Mycobacterium vaccae alleviates allergic airway inflammation and airway hyper-responsiveness in asthmatic mice by altering intestinal microbiota.

Host immunity can influence the composition of the gut microbiota and consequently affect disease progression. Previously, we reported that a Mycobacterium vaccae vaccine could ameliorate allergic inflammation in asthmatic mice by regulating inflammatory immune processes. Here, we investigated the anti-inflammatory effects of M. vaccae on allergic asthma via gut microbiota modulation. An ovalbumin (OVA)-induced asthmatic murine model was established and treated with M. vaccae. Gut microbiota profiles were determined in 18 BALB/c mice using 16S rDNA gene sequencing and metabolomic profiling was performed using liquid chromatography quadrupole time-of-flight mass spectrometry. Mycobacterium vaccae alleviated airway hyper-reactivity and inflammatory infiltration in mice with OVA-induced allergic asthma. The microbiota of asthmatic mice is disrupted and that this can be reversed with M. vaccae. Additionally, a total of 24 differential metabolites were screened, and the abundance of PI(14:1(9Z)/18:0), a glycerophospholipid, was found to be correlated with macrophage numbers (r = 0.52, p = 0.039). These metabolites may affect chemokine (such as macrophage chemoattractant protein-1) concentrations in the serum, and ultimately affect pulmonary macrophage recruitment. Our data demonstrated that M. vaccae might alleviate airway inflammation and hyper-responsiveness in asthmatic mice by reversing imbalances in gut microbiota. These novel mechanistic insights are expected to pave the way for novel asthma therapeutic strategies.

The regulation of expression and splicing of transcription factors are related to the muscle damage caused by radiation in tree shrews.

Radiotherapy-induced muscle injury (RIMI) is a major complication of radiotherapy for nasopharyngeal carcinoma. Transcription factor (TF) expression and alternative splicing are crucial events in transcriptional and posttranscriptional regulation, respectively, and are known to be involved in key signaling pathways contributing to a variety of human disorders, including radiation injury. To investigate the TFs and alternative splicing events involved in RIMI, we constructed a tree shrew model as described previously in which the RIMI group received 20 Gy of irradiation on the tensor veli palatini (TVP) muscles. The irradiated muscles were evaluated by RNA sequencing (RNA-seq) 6 months later, and the results compared with those for normal TVP muscles. The alt5p and alt3p events were the two main types of differentially regulated alternative splicing events (RASEs) identified via the Splice sites Usage Variation Analysis (SUVA) software, and these RASEs were highly conserved in RIMI. According to functional enrichment analysis, the differentially RASEs were primarily enriched in pathways related to transcriptional regulation. Furthermore, we identified 16 alternative splicing TFs (ASTFs) in ASTF-differentially expressed gene (DEG) networks based on co-expression analysis, and the regulatory networks were chiefly enriched in pathways linked to cell proliferation and differentiation. This study revealed that RASEs and ASTF-DEG networks may both play important regulatory roles in gene expression network alteration in RIMI. Future studies on the targeting mechanisms and early interventions directed at RASEs and ASTF-DEG networks may aid in the treatment of RIMI.

Epstein Barr virus infection in tree shrews alters the composition of gut microbiota and metabolome profile.

BACKGROUND: Epstein-Barr virus (EBV) infection is a major global threat; its manifestations range from the absence of symptoms to multiorgan malignancies and various gastrointestinal diseases. Analyzing the composition and metabolomic profile of gut microbiota during acute EBV infection might be instrumental in understanding and controlling EBV. METHODS: Six tree shrews were inoculated with EBV by intravenous injection. Blood was collected at regular intervals thereafter from the femoral vein to detect EBV and inflammatory biomarker. At the same time, tree shrew faeces were collected for 16 S rRNA gene sequencing and Non-targeted metabolomics analysis. RESULTS: 16 S rRNA gene characterization along with ß diversity analysis exhibited remarkable alterations in gut microflora structure with a peak at 7 days post-infection(dpi). Some alterations in the relative richness of bacterial taxon were linked to infectious indicators. Of note, Butyricicoccus relative richness was positively linked to EBV presence in the blood and plasma, the opposite correlation was seen with Variovorax and Paramuribaculum. Non-targeted metabolomics indicated the fecal metabolome profile altered during EBV infection, particularly 7 dpi. The relative abundance of geranic acid and undecylenic acid in stool samples was positively linked to systemic inflammatory biomarkers, and an inverse relationship was reported with the estrone glucuronide, linoleic acid, protoporphyrin IX and tyramine. CONCLUSION: Collectively, EBV infection in this model correlated with changes in the composition and metabolome profile of the gut microbiota.

