Use of acoustic stimulus to determine behavioral changes in zebrafish after Cd exposure in a water quality warning system.

Behavioral changes in zebrafish are an effective early warning system to determine water quality. However, only a few studies have examined the response of zebrafish to non-chemical stimulus after exposure to a contaminant. Therefore, this study investigated the differences in the behavioral responses of zebrafish to acoustic stimuli before and after exposure to cadmium (Cd). Acoustic escape response sensitivity curves were obtained and analyzed, followed by the determination of sensitive stimulus conditions at 100 Hz and 97 dB with a duration of 30 s and an interval of 30 min. Zebrafish exhibit a significant acoustic escape response, which is significantly reduced after exposure to Cd. The results showed that zebrafish stop demonstrating acoustic escape responses when exposed to higher Cd concentrations or longer acoustic exposures. Based on these results, a novel method for detecting abnormal behavior in zebrafish by acoustic stimulation has been proposed, which is expected to reduce the false alarm rate of this type of water quality technology.

Circ_0001402 knockdown suppresses the chemoresistance and development of DDP-resistant cutaneous squamous cell carcinoma cells by functioning as a ceRNA for miR-625-5p.

Cutaneous squamous cell carcinoma (CSCC) is the most common metastatic skin cancer. Circular RNAs (circRNAs) are differentially expressed in CSCC and can sequester and sponge microRNAs. GSE74758 shows that hsa_circ_0001402 (circ_0001402) is the most overexpressed circRNA in CSCC. Expression of circ_0001402, microRNA(miR)-625-5p and karyopherin subunit alpha 4 (KPNA4) was detected by quantitative real-time polymerase chain reaction and/or Western blot. Colon formation, flow cytometry, Transwell assays and xenograft tumour model confirmed the development of CSCC cells. The competing endogenous RNA (ceRNA) interaction was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Circ_0001402 was significantly upregulated in CSCC tissues and cells, and higher expression of circ_0001402 was found in DDP-resistant samples. Functionally, circ_0001402 knockdown induced apoptosis and inhibited half maximal inhibitory concentration of DDP, colony formation, migration and invasion of DDP-resistant CSCC cells, accompanied with the depressed multi-drug resistance-1 (MDR1) and MDR-related protein-1, while miR-625-5p inhibitor could counteract these effects. Mechanically, circ_0001402 mediated the expression regulation of KPNA4 via functioning as a ceRNA for miR-625-5p. KPNA4 re-expression could abate the functions of miR-625-5p. Furthermore, circ_0001402 knockdown could hinder tumour growth of DDP-resistant CSCC. Circ_0001402 knockdown can suppress the development and chemoresistance of DDP-resistant CSCC cells at least partly through targeting miR-625-5p/KPNA4 axis.

Effects of transarterial chemoembolization on the immunological function of patients with hepatocellular carcinoma.

The present study aimed to investigate the effects of transarterial chemoembolization (TACE) on the immune function of patients with hepatocellular carcinoma (HCC). A total of 114 patients with HCC were selected and their peripheral blood was collected before and 1 month after TACE treatment. Flow cytometry and reverse transcription-quantitative PCR were performed to analyze the changes in immune function in patients before and after treatment. Kaplan-Meier curves were plotted for survival analysis. The programmed cell death ligand 1 (PD-L1) and programmed cell death protein 1 (PD1) expression before TACE treatment were significantly higher in patients with poor TACE response compared with those patients with well response. Higher PD-L1 mRNA expression in the peripheral blood mononuclear cells after TACE predicted a superior prognosis. After TACE treatment, the proportion of CD4+/CD8+ cells were decreased while the expression levels of programmed cell death protein 1 (PD1) were significantly increased. To conclude, TACE could reduce the proportion of CD4+/CD8+ cells and improve the mRNA expression levels of PD1 in patients with HCC. The expression levels of PD1 and PD-L1 were closely related to the therapeutic effect of TACE and the prognosis of patients with HCC. TACE combined with immunotherapy may have potential clinical value for patients with HCC.

[Effects of drought stress on the stoichiometric characteristics in different organs of three shrub species]. / 干旱胁迫对3种灌木不同器官化学计量特征的影响.

In order to explore how water stress affects the stoichiometric characteristics in leaves, stems, very fine roots (0-1 mm), fine roots (1-2 mm) and thick roots (>2 mm) of three shrub species, we studied the effects of three water treatments [(75±5)%, (55±5)% and (35±5)% of field water capacity (FC)] on the stoichiometric characteristics of different organs of Syringa oblata, Rosa xanthina and Forsythia suspensa in a pot experiment. The results showed that there were significant differences in nitrogen (N) and phosphorus (P) contents, C:N, C:P and N:P of the same organ among the three species. With the intensification of drought stress, there was no significant change of C content in all organs of the three species. The N content increased in leaves, but decreased gradually in stems. The N content in very fine roots and fine roots increased first and then decreased. The P content decreased in leaves and stems, while increased first and then decreased in very fine roots and fine roots. Under drought stress, leaf C:N decreased, C:P and N:P of leaf and stem increased. There was the strongest effect of drought stress on the C:N of very fine roots and C:P and N:P of leaves. There was the least effect of drought stress on C:N, N:P of thick roots and C:P of very fine roots. There was no significant correlation between the contents of C, N in soil and the contents of C, N and P in shrub organs, but soil P content was significantly correlated with the contents of C, N and P in leaves and roots. It was concluded that the relative P limitation in soil was the most important factor affecting the stoichiometric characteristics of shrub organs. Drought had different effects on the stoichiometry of different organs in different shrub species. The stoichio-metry of leaves and very fine roots was more sensitive to drought stress than that of other organs. Drought might affect the stoichiometric characteristics especially related to P in different organs of shrubs, mainly by affecting plant absorption of soil P and its distribution in different organs.

