Objective:To explore the role of cathepsin S(CTSS) in diabetic vascular injury-induced restenosis.Methods:(1) Dendritic cells(DCs) were stimulated with different concentrations of glucose, and CTSS was either knocked down or upregulated in dendritic cells using adenovirus transfection. The mRNA and protein expression levels of CTSS were detected by RT-qPCR and Western blot, and the changes of interleukin(IL)-6 levels were assessed using RT-qPCR and ELISA in response to CTSS. (2) The extent of Th17 cell differentiation was evaluated with Flow cytometry when CTSS was downregulated or overexpressed. Levels of ROR-γt, IL-17A, IL-17F, IL-22, and IL-23 were measured. (3) Streptozomycin(STZ, 60 mg/kg) was injected into the intraperitoneal cavity of rats fasted for 12 h to obtain a diabetic rat model, and the restenosis model was obtained by balloon catheter and carotid guidewire injury, and the differentiation degree of Th17 cells in different groups of rats was compared when CTSS was up-regulated and down-regulated.Results:(1) DC viability decreased when stimulated with 35 mmol/L glucose for 48 hours. Compared to the control group, glucose treatment led to a concentration-dependent increase in CTSS and IL-6 levels in DCs( P<0.05). Inhibition of CTSS reduced IL-6 protein levels, while its overexpression increased IL-6 protein levels( P<0.05). (2) Compared with the control group, CTSS inhibition in DC decreased the percentage of Th17 cells in T cells, with decreased protein levels of ROR-γt, IL-17A, IL-17F, IL-22, and IL-23, and vice versa ( P<0.050). (3) After carotid artery injury, CTSS expression was increased in perivascular adipose tissue(PVAT) of rats, and levels of ROR-γt, IL-17A, IL-17F, IL-22, and IL-23 in PVAT were significantly elevated. Down-regulation of CTSS eliminated the glucose-induced enhancement. Conclusion:Inhibition of CTSS in DC reduces Th17 cell differentiation and thereby suppresses restenosis following diabetic vascular injury.

Patients with bicuspid aortic valve (BAV) are characterized by asymmetric anatomy, severe calcification and combined aortic dilatation. Compared with tricuspid aortic valve stenosis patients, patients with BAV stenosis confront with greater surgical risks in transcatheter aortic valve replacement (TAVR), including paravalvular leak, aortic valve rupture, coronary artery obstruction, atrioventricular block and so on. However, with the advent of new generation of prosthetic valves and optimization of surgical strategies, several studies have shown that TAVR is safe and effective in the treatment of BAV stenosis. Therefore, we aim to provide an overview of the use of TAVR in patients with BAV stenosis.

Objective To investigate the efficacy and safety of edaravone dexborneol combined with alteplase in the treatment of acute ischemic stroke （AIS）. Methods The data were collected from 124 patients with AIS who were admitted to our hospital from November 2020 to April 2022. The patients were randomly divided into experimental group （intravenous thrombolysis with alteplase + treatment with edaravone dexborneol） and control group （intravenous thrombolysis with alteplase）， and the two groups were compared for efficacy. Results The overall response rate in the experimental group was significantly higher than that in the control group （82.3% vs 64.5%， P < 0.05）. The National Institutes of Health Stroke Scale scores at different stages after thrombolysis were significantly lower in the experimental group （5.40 ± 3.82， 4.14 ± 3.44， and 0.57 ± 0.99） than in the control group （P < 0.05）. No adverse drug reactions were observed in the two groups during the treatment. Conclusion Edaravone dexborneol combined with alteplase has definite clinical efficacy in the treatment of AIS.

