The E protein of dengue virus type Ⅱ was presented on ferritin of Helicobacter pylori to construct a novel dengue nanoparticle vaccine candidate,and the immunological indexes of the vac-cine were evaluated,aiming to provide new ideas for the development of dengue vaccine.The re-combinant plasmid of E-Ferritin was optimized and synthesized,and then transfected into HEK-293F cells.The recombinant protein was expressed,identified,purified and analyzed.Mice were im-munized with E-Ferritin nanoparticle vaccine by intramuscular injection on the hind limbs on the day 0,14 and 28.ELISA,neutralization test,flow cytometry and lymphocyte proliferation test were used to detect the levels of specific antibodies,neutralizing antibodies,CD3+,CD4+and CD8+T lymphocytes in spleen cells and the proliferation of spleen lymphocytes after specific stimulation.The target protein with a size of about 69 kDa was expressed in the cells with a single band.The purified protein concentration was 0.407 g/L,and the purity was 82.32％.The results from transmission electron microscopy showed that E-Ferritin protein could be recombined into a particle structure with a particle size of about 50 nm.The results of mouse immune experiments showed that E-Ferritin protein had good immunogenicity.The average specific antibody titer of E-ferritin protein in serum was 1∶92 160 after immunization 42 d.The main subclass of antibody was IgGl.The results of flow cytometry showed that E-Ferritin as an immunogen could induce higher levels of CD4+and CD8+T lymphocyte immune response.In lymphocyte proliferation test,the level of specific stimulation in the vaccine group was significantly higher than that in the non-specific stimulation group.In conclusion,the dengue virus envelope protein ferritin nanoparticle vaccine constructed in this study has good immunogenicity,which can provide reference for the de-velopment of new dengue vaccine candidates.

Objective To investigate the effect and mechanism of IL-17 on heart failure(HF)in hy-pertensive rats based on NF-κB/sarco-endoplasmic reticulum calcium ATPase 2(SERCA2)signa-ling pathway.Methods Thirty SPF male spontaneously hypertensive rats(SHR)aged 6-8 weeks were divided into control group,model group,phosphate buffer salt(PBS)solution injec-tion group(PBS group),IL-17 protein injection group(IL-17 group)and IL-17 and antibody injec-tion group(IL-17+IgG group),with 6 animals in each group.Hypertensive HF model was estab-lished,and corresponding agents were applied to the PBS group,IL-17 group and IL-17+IgG group intraperitoneally,respectively.The role of IL-17,NF-κB and SERCA2 in hypertensive HF was studied with HE staining,immunohistochemical assay,Western blotting,RT-qPCR and ELISA.Results Significantly higher serum levels of NT-proBNP and IL-17,enhanced myocardial expression of IL-17 mRNA and NF-κB protein,lower serum VEGF level,and down-regulated pro-tein level of SERCA2 in heart tissue were observed in the model group and the PBS group when compared with the control group(P＜0.01).The IL-17 group had obviously higher serum NT-proBNP and IL-17 levels and myocardial expression of IL-17 mRNA and NF-κB protein,and reduced serum VEGF level and SERCA2 protein level in heart tissue than the model group(P＜0.01).IL-17+IgG treatment resulted in notably lower serum IL-17 level and myocardial NF-κB protein level when compared with those of model group(8.98±1.20 vs 11.19±1.22,0.88±0.03 vs 0.93±0.03,P＜0.01),and also resulted in remarkably reduced serum levels of NT-proBNP and IL-17 and myocardial expression of IL-17 mRNA and NF-κB protein but increased serum VEGF level and SERCA2 protein level in heart tissue when compared with the IL-17 group(P＜0.01).The heart rate,SBP,IVSd,LVPWd,LVEDD and LVESD were significantly lower,while LVFS was notably higher in the IL-17+IgG group than the model group and IL-17 group(P＜0.01).The IL-17+IgG group had obviously higher LVEF than the IL-17 group[(70.81±6.50)％vs(62.77±5.43)％,P＜0.01].Conclusion IL-17/NF-κB/SERCA2 signaling pathway is involved in the regulation of inflammatory response after hypertensive HF,and inhibiting IL-17 can effective-ly improve the cardiac dysfunction caused by hypertensive HF.

