Association between premature vascular smooth muscle cells senescence and vascular inflammation in Takayasu's arteritis.

OBJECTIVES: Arterial wall inflammation and remodelling are the characteristic features of Takayasu's arteritis (TAK). It has been proposed that vascular smooth muscle cells (VSMCs) are the main targeted cells of inflammatory damage and participate in arterial remodelling in TAK. Whether VSMCs are actively involved in arterial wall inflammation has not been elucidated. Studies have shown that cellular senescence in tissue is closely related to local inflammation persistence. We aimed to investigate whether VSMCs senescence contributes to vascular inflammation and the prosenescent factors in TAK. METHODS: VSMCs senescence and senescence-associated secretory phenotype were detected by histological examination, bulk RNA-Seq and single-cell RNA-seq conducted on vascular surgery samples of TAK patients. The key prosenescent factors and the downstream signalling pathway were investigated in a series of in vitro and ex vivo experiments. RESULTS: Histological findings, primary cell culture and transcriptomic analyses demonstrated that VSMCs of TAK patients had the features of premature senescence and contributed substantially to vascular inflammation by upregulating the expression of senescence-associated inflammatory cytokines. IL-6 was found to be the critical cytokine that drove VSMCs senescence and senescence-associated mitochondrial dysfunction in TAK. Mechanistically, IL-6-induced non-canonical mitochondrial localisation of phosphorylated STAT3 (Tyr705) prevented mitofusin 2 (MFN2) from proteasomal degradation, and subsequently promoted senescence-associated mitochondrial dysfunction and VSMCs senescence. Mitochondrial STAT3 or MFN2 inhibition ameliorated VSMCs senescence in ex vivo cultured arteries of TAK patients. CONCLUSIONS: VSMCs present features of cellular senescence and are actively involved in vascular inflammation in TAK. Vascular IL-6-mitochondrial STAT3-MFN2 signalling is an important driver of VSMCs senescence.

Deciphering and advancing CAR T-cell therapy with single-cell sequencing technologies.

Chimeric antigen receptor (CAR) T-cell therapy has made remarkable progress in cancer immunotherapy, but several challenges with unclear mechanisms hinder its wide clinical application. Single-cell sequencing technologies, with the powerful unbiased analysis of cellular heterogeneity and molecular patterns at unprecedented resolution, have greatly advanced our understanding of immunology and oncology. In this review, we summarize the recent applications of single-cell sequencing technologies in CAR T-cell therapy, including the biological characteristics, the latest mechanisms of clinical response and adverse events, promising strategies that contribute to the development of CAR T-cell therapy and CAR target selection. Generally, we propose a multi-omics research mode to guide potential future research on CAR T-cell therapy.

4cRNA NEAT1 Sponge Adsorption of miR-378 Modulates Activity of Lipopolysaccharide-treated Articular Chondrocytes and Influences the Pathological Development of Osteoarthritis.

