[Feasibility study on the use of peripheral blood differentially expressed genes for objective classification of chronic subjective tinnitus: a case study on high-frequency tinnitus].

Objective: To explore the feasibility of constructing an objective tinnitus subtype model based on peripheral blood differentially expressed genes (DEGs) using a combination of Weighted Gene Co-expression Network Analysis (WGCNA) and Random Forest algorithm (RF). Methods: From October 2019 to June 2020, peripheral blood DEGs were obtained from 37 patients (from the Third Affiliated Hospital of Sun Yat-sen University)with chronic subjective high-frequency tinnitus (21 unbothersome type, 16 bothersome type) and 20 healthy volunteers through high-throughput sequencing. WGCNA was used to construct gene modules with different expression patterns and analyze their relationships with tinnitus characteristics. Subsequently, RF was employed to build subtype models, which were evaluated by the area under the receiver operating characteristic curve (AUC), accuracy, and F1-score. Results: A total of 12 351 intergroup DEGs were divided into 9 gene modules. Among them, MEblue, MEgreen, and MEbrown showed significant negative correlations with the healthy volunteer group, while MEpink showed a significant positive correlation with the tinnitus distress group. The "Tinnitus vs. Normal" and "Compensatory vs. Decompensatory" subtype models, based on MEblue and MEpink respectively, both had AUCs greater than 0.80, accuracies above 90%, and F1-scores above 0.90, indicating good performance. Conclusions: Peripheral blood DEGs are potential biological indicators for objective classification of subjective tinnitus. The combined application of WGCNA and the Random Forest algorithm should be a viable approach to constructing an objective tinnitus subtype model. However, further exploration and refinement are needed to validate the model's generalizability, cross-dataset performance, and algorithm optimization.

[The value of high-definition intelligent endoscopy combined with stroboscopy in the diagnosis and treatment of vocal cord leukoplakia].

Objective: To evaluate the clinical value of high-definition intelligent endoscopy (iSCAN) combined with stroboscopy in identifying vocal cord leukoplakia. Methods: Seventy-nine patients with vocal cord leukoplakia who underwent CO2 laser laryngeal microsurgery and diagnosed by histopathology were recruited between October 2020 to August 2021. The morphological features, microvascular morphology and mucosal waves were observed by stroboscope; SPSS 20.0 software was used for statistical analysis. Results: There were 79 patients with a total of 119 lesions (56 on left and 63 on right).Pathological examination showed that 51 sides of the vocal cords were malignant lesions (severe dysplasia, carcinoma in situ and invasive carcinoma), and 68 sides were benign lesions.Under stroboscopy, 69 sides of mucosal wave were normal or slightly decreased, and 50 sides were severely decreased or disappeared.The decrease degree of mucosal wave was positively correlated with malignant lesions (ρ=0.687, P<0.001).Under iSCAN endoscopy, there was a positive correlation between the morphological changes of microvessels at the lesion site (vertical) and the malignant lesion (ρ=0.687, P<0.001).Univariate analysis showed that lesion size, thickness, uneven color, granular elevation, peripheral erythema and asymmetry were positively correlated with malignant lesions (ρ=0.530, 0.401, 0.538, 0.315, 0.497, 0.281, P<0.05).Logistic regression analysis showed that the risk of pathological malignancy with large lesions was 5.437 times higher than those of small lesions, the vertical vascular changes under iSCAN were 8.711 times higher than that of normal vascular morphology, and the severe reduction or disappearance of mucosal waves was 9.12 times higher than that of normal or mild reduction of mucosal waves. Conclusion: ISCAN can be combined with staphyloscopy to comprehensively observe and evaluate the changes of vocal cord morphology, submucosal microvessels and mucosal wave of vocal cord in patients with vocal cord leukoplosis, thus improving the ability to distinguish benign and malignant lesions.

[Significance of aspergillus fumigatus-specific IgG in the diagnosis of allergic bronchopulmonary aspergillosis].

