MicroRNA-185-5p targets tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta to regulate non-small cell lung cancer progression.

BACKGROUND: Lung cancer (LC) is one of the most frequent cancers worldwide, as well as the leading cause of cancer-related death. Non-small cell lung cancer (NSCLC, which accounts for 85% of occurrences) is the main type of LC. MiRNAs appear to play a role in the occurrence and progression of many malignancies, according to mounting data. The underlying mechanism of miRNAs in regulating NSCLC cell biological activity and progression, on the other hand, is still being investigated. METHODS: QRT-PCR were used to detect miR-185-5p expression and YWHAZ mRNA in NSCLC. The CCK-8 assay was used to determine the tumor cells' ability to proliferate. Transwall assay was used to test the migratory and invasive properties of cells. Cell apoptosis was detected using flow cytometry. Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ), E-Cadherin, N-Cadherin and cleaved-caspase3 protein expression were assessed using Western Blot. The bioinformatics analysis software StarBase2.0 predicted miR-185-5p downstream targets. To confirm the target association between miR-185-5p and YWHAZ, a luciferase experiment was used. In addition, an NCl-H1299 xenograft model was created to assess the anti-tumor impact of miR-185-5p in vivo. The expression level of YWHAZ in tumor tissues of small xenograft tumor model was detected by immunohistochemistry assay. RESULTS: Decreased miR-185-5p expression levels were observed in NSCLC. In vitro, over-expressed miR-185-5p decreased cell viability, proliferation, invasion/migration, and induced cell apoptosis, while inhibiting tumor growth in vivo. Dual-luciferase gene experiments confirmed that YWHAZ binds to miR-185-5p. Overexpression of YWHAZ partially restored the inhibitory effects of miR-185-5p on cell behaviors. CONCLUSION: MiR-185-5p was down-regulated in NSCLC, and that overexpressed miR-185-5p inhibited malignant behaviors of cells and tumor growth by negatively regulating YWHAZ.

Bariatric surgery reduces sleep apnea in obese patients with obstructive sleep apnea by increasing pharyngeal cross-sectional area during the early postoperative period.

OBJECTIVE: Bariatric surgery (BS) is considered one of the most effective treatments for obese individuals with Obstructive Sleep Apnea (OSA). However, otolaryngologists have raised concerns about the structural alterations caused by BS on the upper respiratory tract, especially, on the pharyngeal cavity. METHODS: In this study, we recruited 42 individuals who underwent BS at our hospital. They were divided into two groups based on apnea-hypopnea index (AHI): mild group (5 ≤ AHI < 15) and moderate-severe group (AHI ≥ 15). The participants were followed up for 12 months and several indicators, including body mass index (BMI), polysomnography (PSG), and acoustic pharyngometry (APh), were assessed repeatedly before surgery and at 3, 6, and 12 months (m) after surgery. RESULTS: Participants exhibited significant decreases in BMI (F = 128.1, P = 0.001) and total weight loss (F = 176.7, P < 0.001) after BS. The AHI value among obese patients with mild OSA decreased significantly within three months after surgery (0 day vs. 3 months, P < 0.01), and decreased significantly more than 12 months with moderate-to-severe patients (0 day vs. 3 months, 3 months vs. 6 months, 6 months vs. 12 months, P < 0.01). The therapeutic effect of OSA of the mild group was significantly better compared with that of the moderate-severe group at 6 months (mean rank = 28.13 vs. 14.21, P < 0.001) and 12 m (mean rank = 26.75 vs. 15.52, P = 0.001). The APh results revealed that the pharyngeal volume of the two groups increased significantly between 0 day and 6 months after surgery (P < 0.01). The oropharyngeal junction (OPJ) area and the glottal area were increased significantly between 0 day and 6 m after surgery (P < 0.01). CONCLUSION: BS can relieve apnea and OSA symptoms among obese patients with OSA, especially in the early postoperative period. Moreover, OSA severity was closely associated with OPJ and glottal areas, rather than pharyngeal cavity volume.

[Study on the structural changes of pharyngeal cavity after bariatric surgery in obese patients with obstructive sleep apnea].

