Gasdermin E-mediated keratinocyte pyroptosis participates in the pathogenesis of psoriasis by promoting skin inflammation.

BACKGROUND: Psoriasis is a common chronic inflammatory disease with an unclear aetiology. Keratinocytes in psoriasis are susceptible to exogenous triggers that induce inflammatory cell death. OBJECTIVES: To investigate whether gasdermin E (GSDME)-mediated pyroptosis in keratinocytes contributes to the pathogenesis of psoriasis. METHODS: Skin samples from patients with psoriasis and from healthy controls were collected to evaluate the expression of GSDME, cleaved caspase-3 and inflammatory factors. We then analysed the data series GSE41662 to further compare the expression of GSDME between lesional and nonlesional skin samples in those with psoriasis. In vivo, a caspase-3 inhibitor and GSDME-deficient mice (Gsdme-/-) were used to block caspase-3/GSDME activation in an imiquimod-induced psoriasis model. Skin inflammation, disease severity and pyroptosis-related proteins were analysed. In vitro, tumour necrosis factor (TNF)-α-induced caspase-3/GSDME-mediated pyroptosis in the HACAT cell line was explored. RESULTS: Our analysis of the GSE41662 data series found that GSDME was upregulated in psoriasis lesions vs. normal skin. High levels of inflammatory cytokines such as interleukin (IL)-1ß, IL-6 and TNF-α were also found in psoriasis lesions. In mice in the Gsdme-/- and caspase-3 inhibitor groups, the severity of skin inflammation was attenuated and GSDME and cleaved caspase-3 levels decreased after imiquimod treatment. Similarly, IL-1ß, IL-6 and TNF-α expression was decreased in the Gsdme-/- and caspase-3 inhibitor groups. In vitro, TNF-α induced HACAT cell pyroptosis through caspase-3/GSDME pathway activation, which was suppressed by blocking caspase-3 or silencing Gsdme. CONCLUSIONS: Our study provides a novel explanation of TNF-α/caspase-3/GSDME-mediated keratinocyte pyroptosis in the initiation and -acceleration of skin inflammation and the progression of psoriasis.

Psoriasis is chronic and autoinflammatory common skin disease that affects 2­3% of the world's population. The disease is characterized by persistent inflammation in various body systems, including the skin and joints. However, the exact cause of the disease is unclear. In this study from China, we found that in people with psoriasis a protein called 'gasdermin E' (or 'GSDME') is increased in a type of skin cell called keratinocytes. In psoriasis, these keratinocytes are susceptible to a type of cell death called 'pyroptosis'. We aimed to find out whether pyroptosis caused by GSDME in keratinocytes contributes to the development of psoriasis. To do this, we looked at samples of skin from people with psoriasis and compared these to samples from healthy controls (those without psoriasis). Firstly, we investigated the levels of GSDME, another protein called caspase-3 and other inflammatory factors in the skin lesions from patients with psoriasis. Secondly, we analysed previously published data from 24 patients with psoriasis. Finally, we carried out a range of experiments to confirm our findings. We found that keratinocyte pyroptosis was mediated by the messenger proteins TNF-α/caspase-3, and that GSDME played a key role in the initiation and acceleration of skin inflammation and the progression of psoriasis. Targeting the GSDME pathway may be a novel strategy in treating psoriasis.

Meta-analysis of factors influencing anterior knee pain after total knee arthroplasty.

BACKGROUND: Total knee arthroplasty (TKA) is a mature procedure recommended for correcting knee osteoarthritis deformity, relieving pain, and restoring normal biomechanics. Although TKA is a successful and cost-effective procedure, patient dissatisfaction is as high as 50%. Knee pain after TKA is a significant cause of patient dissatisfaction; the most common location for residual pain is the anterior region. Between 4% and 40% of patients have anterior knee pain (AKP). AIM: To investigate the effect of various TKA procedures on postoperative AKP. METHODS: We searched PubMed, EMBASE, and Cochrane from January 2000 to September 2022. Randomized controlled trials with one intervention in the experimental group and no corresponding intervention (or other interventions) in the control group were collected. Two researchers independently read the title and abstract of the studies, preliminarily screened the articles, and read the full text in detail according to the selection criteria. Conflicts were resolved by consultation with a third researcher. And relevant data from the included studies were extracted and analyzed using Review Manager 5.4 software. RESULTS: There were 25 randomized controlled trials; 13 were comparative studies with or without patellar resurfacing. The meta-analysis showed no significant difference between the experimental and control groups (P = 0.61). Six studies were comparative studies of circumpatellar denervation vs non-denervation, divided into three subgroups for meta-analysis. The two-subgroup meta-analysis showed no significant difference between the experimental and the control groups (P = 0.31, P = 0.50). One subgroup meta-analysis showed a significant difference between the experimental and control groups (P = 0.001). Two studies compared fixed-bearing TKA and mobile-bearing TKA; the results meta-analysis showed no significant difference between the experimental and control groups (P = 0.630). Two studies compared lateral retinacular release vs non-release; the meta-analysis showed a significant difference between the experimental and control groups (P = 0.002); two other studies compared other factors. CONCLUSION: Patellar resurfacing, mobile-bearing TKA, and fixed-bearing TKA do not reduce the incidence of AKP. Lateral retinacular release can reduce AKP; however, whether circumpatellar denervation can reduce AKP is controversial.