Context: Osteoarthritis (OA) is a chronic joint disease that can eventually lead to degeneration, fibrosis, fractures, and defects of the articular cartilage. Long non-coding RNA (lncRNA) is a key substance in many processes, such as epigenetic regulation and cell-cycle and cell-differentiation modulation, and its relationship with OA has been repeatedly verified. Objective: The study intended to clarify the influence of lncRNA nuclear enriched abundant transcript 1 (NEAT1), lncRNA NEAT1, on lipopolysaccharide (LPS)-induced OA chondrocytes through sponge adsorption of microRNA-378 (miR-378) and to provide novel insights into future diagnosis and treatment of OA. Design: The research team performed an animal study. Setting: The study took place in the Department of Rehabilitation Medicine at Linyi People's Hospital in Linyi, Shangdong, China. Animals: The study's animals were 10 Sprague Dawley (SD) rats, 3-5 days old and 10-15 g in weight, of the specific-pathogen-free (SPF) grade. Intervention: The rat chondrocytes for the positive control group (the model group) were treated with 500 ng/mL of LPS to induce OA. Chondrocytes treated with the same amount of normal saline were used as the negative control group. The chondrocytes of the LPS-induced rats were into six groups: (1) a positive control group transfected with NEAT1-interfering RNA, the sh-NEAT1 group; (2) a negative control group not transfected with NEAT1-interfering RNA, the NEAT1 empty vector (NC-NEAT1) group; (3) an intervention group co-transfected with NEAT1 interfering RNA and the miR-378 inhibitor sequence (Inh-miR-378 the sh-NEAT1+ Inh-miR-378 group; (4) a negative control group transfected with NEAT1 interfering RNA but not transfected with the miR-378 inhibitor sequence, the sh-NEAT1+ miR-378 negative control (NC-miR-378) group; (5) a negative control group transfected with the miR-378 inhibitor sequence but not transfected with NEAT1 interfering RNA, the NEAT1 empty vector (NC-NEAT1) + Inh-miR-378 group; (6) a negative control group not transfected with either NEAT1 interfering RNA or the miR-378 inhibitor sequence, the NC-NEAT1 + NC-miR-378 group. Outcome Measures: An OA-chondrocyte model was induced by LPS and measurements of NEAT1 and miR-378 expression were made by real-time quantitative reverse transcription (qRT)- polymerase chain reaction (PCR). Then, small NEAT1-interfering RNA (sh-NEAT1), empty vector NEAT1 (NC-NEAT1), inhibitor-sequence-miR-378 (Inh-miR-378), and negative-control-miR-378 (NC-miR-378) were transfected into cells, and cell viability and apoptosis rate were measured. Finally, the study verified the relationship between NEAT1 and miR-378. Results: Compared to the control group, NEAT1 was significantly elevated in the model group, and its miR-378 was significantly decreased. Silencing NEAT1 can enhance OA-chondrocyte activity and decrease apoptosis. When NEAT1 and miR-378 were inhibited together, as shown fort the NC-NEAT1 + NC-miR-378 group, NEAT1 expression, as well as the multiplication and apoptosis ability of the OA-model cells, were the same as those of cells transfected with an empty vector, the NC-NEAT1 group. Also, the NEAT1 + NC-miR-378 group's cell activity was lower than that of the sh-NEAT1+NC-miR-378 group but higher than that of the NC-NEAT1 + Inh-miR-378 group. Finally, higher fluorescence activity occurred for NEAT1-mutant type (MUT) transfected with Inh-miR-378. Conclusions: NEAT1, which is highly expressed in OA, mediates LPS-induced OA-chondrocyte activity through sponge adsorption of miR-378.

Serum IL-6 level trajectory for predicting the effectiveness and safety of tocilizumab in the treatment of refractory Takayasu arteritis.

AIM: To explore the value of serial monitoring of serum interleukin-6 (IL-6) levels for predicting treatment response and occurrence of adverse events during tocilizumab (TCZ) treatment in refractory Takayasu arteritis (TAK). METHODS: TAK patients receiving TCZ treatment were prospectively recruited and followed up at 1 month, 3 months and then every 3-6 months. Serum IL-6 levels were measured at each visit. Overall response was the combination of complete and partial response, requiring resolution of signs and symptoms, hsCRP and ESR level decreased at least by half, no progression on imaging and dose of glucocorticoid <15 mg/d. RESULTS: Thirty-five patients with a median follow up duration of 17 [9-44] months were included. The change of IL-6 after TCZ treatment for 6 months compared to the baseline was significantly lower in patients achieved overall response at 6, 12, 18 and 24 months. The ratio of IL-6 at 6 months to baseline could predict overall response at 12 and 24 months after TCZ treatment. With a cutoff value of 1.6, the sensitivity and specificity were 83.3 % and 87.5 % for 12 months, while 100 % and 88.9 % for 24 months. Patients with the ratio less than 1.6 were also 9 times more likely to achieve sustained improvement without treatment intensification. No correlation between IL-6 dynamics and occurrence of adverse events was found. CONCLUSIONS: The change of IL-6 levels after TCZ treatment for 6 months compared to the baseline can predict the overall treatment response at 12 months, 24 months and sustained improvement.

Baricitinib for refractory Takayasu arteritis: a prospective cohort study in a tertiary referral centre.

OBJECTIVE: To investigate the treatment efficacy and safety of baricitinib in patients with refractory Takayasu arteritis (TAK). METHODS: We performed a prospective cohort study in which baricitinib 4 mg daily was prescribed to patients with refractory TAK, combined with oral glucocorticoids (GCs). RESULTS: 10 patients with refractory TAK were enrolled with a median age of 28 (IQR=22-37) years, median disease duration of 50 (IQR=24-65) months. The median dose of GCs was 10 (IQR=8.1-22.5) mg prednisone or equivalence dosage at baseline. At 6 months of baricitinib treatment, 6/10 (60%) patients had an overall treatment response. During an average follow-up of 15.3 (range 4-31) months, 4/10 (40%) patients maintained overall treatment response. 8/10 (80%) patients tapered or maintained the same dose of GCs with no change of the combined classical synthetic disease-modifying antirheumatic drugs. Two patients discontinued GCs at 18 and 24 months and were in continuous remission till the end of the study. One patient withdrew baricitinib due to liver dysfunction. CONCLUSION: Baricitinib 4 mg daily is effective for refractory TAK and is well tolerated.