Comparison of sonotubometry, impedance, tubo-tympano-aerography, and tubomanometry to test eustachian tube function.

PURPOSE: There is currently no gold standard for the diagnosis of eustachian tube (ET) dysfunction. To provide an objective basis for the clinical diagnosis of ET dysfunction, we explored the characteristics of sonotubometry, impedance, tubo-tympano-aerography (TTAG), and tubomanometry (TMM) in volunteers with healthy ETs. MATERIALS AND METHODS: Sonotubometry, impedance, TTAG, and TMM tests were performed in 110 healthy ears of 55 volunteers, and the characteristics of each ET test were compared and discussed. RESULTS: The ET opening rate was compared between sonotubometry with dry swallowing, impedance with the Valsalva maneuver, TTAG with the Valsalva maneuver, and TMM with a nasopharyngeal pressure of 50 mbar in 100 (90.9%), 102 (92.7%), 99 (90.0%), and 104 (94.5%) ears, respectively; there was no significant difference among the four methods (P = 0.575). In sonotubometry, both dry swallowing and the Valsalva maneuver were superior to wet swallowing in terms of detecting ET opening (P = 0.000). In TMM, both the opening rate and the external auditory canal pressure were positively correlated with the nasopharyngeal pressure. Specifically, the opening rate and external auditory canal pressure increased with an increase in the nasopharyngeal pressure (r = 0.271, P = 0.000; r = 0.315, P = 0.000, respectively). CONCLUSIONS: Sonotubometry, impedance, TTAG, and TMM have their own advantages and disadvantages. In clinical practice, the appropriate ET function test should be chosen on the basis of the patient's specific condition.

Micro-CT study of the human cochlear aqueduct.

OBJECTIVE: The anatomic structure of the cochlear aqueduct (CA) in human temporal bone specimens was observed using micro-computed tomography (CT). MATERIALS AND METHODS: Micro-CT scanning of 18-µm-thick slices was performed on 30 slides of human temporal bone specimens to observe the CA structure and its relationship with its surroundings. The length, internal and external apertures, and the narrowest width of the CA were measured. The differences in CAs were compared between high jugular bulb (HJB) specimens and normal specimens. RESULTS: A large number of CA images were acquired using Micro-CT scanning, which clearly displayed the basic anatomic structures, stereotactic localizations, and adjacent relationships of the CAs. The whole course of a CA was 12.31 ± 3.60 mm, the diameter of the internal aperture was 465 ± 242 µm, the diameter of the external aperture was 2.88 ± 1.06 mm, the narrowest diameter was 601 ± 335 µm, the diameter of the opening of inferior cochlear vein (ICV) was 151 ± 50 µm, the distance between the internal aperture and ICV was 270 ± 197 µm, and the distance between the inferior margin of the internal acoustic meatus (IAM) and the top most part of the external aperture of the CA was 6.783 ± 2.15 mm. No bony obstruction of the CA or CA enlargement was observed in the specimens. A total of 28 CAs had one accompanying bony canal in the surroundings. The length and travelling of the CA were not affected by the level of the jugular bulb (JB). The variation of the travelling of the ICV was larger than that of the CA. CONCLUSION: Micro-CT adequately displayed the bony CA canal and provided a new method for anatomical studies of the CA and a basis for functional studies.

Preoperative evaluation of cochlear implantation through the round window membrane in the facial recess using high-resolution computed tomography.

