colon cancer cell line SW480. pEgr.1-TRAIL transfection combined with radiotherapy caused dramatically elevation of TRAIL expression both in mRNA and protein levels, much lower radiobiological parameters in clonogenic assays, accompanied by remarkably increase in apoptosis ratio. Furthermore, pEgr.1-TRAIL transfected cells displayed higher proportion in G0/G1 phase. Our results suggested that pEgr.1-TRAIL can sensitize SW480 cells to radiation, and the radiosensitization is related to cell cycle changes and apoptosis mediated by up-regulation of TRAIL expression. These findings support the potential future application of genetic radiotherapy against carcinoma.

Aconitine induces cell apoptosis in human pancreatic cancer via NF-κB signaling pathway.

OBJECTIVE: Pancreatic cancer is one of the leading causes of death from cancer in European countries and the United States. This study sought to investigate the effects of aconitine, a well-known aconitum plant-produced toxin, on pancreatic cancer cell growth and apoptosis and to explore the potential mechanisms. MATERIALS AND METHODS: In this study, pancreatic cancer cell lines Miacapa-2 and PANC-1 were cultured, and cell viability was examined in these two cells treated with different doses of aconitine. Moreover, cell apoptosis was also analyzed upon aconitine treatment, and the specific mechanism was examined by Western blot assay and caspase activity detection. RESULTS: The results showed that aconitine inhibited pancreatic cancer cell growth in a dose- and time-dependent manner. The administration of aconitine in Miapaca-2 and PANC-1 cells also induced cell apoptosis by upregulating the expression of pro-apoptotic factors Bax, cl-caspase-3, cl-caspase-9, and cleaved poly (ADP-ribose) polymerase 1 (PARP1), and by decreasing the anti-apoptotic Bcl-2 expression. More importantly, NF-κB was also decreased upon aconitine treatment. In a xenograft mouse model of pancreatic cancer, aconitine suppressed tumor growth and increased cell apoptosis. CONCLUSIONS: This study is the first report on the effects of aconitine on pancreatic cancer, and it reveals that aconitine may serve as a potent therapeutic strategy for clinical treatment of pancreatic cancer.

Hyperalgesia due to nerve damage: role of nerve growth factor.

The hypothesis that nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) contribute to hyperalgesia resulting from nerve damage was tested in rats in which the sciatic nerve was partially transected on one side. Administration of antisera raised against NGF and BDNF relieved mechanical and thermal hyperalgesia in these animals. It has been suggested that NGF may elicit hyperalgesia by inducing mast cells to release algesic agents such as serotonin (5-HT). We found that degranulation of mast cells with compound 48/80 relieved mechanical and thermal hyperalgesia produced by nerve damage. We also found that local injection of the 5-HT2A and 5-HT3 receptor antagonists ketanserin and ICS 205-930 into the affected hind paw relieved mechanical hyperalgesia in a dose-dependent fashion. These findings support the idea that in this rat model of hyperalgesia due to peripheral nerve damage, NGF acts on mast cells to induce release of 5-HT, which sensitizes nociceptors. Hyperalgesia due to nerve injury and hyperalgesia due to inflammation may share some common features.

Comparison of 9-fluorenylmethoxycarbonyl and 9-fluoreneacetyl-tagged silica-based derivatization reagents in high-performance liquid chromatography.

Two silica reagents based on a 4-hydroxy-3-nitrobenzoyl backbone were synthesized and characterized with 9-fluorenylmethoxycarbonyl (FMOC) and 9-fluoreneacetyl (FA) tags. These reagents were tested by derivatization of primary and secondary amines. Derivatization conditions such as temperature, time and triethylamine catalyst were tested. The FA-tagged silica reagent showed better performance than the FMOC-tagged silica reagent by a comparison of derivatization efficiencies, stabilities of reagents, and blank reagent interferences with derivatization. Finally, cadaverine and an aliphatic amine mixture were analysed using the FA-tagged reagent by pre-column, off-line derivatization and fluorescence detection.

An O(2n) volume molecular algorithm for Hamiltonian path.

