Background/Aims: Ubiquitination is widely involved in the progression of hepatocellular carcinoma (HCC) by regulating various cellular processes. However, systematic strategies for screening core ubiquitin-related genes, clarifying their functions and mechanisms, and ultimately developing potential therapeutics for patients with HCC are still lacking. Methods: Cox and LASSO regression analyses were performed to construct a ubiquitin-related gene prediction model for HCC. Loss- and gain-of-function studies, transcriptomic and metabolomics analysis were used to explore the function and mechanism of UBE2S on HCC cell glycolysis and growth. Results: Based on 1,423 ubiquitin-related genes, a four-gene signature was successfully constructed to evaluate the prognosis of patients with HCC. UBE2S was identified in this signature with the potential to predict the survival of patients with HCC. E2F2 transcriptionally upregulated UBE2S expression by directly binding to its promoter. UBE2S positively regulated glycolysis in a HIF-1α-dependent manner, thus promoting the proliferation of HCC cells. Mechanistically, UBE2S enhanced K11-linkage polyubiquitination at lysine residues 171 and 196 of VHL independent of E3 ligase, thereby indirectly stabilizing HIF-1α protein levels by mediating the degradation of VHL by the proteasome. In particular, the combination of cephalomannine, a small molecule compound that inhibits the expression of UBE2S, and PX-478, an inhibitor of HIF-1α, significantly improved the anti-tumor efficacy. Conclusions UBE2S is identified as a key biomarker in HCC among the thousands of ubiquitin-related genes and promotes glycolysis by E3 enzyme-independent ubiquitination, thus serving as a therapeutic target for the treatment of HCC.

Humans ; COVID-19 ; Consensus ; SARS-CoV-2 ; China

Lung cancer is the most common incident cancer and the leading cause of cancer death. In recent years, the development of tumor immunotherapy especially chimeric antigen receptor T (CAR-T) cell has shown a promising future. Epidermal growth factor receptor variant III (EGFRvIII) is a tumor-specific mutation expressed in various types of tumors and has been detected in non-small cell lung cancer with a mutation rate of 10%. Thus, EGFRvIII is a potential antigen for targeted lung cancer therapy. In this study, CAR vectors were constructed and transfected into virus-packaging cells. Then, activated T cells were infected with retrovirus harvested from stable virus-producing single clone cell lines. CAR expression on the surfaces of the T cells was detected by flow cytometry and Western blot. The function of CAR-T targeting EGFRvIII was then evaluated. The EGFRvIII-CAR vector was successfully constructed and confirmed by DNA sequencing. A stable virus-producing cell line was produced from a single clone by limited dilution. The culture conditions for the cell line, including cell density, temperature, and culture medium were optimized. After infection with retrovirus, CAR was expressed on more than 90% of the T cells. The proliferation of CAR-T cells were induced by cytokine and specific antigen in vitro. More importantly, EGFRvIII-CART specifically and efficiently recognized and killed A549-EGFRvIII cells with an effector/target ratio of 10:1 by expressing and releasing cytokines, including perforin, granzyme B, IFN-γ, and TNF-α. The in vivo study indicated that the metastasis of A549-EGFRvIII cells in mice were inhibited by EGFRvIII-CART cells, and the survival of the mice was significantly prolonged with no serious side effects. EGFRvIII-CART showed significantly efficient antitumor activity against lung cancer cells expressing EGFRvIII in vivo and in vitro. Therefore, CAR-T targeting EGFRvIII is a potential therapeutic strategy in preventing recurrence and metastasis of lung cancer after surgery.
Animals ; Carcinoma, Non-Small-Cell Lung ; immunology ; therapy ; Cell Line, Tumor ; ErbB Receptors ; immunology ; metabolism ; Female ; Humans ; Immunotherapy, Adoptive ; methods ; Lung Neoplasms ; immunology ; therapy ; Mice ; Mice, Inbred NOD ; Receptors, Chimeric Antigen ; immunology ; T-Lymphocytes ; immunology ; Xenograft Model Antitumor Assays

