OBJECTIVE: To elucidate the role and underlying mechanism of ubiquitin-specific protease 35 (USP35) in ferroptosis of rheumatoid arthritis-fibroblast like synoviocytes (RA-FLS), thereby enhancing our comprehension of the pathogenesis of RA and identifying potential therapeutic targets for its treatment. METHODS: (1) RA-FLS were cultured in vitro and transduced with lentiviral vectors to establish stable cell lines: A USP35-knockdown line (short hairpin ribonucleic acid of USP35, shUSP35) and its control (negtive control of short hairpin ribonucleic acid, shNC), as well as a overexpression of USP35 line (USP35 OE) and its control (Vector). To investigate the role of USP35 in ferroptosis regulation, a ferroptosis model was induced in RA-FLS by treatment with 1 μmol/L Erastin. The cells were divided into six groups: shNC, shNC + Erastin, shUSP35 + Erastin, Vector, Vector + Erastin, and USP35 OE + Erastin. (2) Cell viability was detected using the cell counting kit-8 (CCK-8). (3) Reactive oxygen species (ROS), malondialdehyde (MDA), glutathione/glutathione disulfide (GSH/GSSG) ratios, and Ferrous ion (Fe²⁺) levels were measured using specific assay kits to evaluate oxidative stress, lipid peroxidation, and glutathione redox status in the cells. (4) Protein expression levels of solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) were detected using Western blotting to investigate their potential involvement in USP35-mediated ferroptosis regulation. RESULTS: (1) Compared with the shNC +Erastin group, the cell viability of the shUSP35+Erastin group was significantly decreased (P < 0.001), while it was notably increased in the USP35 OE+Erastin group compared with the Vector+Erastin group (P < 0.001). These findings indicated that USP35 could alleviate the inhibitory effect of Erastin on RA-FLS cell viability. (2) In comparison to the shNC+Erastin group, the levels of ROS (P < 0.001), MDA (P < 0.05), and Fe²⁺ (P < 0.001) were significantly elevated, and the GSH/GSSG ratio was increased (P < 0.05) in the shUSP35+Erastin group. Conversely, the levels of ROS (P < 0.001), MDA (P < 0.05), and Fe²⁺ (P < 0.05) were significantly decreased, and the GSH/GSSG ratio was decreased (P < 0.05) in the USP35 OE+Erastin group compared with the Vector+Erastin group. These results suggested that USP35 could inhibit Erastin-induced oxidative stress and lipid peroxidation in RA-FLS. (3) In Erastin-induced RA-FLS, the expression of USP35 was positively correlated with the protein levels of SLC7A11 and GPX4, indicating a potential mechanism by which USP35 regulated ferroptosis in these cells. CONCLUSION USP35 inhibits ferroptosis in RA-FLS, potentially through the increased expression of SLC7A11 and GPX4.
Ferroptosis ; Humans ; Arthritis, Rheumatoid/metabolism* ; Synoviocytes/pathology* ; Reactive Oxygen Species/metabolism* ; Ubiquitin-Specific Proteases/metabolism* ; Fibroblasts/pathology* ; Cell Survival ; Piperazines/pharmacology* ; Endopeptidases/metabolism* ; Cells, Cultured ; Cell Line ; Amino Acid Transport System y+

To construct recombinant eukaryotic expression plasmid vector of human IL-34 gene, and to study the effects of IL-34 expressed by human bone marrow-derived mesenchymal stem cells (hBM-MSCs) on THP-1 cells. Full-length IL-34 encoding sequence was amplified by PCR. And this fragment was cloned into the plasmid pIRES2-EGFP. Western blotting and ELISA were used to analyze the expression of IL-34 in hBM-MSCs. THP-1 cells were cultured with hBM-MSCs medium containing IL-34 protein. Real-time PCR detected the effects of IL-34 on the expression of IL-10 and TNFα in THP-1 cells. Restrictive enzyme analysis and sequencing demonstrated that IL-34 eukaryotic expression vector was successfully constructed. IL-34 protein expressed by hBM-MSCs could promote IL-10 and TNFα expression in THP-1 cells. Those results show that IL-34 expressed by hBM-MSCs has regulating effect on THP-1 cells.