We design volume-efficient molecular algorithms for all problems in #P, using only reasonable biological operations. In particular, we give a polynomial-time 0(2(n)n2log2n)-volume algorithm to compute the number of Hamiltonian paths in an n-node graph. This improves Adleman's celebrated n!-volume algorithm for finding a single Hamiltonian path.

Ultrasonographic diagnosis of achalasia.

A perspective ultrasonographic study on 10 cases of achalasia showed characteristic ultrasonographic features: dilation and persistent water retention of the gastroesophageal vestibule, symmetrical parietal thickening, and delayed or intermittent opening of the cardiac orifice after drinking. We suggest that ultrasonography should play an important role in clinical management of achalasia. If the ultrasonographic features of achalasia were known, the misdiagnosis of achalasia for cardiac carcinoma could be avoided. When an infiltrating cardiac carcinoma found to be smoothly narrowing and difficult to distinguish from achalasia radiologically, an ultrasonogram may be helpful to make a correct diagnosis.

[Studies on the chemical constituents of Acronychia pedunculata (L.) Mig].

Two crystalline substances were isolated from the stem bark of Acronychia pedunculata and identified as acrovestone and bauerenol on the basis of chemical studies and spectrometric analysis.

CTC1 increases the radioresistance of human melanoma cells by inhibiting telomere shortening and apoptosis.

Melanoma has traditionally been viewed as a radioresistant cancer. However, recent studies suggest that under certain clinical circumstances, radiotherapy may play a significant role in the treatment of melanoma. Previous studies have demonstrated that telomere length is a hallmark of radiosensitivity. The newly discovered mammalian CTC1­STN1-TEN1 (CST) complex has been demonstrated to be an important telomere maintenance factor. In this study, by establishing a radiosensitive/radioresistant human melanoma cell model, MDA-MB-435/MDA-MB­435R, we aimed to investigate the association of CTC1 expression with radiosensitivity in human melanoma cell lines, and to elucidate the possible underlying mechanisms. We found that CTC1 mRNA and protein levels were markedly increased in the MDA-MB­435R cells compared with the MDA-MB­435 cells. Moreover, the downregulation of CTC1 enhanced radiosensitivity, induced DNA damage and promoted telomere shortening and apoptosis in both cell lines. Taken together, our findings suggest that CTC1 increases the radioresistance of human melanoma cells by inhibiting telomere shortening and apoptosis. Thus, CTC1 may be an attractive target gene for the treatment of human melanoma.

Rapid extraction of layer relative humidity, geopotential thickness, and atmospheric stability from satellite sounding radiometer data.

Layer moisture and temperature are important variables for forecasting local weather phenomena. The purpose of ths paper is to present a method to rapidly compute layer average relative humidities and geopotential thicknesses from satellite measurements of atmospheric water vapor and oxygen radiation emission to space. Both polar-orbiting and geostationary satellite data are considered. Analyses show that the relative humidity and geopotential thickness patterns from satellite data are consistent with radiosonde data. The relative humidity calculation is insensitive to the assumed temperature profile condition. It is shown that time variations of the atmospheric moisture can be monitored from a GOES-4 VAS sequential image. Also an attempt is made to derive the atmospheric stability index, total-totals, by use of satellite derived relative humidities and geopotential thicknesses. The results show good correspondence between the derived stability patterns and subsequent convective weather.

Direct determination of adamantanamine in plasma and urine with automated solid phase derivatization.

A simple, highly sensitive and selective method is described for adamantanamine determination in plasma and urine by high-performance liquid chromatography with fluorescence detection. The method involved a simultaneous extraction and derivatization of biological fluids with a 9-fluoreneacetate (9-FA) solid-phase derivatization reagent. This approach eliminated tedious sample preparation steps and provided automatic derivatization with selective and efficient sample clean-up for direct injection of biological fluids. Derivatized adamantanamine was separated under conventional reversed-phase conditions and determined by fluorescence detection. The optimization and validation of the derivatization method with the 9-FA solid-phase reagent is described.