Protein expression shift and potential diagnostic markers through proteomics profiling of tuberculous pleurisy biopsy tissues.

OBJECTIVES: Tuberculous pleurisy is a common type of tuberculosis (TB), but its diagnosis is challenging. This study aimed to profile the protein expression of this disease and identify new diagnostic makers. METHODS: Biopsy tissues from patients with tuberculous pleurisy and controls were taken through thoracoscopy, and proteins were extracted for Tandem Mass Tag Mass Spectrometry. Differential protein expression was performed between patients and controls, and the identified proteins were analyzed for pathway enrichment. Selected proteins were further validated in another set of samples using a more quantitative method. RESULTS: A total of 5101 proteins were detected and quantified in a discovery set of patients and controls. Overall protein expression was quite different between patients and controls. Most proteins were down-expressed, while a minority were overly expressed in the patient samples. At p value < 0.05 and absolute fold change >2, 295 proteins were found to be up-expressed and 608 down-expressed. The top enriched pathways included ECM-receptor interaction, complement and coagulation cascades and focal adhesion. All 19 selected candidates were validated in an independent set of patient and control samples. CONCLUSION: This unbiased proteomics approach not only provided unique insights into protein expression and pathways, but also discovered potential diagnostic markers for tuberculous pleurisy.

Apatinib preferentially inhibits PC9 gefitinib-resistant cancer cells by inducing cell cycle arrest and inhibiting VEGFR signaling pathway.

BACKGROUND: Lung cancer is one of the most common and deadly tumors around the world. Targeted therapy for patients with certain mutations, especially by use of tyrosine kinase inhibitors (TKIs) targeting epidermal growth factor receptor (EGFR), has provided significant benefit to patients. However, gradually developed resistance to the therapy becomes a major challenge in clinical practice and an alternative to treat such patients is needed. Herein, we report that apatinib, a novel anti-angiogenic drug, effectively inhibits obtained gefitinib-resistant cancer cells but has no much effect on their parental sensitive cells. METHODS: Gefitinib-resistant lung cancer cell line (PC9GR) was established from its parental sensitive line (PC9) with a traditional EGFR mutation after long time exposure to gefitinib. Different concentrations of apatinib were used to treat PC9, PC9GR, and other two lung cancer cell lines for its anti-growth effects. RNA sequencing was performed on PC9, PC9GR, and both after apatinib treatment to detect differentially expressed genes and involved pathways. Protein expression of key cycle regulators p57, p27, CDK2, cyclin E2, and pRb was detected using Western blot. Xenograft mouse model was used to assess the anti-tumor activity of apatinib in vivo. RESULTS: The established PC9GR cells had over 250-fold increased resistance to gefitinib than its sensitive parental PC9 cells (IC50 5.311 ± 0.455 µM vs. 0.020 ± 0.003 µM). The PC9GR resistance cells obtained the well-known T790M mutation. Apatinib demonstrated much stronger ( ~ fivefold) growth inhibition on PC9GR cells than on PC9 and other two lung cancer cell lines, A549 and H460. This inhibition was mostly achieved through cell cycle arrest of PC9GR cells in G1 phase. RNA-seq revealed multiple changed pathways in PC9GR cells compared to the PC9 cells and after apatinib treatment the most changed pathways were cell cycle and DNA replication where most of gene activities were repressed. Consistently, protein expression of p57, CDK2, cyclin E2, and pRb was significantly impacted by apatinib in PC9GR cells. Oral intake of apatinib in mouse model significantly inhibited establishment and growth of PC9GR implanted tumors compared to PC9 established tumors. VEGFR2 phosphorylation in PC9GR tumors after apatinib treatment was significantly reduced along with micro-vessel formation. CONCLUSIONS: Apatinib demonstrated strong anti-proliferation and anti-growth effects on gefitinib resistant lung cancer cells but not its parental sensitive cells. The anti-tumor effect was mostly due to apatinib induced cell cycle arrest and VEGFR signaling pathway inhibition. These data suggested that apatinib may provide a benefit to patients with acquired resistance to EGFR-TKI treatment.

Morphological evaluation using MRI of the olfactory filaments (fila) in a post-traumatic olfactory rat model.