Objective:To study whether and how bariatric surgery changes the structure of the pharyngeal cavity in obese patients with obstructive sleep apnea（OSA）. Methods:Forty-two patients who underwent laparoscopic sleeve gastrectomy were recruited. Morphological indicators（BMI, neck and waist circumference）, PSG and acoustic pharyngometry indicators were evaluated pre-operatively and 3, 6, and 12 months post-operatively. Results:All indicators including morphology, pharyngeal cavity structure and OSA severity changed significantly after surgery. Among them, BMI, neck circumference, waist circumference and AHI value were significantly reduced（P<0.001）, while pharyngeal cavity volume, pharynx volume, oropharyngeal junction area, glottis area and LSaO2 increased significantly（P<0.001）. The results of multiple comparisons showed that BMI, neck and waist circumference decreased significantly in the first 6 months, and no further decline occurred during 6 to 12 months postoperatively. The decrease in AHI and LSaO2 mainly occurred within the first 3 months postoperatively, while there was no statistically significant difference in these two indicatiors between 3 months vs. 6 months, 6 months vs. 12 months postoperatively. The area of the oropharyngeal junction increased significantly within 0 to 3 months after surgery, while the volume of the pharyngeal cavity and the area of the glottis increased at 6 months and 12 months after surgery. Conclusion:Bariatric surgery can significantly reduce body weight and reduce fat accumulation in the neck. It can also enlarge the volume and cross-sectional area of the pharyngeal cavity, and improve upper airway obstruction, therefore reduce the symptoms of sleep apnea in obese patients with OSA to a certain extent.

Development of a TEM-cell-integrated CO2 incubator for cell-based transient electromagnetic field bioeffect study.

To facilitate the cell-based experiment for pulsed electromagnetic field biological effect study, a novel TEM-cell-integrated CO2 incubator was developed. The integrated experimental system could simultaneously meet the requirement of standard cell culture condition and the various Transient Electromagnetic Field (TEF) exposure, which made it possible to study the relationship between different electromagnetic pulse exposure and the cellular responses in a reliable way. During the research, a comparison experiment was carried out to evaluate the necessity of the integrated incubator system: firstly, two different types of cell lines, which are the human prostate cancer cell line (PC3) and the pancreatic ß cell line (MIN6) were chosen and exposed in the TEM-cell which located in the open area and the integrated system, respectively, with the same EFT radiation conditions; then, the cells' viability, the cellular ROS level and the mitochondrial membrane potential (MMP) were detected, respectively. The results showed that in the same parameter of the EFT radiation, the processes of the cells had a significant difference and even opposite in the incubator and open area, and all the results could be reproducible. The phenomenon indicated the stability of the TEM-cell-integrated CO2 incubator, and also demonstrated the necessity to strictly control the cell culture condition when carrying out the precise mechanism study of the TEF bioresponse at the cellular levels.

Bioactive Peptides from Walnut Residue Protein.

Walnut residue is a kind of high-quality plant protein resource. The bioactive peptide prepared from walnut residue has excellent health care functions such as antioxidation and antihypertensive activity, but at present, walnut residue is often regarded as waste or low value feed, fertilizer and other materials. The uneconomical use of walnut residue has hindered the development of the walnut industry to some extent. Effective utilization of walnut residue protein to develop bioactive peptides and other products is of great significance to realize the comprehensive utilization of walnut residue, improve the added value of by-products, and change the current low utilization rate of walnut residue. In this paper, the preparation, purification and structure identification of walnut protein bioactive peptides are reviewed, and different functional walnut active peptides (WBPs) are introduced. The potential effects of these bioactivities on human health and their different uses in food, medicine and other industries are discussed. The purpose is to provide reference information for the effective utilization of walnut residue resources and the development of walnut industry.

Newcastle disease virus selectively infects dividing cells and promotes viral proliferation.

Newcastle disease virus (NDV) can select cells to infect, but the mechanism of its cell selectivity has not been comprehensively investigated. Here, we use HeLa cells to establish that NDV can selectively infect cells at the single-cell level. We labeled proliferating cells with 5'-bromo-2-deoxyuridine (BrdU) and examined the colocalization of BrdU with NDV in cells to clarify the relationships between NDV infection and cell proliferation. Receptors at the plasma membrane mediate NDV entry into host cells. We labeled sialic acid receptor isoforms, compared their densities between different cell types and measured the sialic acid receptor densities in different cell phases. Our results suggest that NDV displays host tropism to HeLa cells compared to BHK cells and that the differences in the receptor isoform expression patterns between cell types contribute to the selection of HeLa by NDV. At the single-cell level, the dynamics of receptor expression changes during different cell phases contributing to the selection of cells in S/G2 phase for NDV infection. Furthermore, cell proliferation benefits viral replication, and enhanced virus replication leads to increased damage to cells. The elucidation of the mechanisms underlying host cell selection by NDV may help in the screening and characterizing of additional candidate oncolytic virus strains.

