Anticoagulants are the cornerstone of the prevention and treatment of thromboembolic diseases. Existing parenteral and oral anticoagulants achieve effective control of thrombosis by interfering with key aspects of the coagulation cascade reaction， but this is accompanied by an increased risk of bleeding. FⅪ inhibitors， anticoagulants targeting coagulation factor Ⅺ （FⅪ）， can block the amplification phase of the thrombin generation process by inhibiting FⅪ， reducing thrombogenesis with less impact on normal hemostatic effects， and have become one of the most promising new anticoagulants. There are currently no marketed FⅪ inhibitor drugs， while FⅪ inhibitors in phase Ⅱ or phase Ⅲ clinical trials include 3 classes：antisense oligonucleotide， monoclonal antibody and small molecule inhibitors. In addition， most of the natural inhibitors and nucleic acid aptamers targeting FⅪ are under preclinical development. As new target drugs for anticoagulation therapy， FⅪ inhibitors are expected to become a safer and more effective therapeutic option， compensating for the limitations of current anticoagulants and providing patients with more effective thromboprophylaxis and therapeutic options while reducing the risk of bleeding.

Objective To investigate the fluorescence resonance energy transfer (FRET) effect between dylight (DL) and AuNP (AuNP), and to construct a new fluorescence immunoassay for insulin in combination with the immunocompetition method. Methods Insulin antigen (Ag) and insulin antibody (Ab) were conjugated with DL and AuNP respectively to form DL-Ag conjugate and AUNp-AB conjugate. A novel fluorescence immunoassay for insulin was developed on the basis of FRET effect and the immune competition response between them. Then the performance of the method was evaluated and its application in actual samples was explored. Results The fluorescence immunoassay showed high sensitivity (0.015 ng/mL), short measurement time (4 min) and good specificity. It was successfully used in the measurement of serum insulin, and the recovery was between 96.9% and 121.1%. Conclusion FRET effect between AuNP and DL can be applied to develop a fluorescence immunoassay for the measurement of serum insulin.
Fluorescence Resonance Energy Transfer ; Insulin ; Immunoassay

Objective : To investigate whether NaHS，a hydrogen sulfide donor，can improve myocardial injury in sepsis by inhibiting oxidative stress and activating the Xc －/ GPX4 signaling pathway in ferroptosis． Methods : Lipopolysacc-haride(LPS) induced H9c2 in rat cardiomyocytes to form an in vitro model of myocardial injury in sep- sis，which was divided into Control group，LPS group and LPS + NaHS group．The kits were applied to detect the changes of cardiomyocyte viability，Fe2 + ，LDH and CK-MB，determine the levels of oxidative stress indexes GSH and MDA，detect the changes of cellular ROS and mitochondrial membrane potential levels by fluorescent probes， and detect the expression levels of ferroptosis regulatory proteins SLC7A11 and GPX4 by Western blot. Results: Compared with the Control group，H9c2 cell viability decreased，Fe2 + concentration increased ，GSH ，MDA and ROS levels increased，mitochondrial JC-1 monomer increased ，expression levels of ferroptosis regulatory proteins SLC7A11 and GPX4 decreased，and cell damage increased after LPS stimulation (P＜0. 05) ．Compared with the LPS group，NaHS attenuated LPS-induced H9c2 cell injury and elevated Fe2 + concentration，decreased the level of LPS-induced oxidative stress in H9c2 cells ，and increased the expression levels of ferroptosis regulatory proteins SLC7A11 and GPX4 (P＜0. 05 ) ． Conclusion The mechanism by which NaHS attenuates myocardial injury in sepsis may be related to the inhibition of oxidative stress and activation of the Xc －/ GPX4 signaling pathway in fer- roptosis.

Objective:To investigate whether cachexia affects the treatment effect of immune checkpoint inhibitors for non-small cell lung cancer (NSCLC).Methods:The prognosis of 62 patients with advanced NSCLC who received anti-programmed cell death-1 (PD-1) in Henan Provincial People′s Hospital from 2019 to 2021 were retrospectively analyzed. The cachexia was evaluated before and after the second course of immunotherapy. Kaplan-Meier and Log rank methods were used for survival analysis, Cox regression model was used for multivariate analysis, and Spearman′s correlation analysis was used for correlation analysis.Results:After the second course of immunotherapy, psoas major muscle area (PMMA) values of the cachexia group and the control group were (14.10±4.09) and (11.66±3.22) cm 2 respectively, with statistics significance ( P=0.001). The level of Prealbumin and body weight were correlated with cachexia ( P<0.05). The 6-month and 1-year survival rates of 62 cases in the whole group were 58.6% and 42.5%, respectively. The progression-free survival (PFS) in the control group (7.6 months) was higher than that in the cachexia group (3.8 months, P=0.006). The PFS in patients with high expression of PD-L1 (7.1 months) was longer than that of patients with low expression (3.8 months, P=0.009). The overall survival (OS) in the cachexia group (6.3 months) was lower than that in the control group (18.2 months, P=0.006). The OS in patients with high expression of PD-L1 (14.5 months) was longer than that of patients with low expression (1 months, P=0.038). The level of Prealbumin, the level of PD-L1 expression and the change rate of PMMA were related to the OS of the patients ( P<0.05). The level of Prealbumin and the change rate of PMMA were the independent influencing factors of the OS ( P<0.05). The PMMA and the level of Prealbumin were negatively correlated ( r=-0.003 8, P<0.05). Conclusion:Cachexia has a negative impact on the outcomes of patients who received anti-PD-1 immune checkpoint inhibitor therapy.