East-Asian lupus nephritis in the Hopkins Lupus Cohort.

Background and Objective: East Asian systemic lupus erythematosus (SLE) is under represented in lupus cohorts outside of East Asia. We asked whether lupus nephritis was more common and more severe in East Asians than in other ethnicities in a large United States SLE cohort. Methods: The Hopkins Lupus Cohort, a longitudinal cohort of 2802 patients (53.5% Caucasian, 39.2% African-American, 3.2% East Asian) was studied. The SLICC/ACR Damage Index was used to assess renal outcomes. Results: East Asian patients had the same prevalence of lupus nephritis as African-Americans and both were higher than Caucasians. East Asians were not significantly different in frequency of end stage kidney disease compared with African-Americans. East Asians were more likely than Caucasians to have anti-Sm, low C3 and low C4. East Asians were more likely than African-Americans to have low C3 and low C4. Conclusion: East Asians living in the United States were more likely to have lupus nephritis than Caucasians. Poor outcomes such as end stage kidney disease occurred at an equal frequency in East Asians as in African-Americans. Lupus nephritis was both more frequent and more severe in East Asians than in African-Americans.

Has-miR-300-GADD45B promotes melanoma growth via cell cycle.

Response to oncogenic factors like UV, GADD45 family in skin participates in scavenging ROS, DNA repair and cell cycle control. Because of this, the previous study of the chronic UVB injury model has found that hsa-miR-300 can conduct intercellular transport by exosomes and target regulation of GADD45B. Whether the hsa-miR-300-GADD45B still regulates tumor development by cell cycle pathway is unclear. Through transcriptomic analysis of primary (n=39) and metastatic (n=102) melanoma, it was confirmed that in metastatic samples, some of the 97 down-regulated genes participate in maintaining skin homeostasis while 42 up-regulated genes were enriched in cancer-related functions. Furthermore, CDKN1A, CDKN2A, CXCR4 and RAD51 in the melanoma pathway, were also differentially expressed between normal skin and melanoma. CDKN1A and CDKN2A were also found to be involved in TP53-dependent cell cycle regulation. In conclusion, it was speculated that CDKN1A, CDKN2A, TP53, GADD45B and hsa-miR-300 may have regulatory relationships. It was demonstrated that there is a bidirectional regulation between hsa-miR-300 and TP53. In addition, miR-300 can regulate CDKN1A by GADD45B/TP53 and promote melanoma growth by accelerating the cell cycle transition from G1/S to G2 phase.

Donor-derived and off-the-shelf allogeneic anti-CD19 CAR T-cell therapy for R/R ALL and NHL: A systematic review and meta-analysis.

Allogeneic anti-CD19 chimeric antigen receptor (CAR) T-cell therapy has the potential for extensive clinical applications. This study aimed to evaluate its efficacy and safety in treating relapsed or refractory (R/R) acute lymphoblastic leukemia (ALL) and non-Hodgkin lymphoma (NHL). Four databases were searched for relevant studies. Among patients treated with donor-derived CAR T-cell therapy, ALL patients had a complete remission (CR) rate of 80 % and a 1-year overall survival rate of 51 %. The graft-versus-host disease (GvHD) rate was 4 %, cytokine release syndrome was 69 %, and immune effector cell-associated neurotoxicity syndrome was 8 %. For off-the-shelf CAR T-cell therapy, the CR rate for ALL was 70 %, and for NHL, it was 52 %. The objective response rate for NHL was 72 %. The pooled GvHD of off-the-shelf CAR T-cell therapy for ALL and NHL combined was 0 %. Allogeneic anti-CD19 CAR T-cell therapy are effective and safe for treating R/R ALL and NHL. AVAILABILITY OF DATA AND MATERIALS: All datasets generated in this study are included in the article/Supplementary Material.

Layer-specific strain and dyssynchrony index alteration in new-onset systemic lupus erythematosus patients without cardiac symptoms.

