[Comparative analysis of population distribution, severity, emotional, and cognitive characteristics among different side idiopathic tinnitus patients].

To provide basis for prevention and treatment by analyzing the clinical features, emotional and cognitive states and their correlations of idiopathic tinnitus. Cross-sectional study was used. Thirty-six right, 44 left, and 46 bilateral idiopathic tinnitus patients diagnosed in Beijing Tongren Hospital were prospectively enrolled from October, 2020 to February, 2022. The clinical data was recorded and the THI, DBI, STAI, and MoCA were evaluated. The clinical features and the incidence of severe tinnitus, hearing lose, anxiety, and cognitive impairment were compared by one-way ANOVA, Kruskal-Wallis H, and chi-square test. The correlation between tinnitus or hearing and emotional and cognitive states were evaluated by multivariable correlation analysis. There was no significant difference in age, BMI, years of education, tinnitus duration, and the incidence of hearing loss among groups (F=0.730,P=0.484;F=1.535,P=0.219;F=1.506,P=0.226;χ²=4.242,P=0.120;χ²=6.672,P=0.083). In right, left, and bilateral tinnitus patients, the number of severe tinnitus was 12, 7, and 20 cases and the incidence was 33.3%, 15.9%, and 43.5%; the number of depression was 13, 14, and 26 cases and incidence was 36.1%, 31.8%, and 53.5%; the number of trait anxiety was 3, 2, and 10 cases and the incidence was 8.3%, 4.5%, and 21.7%. Compared with left tinnitus patients, the incidence of severe tinnitus, depression, and trait anxiety was higher in bilateral tinnitus patients (χ²=8.139,P=0.004;χ²=5.558,P=0.018;χ²=5.753,P=0.007). The incidence of state anxiety and cognitive impairment were no significant difference among groups (χ²=0.142,P=0.931;χ²=1.338,P=0.512). The overall incidence of state anxiety and cognitive impairment were 16.7%(21/126) and 37.3%(47/126) respectively. There was positive correlation between THI score and BDI, S-AI, and T-AI scores (r=0.529,P=0.001; r=0.649,P<0.001; r=0.483,P=0.003) and negative correlation between THI and MoCA scores (r=-0.364,P=0.029) in right tinnitus group. The positive correlation was found between THI score and BDI, S-AI, and T-AI scores in left tinnitus group (r=0.508,P<0.001; r=0.506,P<0.001; r=0.357,P=0.017). The positive correlation between THI score and BDI, S-AI, and T-AI scores (r=0.753,P<0.001; r=0.527,P<0.001; r=0.536,P<0.001) and the positive correlation between tinnitus duration and MoCA score(r=0.334,P=0.023) were also found in bilateral tinnitus group.

[Application of lymphocytes test in peripheral blood of patients with systemic sclerosis during the treatment].

OBJECTIVE: To explore the significance of lymphocytes in systemic sclerosis (SSc), by detecting the levels of T lymphocytes, B lymphocytes and natural killer (NK) cells, and analyzing the correlation between the lymphocytes and clinical laboratory indexes. METHODS: The numbers and proportion of T, CD4+T, CD8+T, B, and NK cells were detected by flow cytometry in peripheral blood of 32 SSc patients who had taken immunosuppressive drugs and 30 healthy controls (HC). The comparison of the lymphocyte subsets in SSc with them in the HC groups, and the correlation between the lymphocytes and other clinical and laboratory indicators were analyzed by the relevant statistical analysis. RESULTS: Compared with the HC group, the numbers of T, CD4+T, CD8+T, and NK cells in peripheral blood of SSc group, who had taken immunosuppressive drugs, were significantly decreased (P < 0.05). More-over, the proportion of NK cells in peripheral blood of the SSc group was also significantly lower than that in the HC group (P=0.004). In addition, all the lymphocyte subsets were decreased in peripheral blood of more than 65% of the SSc patients who had taken immunosuppressive drugs. Compared with CD4+T normal group, the positivity of Raynaud's phenomenon, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) was significantly increased in CD4+T reduction group, respectively (P＝0.024, P < 0.001, P=0.018). ESR was higher in CD8+T reduction group than CD8+T normal group (P＝0.022). The prevalence of fingertip ulcer was significantly increased in B cell decrease group (P=0.019). Compared with NK cell normal group, the prevalence of fingertip ulcer was significantly increased in NK cell lower group (P=0.033), IgM was remarkablely decreased yet (P=0.049). The correlation analysis showed that ESR was negatively correlated with the counts of T lymphocytes (r＝－0.455, P＝0.009), CD4+T lymphocytes (r＝－0.416, P＝0.018), CD8+T lymphocytes (r＝－0.430, P=0.014), B cells (r＝－0.366, P＝0.039). CONCLUSION: The number of CD4+T, CD8+T, B, and NK cells significantly decreased in peripheral blood of SSc patients who had used immunosuppressive drugs, some lymphocyte subsets might be related with Raynaud's phenomenon and fingertip ulcer, and reflected the disease activity by negatively correlated with ESR and CRP; the numbers of lymphocyte subsets in peripheral blood should be detected regularly in SSc patients who had taken immunosuppressive drugs.