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. This malignancy is associated with poor prognosis and high mortality. Novel approaches for prolonging the overall survival of patients with advanced HCC are urgently needed. The antitumor activities of adoptive cell transfer therapy (ACT), such as strategies based on tumor-infiltrating lymphocytes and cytokine-induced killer cells, are more effective than those of traditional strategies. Currently, chimeric antigen receptor T-cell (CAR-T) immunotherapy has achieved numerous breakthroughs in the treatment of hematological malignancies, including relapsed or refractory lymphoblastic leukemia and refractory large B-cell lymphoma. Nevertheless, this approach only provides a modest benefit in the treatment of solid tumors. The clinical results of CAR-T immunotherapy for HCC that could be obtained at present are limited. Some published studies have demonstrated that CAR-T could inhibit tumor growth and cause severe side effects. In this review, we summarized the current application of ACT, the challenges encountered by CAR-T technology in HCC treatment, and some possible strategies for the future direction of immunotherapeutic research.
Adoptive Transfer ; methods ; Carcinoma, Hepatocellular ; immunology ; therapy ; Humans ; Immunotherapy, Adoptive ; methods ; Liver Neoplasms ; immunology ; therapy ; Lymphocytes, Tumor-Infiltrating ; cytology ; Randomized Controlled Trials as Topic ; Receptors, Chimeric Antigen ; T-Lymphocytes ; cytology

Objective To explore whether Rho kinase inhibitor protects endotoxemia mice from kidney injury,and to investigate the mechanism underlying this effect. Methods Adult male C57BL/6 mice were randomly divided into three groups (n = 8 for each group): control,lipopolysaccharide (LPS),and LPS+ Y-27632 (Rho kinase inhibitor). For induction of acute kidney injury,mice were administered 30 mg/kg LPS intraperitoneally. Y-27632 (10 mg/kg body weight) was injected intraperitoneally 18 h and 1 h before injection of LPS,and an equal volume of sterile saline was administered at the corresponding time point in each group. The mice were killed 8 h after LPS administration. Blood samples and kidney tissues were taken and preserved for subsequent analysis. Results Pretreatment with Y-27632 significantly attenuated LPS-induced kidney injury;pretreatment with Y-27632 markedly reduced renal expression of inflammatory cytokines (TNF-α and IL-1β) in endotoxemia mouse,and also significantly inhibited LPS-induced caspase-3 expression in the kidney; and Y-27632 pretreatment dramatically reduced TLR4 protein expression and NF-κBp65 phosphorylation in kidney tissues of endotoxemia mouse. Conclusion Rho kinase inhibitor may inhibit TLR4 and NF-κB signaling pathway to reduce the inflammatory response in the kidneys of endotoxemia mice and alleviate acute renal injury induced by LPS.