PURPOSE: We evaluated the risk of cochlear implantation through the round window membrane in the facial recess through a preoperative analysis of the angle between the facial nerve-round window and the cranial midline using high-resolution temporal bone CT. METHODS: Temporal bone CT films of 176 patients with profound sensorineural hearing loss at our hospital from 2013 to 2015 were reviewed. The preoperative temporal bone CT scans of the patients were retrospectively analysed. The vertical distance (d value) from the leading edge of the facial nerve to the posterior wall of the external auditory canal and the angle (α value) between the line from the leading edge of the facial nerve to the midpoint of the round window membrane and the median sagittal line on the round window membrane plane were measured. Based on intraoperative observation, the round window membrane was divided into complete round window membrane exposure (group A), partial exposure (group B), and unexposed (group C) groups, and statistical analysis was performed. RESULTS: The α value could be effectively measured for all 176 patients (62.60 ± 7.12), and the d value could be effectively measured for 95 cases (5.53 ± 1.00). An analysis of the correlation between the α and d values of these 95 cases found a negative correlation. Of the 176 cases, one-way analysis of variance (ANOVA) showed that the differences among the groups were significant [P = 0.000 (< 0.05)]. CONCLUSION: The angle (α value) between the line connecting the leading edge of the facial nerve to the midpoint of the round window and the median sagittal line measured in preoperative CT scans was associated with the difficulty of intraoperatively exposing the round window membrane. When the α value was larger than a certain degree, the difficulty of exposing the round window membrane was increased. In such cases, the surgeon should fully expose the round window membrane during surgery, which could result decrease the likelihood of complications.

A study of Epstein-Barr virus infection in the Chinese tree shrew(Tupaia belangeri chinensis).

BACKGROUND: Epstein-Barr virus (EBV) is closely associated with many human diseases, including a variety of deadly human malignant tumours. However, due to the lack of ideal animal models,the biological characteristics of EBV, particularly its function in tumourigenesis, have not been determined. Chinese tree shrews (Tupaia belangeri chinensis), which are similar to primates, have been used to establish a variety of animal models and have recently received much attention. Here, we established tree shrews as a model for EBV infection by intravenous injection. METHODS: Ten tree shrews were inoculated with EBV by intravenous injection,and blood was collected at regular intervals thereafter from the femoral artery or vein to detect EBV markers. RESULTS: Eight of 10 tree shrews showed evidence of EBV infection. In the 8 EBV-infected tree shrews, EBV copy number increased intermittently or transiently, EBV-related gene expression was detected, and anti-EBV antibodies increased to varying degrees. Macroscopic hepatomegaly was observed in 1 tree shrew, splenomegaly was observed in 4 tree shrews, and enlarged mesenteric lymph nodes were observed in 3 tree shrews. Haematoxylin and eosin (HE) staining showed splenic corpuscle hyperplasia in the spleens of 4 tree shrews and inflammatory cell infiltration of the liver of 1 tree shrew and of the mesenteric lymph nodes of 3 tree shrews. EBER in situ hybridization(ISH) and immunohistochemical (IHC) staining showed that EBER-, LMP1- and EBNA2- positive cells were present in the spleens and mesenteric lymph nodes of some tree shrews. Western blotting (WB) revealed EBNA1-positive cells in the spleens of 4 tree shrews. EBV markers were not detected by HE, EBER-ISH or IHC in the lung or nasopharynx. CONCLUSIONS: These findings suggest that EBV can infect tree shrews via intravenous injection. The presented model offers some advantages for exploring the pathophysiology of EBV infection in humans.

Auditory brainstem responses after electrolytic lesions in bilateral subdivisions of the medial geniculate body of tree shrews.