OBJECTIVE: To investigate the classification of idiopathic inflammatory myopathies (IIM) based on clinical manifestations and myositis- specific antibodies using cluster analysis. METHODS: We retrospectively analyzed the data of patients with IIM admitted in Nanfang Hospital in 2015-2019. The clinical data of the patients including serum creatine kinase (CK), interstitial lung disease (ILD), cancer, and myositis-specific antibodies were collected for two-step cluster analysis to identify the distinct clusters of patients, whose clinical characteristics were subsequently analysed. RESULTS: A total of 71 patients with IIM were included in this study, including 30 (42.3%) with polymyositis (PM), 20 (28.2%) with classic dermatomyositis (DM), 16 (22.5%) with amyopathic dermatomyositis (CADM), and 5 (7.0%) with immune-mediated necrotizing myopathy (IMNM). Two-step cluster analysis identified 3 distinctive subgroups: Cluster 1 of 15 (51.7%) patients characterized by rash, positive anti-MDA5 antibody and hypoproteinemia ( < 0.05) with normal or slightly elevated CK level, mainly corresponding to CADM; Cluster 2 of 4 (57.1%) patients with significantly elevated CK and positive anti-SRP antibody ( < 0.001) corresponding to IMNM; and Cluster 3 of 17 (48.6%) patients consisting primarily of patients with PM, characterized by positivity for anti- aminoacyl transfer RNA synthetases antibodies (=0.022) corresponding to antisynthetase syndrome (ASS). CONCLUSIONS Patients with IIM can be divided into 3 subgroups based on their clinical and serological characteristics (especially myositis-specific antibodies), and among them ASS may represent an independent IIM subgroup with unique clinical characteristics.
Antibodies ; metabolism ; Humans ; Myositis ; classification ; physiopathology ; Retrospective Studies

OBJECTIVE: To investigate the classification of idiopathic inflammatory myopathies (IIM) based on clinical manifestations and myositis- specific antibodies using cluster analysis. METHODS: We retrospectively analyzed the data of patients with IIM admitted in Nanfang Hospital in 2015-2019. The clinical data of the patients including serum creatine kinase (CK), interstitial lung disease (ILD), cancer, and myositis-specific antibodies were collected for two-step cluster analysis to identify the distinct clusters of patients, whose clinical characteristics were subsequently analysed. RESULTS: A total of 71 patients with IIM were included in this study, including 30 (42.3%) with polymyositis (PM), 20 (28.2%) with classic dermatomyositis (DM), 16 (22.5%) with amyopathic dermatomyositis (CADM), and 5 (7.0%) with immune-mediated necrotizing myopathy (IMNM). Two-step cluster analysis identified 3 distinctive subgroups: Cluster 1 of 15 (51.7%) patients characterized by rash, positive anti-MDA5 antibody and hypoproteinemia ( < 0.05) with normal or slightly elevated CK level, mainly corresponding to CADM; Cluster 2 of 4 (57.1%) patients with significantly elevated CK and positive anti-SRP antibody ( < 0.001) corresponding to IMNM; and Cluster 3 of 17 (48.6%) patients consisting primarily of patients with PM, characterized by positivity for anti- aminoacyl transfer RNA synthetases antibodies (=0.022) corresponding to antisynthetase syndrome (ASS). CONCLUSIONS Patients with IIM can be divided into 3 subgroups based on their clinical and serological characteristics (especially myositis-specific antibodies), and among them ASS may represent an independent IIM subgroup with unique clinical characteristics.
Antibodies ; Autoantibodies ; Dermatomyositis ; Humans ; Lung Diseases, Interstitial ; Myositis ; Retrospective Studies