[Correlation analysis of Sema4D with rheumatoid arthritis disease activity, bone destruction and rheumatoid arthritis-related interstitial lung disease].

Objective: To explore the correlation between Semaphorin 4D (Sema4D) and rheumatoid arthritis (RA) clinical manifestations, laboratory indexes, bone destruction and rheumatoid arthritis related interstitial lung disease(RA-ILD), and to analyze its significance in evaluating the severity of patients with rheumatoid arthritis. Methods: A total of 108 RA patients and 50 healthy controls from September 2018 to October 2019 were collected from the Department of Rheumatology and Immunology, Peking University People's Hospital and Beijing Haidian Hospital. According to the DAS 28 score, RA patients were divided into active disease group (DAS28>2.6) and stable disease group (DAS28 ≤ 2.6). Fifty healthy controls. The levels of Sema4D in serum were detected by enzyme-linked immunoassay method (ELISA), and their correlation with clinical manifestations of RA, laboratory indicators, degree of bone damage and RA-ILD were analyzed. Results: The level of serum Sema4D in RA active group was significantly higher than that in stable group and healthy control group (P<0.05). The concentration serum Sema4D was positively correlated with C-reactive protein(CRP) (r=0.28, P<0.05), rheumatoid factor(RF) (r=0.25, P<0.05) and the 28-joint disease activity score (DAS28) (r=0.45, P<0.01). The concentration serum Sema4D was positively correlated with ß-Crosslaps(r=0.20, P<0.05) and Sharp-van der Heijde score (SHS)(r=0.13, P<0.05). The concentration serum Sema4D was positively correlated with Vascular endothelial growth factor (VEGF)(r=0.25, P<0.05) and Warrick score of chest CT in RA patients(r=0.27, P<0.05). The anti-cyclic citrullinated peptid(CCP) antibody, DAS28, VEGF and the incidence of RA-ILD were significantly higher than that in Sema4D negative group (P<0.05). Conclusions: Serum Sema4D level in RA patients is closely related to the disease activity, bone destruction and RA-ILD. Serum Sema4D can be used as an indicator of disease monitoring and prognosis evaluation in RA patients.

A decision tree model to distinguish between benign and malignant pulmonary nodules on CT scans.

OBJECTIVE: Chest computed tomography (CT) is increasingly being used to screen for lung cancer. Machine learning models could facilitate the distinction between benign and malignant pulmonary nodules. This study aimed to develop and validate a simple clinical prediction model to distinguish between benign and malignant lung nodules. PATIENTS AND METHODS: Patients who underwent a video thoracic-assisted lobectomy between January 2013 and December 2020 at a Chinese hospital were enrolled in the study. The clinical characteristics of the patients were extracted from their medical records. Univariate and multivariate analyses were used to identify the risk factors for malignancy. A decision tree model with 10-fold cross-validation was constructed to predict the malignancy of the nodules. The sensitivity, specificity, and area under the curve (AUC) of a receiver operatic characteristics curve were used to evaluate the model's prediction accuracy in relation to the pathological gold standard. RESULTS: Out of the 1,199 patients with pulmonary nodules enrolled in the study, 890 were pathologically confirmed to have malignant lesions. The multivariate analysis identified satellite lesions as an independent predictor for benign pulmonary nodules. Conversely, the lobulated sign, burr sign, density, vascular convergence sign, and pleural indentation sign were identified as independent predictors for malignant pulmonary nodules. The decision tree analysis identified the density of the lesion, the burr sign, the vascular convergence sign, and the drinking history as predictors of malignancy. The area under the curve of the decision tree model was 0.746 (95% CI 0.705-0.778), while the sensitivity and specificity were 0.762 and 0.799, respectively. CONCLUSIONS: The decision tree model accurately characterized the pulmonary nodule and could be used to guide clinical decision-making.

Morphological variation of mutant sunflowers (Helianthus annuus) induced by space flight and their genetic background detection by SSR primers.

After sunflower seeds were exposed to space conditions, various mutant plants were screened from the descendent plants. The morphological characters of plants changed in flower color from golden to yellow, light yellow, or even to yellowish green. The ligulate petals of the unisexual floret broadened, or became thin, while the short tubular petals of bisexual floret elongated to some extent, or even turned into semi-ligulate petals or ligulate petals, making the phenotype of the whole inflorescence like a chrysanthemum. The shape and thickness of leaves varied in some of these plants. Absolute sterile plants in mutant plants were found to possess neither normal bisexual florets nor unisexual florets, but the "pseudo-floret" only consisted of pieces of shield-like bracts on protuberant floral disc. Thirty-five pairs of simple sequence of repeat primers were used to detect the genetic variation of the plants, and the results showed that only a variation was tested in the mutant plants from 4 primers. The different PCR products obtained were extracted for sequencing and alignment analysis, and the aligned results showed that the DNA sequence changed by deletion, insertion and replacement that occurred at some sites. The results proved the high mutagenic efficacy of space flight, and ways of DNA transformation due to space conditions.