The Charlson Comorbidity Index and depression are associated with satisfaction after short-segment lumbar fusion in patients 75 years and older.

Background: The rate and volume of lumbar spinal fusion (LSF) surgery performed for patients aged 75 years and older increased in recent years. The purposes of our study were to identify factors associated with postoperative dissatisfaction and evaluate the predictive value of comprehensive geriatric assessment (CGA) for dissatisfaction at 2 years after elective short-segment (one- or two- level) LSF in patients aged 75 and older. Methods: This was a retrospective study using a prospectively collected database of consecutive patients (aged 75 and older) who underwent elective short-segment transforaminal lumbar interbody fusion surgery for degenerative diseases from June 2018 to May 2020. Preoperative CGA consisting six domains was performed for each patient 1 day before the operative day. Univariate and multivariate analyses were performed to identify factors that predict for dissatisfaction with surgical treatment. The primary outcome was patient satisfaction with LSF surgery, as measured by the North American Spine Society (NASS) satisfaction scale. Secondary outcomes included postoperative complications, the length of stay, visual analog scale (VAS), and Oswestry Disability Index. Results: A total of 211 patients were available for a follow-up at 2 years and included in our final study cohort with a mean age of 80.0 years. A total of 175 patients (82.9%) were included in the satisfied group, and 36 patients (17.1%) were included in the not dissatisfied group. In the dissatisfied group, there was a higher incidence of postoperative complications (30.6% vs. 14.3%, p = 0.024) and greater VAS scores for lower back (4.3 ± 1.9 vs. 1.3 ± 1.4, p = 0.001) and leg (3.9 ± 2.1 vs. 0.9 ± 1.3, p = 0.001). Multivariate regression analysis revealed that patients with greater CCI score [odd ratio (OR) 2.56, 95% CI, 1.12-5.76; p = 0.030 for CCI 1 or 2 and OR 6.20, 95% CI, 1.20-28.69; p = 0.024], and depression (OR 3.34, 95% CI, 1.26-9.20; p = 0.016) were more likely to be dissatisfied compared with patients with the CCI score of 0 and without depression. Conclusions: Satisfaction after LSF in older patients (aged 75 and older) was similar to that of previously reported younger patients. Preoperative depression and higher CCI scores were independent risk factors for postoperative dissatisfaction two years after LSF surgery. These results help inform decision-making when considering LSF surgery for patients aged 75 and older.

Optimized cytotoxicity assay for co-suspended effector and target cells.

In recent years, adoptive cell therapy of immune effector cells, such as chimeric antigen receptor-T (CAR-T) cells, natural killer (NK) cells, and epitope-specific cytotoxic T lymphocyte (CTL) cells have been employed in clinical trials. In addition, CD19 CAR-T cells have been approved by the FDA for treatment of non-Hodgkin lymphoma and diffuse large B-cell lymphoma. In this context, it is vital to detect cellular cytotoxicity and monitor the quality of ex vivo expanded immune cells before product release and patient infusion. Target cells could proliferate in parallel with effector cells during the cytotoxicity assay, making it difficult to estimate the death ratio using conventional approaches. Meanwhile, non-specific dyes or non-homogeneous biomarkers for target cells may interfere with the final readout post addition of effector cells. Here, we modified a component of the coincubation medium to suppress the spontaneous release of bis(acetoxymethyl)2,2':6',2″-terpyridine-6,6″-dicarboxylate and sustained the window at a stable range (~70%). Further, the optimized Eu-TDA method presented reliable outcomes compared with lactate dehydrogenase detection and was compatible with cytotoxicity tests for NK cells and specific CTLs. Finally, the reported assay can accurately detect death of target cells depending on the amount of hydrophilic complex and can be reliably applied in quality control and cell activity evaluation tests on co-suspended effector and target cells. SUMMARY: A medium component for cellular coincubations (and associated protocols) have been optimized and validated for cytotoxicity assays, which can reliably evaluate the potency of engineered CD19 CAR-T cells, NK cells, and specific CTLs. In particular, the reported method can be applied widely in routine assays for bi-suspended effector and target cells with a stable window.