Objective　To investigate the prognosis and influencing factors of intravascular interventional therapy for posterior circulation infarction. Method　For selecting our department admitted in October 2019 and October 2017-two years hospitalized with a total of 42 cases of clinically diagnosed patients with posterior circulation infarction，patient gender，age，on admission，the main symptoms and signs，pathogenic factors，pathogenesis time interval，when to see a doctor to vascular disease opening time，lesion blood vessel parts，collateral compensatory，operation method choice and recanalization，vascular interventional treatment and postoperative 7 days before the NIHSS score，GCS score，mRS score when discharged from hospital，postoperative complications and prognosis of 90 days. According to the improved Rankin score of patients with 90-day prognosis，the patients were divided into a good prognosis group （mRS score <4 points） and a poor prognosis group （mRS score ≥ 4 points），and statistical analysis was conducted. Results　（1） For posterior circulation infarction，multiple interventional therapies were used to treat the lesion vessel opening rate of 71.4%，the 3-month mortality rate of posterior circulation infarction was 28.6%. And the good prognosis rate was 50%. （2）Univariate analysis suggested a statistically significant difference in the prognosis of patients with smoking P<0.01）. （3）Logistic regression analysis showed that after adjusting for cerebrovascular risk factors such as smoking，lower basilar artery occlusion was associated with poor prognosis. （4）Good intraoperative lesion vessels （Ⅱb level or Ⅲ level） and is related to good prognosis. Conclusion　Endovascular interventional therapy is an efficient and rapid method for posterior circulation infarction to open blood vessels. Good recanalization of diseased blood vessels during operation is positively correlated with good prognosis，while smoking and lower basilar artery occlusion are correlated with poor prognosis.

Objective To explore the role of TLR4/NF-kB signal pathway in rat nerve injury after traumatic brain injury coupled with seawater drowning.Methods A total of 96 male Sprague-Dawley rats were randomly divided into 3 groups:the sham surgery group(n =12),the traumatic brain injury (TBI) group(n =42) and the traumatic brain injury coupled with seawater drowning group (or the TBI + SWD group) (n =42).At the timepoints of 3,6,12,24,72 and 168 hours after the establishment of the TBI model,immunohistochemistry was used to detect the expression of TLR4 and NF-kB cells in the rat brain tissue.Real time PCR (rt-PCR) was applied to detect the expression levels of TLR4 and NF-kB mRNA in the hippocampus of the brain tissue,and enzyme-linked immunosorbent assay (ELISA) was employed to detect the content of TNF-α in the rat brain tissue.Results The number of TLR4 and NF-kB positive cells in the brain tissue of the TBI group and the TBI + searwater drowning group increased.The expressions of TLR4 and NF-kB mRNA in the hippocampus of the brain tissue of the TBI group and the TBI + SWD group significantly increased 3 hours after establishment of the TBI model.For the TBI group,the expressions of TLR4 and NF-kB were respectively (1.63 ±0.52) and (1.52 ±0.41),while in the the TBI + SWD group,the expressions of the above 2 substantce were respectively (4 1.87 ± 0.93) and (1.87 ± 0.93).At hour 24,the expression reached peak.For the TBI group,the expressions of TLR4 and NF-kB were respectively (49.61 ± 0.34) and (4.60 ± 0.51),while for the the TBI + SWD group,the expressions of the above 2 substantces were respectively (16.11 ±0.49) and (5.80 ± 0.52),then they decreased gradually.The expression levels of TNF-α in the TBI group and the TBI + SWD group were found to elevate 3 hours after establishment of the model.TNF-α expression level for the TBI group was (53.68 ± 1.48) ng/L),while the expression level of the same substance for the TBI + SWD group was (60.14 ± 2.06) ng/L.Wave peak appeared at hour 12,and the wave peak for the TBI group was (79.28 ± 2.46) ng/L,while for the TBI + SWD group,it displayed at (103.51 ± 5.53) ng/L.Conclusion Seawater drowning could aggravate nerve inflammatory response mediated by TLR4/NF-kB signal pathway,which might play an important role in the rat brain injury induced by trauma.