Objective:To investigate the role of D-amino acid oxidase (DAAO) in arsenic exposure induced learning memory impairment in offspring mice.Methods:Eighteen Kunming pregnant mice were randomly divided into a non-treatment group (distilled water, n = 6) and an arsenic exposed group [60 mg/L sodium arsenite (NaAsO 2) in drinking water, n = 12] by randomized numerical table method. From the first day of pregnancy until weaning, the mother mice were exposed to arsenic. After weaning, the offspring mice were exposed to arsenic through free drinking water until the end of the experiment. Six weeks after birth, the offspring mice of the arsenic exposed group were divided into a NaAsO 2 group and a NaAsO 2 + DAAO inhibitor 6-chlorobenzo [d] isoxazol-3-ol (CBIO) group according to the group of mother mice. The offspring mice of non-treatment group as the control group. The offspring mice of the control group and NaAsO 2 group were given intraperitoneal injection of 0.5% sodium carboxymethylcellulose, while the NaAsO 2 + CBIO group was given intraperitoneal injection of 60 mg/kg CBIO for two consecutive weeks. At the end of the intervention, the spatial learning and memory abilities of the offspring mice were measured using a Y-maze experiment. After cardiac perfusion, brain tissue was taken from the offspring mice and the hippocampus was isolated. Hematoxylin-eosin (HE) staining was used to observe the morphology of the hippocampal neurons in the offspring mice. D-serine content was determined by ultra-high performance liquid chromatography tandem triple quadrupole mass spectrometry (UHPLC-MS/MS). The mRNA levels of synaptophysin (SYP), synaptosomal-associated protein 25 (SNAP25), postsynaptic density 95 (PSD95), DAAO, and N-methyl-D-aspartate receptor (NMDAR) subunits NR1, NR2A, NR2B were measured by real-time fluorescence quantitative PCR. Results:There was a statistically significant difference in the offspring mice alternating response rate of the Y-maze experiment among the control, NaAsO 2, and NaAsO 2 + CBIO groups [ (58.06 ± 3.78) %, (48.61 ± 5.75)%, (56.25 ± 6.76)%, F = 4.87, P = 0.023], and the NaAsO 2 group was significantly lower than the control and NaAsO 2 + CBIO groups ( P < 0.05). The HE staining results showed that the control group had a higher number of neuronal cells in the CA1 and CA3 regions of the hippocampal tissue, and they were in good condition. The number of neuronal cells in the NaAsO 2 group decreased, with irregular morphology and unclear cell contours. The NaAsO 2 + CBIO group had a higher number of neuronal cells, improved cell morphology and contour, and reduced tissue damage. There were statistically significant differences in D-serine content and mRNA levels of SYP, SNAP25, PSD95, DAAO, NR1, NR2A, and NR2B in the hippocampus of the offspring mice from the control, NaAsO 2, and NaAsO 2 + CBIO groups ( F = 5.41, 4.41, 10.16, 7.60, 6.98, 5.63, 6.53, 4.33, P = 0.017, 0.031, 0.002, 0.005, 0.007, 0.015, 0.009, 0.033). Among them, the D-serine content and mRNA levels of SYP, SNAP25, PSD95, NR1, and NR2B in the hippocampus of the NaAsO 2 group were significantly lower than those in the control group and NaAsO 2 + CBIO group, the mRNA level of NR2A was significantly lower than that in the control group, and the mRNA level of DAAO was significantly higher than that in the control and NaAsO 2 + CBIO groups ( P < 0.05). Conclusions:Arsenic exposure can induce the learning and memory impairment in offspring mice, reduce D-serine content in the hippocampus, as well as the transcription level of NMDAR subunits and synapse-associated proteins, up-regulate the transcription levels of DAAO; and inhibit DAAO, leading to increased D-serine content, the transcription levels of NMDAR subunits and synapse-associated proteins, improving arsenic exposure induced learning and memory impairment in the offspring mice.

Objective To observe the value of deep learning combine with radiomics based on MRI for evaluating H3 K27 status of midline gliomas.Methods Totally 127 patients with diffuse midline glioma H3 K27-altered(H3-DMG)and 127 patients with midline glioblastoma(GBM)without H3 K27 mutation were retrospectively enrolled.The patients were randomly divided into training set(n=204)and test set(n=50)at the ratio of 8:2.U-Net neural network visual and radiomics features of tumors were extracted based on MRI,and a deep learning radiomics model was established,its value for evaluating H3 K27 status was observed.Results Based on training set,0.500 was obtained as the security score partition value for the model classification task.In test set,the median safety score of the obtained deep learning radiomics model for evaluating H3 K27 status of H3-DMG and GBM was 0(0,0)and 0.999(0.616,1.000),respectively,for the former was lower than for the latter(Z=-5.114,P＜0.001).The sensitivity,specificity,accuracy and area under the curve of deep learning radiomics model for evaluating H3 K27 status in training set was 93.14％,81.37％,87.25％and 0.953(95％CI[0.923,0.976]),respectively,while was 88.00％,80.00％,84.00％and 0.922(95％CI[0.829,0.986])in test set,respectively.Conclusion Deep learning radiomics based on MRI could accurately evaluate H3 K27 status of midline gliomas.