Clinical outcome of femoral neck system versus cannulated compression screws for fixation of femoral neck fracture in younger patients.

BACKGROUND: The clinical outcome of a new fixation device (femoral neck system, FNS) for femoral neck fractures remains unclear. The main purpose of this study was to evaluate two different internal fixation methods for the treatment of femoral neck fractures in patients aged under 60 years. METHODS: We retrospectively studied patients who underwent internal fixation surgery in our hospital for femoral neck fractures between January 2017 and January 2020. Cannulated compression screws (CCS) and FNS groups were divided according to different internal fixation methods. General data (such as sex, age, body mass index, type of fracture) of all patienFemoral neck shorteningts were collected, and joint function was evaluated using the Harris Hip Score (HHS) before and 1 year after surgery. We recorded related surgical complications, including femoral head necrosis, nonunion, and femoral neck shortening. RESULTS: There were no significant differences in age, sex, or body mass index between the two groups. There was no statistical difference in HHSs between the two groups before surgery. Patients who underwent FNS treatment had longer surgery time (79.75 ± 26.35 min vs. 64.58 ± 18.56 min, p = 0.031) and more blood loss (69.45 ± 50.47 mL vs. 23.71 ± 28.13 mL, p < 0.001). The degree of femoral neck shortening in the FNS group was significantly lower than that in the CCS group (10.0% vs 37.5%, p = 0.036). Regarding postoperative complications, there was no statistical difference in the incidence of femoral head necrosis and fracture nonunion between the two groups. CONCLUSION: Patients younger than 60 with femoral neck fractures can obtain satisfactory clinical results with CCS or FNS treatment. FNS has excellent biomechanical properties and shows significantly higher overall construct stability.

Circ_0020093 ameliorates IL-1ß-induced apoptosis and extracellular matrix degradation of human chondrocytes by upregulating SPRY1 via targeting miR-23b.

Osteoarthritis (OA) is a chronic disease characterized by articular cartilage degeneration and uncontrolled chondrocyte apoptosis. At present, accumulating evidence introduces that circular RNA (circRNA) is involved in the development of OA. The aim of our study was to explore the role and the functional mechanism of circ_0020093 in OA cell model. Human chondrocytes were treated with interleukin-1 beta (IL-1ß) to construct OA model. The expression of circ_0020093, miR-23b, and Sprouty 1 (SPRY1) mRNA was measured by quantitative real-time polymerase chain reaction (qRT-PCR). Cell apoptosis was assessed by flow cytometry assay. The expression of extracellular matrix (ECM)-associated markers and SPRY1 protein level was detected by qRT-PCR and Western blot. Bioinformatics analysis-predicted relationship between miR-23b and circ_0020093 or SPRY1 was further verified by dual-luciferase reporter assay and RNA pull-down assay. In this study, we found that the expression of circ_0020093 and SPRY1 was declined, while miR-23b expression was elevated in IL-1ß-treated chondrocytes. IL-1ß induced chondrocyte apoptosis and ECM degradation, while these negative effects were alleviated by circ_0020093 overexpression or miR-23b inhibition. MiR-23b was a target of circ_0020093, and SPRY1 was a downstream target of miR-23b. Rescue experiments showed that miR-23b enrichment reversed the role of circ_0020093 overexpression, and SPRY1 knockdown also reversed the effects of miR-23b inhibition. Importantly, circ_0020093 positively regulated SPRY1 expression by targeting miR-23b. In conclusion, circ_0020093 ameliorates IL-1ß-induced apoptosis and ECM degradation of human chondrocytes by regulating the miR-23b/SPRY1 axis.

Antiplatelet therapy for the prevention of atherosclerosis in chronic kidney disease (ALTAS-CKD) patients: study protocol for a randomized clinical trial.