The clinical and prognostic significance of YWHAZ in non-small-cell lung cancer patients: Immunohistochemical analysis.

YWHAZ has been suggested to as an oncogene in various human malignancies, including non-small-cell lung cancer (NSCLC). Our study presents more evidence to confirm the clinical significance and biological function of YWHAZ in NSCLC. In our results, YWHAZ was upregulated in lung squamous cell carcinoma tissues and lung adenocarcinoma tissues through analyzing The Cancer Genome Atlas (TCGA) database, and confirmed high levels of YWHAZ messenger RNA and protein in lung squamous cell carcinoma tissues and lung adenocarcinoma tissues through quantitative real-time polymerase chain reaction and immunohistochemistry. Moreover, YWHAZ overexpression was correlated with advanced clinical stage, more lymph node metastasis and present distant metastasis in NSCLC patients. Survival analysis indicated that high level of YWHAZ protein expression was associated with short overall survival time in NSCLC patients, and YWHAZ expression was independent prognostic factors for overall survival in NSCLC patients. Moreover, Silencing of YWHAZ expression represses NSCLC cell migration and invasion. In conclusion, YWHAZ is a credible prognostic biomarker, and may be a therapeutic target in NSCLC.

[Development of tumor targeting PHBHHx nanoparticles by PhaP mediated immobilization of EGFR-targeting peptide].

PHA granule binding protein phasin (PhaP) has a high affinity for hydrophobic materials and can bind to hydrophobic polymers via strong hydrophobic interaction. In this study, an EGFR-targeting peptide (ETP) was fused with PhaP and the fusion protein ETP-PhaP was produced in recombinant Escherichia coli BL21 (DE3) (pPI-ETP-P) and then purified by Ni affinity purification. The tumor targeting PHBHHx nanoparticles were developed based on PhaP mediated ETP immobilization and the cellular uptake of the ETP-PhaP modified PHBHHx NPs and none modified PHBHHx NPs by cervical cancer cell lines SiHa (EGFR over expressed) and CaSKi (EGFR low expressed) were analyzed. The purified ETP-PhaP could be adsorbed onto the hydrophobic surface of PHBHHx NPs. The ETP-PhaP modified PHBHHx NPs could target to EGFR over expressed cervical cancer cells SiHa more efficiently than to the EGFR low expressed CaSKi cells. These results demonstrated the advantage in effectiveness and convenience of PhaP mediated ETP adsorption on PHBHHx nanoparticles, providing a novel strategy for hydrophobic nanocarrier surface modification.

De novo transcriptome sequencing and analysis of the juvenile and adult stages of Fasciola gigantica.

Fasciola gigantica is regarded as the major liver fluke causing fasciolosis in livestock in tropical countries. Despite the significant economic and public health impacts of F. gigantica there are few studies on the pathogenesis of this parasite and our understanding is further limited by the lack of genome and transcriptome information. In this study, de novo Illumina RNA sequencing (RNA-seq) was performed to obtain a comprehensive transcriptome profile of the juvenile (42days post infection) and adult stages of F. gigantica. A total of 49,720 unigenes were produced from juvenile and adult stages of F. gigantica, with an average length of 1286 nucleotides (nt) and N50 of 2076nt. A total of 27,862 (56.03%) unigenes were annotated by BLAST similarity searches against the NCBI non-redundant protein database. Because F. gigantica needs to feed and/or digest host tissues, some proteases (including cysteine proteases and aspartic proteases), which play a role in the degradation of host tissues (protein), have been paid more attention in the present study. A total of 6511 distinct genes were found differentially expressed between juveniles and adults, of which 3993 genes were up-regulated and 2518 genes were down-regulated in adults versus juveniles, respectively. Moreover, stage-specific differentially expressed genes were identified in juvenile (17,009) and adult (6517) F. gigantica. The significantly divergent pathways of differentially expressed genes included cAMP signaling pathway (226; 4.12%), proteoglycans in cancer (256; 4.67%) and focal adhesion (199; 3.63%). The transcription pattern also revealed two egg-laying-associated pathways: cGMP-PKG signaling pathway and TGF-ß signaling pathway. This study provides the first comparative transcriptomic data concerning juvenile and adult stages of F. gigantica that will be of great value for future research efforts into understanding parasite pathogenesis and developing vaccines against this important parasite.