This study aimed to establish a tree shrew model of bilateral electrolytic lesions in the medial geniculate body (MGB) to determine the advantages of using a tree shrew model and to assess the pattern of sound processing in tree shrews after bilateral electrolytic damage in different parts of the MGB. The auditory brainstem responses (ABRs) of a normal control group (n = 30) and an electrical damage group (n = 30) were tested at 0 h, 24 h, 48 h, 72 h, 7 days, 15 days, and 30 days after surgery. (1) The bilateral ablations group exhibited a significant increase in the ABR threshold of the electrolytic damage group between pre- and post-operation. (2) There were significant increases in the I-VI latencies at 0 h after MGBd and MGBm lesions and at 24 h after MGBv lesion. (3) The amplitudes of wave VI were significantly decreased at 24 h and 48 h after MGBd lesion, at 72 h and 7 days after MGBm lesion, and at 24 h, 48 h, 72 h, and 7 days after MGBv lesion. (1) The electrolytic damage group suffered hearing loss that did not recover and appeared to be difficult to fully repair after bilateral ablation. (2) The latencies and amplitudes of responses in the MGB following bilateral electrolytic lesion were restored to pre-operation levels after 15-30 days, suggesting that a portion of the central nuclei lesion was reversible. (3) The tree shrew auditory animal model has many advantages compared to other animal models, such as greater complexity of brain structure and auditory nuclei fiber connections, which make the results of this experiment more useful for clinical diagnoses compared with studies using rats and guinea pigs.

Follistatin-like protein 1 contributes to dendritic cell and T-lymphocyte activation in nasopharyngeal carcinoma patients by altering nuclear factor κb and Jun N-terminal kinase expression.

Follistatin-like protein 1 (FSTL1) is a newly characterized protein that can regulate the immune response in various ways. Dendritic cells (DCs) are central to immune regulation. In this study, we explored the impact of FSTL1 on DC activity in nasopharyngeal carcinoma (NPC) patients. The surface expression of CD40, CD86, and HLA-DR on DCs was analyzed and showed significantly elevated expression levels, indicating DC maturity. After FSTL1 was added to DCs collected from NPC patients (n = 50), controls (n = 47), and healthy donors (n = 10), interferon γ secretion and T-cell receptor expression in cytotoxic T lymphocytes were also investigated. In the experimental groups, the expression of the critical immune protein nuclear factor (NF)-κb was upregulated, whereas Jun N-terminal kinase (JNK) was downregulated. Our findings demonstrate that FSTL1 plays a critical role in immune regulation, enhancing the antigen presentation ability of DCs by up-regulating NF-κb expression and down-regulating JNK expression.

The effects of the auditory brainstem response before and after fluoro-gold injection in medial geniculate body.

OBJECTIVE: This study aimed to use the tree shrew as an otological model, not only to verify the location of the auditory pathway in tree shrews by fluoro-gold (FG) but also to elucidate the effects of the auditory brainstem response (ABR) before and after FG injection. METHODS: FG was injected into the medial geniculate body (MGB) of experimental group (n=10).The normal group (n=10) was inserted the microsyringe, which was not perfused FG. Hearing was assessed by testing ABRs before and after the operation. RESULTS: FG-labelled neurons were primarily distributed in the ipsilateral MGB, the ipsilateral and contralateral nuclei of the inferior colliculus (NIC), the superior olivary nucleus (SON), the dorsal cochlear nucleus (DCN), and the ventral cochlear nucleus (VCN). The ABR after FG injection caused a significant decrease in the wave amplitudes at 24 h that recovered by 72 h. However, the wave I-VI interpeak latencies in the right ear were shortened at 0 and 24 h post-surgery, whereas after 48 h, the interpeak latencies were prolonged. CONCLUSIONS: The FG retrograde tracing technique accurately displays the anatomical location of the auditory pathway in the tree shrew. The change in ABR waves suggested that there was a functional abnormality in the central auditory pathway after FG injection. The auditory thalamus may have self-regulating properties.

14-Thienyl methylene matrine (YYJ18), the derivative from matrine, induces apoptosis of human nasopharyngeal carcinoma cells by targeting MAPK and PI3K/Akt pathways in vitro.