BACKGROUND: Cardiovascular disease (CVD) is the most common complication and the leading cause of death in patients with chronic kidney disease (CKD). Accelerated atherosclerosis is a pathophysiological process that is vital to the occurrence of cardiovascular complications associated with CKD. Abnormal platelet activation is not only the leading cause of atherosclerosis but also plays a critical role in the occurrence of thrombotic events. Currently, antiplatelet drugs are commonly used as a secondary prevention strategy for high blood pressure, obesity, diabetes, and ischemic heart disease and can reduce the risk of CVD in the susceptible population. However, the benefits and evidence of using antiplatelet agents in patients with CKD remain controversial. This study aimed to determine whether antiplatelet therapy can safely prevent atherosclerosis in patients with CKD in the primary care setting. METHODS/DESIGN: The ALTAS-CKD study is a multicenter, prospective, randomized, double-blind, placebo-controlled trial of 554 adult patients with stage 3-5 non-dialysis-dependent CKD recruited from 10 territory medical centers in China. A secured web-based computer randomization system will be used to administer aspirin 100 mg once daily or a matching inactive placebo for 36 months. The primary endpoint will be the occurrence of atherosclerosis, as measured by carotid ultrasonography. The secondary endpoints will be combined cardiovascular events, all-cause mortality, and 50% decrease in the estimated glomerular filtration rate. TRIAL REGISTRATION {2A}: Current controlled trials number: ChiCTR1900021393 . Registered on 18 February 2019.

Evaluation of Nonviral piggyBac and lentiviral Vector in Functions of CD19chimeric Antigen Receptor T Cells and Their Antitumor Activity for CD19+ Tumor Cells.

Nonviral transposon piggyBac (PB) and lentiviral (LV) vectors have been used to deliver chimeric antigen receptor (CAR) to T cells. To understand the differences in the effects of PB and LV on CAR T-cell functions, a CAR targeting CD19 was cloned into PB and LV vectors, and the resulting pbCAR and lvCAR were delivered to T cells to generate CD19pbCAR and CD19lvCAR T cells. Both CD19CAR T-cell types were strongly cytotoxic and secreted high IFN-γ levels when incubated with Raji cells. TNF-α increased in CD19pbCAR T cells, whereas IL-10 increased in CD19lvCAR T cells. CD19pbCAR and CD19lvCAR T cells showed similar strong anti-tumor activity in Raji cell-induced mouse models, slightly reducing mouse weight while enhancing mouse survival. High, but not low or moderate, concentrations of CD19pbCAR T cells significantly inhibited Raji cell-induced tumor growth in vivo. These CD19pbCAR T cells were distributed mostly in mesenteric lymph nodes, bone marrow of the femur, spleen, kidneys, and lungs, specifically accumulating at CD19-rich sites and CD19-positive tumors, with CAR copy number being increased on day 7. These results indicate that pbCAR has its specific activities and functions in pbCAR T cells, making it a valuable tool for CAR T-cell immunotherapy.

Efficacy and mid/long-term survivorship of mobile-bearing unicompartmental knee arthroplasty for medial compartment knee osteoarthritis combined patellofemoral joint arthritis: a prospective cohort study protocol.

INTRODUCTION: Unicompartmental knee arthroplasty (UKA) is one of the most effective surgical procedures for treating isolated medial compartment knee osteoarthritis. However, previous studies have regarded patellofemoral osteoarthritis as a contraindication for UKA. In contrast, most current research shows that damage to the articular cartilage of the patellofemoral joint, even to the extent of full-thickness cartilage loss, has no influence on the outcome of UKA. METHODS AND ANALYSIS: Study settings: This study is a prospective cohort study that will compare the Forgotten Joint Score and Lonner patellofemoral joint score of patients who have undergone UKA; the patients will be divided into two groups (with and without patellofemoral joint osteoarthritis (PFJOA)). PRIMARY OBJECTIVE: Long-term follow-up will be used to evaluate the effect of the operation on the above-mentioned scores in both the groups. SECONDARY OBJECTIVE: We will divide the patients from the with PFJOA group into three subgroups according to the localisation of patellofemoral cartilage lesions (medial zone, lateral zone and central zone). We aim to compare knee joint scores among these groups and clarify the impact of different wear sites on clinical efficacy. We will use CT to explore the potential mechanism through which UKA affects patellofemoral joint-related parameters (lateral patellar tilt, lateral patellar shift and tibia tuberosity-trochlear groove distance). We will also record mid-term/long-term post-surgery complications. ETHICS AND DISSEMINATION: This study's protocol is in accordance with the Declaration of Helsinki. This study was approved by the Ethics Committee of Xuanwu Hospital. The results of this study will be disseminated in international peer-reviewed journals. TRIAL REGISTRATION NUMBER: ChiCTR2000030310.

Patient outcomes in Anteromedial osteoarthritis patients over 80 years old undergoing Oxford Unicompartmental knee Arthroplasty in China.