BACKGROUND: Layer-specific speckle-tracking echocardiography (STE) is a noninvasive approach that assesses subclinical left ventricular dysfunction. We aimed to investigate the (I) alteration of layer-specific STE parameters and the dyssynchrony index; and (II) the disease parameters associated with layer-specific STE change in drug-naïve patients with new-onset systemic lupus erythematosus (SLE) without cardiac symptoms. METHODS: Thirty-five drug-naïve patients with new-onset SLE and twenty-five healthy controls were enrolled. All individuals received both conventional echocardiographic and two-dimensional STE assessment. The data of layer-specific global longitudinal strain (GLS), global circumferential strain (GCS), and peak systolic dispersion (PSD) were acquired in layer-specific STE. RESULTS: All patients had a normal left ventricular ejection fraction (LVEF)(mean LVEF: 58%) and conventional echocardiographic parameters were comparable between patients and controls. Decreased layer-specific GLS and elevated PSD were observed in SLE patients (whole layer GLS: -17.6%±3.0% versus -19.3%±2.6%, P=0.02; endocardial GLS: -20.0%±3.2% versus -22.1%±3.0%, P=0.01; epicardial GLS: -15.6%±2.7% versus -16.8%±2.4%, P=0.04; PSD: 41.0±18.9 versus 28.8±10.1 msec, P=0.007). In contrast, there was no difference in layer-specific GCS at three different levels between patients and controls (P>0.05). More severely impaired GLS was observed in patients with higher disease activity, high-risk antiphospholipid antibody (aPL) profile, or renal involvement. The PSD was increased in patients with higher disease activity or a high-risk aPL profile. Correlational analysis showed that GLS at three layers and PSD correlated with high-sensitivity C-reactive protein (hsCRP) levels (whole GLS: r=0.662, P<0.001; endocardial GLS: r=0.637, P<0.001; epicardial GLS: r=0.658, P<0.001; PSD: r=0.390, P=0.021). PSD correlated with epicardial GLS (r=0.360, P=0.047), when treating the hsCRP level, renal involvement, aPL profile, and disease activity as control variables. Multivariate regression showed the hsCRP level and epicardial GLS were predictors of layer-specific GLS impairment and elevated PSD, respectively. CONCLUSIONS: Drug-naive patients with new-onset SLE are likely to have subclinical GLS impairment and left ventricular dyssynchrony, even in the presence of normal LVEF. SLE-related risk factors are associated with these dysfunctions.

Effects of acupoint catgut embedding on the postmenopausal osteoporosis patients and related mechanism.

OBJECTIVE: To study the effect and mechanism of acupoint catgut embedding on patients of postmenopausal osteoporosis (PMOP). METHODS: In this prospective study, 90 patients with PMOP who received treatment in our hospital were randomly divided into the drug treatment group (n=45) and drug treatment + catgut embedding group (n=45) according to SPSS random table method. The drug treatment group was given conventional western medicine treatment, and the drug treatment + catgut embedding group was given point embedding therapy. Bone mineral density (BMD), calcium (Ca2+), osteoprotegerin (OPG), estrogen (E2), receptor activator of nuclear factor-kB ligand (RANKL), liver and kidney function and blood lipids were detected before treatment in the two groups, and visual analogue score (VAS) and PMOP symptom score were evaluated. The above-mentioned indexes were detected again 3 months and 6 months after treatment. RESULTS: At 3 and 6 months after treatment, the BMD and the levels of Ca2+ and E2 in the two groups were increased, while the levels of OPG and RANKL were decreased, and the improvement in the drug treatment + catgut embedding group was significantly better than that in the drug treatment group (P<0.05). The symptom scores of VAS and PMOP in the drug treatment + catgut embedding group were significantly lower than those in the drug treatment group (all P<0.001). There was no significant difference in the levels of alanine aminotransferase (ALT), total bilirubin (TBil), albumin (ALB), blood urea nitrogen (BUN), serum creatinine (SCr) and serum uric acid (SUA) between the two groups, but the levels of total cholesterol (TC) and triglyceride (TG) in the drug treatment + catgut embedding group were significantly lower than those in the drug treatment group (all P<0.001). CONCLUSION: Acupoint catgut embedding has a good effect on PMOP, and it can increase BMO and improve the clinical symptoms of patients, which is worthy of clinical promotion.

Elevation of serum proprotein convertase subtilisin/kexin type 9 (PCSK9) concentrations and its possible atherogenic role in patients with systemic lupus erythematosus.