BACKGROUND/AIMS: Nasopharyngeal carcinoma (NPC) is a distinctive type of head and neck cancer with the highest incidence in South China. Previous studies have proved that matrine, a main alkaloid isolated from Sophora flavescens Ait, has antitumor activity against NPC. However, the effect is not so pronounced and the underlying mechanism remains largely unclear. Here we investigated whether 14-thienyl methylene matrine (YYJ18) that was derived from matrine could exert more effective suppression activity on NPC, along with the underlying mechanism. METHODS: NPC cell lines CNE1, CNE2 and HONE1 were treated with YYJ18. Cell proliferation and apoptosis were determined by MTT assay and flow cytometry. Activation of mitogen-activated protein kinases (MAPK) and phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathways were determined by Western blotting and quantitative RT-PCR. RESULTS: YYJ18 remarkably inhibited proliferation and induced apoptosis of all three NPC cell lines in a dose-dependent manner, especially in CNE2 cells. Furthermore, YYJ18 treatment significantly suppressed phosphorylation of p38 in CNE2 cells, but upregulated phosphorylation of extracellular signal-regulated kinase1/2 (ERK1/2) and Akt. Next, alterations in downstream signaling were found, including activation of BCL2-associated X protein (Bax), caspase-3 and inactivation of B-cell CLL/lymphoma 2 (Bcl-2). CONCLUSION: We demonstrate the potent inhibitory effects of 14-thienyl methylene matrine on NPC cells for the first time, which could be mediated by modulation of MAPK and PI3K/Akt pathways.

The effect of nitidine chloride on the proliferation and apoptosis of nasopharyngeal carcinoma cells.

PURPOSE: This study was conducted to investigate the effect of nitidine chloride (NC) on the proliferation and apoptosis of nasopharyngeal carcinoma cell line CNE1, CNE2, TWO3, and C666-1, and to explore its antitumor mechanism. METHODS: NC was dissolved in IMDM medium and cultured with nasopharyngeal carcinoma cell line CNE1, CNE2, TWO3 and C666-1. Cell morphology, cell proliferation, cell apoptosis, p53 mRNA and p53 protein levels were assessed. RESULTS: After incubation with NC for 24 h, typical apoptotic morphology was observed. NC inhibited the proliferation and induced apoptosis of all 4 cell lines in a time-dose dependent manner. p53 mRNA and p53 protein levels were significantly increased. CONCLUSIONS: NC inhibited proliferation and induced apoptosis of nasopharyngeal carcinoma cells with upregulation of p53 gene.

RNA-Seq and miRNA-Seq data from Epstein-Barr virus-infected tree shrews reveal a ceRNA network contributing to immune microenvironment regulation.

Epstein-Barr virus (EBV) infection in humans is ubiquitous and associated with various diseases. Remodeling of the immune microenvironment is the primary cause of EBV infection and pathogenesis; however, the underlying mechanism has not been fully elucidated. In this study, we used whole-transcriptome RNA-Seq to detect mRNAs, long non-coding RNAs (lncRNA), and microRNA (miRNA) profiles in the control group, 3 days, and 28 days after EBV infection, based on the tree shrew model that we reported previously. First, we estimated the proportion of 22 cell types in each sample using CIBERSORT software and identified 18 high-confidence DElncRNAs related to immune microenvironment regulation after EBV infection. Functional enrichment analysis of these differentially expressed lncRNAs primarily focused on the autophagy, endocytosis, and ferroptosis signalling pathways. Moreover, EBV infection affects miRNA expression patterns, and many miRNAs are silenced. Finally, three competing endogenous RNA regulatory networks were built using lncRNAs that significantly correlated with immune cell types, miRNAs that responded to EBV infection, and potentially targeted the mRNA of the miRNAs. Among them, MRPL42-AS-5 might act as an hsa-miR-296-5p "sponge" and compete with target mRNAs, thus increasing mRNA expression level, which could induce immune cell infiltration through the cellular senescence signalling pathway against EBV infection. Overall, we conducted a complete transcriptomic analysis of EBV infection in vivo for the first time and provided a novel perspective for further investigation of EBV-host interactions.

Utility of deep learning for the diagnosis of cochlear malformation on temporal bone CT.

OBJECTIVE: Diagnosis of cochlear malformation on temporal bone CT images is often difficult. Our aim was to assess the utility of deep learning analysis in diagnosing cochlear malformation on temporal bone CT images. METHODS: A total of 654 images from 165 temporal bone CTs were divided into the training set (n = 534) and the testing set (n = 120). A target region that includes the area of the cochlear was extracted to create a diagnostic model. 4 models were used: ResNet10, ResNet50, SE-ResNet50, and DenseNet121. The testing data set was subsequently analyzed using these models and by 4 doctors. RESULTS: The areas under the curve was 0.91, 0.94, 0.93, and 0.73 in ResNet10, ResNet50, SE-ResNet50, and DenseNet121. The accuracy of ResNet10, ResNet50, and SE-ResNet50 is better than chief physician. CONCLUSIONS: Deep learning technique implied a promising prospect for clinical application of artificial intelligence in the diagnosis of cochlear malformation based on CT images.