BACKGROUND: The use of Oxford Unicompartmental Knee Arthroplasty (UKA) has increased rapidly in both Western and Asian populations, with excellent functional outcomes and high patient satisfaction. While previous evidence regarding clinical outcomes and survival rates after Oxford UKA was based on studies in Western populations, the results may be different in Asian patients. The relevance of age for postoperative function after Oxford UKA also remains unclear. Hence, the aim of our study was to clarify the effectiveness and safety of Oxford UKA in Asian patients aged over 80 years. METHODS: A retrospective review was performed and included 195 patients (209 knees) who underwent an Oxford UKA between June 2015 and July 2018. We divided the patients into three groups by age: Group 1, 60-69 years; Group 2, 70-79 years; and Group 3, over 80 years. We used the Hospital for Special Surgery (HSS) score and Western Ontario and McMaster (WOMAC) Universities Osteoarthritis Index score to evaluate the general condition of the patients' knees before surgery and at last follow-up. We also recorded perioperative and short-term complications. RESULT: Group 1 consisted of 60 patients (60 knees); Group 2, 70 patients (79 knees); and Group 3, 65 patients (70 knees). The mean follow-up was 21.34 ± 12.04, 22.08 ± 11.38, and 21.76 ± 10.20 months in groups 1, 2, and 3, respectively. At last follow-up, the patients in Group 3 showed lower function scores compared to groups 1 and 2 (P < 0.05), but the HSS scores and the WOMAC scores were significantly improved in all three groups. In terms of perioperative and other complications, the three age groups did not differ significantly. CONCLUSION: Oxford UKA is an effective and safe treatment for osteoarthritis, even in elderly patients in China. Elderly patients have lower knee function scores than younger patients. However, the knee joint pain of the elderly patients was relieved and function improved compared to the preoperative condition.

Immunotherapy of patient with hepatocellular carcinoma using cytotoxic T lymphocytes ex vivo activated with tumor antigen-pulsed dendritic cells.

Purpose The aim of this study was to evaluate the clinical response of immunotherapy with dendritic cell-cytotoxic T lymphocytes (DC-CTLs) in patients with hepatocellular carcinoma (HCC). Method Sixty-eight patients with a confirmed diagnosis of HCC and who received follow-up until December 2015 were enrolled. We measured immune phenotypes of DCs and activated T cells using flow cytometry and clinical indexes using an electrochemiluminescence method. Results DCs exhibited up-regulation of the maturation markers CD83, CD80, CD11c, and CD86 on day8. Levels of IFN-γ and TNF-α were higher in the DCs pulsed with tumor-associated antigens (TAAs) than in DCs with a non-proliferative recombinant adenovirus. The percentage of regulatory T cells (Tregs) decreased in patients after DC-CTLs therapy. In addition, serum levels of AFP, AFP-L3, ALT, and CA19-9 were significantly reduced in these patients. Quality of life was improved, especially on physical functioning scales. Median overall survival (OS) and progression-free survival (PFS) were 8.2 months and 4.3 months, respectively, for the control group and 12.8 months and 9 months, respectively, for the DC-CTL group. Patients treated with DC-CTLs therapy showed a statistically significant PFS and OS curve (OS: p=0.016; PFS: p<0.0001). In addition, no serious adverse reactions were observed. Conclusion This study indicated that Tregs, as well as serum levels of AFP, AFP-L3, ALT, and CA19-9, which were correlated with a poor prognosis, decreased after DC-CTL treatments. The OS, PFS and the quality of life of HCC patients partially improved.

Establishment of mouse teratocarcinomas stem cells line and screening genes responsible for malignancy.

The sequential transplantation of embryonal carcinoma cells in vivo can accelerate the growth and malignancy of teratocarcinomas. However, the possible molecular mechanisms in this process that reflect cancer formation in the early stage are largely unknown and. To identify which genes are associated with the changes of malignancy of teratocarcinomas, we established a tumorigenesis model in which teratocarcinoma were induced via injecting embryonic stem cells into immuno-deficiency mice, isolating teratocarcinoma stem cell from a teratocarcinoma in serum-free culture medium and injecting teratocarcinoma stem cells into immune-deficient mice continuously. By using high-throughput deep sequence technology, we identified 26 differentially expressed genes related to the changes of characteristics of teratocarcinoma stem cell in which 18 out of 26 genes were down-regulated and 8 genes were up-regulated. Among these genes, several tumor-related genes such as Gata3, Arnt and Tdgf1, epigenetic associated genes such as PHC1 and Uty were identified. Pathway enrichment analysis result revealed that Wnt signaling pathway, primary immunodeficiency pathway, antigen processing and presentation pathway and allograft rejection pathway were involved in the teratocarcinoma tumorigenesis (corrected p value<0.05). In summary, our study established a tumorigenesis model and proposed some candidate genes and signaling pathways that may play a key role in the early stage of cancer occurrence.