The data of 18 patients with allergic bronchopulmonary aspergillosis (ABPA) who received aspergillus fumigatus-specific IgG detection from 2015 to 2021 in Peking University Third Hospital were retrospectively analyzed. Among them, 11 were male and 7 were female, aged 18-79 years. All patients had a history of asthma or symptoms of cough and asthma, and aspergillus fumigatus-specific IgE was positive; 16 patients had total serum IgE>500 U/ml, of which 13 patients had total serum IgE>1 000 U/ml. Among other diagnostic indicators, peripheral blood eosinophils were >0.5×109/L in 16 cases; lung CT showed bronchiectasis in 15 cases; serum aspergillus fumigatus-specific IgG was positive (>120 AU/ml) in 10 cases. There was no significant difference in serum total IgE level, peripheral blood eosinophil count, and bronchiectasis ratio between positive and negative cases of aspergillus fumigatus-specific IgG (all P>0.05). In this study, the positive rate of aspergillus fumigatus-specific IgG in patients with ABPA was more than 50%, which has auxiliary value in the diagnosis of ABPA.

[Analysis of syphilis antibody screening results of outpatients and inpatients in a hospital in Beijing from 2016 to 2020].

Objective: The results of syphilis antibody screening in Peking University Third Hospital from 2016 to 2020 were analyzed,to explore the characteristics of sex, age and distribution of patients with positive syphilis antibody. Methods: A retrospective study was conducted to collect the results of syphilis antibody in outpatients and inpatients of Peking University Third Hospital from 2016 to 2020. Syphilis antibodies were screened in 626 528 patients aged 1-98 years, 4 232 were retested positive by TPPA test, including 2 132 males (50.4%) and 2 100 females (49.6%). Chemiluminescence immunoassay (CMIA) was used for syphilis antibody screening, and Treponema pallidum particle agglutination (TPPA) test was used for reexamination. SPSS20.0 data statistical analysis software was used to analyze the detection rate, age, sex, department and clinical diagnosis of patients with positive syphilis antibody by χ² test. Results: Among 626 528 patients who were positive for treponema pallidum antibody screening, 4 232 were retested positive by TPPA test, accounting for 0.68% of the total number of patients tested. The number of syphilis tests increased year by year, however the positive detection rate decreased. The positive detection rate of syphilis antibody in 2020 decreased by 18.9% compared with 2016. The positive rate of syphilis antibody in male patients was higher than that in female patients, accounting for 0.80% and 0.59% of the total number of patients tested respectively. The positive rate of syphilis antibody of different genders increased with age, the total positive rate of 0-20, 21-40, 41-60, 61-80,>80 years old were 0.15%, 0.45%, 0.95%, 1.07% and 1.41%, respectively. While the increase rate of males was higher than that of females. The positive rate of male over 80 years old was 18.13 times of the group of 0-20 years old, and 5.54 times in women. The top 6 departments with positive syphilis antibody detection rate were emergency department, oncology department, respiratory department, geriatrics department, endocrinology department and neurology department, and the positive rates were 1.79% (104/5 810),1.46% (55/3 767),1.20% (74/6 167),1.20% (22/1 833),1.10% (32/2 909),1.09% (94/8 624), respectively. From the analysis of clinical diagnosis, the proportion of positive syphilis antibody in infertile patients (0.64%, 672/104 911) was higher than that in naturally conceived patients (0.10%, 24/23 969). Conclusions: From 2016 to 2020, the positive detection rate of syphilis antibody in Peking University Third Hospital decreased year by year. However, the positive detection rate increased with the age of patients. The positive rate of syphilis antibody in male was higher than that in female. The positive rate of syphilis antibody in pregnant women was lower than that in infertile patients.

MicroRNA-4443 regulates monocyte activation by targeting tumor necrosis factor receptor associated factor 4 in stroke-induced immunosuppression.

BACKGROUND AND PURPOSE: MicroRNAs (miRNAs) have been demonstrated to play crucial roles in the early stage of acute ischaemic stroke (AIS). The purpose of this study was to investigate the expression patterns of miRNAs in peripheral blood mononuclear cells (PBMCs) from AIS patients and further explore related molecular mechanisms in stroke-induced immunodeficiency syndrome (SIDS). METHODS: The miRNA expression patterns of PBMCs were detected by miRNA microarray and validated by quantitative real-time polymerase chain reaction (qRT-PCR) in AIS patients and healthy controls. Bioinformatics methods and luciferase reporter assays were used to detect the downstream target genes. Following stimulation with lipopolysaccharide and interleukin-4, the expression of miR-4443, tumor necrosis factor receptor associated factor 4 (TRAF4) and the nuclear factor kappa B (NF-κB) pathway were evaluated. Furthermore, transfection with miR-4443 mimic or inhibitor in the monocytes was carried out to gain insight into the mechanisms in SIDS. RESULTS: Interleukin-10 in AIS patients was significantly higher than that of healthy controls. The miRNA microarray analysis and qRTPCR validation showed that only miR-4443 was upregulated expressed in PBMCs from AIS patients, especially in monocytes. miR-4443 was shown to directly interact with the 3' untranslated regions of TRAF4, resulting in suppression of TRAF4 protein expression. Furthermore, the expression of miR-4443 and TRAF4 was regulated by stimulation with lipopolysaccharide or interleukin-4. Additionally, overexpression of miR-4443 suppressed the TRAF4/Iκα/NF-κB signaling pathway to activate the expression of anti-inflammatory cytokines in monocytes. CONCLUSIONS: The increased expression of miR-4443 induced monocyte dysfunction by targeting TRAF4, which may function as a crucial mediator in SIDS.