Nomogram for prediction of hearing rehabilitation outcome in children with congenital sensorineural hearing loss after cochlear implantation.

Background: Reliable predictors for rehabilitation outcomes in patients with congenital sensorineural hearing loss (CSNHL) after cochlear implantation (CI) are lacking. The purchase of this study was to develop a nomogram based on clinical characteristics and neuroimaging features to predict the outcome in children with CSNHL after CI. Methods: Children with CSNHL prior to CI surgery and children with normal hearing were enrolled into the study. Clinical data, high resolution computed tomography (HRCT) for ototemporal bone, conventional brain MRI for structural analysis and brain resting-state fMRI (rs-fMRI) for the power spectrum assessment were assessed. A nomogram combining both clinical and imaging data was constructed using multivariate logistic regression analysis. Model performance was evaluated and validated using bootstrap resampling. Results: The final cohort consisted of 72 children with CSNHL (41 children with poor outcome and 31 children with good outcome) and 32 healthy controls. The white matter lesion from structural assessment and six power spectrum parameters from rs-fMRI, including Power4, Power13, Power14, Power19, Power23 and Power25 were used to build the nomogram. The area under the receiver operating characteristic (ROC) curve of the nomogram obtained using the bootstrapping method was 0.812 (95 % CI = 0.772-0.836). The calibration curve showed no statistical difference between the predicted value and the actual value, indicating a robust performance of the nomogram. The clinical decision analysis curve showed a high clinical value of this model. Conclusions: The nomogram constructed with clinical data, and neuroimaging features encompassing ototemporal bone measurements, white matter lesion values from structural brain MRI and power spectrum data from rs-fMRI showed a robust performance in predicting outcome of hearing rehabilitation in children with CSNHL after CI.

Isolation, identification, and induced differentiation of satellite cells from skeletal muscle of adult tree shrews.

A method for the in vitro isolation, purification, identification, and induced differentiation of satellite cells from adult tree shrew skeletal muscle was established. The mixed enzyme digestion method and differential adhesion method were used to obtain skeletal muscle satellite cells, which were identified and induced to differentiate to verify their pluripotency. The use of a mixture of collagenase II, hyaluronidase IV, and DNase I is an efficient method for isolating adult tree shrew skeletal muscle satellite cells. The P3 generation of cells had good morphology, rapid proliferation, high viability, and an "S"-shaped growth curve. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence staining indicated that marker genes or proteins were expressed in skeletal muscle satellite cells. After myogenic differentiation was induced, multiple-nucleated myotubes were observed, and the MyHC protein was expressed. The expression of myogenic marker genes changed with the differentiation process. After the induction of adipogenic differentiation, orange-red lipid droplets were observed, and the expression of adipogenic marker genes increased gradually with the differentiation process. In summary, satellite cells from adult tree shrew skeletal muscle were successfully isolated using a mixed enzyme digestion method, and their potential for differentiation into myogenic and adipogenic cells was confirmed, laying a foundation for further in vitro study of tree shrew muscle damage.

Role of microRNA-21 in regulating 3T3-L1 adipocyte differentiation and adiponectin expression.