Objective To explore the role of TLR4/NF-kB signal pathway in rat nerve injury after traumatic brain injury coupled with seawater drowning.Methods A total of 96 male Sprague-Dawley rats were randomly divided into 3 groups:the sham surgery group(n =12),the traumatic brain injury (TBI) group(n =42) and the traumatic brain injury coupled with seawater drowning group (or the TBI + SWD group) (n =42).At the timepoints of 3,6,12,24,72 and 168 hours after the establishment of the TBI model,immunohistochemistry was used to detect the expression of TLR4 and NF-kB cells in the rat brain tissue.Real time PCR (rt-PCR) was applied to detect the expression levels of TLR4 and NF-kB mRNA in the hippocampus of the brain tissue,and enzyme-linked immunosorbent assay (ELISA) was employed to detect the content of TNF-α in the rat brain tissue.Results The number of TLR4 and NF-kB positive cells in the brain tissue of the TBI group and the TBI + searwater drowning group increased.The expressions of TLR4 and NF-kB mRNA in the hippocampus of the brain tissue of the TBI group and the TBI + SWD group significantly increased 3 hours after establishment of the TBI model.For the TBI group,the expressions of TLR4 and NF-kB were respectively (1.63 ±0.52) and (1.52 ±0.41),while in the the TBI + SWD group,the expressions of the above 2 substantce were respectively (4 1.87 ± 0.93) and (1.87 ± 0.93).At hour 24,the expression reached peak.For the TBI group,the expressions of TLR4 and NF-kB were respectively (49.61 ± 0.34) and (4.60 ± 0.51),while for the the TBI + SWD group,the expressions of the above 2 substantces were respectively (16.11 ±0.49) and (5.80 ± 0.52),then they decreased gradually.The expression levels of TNF-α in the TBI group and the TBI + SWD group were found to elevate 3 hours after establishment of the model.TNF-α expression level for the TBI group was (53.68 ± 1.48) ng/L),while the expression level of the same substance for the TBI + SWD group was (60.14 ± 2.06) ng/L.Wave peak appeared at hour 12,and the wave peak for the TBI group was (79.28 ± 2.46) ng/L,while for the TBI + SWD group,it displayed at (103.51 ± 5.53) ng/L.Conclusion Seawater drowning could aggravate nerve inflammatory response mediated by TLR4/NF-kB signal pathway,which might play an important role in the rat brain injury induced by trauma.

CAR-T cell therapy that targets surface antigens to kill tumor cells specifically has recently become another cornerstone in tumor immunotherapy. In this study, a lentiviral expression plasmid of CAR targeting human epidermal growth factor receptor 2 (HER2) was constructed by genetic engineering. The recombinant plasmid was co-transfected with other packaging plasmids into HEK293T cells by calcium phosphate precipitation to generate lenti-car, which are CAR lentiviral particles. HER2-specific CAR-T cells were obtained by transducing human peripheral blood mononuclear cells with lenti-car. Their specific inhibitory effects on HER2-positive and HER2-negative tumor cells were analyzed in vitro. The constructed CAR-T cells were specifically activated by HER2-expressing tumor cells as indicated by secretion of IFN-γ and IL-2. The inhibitory rate on HER2-positive SK-OV-3 cell line was (58.47±1.72)%, significantly higher than that on the mock-treated control group (P<0.05). The inhibitory rate on HER2-negative K562 cell lines was (11.74±2.37)%, which was not significantly different from that on the control group (P>0.05). Furthermore, when we transfected a HER2-expressing vector into K562, the inhibitory rate increased to (30.41±7.59)%, which was higher than that on HER2-negative K562 (P<0.05). Thus, the constructed second-generation HER2-specific CAR-T cells specifically suppressed growth of tumor cells overexpressing HER2 protein, suggesting that HER2-specific CAR-T cells might prove useful for immunotherapy of HER2-positive cancer.

Various genetic switches have been developed to let engineered cells perform designed functions. However, a sustained input is often needed to maintain the on/off state, which is energy-consuming and sensitive to perturbation. Therefore, we developed a set of transcriptional switches for cell states control that were constructed by the inversion effect of site-specific recombinases on terminators. Such a switch could respond to a pulse signal and maintain the new state by itself until the next input. With a bottom-up design principle, we first characterized the terminators and recombinases. Then the mutual interference was studied to select compatible pairs, which were used to achieve one-time and two-time state transitions. Finally, we constructed a biological seven-segment display as a demonstration to prove such switch's immense potential for application.
Recombinases ; metabolism