Regulatory role of B-cell maturation antigen on the toxic effect of chromium ions on human SaOS-2 osteoblasts.

Metal prostheses of artificial joints undergo wear, producing numerous metal particles and ions, such as Cr3+ . Cr3+ is considered a key factor leading to aseptic loosening. Many studies focus on the effect of Cr3+ on osteoblasts; however, little is known about the effect of Cr3+ on the B-cell maturation antigen (BCMA) in the osteoblasts. In this study, we first demonstrated the BCMA expressed in human SaOS-2 osteoblasts through reverse transcriptase-PCR, Western blot, and immunocytochemical analyses. Cr3+ decreased alkaline phosphatase (ALP), osteocalcin (OC), cell mineralization, and collagen type I mRNA and protein expression. Moreover, Cr3+ has an inhibitive effect on the expression of the BCMA in human SaOS-2 osteoblasts. However, after we upregulated the expression of the BCMA, ALP, OC, cell mineralization, and collagen type I mRNA and protein expression were increased. Overall, this study demonstrates that the BCMA is involved in human SaOS-2 osteoblast osteogenetic metabolism and plays a regulatory role on the toxic effect of chromium ions on human SaOS-2 osteoblasts.

Prognostic Value of Levels of Urine Neutrophil Gelatinase-associated Lipocalin and Interleukin-18 in Patients With Delayed Graft Function After Kidney Transplantation.

OBJECTIVE: The aim of our study was to examine how serial urine neutrophil gelatinase-associated lipocalin (uNGAL) and interleukin (IL)-18 concentrations change over time after kidney transplantation and whether we can use them to predict delayed graft function (DGF). METHODS: Spot urine samples for the NGAL and IL-18 tests were taken at 4, 12, 24, 48, and 72 hours after transplantation from every patient at hospital presentation. Urine samples were tested for NGAL by a chemiluminescence assay kit on the ARCHITECT I2000 immunology analyzer. IL-18 were measured by a quantitative immunoenzymatic assay kit. Serum samples for the creatinine measurement were taken at 24, 48, and 72 hours after kidney transplantation. Serum samples were tested for creatinine on the Olympus analyzer 5821 by alkaline picric acid method. The patients were divided into 2 groups: DGF group and non-DGF group. RESULTS: The urine NGAL levels were increased in DGF group at all points over the follow-up period. There are differences (P < .05) in NGAL concentrations between DGF (n = 21) and non-DGF groups (n = 102). However, urine samples from the DGF group (n = 21) had increased IL-18 concentrations at 4, 12, 24, and 48 hours postoperatively compared with non-DGF group samples (n = 102) (P < .05). There were obvious distinctions (P < .05) of serum creatinine (SCr) levels in 24 hours between the DGF (n = 21) and non-DGF groups (n = 102). The specificity and positive predictive value of NGAL in the DGF diagnosis increased with time, but the sensitivity and negative predictive value do not change. The specificity, sensitivity, positive predictive, value and negative predictive value of IL-18 in the DGF diagnosis changed irregularly at multiple time points after transplant. The positive predictive value and negative predictive value of 24-hour SCr were 47.4% and 95.7%, respectively. The positive predictive value and negative predictive value of combination of NGAL, IL-18, and SCr (area under the receiver-operating characteristic curve = 0.984; 95% CI, 0.887-0.994) were 90.9% and 100%, respectively. Overall, the combination of NGAL, IL-18, and SCr was found to have a significantly better positive predictive value than all the other combination assays (P < .05). In addition, there were obvious distinction of the negative predictive value of NGAL and IL-18 combination compared with those of other combinations (P < .05). CONCLUSIONS: The combination of NGAL, IL-18, and SCr measurements after initiation of treatment may be highly effective for risk stratification in patients with DGF. The combination may be useful to cover the complete diagnostic window of patients presenting with DGF.