Objective To retrospectively analysis the clinical effect of temporary hemiepiphysiodesis for treatment of genu varus and valgus with pathologic physis in children.Methods All of 31 children (52 knees) were included in the study from January 2008 to December 2014,20 boys and 11 girls,the age at the time of surgery from 2 year 4 month to 13 year 4 month,mean 6 year 5 month.12 varus and 19 valgus,10 unilateral and 21 bilateral,41 femurs and 44 tibias.The Mechanichal Axis Deviation (MAD) was evaluated and the mechanical lateral distal femur angle (mLDFA) and medial proximal tibia angle (mMPTA) pre-operation and post-surgery was measured.Judging the efficacy with zone system for assessing mechanical axis based on Stevens,we think the results was satisfactory if the mechanical axis falls in zone ± 1 when removed the eightplate and other was unsatisfactory.We made statistical analysis of children who were corrected satisfactory and observed the change of the width and appearance of the physis.We also made the univariate analysis and Logistical multivariate regression analysis about sex,unilateral or bilateral,surgery age,femur or tibia,varus or valgus and severity of deformity to judge the influencing factors between satisfactory and unsatisfactory.We recorded the knee function and deformity recurrence at last follow up.Results The follow-up period was 2 year 6 month to 9 year 5 month,mean 4 year 1 month,44 knees (84.6％) were corrected satisfactory.The mean value of MAD at pre-surgery and at the time when the plate was removed were (-33.3t7.2) mm and (2.1 ±4.3) mm,mean mLDFA were 102.9°±4.9° and 85.3°±3.8°,mean mMPTA were 81.2°±3.4° and 90.5° ±4.4° in genu varus (21 knees);The meam value of MAD at pre-surgery and at the time when the plate was removed were 29.3±6.8 mm and-4.1±6.5 mm,and the mean mLDFA were 79.5o±5.7° and 88.1°±3.5°,mean mMPTA was 97.0°±4.3° and 87.1°±5.2° in genu valgus (23 knees),which were significant difference.There were 28 knees with physis width asymmetry in the 44 knees pre-operation and improved obviously after-surgery and the appearance became normally.8 knees corrected unsatisfactory included 2 varus and 6 valgus.The severity of deformity was a risk factor that affects efficacy with univariate analysis(t=5.124,P=0.000).Regarding the range of the age,we also did the logistic multivariate regression analysis with results showing that surgery age (OR=1.032) and se verity of deformity (OR=1.335) were related to the outcome.There were 8 knees in the 44 knees which had deformity recurrence and the rate was 19.5％ (8/41,3 knees excluded due to doing the limb lengthening surgery when removing the plate) at last follow up,and the knees function were normal.Conclusion Temporary hemiepiphysiodesis is a minimally invasive and effective method for treatment of genu varus or valgus with pathologic physis in children and could improve the appearance of the physis.The age at surgery and severity of deformity are the risk factors that affect efficacy.The pathological changes persisted could cause complication and deformity recurrence which need follow-up after removal of the plate.

Objective To explore the effect of azelastine hydrochloride nasal spray combined with desloratadine to inflammatory factors, cell function and IgE of patients with allergic rhinitis.Methods 92 cases of allergic rhinitis patients treated in the first affiliated hospital of wenzhou medical college hospital from June 2014 to December 2015 were divided into experimental group(n=46) and control group(n=46) according to the random number table method.The control group was given oral loratadine tablets, one piece per time, one time per day, while the experimental group was given azelastine hydrochloride nasal spray on the basis of the control group,each nostril one spray, one time in the morning and night.The clinical efficacy of two groups of patients would be observed after 4 weeks,ELISA would be used to detect serum levels of IFN-γ, IL-4, IL-8 and IgE level, and IFN-γ/IL-4 was the value of Thl/Th2,flow cytometry instrument was used to the determination of T cell subgroup CD4 +,CD8 + cells.Results 4 weeks after treatment,stuffy nose, nasal itching, runny nose, sneezing and nasal cavity change points are lower than before the treatment in both the two groups.Experimental group obviously lower than the control group, the difference was statistically significant ( P <0.05 ).The total effective rate of treatment group is higher than the control group,the difference was statistically significant(χ2 =4.389,P=0.036).The serum level of IFN-γis higher than before treatment in both the two groups.IL-4, IL-8 inflammatory factor levels were lower than before treatment,the experimental group was better than control group,the difference was statistically significant(P<0.05).CD4 +,CD8 +of T cells and Thl/Th2 values are higher than before the treatment in both the two groups,the experimental group was higher than control group, the difference was statistically significant ( P<0.05 ).Serum IgE levels were lower than before the treatment in both the two groups,the experimental group was lower than control group,the difference was statistically significant (P<0.05).Conclusion The therapy of azelastine hydrochloride nasal spray combined with desloratadine can improve the clinical effect of the treatment of allergic rhinitis,reduce inflammation,strengthen the body's immune function, improve thelevel of serum IgE.