MicroRNAs are endogenous small RNAs with a high degree of conservation, participating in a variety of vital activities. In present study, to explore the effect of microRNAs on 3T3-L1 adipocyte differentiation and adiponectin expression, the adipo-related microRNAs were screened and identified by micorRNA microarray. The highly expression plasmid of microRNA-21 with obvious expression up-regulation (miR-21) and its anti-sense (miR-21 inhibitor) were constructed and transfected into 3T3-L1 preadipocytes. The effect of miR-21 on 3T3-L1 adipocyte differentiation was observed, and the protein and mRNA expression level of adiponectin and AP-1 were analyzed. Results showed that, the expression profiles of microRNAs significantly changed during 3T3-L1 adipocyte differentiation. The expression of miR-21 was obviously up-regulated. miR-21 could significantly promote adipocyte differentiation, increase adiponectin mRNA and protein expression, while decrease AP-1 protein level. Meanwhil, miR-21 inhibitor blocked the effects of miR-21 mentioned above. The overexpression of AP-1 could absolutely reverse the stimulatory effect of miR-21 on adiponectin. miR-21 plays an important role in regulating adipocyte differentiation and adiponectin expression by inhibiting AP-1 expression.

Effect of trichosanthin on apoptosis and telomerase activity of nasopharyngeal carcinomas in nude mice.

PURPOSE: To evaluate the effect of trichosanthin (TCS) on telomerase activity and apoptosis in nasopharyngeal carcinoma cells implanted into nude mice. METHODS: Nude mice implanted with CNE1 and CNE2 nasopharyngeal carcinoma cell lines were randomly divided into 3 groups, including TCS group, control group (treatment with saline only), and cyclophosphamide (CTX) group. The weight of mice and the tumor volume were measured. The ultra-microstructural changes were observed by transmission electron microscope. Expression of Bcl-2 and Bax were investigated by immunohistochemistry. Apoptosis of CNE1 and CNE2 cells was determined by the TUNEL method, and telomerase activity by TRAP-ELISA. RESULTS: Compared with the normal control group (saline group), TCS significantly inhibited the growth of both CNE1 and CNE2 tumor cells. CTX also inhibited the growth of both CNE1 and CNE2 tumor cells but this inhibition was slightly slower compared with the TCS. CONCLUSION: TCS suppresses the growth of CNE1 and CNE2 cells lines in vivo. Its anticancer effects were associated with induction of apoptosis and, at least partially, with suppression of telomerase activity.

[Research on the effect of biyan qingdu granula drug-containing serum on inhibition of nasopharyngeal carcinoma cells growth in vitro].

OBJECTIVE: To investigate the effect of Biyan Qingdu Granula drug-containing serum (BQG-DS) on cell growth and apoptosis in nasopharyngeal carcinoma cell lines CNE1, CNE2, TWO3, C666-1, and explore the antineoplastic mechanism of Biyan Qingdu Granula. METHODS: SD rats were randomly divided into three groups: experimental (Biyan Qingdu Granula) group, positive control (cytoxan) group and negative control group. After administration of drug, the serum was collected from the treated animals. MTT assay was used to examine the effect of BQG-DS on the proliferation of CNE1, CNE2, TWO3, C666-1 cell, and flow cytometry was used to observe the cell cycle distribution. Apoptosis of CNE1, CNE2, TWO3, C666-1 cell was further investigated by inverted microscope. RESULTS: BQG-DS inhibited the proliferation of CNE1, CNE2, TWO3, C666-1 cell and the effects were in a time-and concentration-dependent manner. BQG-DS could also induce apoptosis while the G1 phase was arrested. CONCLUSION: BQG-DS inhibits proliferation of nasopharyngeal carcinoma cells via induction of apoptosis and arrest of cell cycle.

Application of UNETR for automatic cochlear segmentation in temporal bone CTs.

OBJECTIVE: To investigate the feasibility of a deep learning method based on a UNETR model for fully automatic segmentation of the cochlea in temporal bone CT images. METHODS: The normal temporal bone CTs of 77 patients were used in 3D U-Net and UNETR model automatic cochlear segmentation. Tests were performed on two types of CT datasets and cochlear deformity datasets. RESULTS: Through training the UNETR model, when batch_size=1, the Dice coefficient of the normal cochlear test set was 0.92, which was higher than that of the 3D U-Net model; on the GE 256 CT, SE-DS CT and Cochlear Deformity CT dataset tests, the Dice coefficients were 0.91, 0.93, 0 93, respectively. CONCLUSION: According to the anatomical characteristics of the temporal bone, the use of the UNETR model can achieve fully automatic segmentation of the cochlea and obtain an accuracy close to manual segmentation. This method is feasible and has high accuracy.