Objective:To investigate whether cachexia affects the treatment effect of immune checkpoint inhibitors for non-small cell lung cancer (NSCLC).Methods:The prognosis of 62 patients with advanced NSCLC who received anti-programmed cell death-1 (PD-1) in Henan Provincial People′s Hospital from 2019 to 2021 were retrospectively analyzed. The cachexia was evaluated before and after the second course of immunotherapy. Kaplan-Meier and Log rank methods were used for survival analysis, Cox regression model was used for multivariate analysis, and Spearman′s correlation analysis was used for correlation analysis.Results:After the second course of immunotherapy, psoas major muscle area (PMMA) values of the cachexia group and the control group were (14.10±4.09) and (11.66±3.22) cm 2 respectively, with statistics significance ( P=0.001). The level of Prealbumin and body weight were correlated with cachexia ( P<0.05). The 6-month and 1-year survival rates of 62 cases in the whole group were 58.6% and 42.5%, respectively. The progression-free survival (PFS) in the control group (7.6 months) was higher than that in the cachexia group (3.8 months, P=0.006). The PFS in patients with high expression of PD-L1 (7.1 months) was longer than that of patients with low expression (3.8 months, P=0.009). The overall survival (OS) in the cachexia group (6.3 months) was lower than that in the control group (18.2 months, P=0.006). The OS in patients with high expression of PD-L1 (14.5 months) was longer than that of patients with low expression (1 months, P=0.038). The level of Prealbumin, the level of PD-L1 expression and the change rate of PMMA were related to the OS of the patients ( P<0.05). The level of Prealbumin and the change rate of PMMA were the independent influencing factors of the OS ( P<0.05). The PMMA and the level of Prealbumin were negatively correlated ( r=-0.003 8, P<0.05). Conclusion:Cachexia has a negative impact on the outcomes of patients who received anti-PD-1 immune checkpoint inhibitor therapy.

Objective:To investigate the mechanism of cathepsin G(CatG) in improving the treatment efficacy of T cells in gliomas.Methods:(1) Clinical data of 397 glioma patients in the glioma database were collected, Kaplan-Meier method was used to perform survival analysis, and the correlation between CTSG and β2-microglobulin ( β2M) mRNA expressions in glioma tissues was analyzed. (2) Glioma stem cell (GSC) 387 and GSC3565 were isolated from glioblastoma and differentiated into differentiated glioma cell (DGC) 387 and DGC3565, respectively; GSC387 was divided into CatG group and CatG inhibitor group, and cells in the CatG group and CatG inhibitor group were cultured with 0.1 μg/μL recombinant human leukocyte antigen (HLA)-A*02:01 and HLA-B*15:01 combined with 4 ng/μL CatG or 10 mol/L CatG inhibitor for 10 min, respectively; the expressions of HLA-A*02:01 and HLA-B*15:01 were detected by Thomas bright blue staining, and the protein expressions of major histocompatibility complex (MHC)-I and MHC-DR were detected by Western blotting. (3) GSC387, GSC3565, DGC387, and DGC3565 were divided into 4 groups, including CatG group, CatG inhibitory group, blank antibody group 1 and blank antibody group 2, respectively; 4 ng/μL CatG, 10 μmol/L CatG inhibitor, blank antibody 1 and blank antibody 2 were added into the cells from the 4 groups for 24 h, and the expression of HLA-ABC was detected by flow cytometry. (4) GSC387, GSC3565, DGC387, and DGC3565 were divided into CatG group and CatG inhibitory group, respectively; luciferase assay was used to detect the influence of CatG in the killing effects of T cells and natural killer cells. Results:(1) The survival rate in patients from CTSG mRNA high expression group was significantly higher than that in patients from CTSG mRNA low expression group, and the survival rate in patients from β2M mRNA low expression group was statistically higher than that in patients from β2M mRNA high expression group ( P<0.05); a negative correlation between CTSG mRNA and β2M mRNA expressions was noted in glioma tissues ( r=-9.160, P=0.000). (2) Thomas bright blue staining showed that the expressions of HLA-A*02:01 and HLA-B*15:01 obviously increased in the CatG group as compared with those in the CatG inhibitor group; Western blotting showed that as compared with the CatG inhibitor group, the CatG group had increased MHC-I expression, and decreased expressions of α and β chains of MHC-DR. (3) Flow cytometry showed that the HLA-ABC expressions in GSC387 and GSC3565 of the CatG group were statistically higher than those in the CatG inhibitor group ( P<0.05). (4) Luciferase assay showed that, as compared with the CatG inhibitor group, the CatG group had statistically higher proportion of T cells killing GSCs ( P<0.05). Conclusion:CatG can improve the immunotherapy efficacy in GSCs, mainly by increasing the MHC-I expression on the cell surface.