OBJECTIVE: To investigate whether magnetic resonance imaging (MRI) can be used to directly assess olfactory bulb (OB) lesions and quantify the associated morphological changes of olfactory filaments (OF), also known as fila, in an in vivo OB-lesion rat model of the brain. METHODS: A surgical group (n = 5) of male Sprague-Dawley rats was subjected to the unilateral damage of the OB by a steel needle. The control group (n = 5) did not receive surgery. To assess olfactory system injury in vivo, T2-weighted MRI images were acquired in an oblique plane at a 30° angle from transverse plane one day after surgery. These brain regions were also assessed in the controls. The olfactory function was evaluated using the buried food pellet test (BFPT) 5 days before and after surgery. RESULTS: The OF could be clearly observed on the MRI images from all animals. The left and right OF mean lengths (mm) were similar in the control group (0.81 ± 0.18 vs 0.89 ± 0.17, P > 0.05). In the surgical group, the OB was partially injured in all rats. These rats did not show differences in OF length between left- and right-side (0.83 ± 0.18 vs 0.93 ± 0.24, P > 0.05) at the time of measurement. The time (sec) required to find the food pellets in the BFPT was longer after than before the surgery (83.80 ± 34.37 vs 231.44 ± 53.23, P < 0.05). CONCLUSIONS: MicroMRI may be a feasible tool to evaluate the OF and OBs in rat models. The unilateral partial OB lesion model appears to be an effective post-traumatic olfactory dysfunction model.

[Preliminary study of DTI on cerebral white matter micro-structure of patients with idiopathic olfactory loss].

OBJECTIVES: To investigate the cerebral white matter micro-structure in patients with idiopathic olfactory loss using diffusion tensor imaging (DTI). METHODS: Sixteen patients with idiopathic olfactory loss and sixteen normal subjects matched by age and sex were recruited in this study. Sniffin'Stick olfactory test was performed to evaluate the olfactory function of all subjects. We acquired diffusion tensor images with an echo planar imaging (EPI) sequence from all subject on a 3T scanner. The fractional anisotropy (FA) images were performed using DTI-studio, and bilateral Piriform cortex, Orbitofrontal cortex, Hippocampus and Insula cortex adjacent white matter and Capsula interna were delineated as the region of interesting (ROI), the FA for each ROI was calculated. Independent sample t test analysis was used to compare the FA value of all ROIs between the controls and patients. In addition, correlation analysis between FA value and MMSE score in patients were conducted. RESULTS: Compared with the controls, patients showed significantly decreased FA value in the adjacent white matter of bilateral Piriform cortex, Orbitofrontal cortex, Hippocampus and Insula cortex (P<0.05). There is no significant difference of FA value in bilateral Capsula interna between two groups (P>0.05). CONCLUSIONS: The patients with idiopathic olfactory loss show the damage of white matter micro-structure in the olfactory center, which could be important for the pathogenesis study and early intervention of idiopathic olfactory loss.

Unique variations of Epstein-Barr virus-encoded BARF1 gene in nasopharyngeal carcinoma biopsies.

The Epstein-Barr virus (EBV) BamHI-A rightward frame 1 (BARF1) gene is frequently expressed in EBV-associated epithelial malignancies and involves in oncogenicity and immunomodulation. To characterize the variations of BARF1 gene in different populations, the sequences of BARF1 gene in Northern Chinese nasopharyngeal carcinoma (NPC), EBV-associated gastric carcinoma (EBVaGC) and healthy donors were analyzed. The correlation of BARF1 variation with polymorphisms of BamHI F fragment (type F and f variants) and EBV-coded viral interleukin-10 (vIL-10) gene (B95-8 and SPM patterns) was also explored. Two major subtypes of BARF1 gene, designated as B95-8 and V29A, were identified. B95-8 subtype had identical amino acid sequence to B95-8 and was the dominant subtype among the EBV isolates from Northern China. V29A subtype, with one consistent amino acid change at residue 29 (V→A) and several nucleotide changes, showed higher frequency in NPC cases (25.3%, 20/79) than in EBVaGC cases (0/45) or healthy donors (4.3%, 2/46) (NPC vs. EBVaGC: P=0.0001; NPC vs. healthy donor: P=0.004). A preferential linkage between BamHI F and BARF1/vIL-10 polymorphisms was found. Type f isolates was specially correlated with the V29A/SPM genotype in NPC isolates and type f/V29A/SPM was preferentially found in NPC. BARF1/c-fms homology domain, transforming domain and cytotoxic T lymphocyte (CTL) epitopes of BARF1 were highly conserved in most isolates, suggesting the important role of BARF1 in virus infection and the potential usefulness in EBV-targeting immunotherapy of EBV-associated tumors. The relatively higher prevalence of type f/V29A/SPM strains in NPC may also suggest the association between these variations in multiple viral genes and NPC.