First Report of Bovine Leukemia Virus Infection in Yaks (Bos mutus) in China.

Enzootic bovine leukosis (EBL) is a chronic lymphosarcoma disease of cattle caused by bovine leukemia virus (BLV). No information is available concerning the epidemiology of BLV infection in yaks (Bos mutus). One thousand five hundred and eighty-four serum samples from 610 black yaks and 974 white yaks from Gansu province, northwest China, were collected between April 2013 and March 2014 and tested for BLV antibodies using a commercially available ELISA kit. The overall BLV seroprevalence in yaks was 21.09% (334/1584), with 24.26% (148/610) black yaks and 19.10% (186/974) white yaks yielding positive results. Risk factor analysis indicated that with the exception of breed (OR = 1.36, 95% CI = 1.06-1.73, P < 0.05), the age, region, gender, farm, and the numbers of pregnancies were not considered as risk factors for the presence of BLV in yaks included in this study. This is the first report of BLV infection in yaks in China, which provides information for controlling BLV infection in yaks.

Microbial synthesized biodegradable PHBHHxPEG hybrid copolymer as an efficient intracellular delivery nanocarrier for kinase inhibitor.

BACKGROUND: Protein Kinases are key regulators of cell function and play essential roles in the occurrence and development of many human diseases. Many kinase inhibitors have been used for molecular targeted treatment of those diseases such as cancer and inflammation. However, those highly hydrophobic kinase inhibitors shared the common features of poor bioavailability and limited in vivo half-life, which strongly impeded their practical applications. Our previous study demonstrated that microbial synthesized biodegradable polyester poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx), a member of polyhydroxyalkanoates (PHAs) family, could serve as a promising delivery nanocarrier for those hydrophobic kinase inhibitors. Recently, a novel natural synthesized hybrid copolymer, PEG200 end-capped PHBHHx (PHBHHxPEG) was produced by Aeromonas hydrophila fermentation. In this study, the novel PHBHHxPEG NPs were prepared and investigated to serve as intracellular delivery nanocarriers for sustained release of hydrophobic kinase inhibitors. RESULTS: PHBHHxPEG nanoparticles (NPs) prepared by an emulsification-solvent evaporation method were spherical with a diameter around 200 nm. The entrapment efficiency on rapamycin in PHBHHxPEG NPs was 91.9% and the sustained release of rapamycin from PHBHHxPEG NPs could be achieved for almost 10 days. The cellular uptake of PHBHHxPEG NPs was significant higher than that of PHBHHx NPs. The anti-proliferation effect and mTOR inhibition ability of rapamycin-loaded PHBHHxPEG NPs was stronger than that of drug-loaded PHBHHx NPs and free rapamycin. CONCLUSIONS: PHBHHxPEG NPs could achieve the efficient entrapment and sustained release of rapamycin. The novel biodegradable PHBHHxPEG appeared a promising nanocarrier for sustained delivery of hydrophobic kinase inhibitors with improved cellular uptake and kinase inhibition efficiency.

[Expression of heparanase on the squamous cell carcinoma of head and neck and its clinical significance].