A Patient With Sporotrichosis Diagnosed By Molecular Biology Combined With Traditional Methods.

We present a case of sporotrichosis diagnosed by molecular biology combined with traditional methods. Fungal DNA was extracted from skin tissues of the patient by fungal DNA extraction kit. The PCR amplified fragments were compared on GenBank website, and the pathogenic fungi were identified. This case was diagnosed by molecular biology, clinical manifestations, fungal morphology and pathology. The pathogenic fungi of the patient was identified as Sporothrix globosa by comparison, combined with typical clinical manifestations, typical filamentous colonies by fungi culture and pathological findings of infectious granuloma. The case could be diagnosed as Sporothrix globosa-induced sporotrichosis. Molecular biology combined with traditional detection methods for diagnosis of sporotrichosis is helpful to improve the accuracy of diagnosis.

CARD9 deficiencies linked to impaired neutrophil functions against Phialophora verrucosa.

Caspase recruitment domain-containing protein 9 (CARD9) is an adaptor molecule that is critical for NF-κB activation and forms a complex with B cell lymphoma 10 and mucosa-associated lymphoid tissue lymphoma translocation gene 1 that mediates C-type lectin receptors (CLRs)-triggered intracellular signaling during antifungal immunity. However, the role of CARD9 in the host defense against Phialophora verrucosa (P. verrucosa) infection remains to be elucidated. In the present study, we investigated the functions of polymorphonuclear neutrophils (PMNs) from patients with CARD9 deficiencies against P. verrucosa. By isolating PMNs from patients and healthy blood donors and subsequently challenging the cells with P. verrucosa, we demonstrated that, compared with healthy donors, CARD9-deficient PMNs exhibited defects in P. verrucosa killing and pro-inflammatory cytokine productions, which can be rescued in the presence of serum; however, the CARD9-deficient PMNs exhibited normal reactive oxygen species generation and phagocytotic ability. In conclusion, our results indicate that CARD9 is indispensable for P. verrucosa killing by PMNs, and serum opsonization acts as a CARD9-independent way, which could be a promising immunotherapy in the future.

Th and Treg response induced by Aspergillus fumigatus pulsed dendritic cells in vitro.

BACKGROUND: Dendritic cells (DCs) can recognize the pathogen-associated molecular patterns (PAMP) of Aspergillus fumigatus (A. fumigatus), activating the immune response. During A. fumigatus infection, a Th and Treg response induced in the fungi-pulsed DCs is not yet well understood. METHODS: In this study, bone marrow-derived dendritic cells (BMDCs) were separated and proliferated from C57BL/6 mice. A. fumigatus pulsed DCs were generated and cultured with CD4(+) T cells derived from the spleen of C57BL/6 mice in vitro. CD4(+) T cells differentiation after co-culture were analyzed by flow cytometry, ELISA, and real-time PCR analysis. RESULTS: The A. fumigatus pulsed DCs exhibited increased Th1 and Treg frequency, Th1-related cytokines (IFN-γ and IL-12), Treg-related cytokines (TGF-ß) and T-bet, and Foxp3 mRNA levels compared with the control group. There was no significant difference between A. fumigatus pulsed DCs group and the control group about Th17 and Th2 frequency. CONCLUSIONS: The inactivated conidia of A. fumigatus were able to activate BMDCs and made them capable of triggering T cell responses in vitro. A. fumigatus loaded DCs was a weak inducer of Th17 and Th2, but induced a strong Th1 and Treg response.

Micronucleus expression and acute leukemia prognosis.

The micronucleus frequency (MNF) in peripheral blood lymphocytes (PBL) is a biomarker of chromosomal damage and genome instability in human populations.The relationship of micronucleus frequency with prognosis of patients with acute leukemia is not clear. We therefore investigated MNF in mitogen-activated peripheral blood lymphocytes from patients with hematologic diseases and solid tumours. Patients included 50 with acute leukemia, 49 diagnosed with myelodysplastic syndrome (MDS), 54 with benign blood diseases, and 45 with solid tumours, examined with 50 healthy controls. The mean MNF was significantly higher in cases of hematologic diseases and solid tumor patients than in healthy controls (P<0.001). There was no evident difference between MNF in the acute leukemia (7.15 ± 2.18) and solid tumor groups (7.11 ± 1.47), but both were higher than in the MDS group (5.12 ± 1.29) and benign blood diseases group (3.08 ± 1.08). Taking 7.15%, the average MNF of the acute leukemia group as standard, and dividing 50 cases of acute leukemia patients into high MNF group (MNF ≥ 7.15%) and low MNF group (MNF<7.15%). The overall response (complete remission + partial remission) rates of the low MNF group were significantly higher than in the high MNF group (P=0.001). The high MNF group further showed lower overall survival rates than the low MNF group. MNF expression and progression-free survival seemed to have a opposite relationship, with a correlation coefficient of -0.702. These data indicate that MNF in peripheral blood lymphocytes is important for evaluation of prognosis of acute leukemia patients, and it can reflect progression of disease to a certain degree.