BACKGROUND: Systemic lupus erythematosus (SLE) patients have tendencies of accelerated atherosclerosis (AS) which can only partly be explained by traditional cardiovascular disease (CVD) risk factors. Imbalanced inflammation also plays a vital role. Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a new therapeutic target for AS for its dual mechanisms in lipids and inflammation. We aimed to assess serum PCSK9 concentrations in SLE patients and its possible role in atherogenesis of SLE. METHODS: Ninety SLE patients and 50 healthy controls were included. SLE patients were further divided into SLE-AS and SLE-NonAS subgroups, according to the carotid intima-media thickness (cIMT). Traditional CVD risk factors, inflammatory biomarkers and PCSK9 concentrations were compared between: (I) SLE patients and controls; (II) SLE-AS subgroup and SLE-NonAS subgroup; (III) SLE patients with and without lupus nephritis (LN). Correlational analysis, univariate and multivariate linear regression analysis were applied to analyze the association between PCSK9 levels and disease parameter in SLE patients. Effects on PCSK9 concentrations by monotherapy with hydroxychloroquine (HCQ), which is thought having protective effects against AS in SLE, were investigated by follow-up analysis in 15 SLE patients. RESULTS: We found that SLE patients had significantly elevated serum PCSK9 levels than controls, especially in SLE-As subgroup or those with LN, accompanied with higher ratio of cIMT thickening. Correlational analysis showed PCSK9 concentrations correlated with C-reactive protein (CRP) levels, age and erythrocyte sedimentation rate (ESR). Univariate and multivariate linear regression revealed that only CRP, but not age or ESR was positive predictors of PCSK9. Interestingly, monotherapy with HCQ for three months significantly reduced PCSK9 and CRP levels in inactive SLE patients. CONCLUSIONS: Our results suggested that elevated PCSK9 levels in SLE are probably associated with atherogenic inflammation in SLE. HCQ, which is thought having protective effects against AS in SLE, can effectively reduce PCSK9 levels in SLE patients.

[Elevated level of serum PCSK9 in patients with systemic lupus erythematosus].

Objective To detect the level of serum proprotein convertase subtilisin/kexin type 9 (PCSK9) in patients with systemic lupus erythematosus (SLE) and investigate the correlation between PCSK9 level and disease parameters. Methods Forty-seven SLE patients and 30 age, sex-matched healthy controls (HCs) were included in our research. Traditional cardiovascular disease (CVD) risk factors were compared between the two groups. The level of serum PCSK9 was examined by ELISA. Carotid intima-media thickness (cIMT) was measured by carotid ultrasound. According to the measured value of cIMT, SLE patients were divided into SLE-AS (cIMT≥1.0 mm) and SLE-NonAS (cIMT<1.0 mm) subgroups. Atherogenic factors and PCSK9 levels were compared between the two subgroups. Univariate correlational analysis of PCSK9 levels and disease parameters was conducted in the SLE patients. Results No difference was found in the total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), ApoA1, ApoB, triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), fasting blood-glucose (FBG), body mass index (BMI) or uric acid (UA) between the SLE patients and HCs. However, the higher ratio of cIMT thickening and the elevation of serum PCSK9 levels were observed in the SLE patients as compared with the HCs. No significant difference in the traditional risk factors for CVD was found, but significant difference in the level of C-reactive protein (CRP) existed between the SLE-AS subgroup and SLE-NonAS subgroup. The level of serum PCSK9 in the SLE-AS subgroup was significantly higher than that in the SLE-NonAS subgroup. PCSK9 concentrations were positively correlated with CRP levels, but not correlated obviously with the age, SLEDAI, lipids parameters (TC, LDL-C, ApoA1, ApoB, TG, HDL-C), BMI or UA levels. This tendency seemed to be more significant in the female patients. Conclusion Elevated level of serum PCSK9 can be observed in SLE patients, especially in those with thickening of cIMT. PCSK9 may be associated with atherogenic inflammation in SLE.

The correlational research among serum CXCL13 levels, circulating plasmablasts and memory B cells in patients with systemic lupus erythematosus: A STROBE-compliant article.

We investigated whether serum CXC ligand 13 protein (CXCL13) levels correlate with the circulating plasmablasts and memory B-cells alteration in systemic lupus erythematosus (SLE) patients. The diagnostic use of CXCL13 concentrations in active lupus was also analyzed.A total of 36 SLE patients and 18 healthy controls were included. Serum CXCL13 levels were examined by enzyme-linked immunosorbent assay. The frequency and absolute count of circulating plasmablasts and memory B cells were analyzed by flow cytometry. Receiver operating characteristic curves (ROC curves) were generated to analyze the utility of serum CXCL13 level and plasmablasts frequency as tools for the recognition of active SLE.Elevation of serum CXCL13 levels, higher plasmablasts frequency, and reduction of memory B-cells count were observed in SLE patients, compared with healthy controls. Interestingly, correlational analyses showed not only significantly positive association between CXCL13 levels and SLE Disease Activity Index (SLEDAI) or plasmablasts frequency, but an inverse correlation between CXCL13 concentration and memory B-cell count. ROC curves showed that serum CXCL13 level and plasmablasts frequency were practical in identifying active disease from overall SLE patients, with considerable accuracy.Serum CXCL13 levels correlate with the alteration of plasmablasts and memory B cells in SLE. CXCL13 may be used as a practical tool in judgment of active SLE.