To explore the evaluation method of evidence body for acupuncture and moxibustion clinical practice guidelines (CPGs), and to provide methodological support for the development of acupuncture and moxibustion CPGs and to promote the establishment and application of acupuncture and moxibustion standard. Based on the academic characteristics of TCM acupuncture, the stratified evidence evaluation method was proposed. Firstly, evidences were collected from the ancient literature, modern literature and clinical experience, and were scientifically distinguished and evaluated. Secondly, all the evidences were integrated as body of evidence by analytic hierarchy process (AHP). Lastly, based on the general index (GI), the clinical recommendations for acupuncture were proposed. Based on the stratified evidence evaluation method, acupuncture and moxibustion CPGs were developed in a more scientific and objectiveway with the comprehensive and sufficient evidences.
Acupuncture Therapy ; Moxibustion

Objective To investigate the clinical features, electrophysiological characteristics and treatment of stiff-person syndrome (SPS). Methods Medical records were retrospectively collected from 8 SPS patients to analysis their clinical features, laboratory studies, electromyography characteristics and treatment effect. Results All 8 patients presented with classic SPS, experienced progressive muscle stiffness, rigidity and spasm with paroxysmal exacerbation, which most frequently involved the thoracolumbar paraspinal muscles and bilateral lower limbs and other parts of body including thoracic and abdominal wall, upper limbs, neck, head and face. Five patients underwent electromyography and the results showed continuous motor unit activity (CMUA) in the involved muscles at rest. CMUA reduced markedly in 2 cases after intravenous diazepam. Anti-glutamic acid decarboxylase (GAD) antibody testing was positive in one of 5 tested cases. All 8 patients experienced partially symptomatic relief for their muscle rigidity and spasm after benzodiazepines. Combined immunotherapy further attenuated the symptoms in two cases receiving intravenous immunoglobulin (IVIG) and one case receiving glucorticosteroids, respectively. Symptoms were completely relieved following thymectomy in 2 cases with thymoma. Conclusion SPS is characterized by progressive muscle stiffness, rigidity and spasm with paroxysmal exacerbation affecting the axial trunk and bilateral lower limbs most frequently. Electromyography indicates CMUA in these involved muscles at rest. Treatment with benzodiazepines combined with immunotherapy can improve the neurological manifestations. Thymectomy can completely relieve symptoms of SPS in patiens with thymoma.

Objective To investigate the effects of Epimedium ,Astragalus ,Radix Puerariae on DMT1 expression in the cere‐bral cortex of APPswe/PS1ΔE9 double transgenic mice model of AD .Methods A total of 60 specific‐pathogen‐free male APPswe/PS1ΔE9 double transgenic mice aged 6 months were equally and randomly assigned to model ,Epimedium ,Astragalus ,Radix puerari‐ae ,compound and DFO groups .An additional 10 6‐month‐old C57BL/6J mice served as negative control group .Using immunohisto‐chemistry and molecular biology methods to investigate the effects of a compound combining the effective components of Epimedi‐um ,Astragalus ,Radix puerariae on DMT1 expression in the cerebral cortex of APPswe/PS1ΔE9 double transgenic mice model of AD . Results Immunohistochemical staining results revealed that DM T 1 positive cell did not show in negative control group .DM T1 ex‐pression was higher in model group compared with the negative control group .DMT1 expression was lower in the compound and deferoxamine groups than in the model group .No significant difference was detected in DM T 1 expression between deferoxamine and compound groups .RT‐PCR ,Western blot and immunohistochemical staining results showed no significant difference .Conclusion These compounds can downregulate DMT1 expression and inhibit iron overload in the cerebral cortex of mice with Alzheimer′s dis‐ease ,reduce iron overload induced impairment of the central nervous system .

Primary immunodeficiency diseases (PIDs) represent a typical group of rare diseases with high mortality and disability with regard to long-term outcome.Saving life and improving quality of life of the patients with PID remain to be the responsibility of the stakeholders from all aspects of the society.As the central player in the long run of PID campaign,pediatric immunologists should play a critical role in providing key epidemiological data in favor of political decision-making and social resource distribution.