Bone Morphogenetic Protein-2 regulates in vitro osteogenic differentiation of mouse adipose derived stem cells.

OBJECTIVE: The aim of this study is to investigate the feasibility and efficiency of Bone Morphogenetic Protein-2 (BMP-2) in regulating in vitro osteogenic differentiation of mouse adipose derived stem cells (ADSCs). MATERIALS AND METHODS: Mouse ADSCs were isolated from adipose tissues of C57/BL6 mice (age of 4-6 w) and cultured. Surface antigens of passage 3 (P3) ADSCs, including CD31, CD34, CD90, CD105 and CD133, were analyzed using flow cytometry. Overexpression of BMP-2 was achieved through gene transfection of ADSCs. In vitro osteogenic differentiation of transfected and non-transfected ADSCs cultured in specific induction media was evaluated by Alizarin Red staining. In addition, expression of osteoblast-specific gene, Runx2, was analyzed by quantitative RT-PCR (qRT-PCR). RESULTS: Abundant ADSCs could be isolated from adipose tissue. P3 ADSCs expressed stem cell-specific molecular markers, CD90 and CD105 but did not express CD31, CD34 or CD133. BMP-2 could efficiently transfect mouse ADSCs. Alizarin Red staining revealed that more calcified nodules were formed in BMP-2 transfected ADSCs. qRT-PCR further confirmed higher level of Runx2 expression in BMP-2 transfected ADSCs (p < 0.05). CONCLUSIONS: BMP-2 can promote in vitro osteogenic differentiation of mouse adipose stem cells.

Direct effects of IL-4 on mast cells drive their intestinal expansion and increase susceptibility to anaphylaxis in a murine model of food allergy.

Interleukin (IL)-4 has critical roles in allergic disorders, including food hypersensitivity. The direct effects of the cytokine on the survival and function of mast cells, the key effectors of food anaphylaxis, have not been established. In this study, we demonstrate that IL-4 induces a marked intestinal mastocytosis in mice. This phenotype is reproduced in animals expressing Il4rαF709, an activating variant of the IL-4 receptor α-chain (IL-4Rα). Il4rαF709 mice exhibit enhanced anaphylactic reactions but unaltered physiological responses to vasoactive mediators. IL-4 induces Bcl-2 and Bcl-X(L) and enhances survival and stimulates proliferation in cultured bone marrow-derived mast cells (BMMC). These effects are STAT6 (signal transducer and activator of transcription factor 6)-dependent and are amplified in Il4rαF709 BMMC. In competitive bone marrow chimeras, Il4rαF709 mast cells display a substantial competitive advantage over wild-type mast cells, which, in turn, prevail over IL-4Rα⁻/⁻ mast cells in populating the intestine, establishing a cell-intrinsic effect of IL-4 in intestinal mast cell homeostasis. Our results demonstrate that IL-4-signaling is a key determinant of mast cell expansion in food allergy.

Immunostimulatory probiotic Lactobacillus rhamnosus HN001 and Bifidobacterium lactis HN019 do not induce pathological inflammation in mouse model of experimental autoimmune thyroiditis.

The possibility that intestinal microflora contribute to the pathogenesis of autoimmune diseases has raised issues regarding the safety of probiotic organisms, especially those with immunostimulating properties, in individuals with such immune dysfunctions. In this study, the effect of consumption of probiotic lactic acid bacteria (LAB) strains Lactobacillus rhamnosus HN001(HN001) and Bifidobacterium lactis HN019 (HN019) on the induction and progression of experimental autoimmune thyroiditis (EAT) was investigated in CBA/CaH (H-2k) mice. HN001 or HN019 in skim milk were fed to mice daily (1-1.5 x 10(8) cfu/mouse/day) for 5 to 9 weeks. A mild form of EAT was induced by subcutaneous injection of mouse thyroglobulin (MTg) with either Freund's adjuvant (complete and incomplete, CFA and IFA) or lipopolysaccharide (LPS). The proliferative responses of spleen lymphocyte to MTg stimulation in vitro and the presence (and degree) of mononuclear cell infiltration in thyroid gland tissues were examined to assess the development and severity of EAT. The levels of serum anti-MTg antibodies (IgG1 and IgG2a) and spleen weight index were determined to detect the presence of autoimmune responses of mice receiving MTg. Results showed that 8 weeks after immunization, 16.67-50% of the mice developed mild EAT with lymphocyte infiltration in the thyroid glands. Probiotic feeding did not induce full-blown EAT. There were no differences in spleen weight index or the proliferative spleenocytes in response to PMA between mice that received MTg alone and mice that received MTg and probiotic LAB strains.