OBJECTIVE: To investigate the relationship among expression of heparanase (HPSE), the clinical and pathologic characteristics of squamous carcinoma on head and neck and the patients' prognosis. METHODS: Sixty-two cases of postoperative tumor specimens were verified by immunohistochemistry S-P method and computer-assisted image analysis method was used. RESULTS: The expression of HPSE in normal epithelium mucosae of head and neck was negative or very weak; in tumor tissue was positive, mainly in cytoplasm and the positive rate was 69.3%. The expression of HPSE hadn't significant difference with the age of patients and pathologic grades of tumors (chi2 = 0.05, chi2 = 3.84, P > 0.05), but had it with clinical stages and metastatic lymph node lesions (chi2 = 3.96, chi2 = 8.06, P < 0.05). The relationship between expression of HPSE in primary tumors and that in metastatic lymph node lesions showed significantly positive correlation (r = 0.9162, P = 0.001). Both HPSE and TNM clinical stage of tumor had significant correlation with the prognosis of patients respectively (P < 0.05). Calculated by Kaplan-Meier method, the accumulative survival rate of 3 years in positive HPSE expression group (25.9%) was much lower than that in negative group (72.7%), there was a significant difference between them by Log-Rank test (chi2 = 11.607, P < 0.001). CONCLUSIONS: The expression of HPSE is significantly increased in squamous carcinomas of head and neck, mainly expressed in cytoplasm. The expression of HPSE has a close relationship with clinical stages and lymph node metastasis of squamous carcinoma on head and neck. The higher the clinical stage, the more manifest the expression of HPSE. The expression of HPSE and TNM clinical stage of tumor are independent factors affecting prognosis.

[Expression of CD44 and nm23-H1 protein in the primary and metastatic lymph node lesions in supraglottic and hypopharyngeal cancer].

OBJECTIVE: To study the metastasis feature of the primary and metastatic lymph node lesions in supraglottic or hypopharyngeal cancer. METHODS: The expression of CD44 and nm23-H1 in specimens from the primary and metastatic lymph node lesions of the 41 cases with supraglottic or hypopharyngeal cancer were studied with immunohistochemistry method and flow cytometry. RESULTS: No correlation was found between the expression of CD44, nm23-H1 and the tumor differentiation of the supraglottic or hypopharyngeal cancer, but their expression related with the clinical staging. The CD44 and nm23-H1 positive expression rates in the primary and metastatic lymph node lesions were 75.6% (31/41), 85.4% (35/41) and 34.1% (14/41), 26.8% (11/41) respectively (P >0.05). The average fluorescence index of CD44 and nm23-H1 in the primary and metastatic lymph node lesions were 1.27 +/- 0.18, 1.33 +/- 0.16 and 1.11 +/- 0.19, 1.08 +/- 0.15 (x +/- s) respectively (P >0.05). CONCLUSIONS: The expressions of CD44 and nm23-H1 in the metastatic lymph node tumor had no difference compared with that in primary tumor of the supraglottic or hypopharyngeal cancer. The difference of metastasis potentials between the primary and metastatic lymph node lesions in the same patient was not proved in this study and should be further investigated from multiple oncogens markers.

[Effects of hypoxia on the expression of heparanase and invasiveness of Hep-2 laryngeal carcinoma cell].

OBJECTIVE: To evaluate the effects of hypoxia on the expression of heparanase and the invasiveness of Hep-2 laryngeal carcinoma cell line. METHODS: Hep-2 cells were incubated at either normoxia (37 degrees C, 5% CO2, 20% O2) or hypoxia (37 degrees C, 5% CO2, 2% O2) condition for 6 h,12 h,24 h,36 h. Flow cytometry was used to detect the protein expression of heparanase under different hypoxia conditions. Fluorescence Index represents the relative content of heparanase protein. Cell invasiveness was measured by matrigel invasion assay. RESULTS: Compared with normoxia group, the heparanase protein expression level in 6 h hypoxia group was increased (P <0.05) and the heparanase protein expression levels in other hypoxia groups were significantly increased (P < 0.01). The heparanase protein expression level was increased (P < 0.05) among 6 h, 12 h, 24 h hypoxia groups; Compared with normoxia group, there was no significant change in the invasion cells at 6 h (P > 0.05) in hypoxia group; whereas the level of cell invasion was significantly increased in 12 h(P < 0.05), 24 h and 36 h groups (P < 0.01). During 6 to approximately 36 h hypoxia period, the increase of hypoxia-induced invasiveness in Hep-2 cell line show time-dependent manner. Meanwhile, there was a positive correlation between the expression of HPSE and the invasiveness of Hep-2 cells. CONCLUSION: Invasion of Hep-2 cells in hypoxia condition correlates with heparanase level. Hypoxia plays an important role in the augmentation of the heparanase expression and the invasiveness of human laryngeal carcinomas.