Objective To investigate the antitumor effects and the possible mechanisms of dendrit-ic cells co-cultured with cytokine-induced killer cells(DC-CIK)in combination with sorafenib on two lung adenocarcinoma cell lines,A549 cells(harboring KRAS gene mutation)and PC-9 cells(harboring EGFR gene mutation). Methods DC and CIK cells were routinely generated in vitro by stimulating PBMCs isola-ted from lung cancer patients with different cytokines and then co-cultured after a week of culturing. Flow cy-tometry analysis(FCM)was used to analyze the phenotype of DC-CIK cells after 7 days of co-culturing. The 50% inhibitory concentration(IC50 )of sorafenib against tumor cells was detected by MTT assay. The tumor cells were treated with DC-CIK cells alone or in combination with sorafenib. The proliferation of tumor cells was tested by CCK-8 kit and dynamically monitored by real-time cellular analysis(RTCA). Annexin-V/ PI staining was used to examine the apoptosis rates in each group. Real-time fluorescent quantitative PCR and FCM were respectively performed to detect the expression of natural killer group 2 member D ligands (NKG2DLs)at mRNA and protein levels after the treatment with sorafenib for 24 h. Results There was no significant difference between the IC50 of sorafenib against A549 and PC-9 cells after a 24-hour exposure(P﹥0. 05). Compared with the DC-CIK biotherapy,treating the tumor cells with DC-CIK cells in combination with sorafenib significantly inhibited the cell proliferation and increased the total apoptosis rates of tumor cells(P﹤0. 05). Moreover,the inhibition rates to tumor cell proliferation were enhanced along with the in-crease of effect-to-target ratio(E/ T). Compared with the single-factor treatment groups,the normalized cell index(NCI)in the combined treatment group was significantly decreased. Blocking NKG2D could abate the inhibitory effect of DC-CIK cells on tumor cell proliferation(P﹤0. 05). The expression of NKG2DLs(inclu-ding ULBP1,UBLP2 and ULBP3)on tumor cells at mRNA and protein levels were increased to different ex-tent after treating with 5 μmol/ L of sorafenib for 24 h. Conclusion There was no significant different be-tween the inhibitory effects of sorafenib on the proliferation of lung adenocarcinoma cancer cells harboring KRAS or EGFR gene mutation. The antitumor effects of DC-CIK cells combined with sorafenib on lung ade-nocarcinoma cells might be induced by regulating the NKG2D-NKG2DLs pathway and enhancing apoptosis. Moreover,the antitumor effects of the combined treatment were better than those of single-factor treatments.

OBJECTIVE: Aimed to analyse the clinical features of the patients with sphenoid sinus mucocele, achieve earlier diagnosis and more timely intervention and decrease the occurrence of misdiagnoses. METHOD: A retrospective study was first conducted in patients with sphenoid sinus mucoele treated in Xiangya hospital from Jan 2000 to Jan 2015. Then literature reports on this disease were collected and analyzed from China National Knowledge Infrastructure (CNKI) and Wan Fang database. RESULT: We collected 82 patients with sphenoid sinus mucocele treated in Xiangya hospital. There were 52 patients presented with headache, 31 patients presented with visual impairment, 10 patients presented with cranial nerve palsy, 2 patients presented with exophthalmos, 15 patients presented with nasal symptoms, and 5 patients with no obvious symptoms. There was no significant difference for symptoms distribution between male and female patients (P > 0.05). Among 45 patients with headache as first symptom and 10 patients with ethmoid sinus mucocele, there were 18 patients and 8 patients subsequently suffering from visual impairment, respectively. We also collected 161 patients in literature except for enrolling, the 82 patients treated in Xiangya hospital, and found that headache was the most common symptom, followed by visual impairment, in the two independent cohorts. CONCLUSION To the best of our knowledge, this is the study of maximum sample for sphenoid sinus mucocele in China. Headache and visual impairment are the most common symptoms for sphenoid sinus mucocele. Surgical treatment should be early performed when the desease accompanied with headache or ethmoid sinus mucocele, to avoid other complications such as visual impairment and even blindness.
China ; Cranial Nerve Diseases ; etiology ; Databases, Factual ; Diagnostic Errors ; Ethmoid Sinus ; Exophthalmos ; etiology ; Female ; Headache ; etiology ; Humans ; Male ; Mucocele ; complications ; diagnosis ; pathology ; Paranasal Sinus Diseases ; Retrospective Studies ; Sphenoid Sinus ; pathology ; Vision Disorders ; etiology