[Repair of bone defect with benign tumor by embryonic bone transplantation].

OBJECTIVE: To evaluate the clinical results of repair of bone defect by embryonic bone transplantation. METHODS: From January 1994 to June 1999, 148 cases of bone defect were repaired by embryonic bone transplantation following alcohol treatment, there were 63 cases with bone cyst, 42 cases with fibrous dysplasia of bone, 26 cases with giant cell tumor of bone, and 17 cases with enchondroma among them. The maximal bone defect was 3.5 cm x 10.0 cm, while the minimal defect was 0.5 cm x 1.0 cm. RESULTS: All of those bone defect with benign tumor were bone union used by embryonic bone transplantation after 3 months to 1 year of operation, the average healing course was 6.2 months, followed up 1 to 6 years, averaged 14 months, no tumor recurrence and no obvious local or system response were observed. CONCLUSION: Embryonic bone can be used as a good repairing material of postoperative bone defect of benign tumors, the clinical results are satisfactory.

[Relationship between exposure time and cryopreservation of articular cartilage].

OBJECTIVE: To search an optimal method for improving viability of cryopreserved articular cartilage. METHODS: Articular cartilage which was sampled from the rabbits were randomly divided into 5 groups. Fresh cartilage was group I, other groups were frozen. Before frozen, other cartilage was exposured in 10% DMSO at 4 degrees C for 30 minutes(group II), 1 hour(group III), 2 hours (group IV), 4 hours(group V), then were stored in liquid nitrogen for 1 week. Viabilities of the chondrocytes were detected by Typan-blue staining, electron transmission microscope, and determination of incorporation 3H-TdR after the temperature returned to normal. RESULTS: 1. The cells were injuried at different extent after the cartilage was frozen. In group I, survival rate of cells was 96% and incorporation of 3H-TdR was (4,953.13 +/- 583.27)%, statistic difference was significant between group I and other groups(P < 0.01). The microstructure of group I was normal while other groups all had damage of the organella, 2. Structures and functions of cells in group IV were best among frozen groups. Organella were less damaged than group II, III, V, survival rate of cells was 56% and incorporation of 3H-TdR was (1,139.88 +/- 146.39)%, statistic difference was significant between group IV and group II, III, V(P < 0.01). CONCLUSION: If cartilage are exposured in 10% DMSO at 4 degrees C for 2 hours before frozen, optimal cryopreservation can be achieved.

Cytochrome c activation of CPP32-like proteolysis plays a critical role in a Xenopus cell-free apoptosis system.

In a cell-free system based on Xenopus egg extracts, Bcl-2 blocks apoptotic activity by preventing cytochrome c release from mitochondria. We now describe in detail the crucial role of cytochrome c in this system. The mitochondrial fraction, when incubated with cytosol, releases cytochrome c. Cytochrome c in turn induces the activation of protease(s) resembling caspase-3 (CPP32), leading to downstream apoptotic events, including the cleavage of fodrin and lamin B1. CPP32-like protease activity plays an essential role in this system, as the caspase inhibitor, Ac-DEVD-CHO, strongly inhibited fodrin and lamin B1 cleavage, as well as nuclear morphology changes. Cytochrome c preparations from various vertebrate species, but not from Saccharomyces cerevisiae, were able to initiate all signs of apoptosis. Cytochrome c by itself was unable to process the precursor form of CPP32; the presence of cytosol was required. The electron transport activity of cytochrome c is not required for its pro-apoptotic function, as Cu- and Zn-substituted cytochrome c had strong pro-apoptotic activity, despite being redox-inactive. However, certain structural features of the molecule were required for this activity. Thus, in the Xenopus cell-free system, cytosol-dependent mitochondrial release of cytochrome c induces apoptosis by activating CPP32-like caspases, via unknown cytosolic factors.