EZH2 silencing by RNA interference inhibits proliferation in bladder cancer cell lines.

The Enhancer of Zeste homologue2 gene (EZH2) is frequently expressed at high levels in malignant tumours, including bladder cancer. It functions as a transcriptional regulator to the maintenance of cell identity, cell cycle regulation and oncogenesis. In the study, we detected EZH2 expression in bladder cancer tissues. These results showed EZH2 high expression in bladder cancer tissue at level of transcript and protein compared with normal bladder tissue and EZH2 expression correlated positively with tumour stage and grade. Then, we used RNA interference to inhibit EZH2 expression in bladder cancer EJ cell line. Efficient downregulation of EZH2 resulted in significantly decreased cell proliferation in EJ cells and retarded transition of G(1) phase to S phase. Our data suggest that EZH2 is involved in the tumourigenesis of bladder cancer and EZH2 downregulation contributes to inhibiting malignant growth by retarding cell entrance to S phase.

An optimized preparation method to obtain high-quality RNA from dry sunflower seeds.

In an attempt to isolate high-quality, intact total RNA from sunflower (Helianthus annuus) seeds for investigation of the molecular mechanisms of mutations, we tested various procedures, using kits, including RNAiso Plus, RNAiso Plus+RNAiso-mate for Plant Tissue, Trizol, and the Qi method, but no high-quality total RNA of high integrity was obtained with any of these methods, probably due to the high content of polyphenols, polysaccharides, and secondary metabolites in mature sunflower seeds. Modifications were made to the Qi method. To avoid polyphenol oxidation, frozen dry seeds free of the seedcase were ground in a mortar with an equal amount of PVP30, and the fine ground powder was transferred to an extraction buffer with 2% PVP30 (w/v), 5% ß-mercaptoethanol (v/v) and LiCl (8 M). A sample homogenate was extracted with chloroform prior to acidic phenol-chloroform extraction. The total RNA was precipitated with 1/4 volume of NaAc and 2 volumes of absolute ethanol to prevent contamination by polysaccharides. The yield of total RNA was 29.95 µg/100 mg husked dry seeds; the ratios of A260/A230 and A260/A280 were 2.44 and 2.09, respectively. Electrophoretic analysis clearly showed 28S and 18S ribosomal RNA bands. Using the extracted RNA, a fragment of the actin gene was successfully expressed by RT-PCR. This modified protocol is suitable for isolating high-quality total RNA from sunflower seeds for molecular research.

Low expression of CircRNA HIPK3 promotes osteoarthritis chondrocyte apoptosis by serving as a sponge of miR-124 to regulate SOX8.

OBJECTIVE: CircRNA, a type of circular RNA, has recently been shown to be a potential target for osteoarthritis (OA). Circular RNA HIPK3 (CircHIPK3) is reported to be abnormally expressed in various disease tissues and affects the occurrence and development of the disease. However, the role and underlying mechanism of CircRNA HIPK3 in osteoarthritis are still unclear. The purpose of this study is to explore the effect of CircRNA HIPK3 on osteoarthritis and analyze its regulatory mechanism. PATIENTS AND METHODS: We took human OA tissues, normal knee cartilage, human OA chondrocytes and normal chondrocytes as the research objects. Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) was used to detect CircHIPK3 expression level, its target gene miRNA 124 (miR-124) and downstream target molecule SOX8. Flow cytometry analysis was applied to discover the apoptosis of CircHIPK3 and miR-124 on OA cartilage in different transfection situations. Moreover, Western blot and RT-qPCR were used to detect the expression of caspase-3 in OA chondrocytes. The binding site of CircRNA HIPK3 and miR-124, miR-124, and SOX8 were verified by using Dual-Luciferase assay. RESULTS: High expressed CircHIPK3 and low expressed miR-124 were found in OA tissues and OA chondrocytes. In addition, the Dual-Luciferase report showed CircHIPK3 acted as a sponge of miR-124 in OA chondrocytes. CircHIPK3 and miR-124 expression in OA tissue were confirmed to be negatively correlated. To our surprise, knocking down CircHIPK3 and transfected miR-124 mimics both inhibited the apoptosis of OA chondrocytes. Further experiments verified that the downstream target molecule of miR-124 was SOX8 in OA chondrocytes. Besides, miR-124 inhibitors reversed the knockdown of CircHIPK3 while si-SOX8 reversed the miR-124 inhibitors effect of apoptosis on OA chondrocytes. CONCLUSIONS: Our results demonstrated that CircHIPK3 was significantly upregulated in OA cartilage tissue and cells. Low expression of CircHIPK3 promoted the apoptosis of OA chondrocytes by promoting miR-124 to suppress SOX8 expression. The molecular mechanism of CircHIPK3 in present study is expected to provide new ideas for the treatment of osteoarthritis.