Objective:To retrospectively analyze the treatment effect and the patterns of failure associated with different clinical target volume on patients with esophageal cancer treated with three dimensional conformal or intensity modulated radiotherapy, and to determine whether involved field radiotherapy is practicable in these patients. Methods:A total of 68 patients with esophageal squa-mous cell carcinoma between January 2007 to June 2011 in our hospital underwent three dimensional conformal or intensity modulated radiotherapy, according to the CTV range is divided into lymph involved-field group (involved field group) and lymph extended field group (extended field group). Results:In Involved field group and expand field group the survival rate of 1, 2 years were 59%, 41%and 61%, 39% respectively (P=0.56), and local control rates were 66%, 48% and 68%, 49% respectively(P=0.78). The total failure rates of involved field and the expand field were 63%and 66%(P=0.89). The local failure rate was 53%and 59%, distant metastasis failure rates were 47%and 44%, the regional failure rates were 11.8%and 7.5%in Involved field and the expand field, there were no difference in Statistics (P=0.39). The lung V10, V20, V30 and mean lung dose of extended field group were greater than that of the in-volved field group, while the mean lung dose and V10 has statistical difference. Conclusion:The involved field group was similar as the extended field group in the survival rate and local control rate, the regional recurrence and distant metastasis are the main cause of treatment failure, so the involved field radiotherapy is feasible for locally advanced esophageal carcinoma.

OBJECTIVE: To investigate the anatomical influence of the hypertrophic inferior turbinate on computational fluid dynamics (CFD) model of unilateral hypertrophic inferior turbinate nasal cavity, and to analyze the bilateral detailed nasal airflow simulations under both inspiratory and expiratory phases in CFD model. METHOD: One male volunteer troubled with unilateral hypertrophic inferior turbinate accepted CT scan. CFD model was built by CT scans through Simplant 10.0 and ANSYS ICEM. Fluent 6.3.26 simulated the airflow of both nasal cavity in breathing rates 200 ml/s. RESULT: 1) In infraturbinal region, the cross-section area (CSA) of the nasal cavity with hypertrophic inferior turbinate was smaller than that in healthy side and the average area difference between two sides was 1.62 cm2. 2) In both inspiration and expiration phases, the hypertrophic infraturbinal produced a markable reduction in intranasal pressures drop along the full length of the infraturbinal region. The volumetric flow rate in the hypertrophic infraturbinal side was 50 ml/s, which equalled to one third of that in healthy side; Mean air speed in the anterior valve region was estimated to be 0.57 m/s at hypertrophic infraturbinal side and 1.83 m/s at healthy side during inspiration; More vortices happened in the hypertrophic infraturbinal side. CONCLUSION The unilateral hypertrophic infraturbinal change the normal anatomy and influence the aerodynamic of nasal cavity, which is harmful to the functions of human nasal in ventilation, temperature accommodation and olfactory sensation.
Adult ; Computer Simulation ; Humans ; Hydrodynamics ; Hypertrophy ; physiopathology ; Male ; Models, Anatomic ; Nasal Cavity ; physiology ; physiopathology ; Nasal Obstruction ; physiopathology ; Tomography, X-Ray Computed ; Turbinates ; physiology ; physiopathology

0 05) CONCLUSION:The clinical efficacy and adverse reaction of domestic cetirizine tablet are similar to that of imported one in treatment of PAR