BACKGROUND:The impact of the endometrium's structure and spiral artery blood flow parameters on the pregnancy rate of artificial insemination by husband remains unclear.This study identified the independent factors and constructed a prediction model with good clinical application efficacy after calibration of other confounding factors. OBJECTIVE:To construct and validate a clinical pregnancy prediction model for artificial insemination by husband based on endometrial structure and uterine spiral artery blood flow parameters. METHODS:A retrospective analysis was conducted on 1 299 patients who underwent artificial insemination by husband treatment at Changzhou Maternal and Child Health Hospital from January 2017 to January 2021.The non-pregnancy group consisted of 1 182 patients,while the pregnancy group included 117 patients.Out of these patients,93 cases were successfully matched between the pregnancy and non-pregnancy groups using a 1∶1 propensity score matching method.Single-factor and multi-factor analyses were used to screen the endometrial structure and uterine spiral artery blood flow parameters to determine their influence on artificial insemination by husband outcomes.The optimal cutoff value was established for each independent influencing factor through receiver operating curve analysis and their risk trend affecting artificial insemination by husband pregnancy outcomes was analyzed using a restricted cubic spline.The clinical efficacy of this combined forecast model was tested by using clinical decision curve and clinical influence curve methods. RESULTS AND CONCLUSION:(1)There was no statistical significance in non-endometrial factors between the pregnancy group and the non-pregnancy group,and the data had a good balance by propensity score matching(P>0.05).(2)Single-factor analysis identified several subendometrial parameters as significant influencing factors of artificial insemination by husband pregnancy outcomes,including vascularization index,flow index,vascular flow index,resistance index,pulsatility index,maximum systolic velocity/end-diastolic velocity,thickness of average junction zone and maximum junction zone from the basal endometrium to the outer myometrium inner layer(P<0.05).(3)Multivariate logistic regression analysis revealed that thickness of average junction zone,pulsatility index,and vascular flow index were independent influencing factors of pregnancy outcomes of artificial insemination by husband,vascular flow index>thickness of average junction zone>pulsatility index.(4)Receiver operating characteristic curve analysis indicated that the area under receiver operating characteristic curve of vascular flow index was 0.704(0.629,0.779),and the optimal cutoff value was 6.26;the area under receiver operating characteristic curve of thickness of average junction zone was 0.660(0.582,0.739),and the optimal cutoff value was 6.38;the area under receiver operating characteristic curve of pulsatility index was 0.642(0.563,0.721),and the optimal cutoff value was 1.18.(5)The restricted cubic spline analysis revealed that artificial insemination by husband pregnancy outcomes were significantly positively affected when the vascular flow index was>6.24 or the thickness of average junction zone was≤6.55 mm,while a negative risk was associated with pulsatility index>1.27.(6)The clinical decision curve and clinical influence curve analyses exhibited that the combined prediction model had the maximum clinical net benefit at the threshold probability value of 0.17-0.93,and the ratio of loss to benefit was consistently less than 1 in the threshold probability range,indicating that the model had good clinical efficacy.(7)It is concluded that after adjusting for other confounding factors outside of the endometrium using propensity score matching and multifactorial logistic regression,the thickness of average junction zone,pulsatility index and vascular flow index were independent factors that influenced pregnancy outcomes of artificial insemination by husband.Through determining their optimal cutoff values and assessing their risk trends,it was confirmed that the combined prediction model had good predictive value and clinical efficacy.

Systemic lupus erythematosus(SLE)is characterized by a large number of anti-autoantibodies produced by abnormally activated B cells,which form immune complexes with autoantigens to induce systemic inflammation,leading to the involvement of multiple systems and organs.Existing treatment strategies targeting B cells(such as belimumab,telitacicept,and rituximab)had limited efficacy.Recently,CD 19 targeting chimeric antigen receptor(CAR)-T cell has shown excellent efficacy in the treatment of SLE.However,autologous CAR-T cell therapy has potential risks such as cytokine release syndrome(CRS),T cell tumors,and infections,and it is expensive.CAR-natural killer(NK)cell therapy,on the other hand,is an off-the-shelf cellular therapy with high safety,no neurotoxicity or tumorigenic risk,and with low treatment costs.Currently,CAR-NK cell has achieved remarkable research results in treating hematological malignancies,and preliminary clinical studies of CAR-NK cell therapy for SLE have showed good efficacy,excellent safety,and predictable durability.This article focuses on the characteristics of the new generation of universal CAR-NK cell and the latest clinical research results for the treatment of relapsed and refractory SLE,offering insights into its promising future in the treatment of SLE and B-cell-related autoimmune diseases.

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease affecting both upper and lower motor neurons (MNs) with large unmet medical needs. Multiple pathological mechanisms are considered to contribute to the progression of ALS, including neuronal oxidative stress and mitochondrial dysfunction. Honokiol (HNK) has been reported to exert therapeutic effects in several neurologic disease models including ischemia stroke, Alzheimer's disease and Parkinson's disease. Here we found that honokiol also exhibited protective effects in ALS disease models both in vitro and in vivo. Honokiol improved the viability of NSC-34 motor neuron-like cells that expressed the mutant G93A SOD1 proteins (SOD1-G93A cells for short). Mechanistical studies revealed that honokiol alleviated cellular oxidative stress by enhancing glutathione (GSH) synthesis and activating the nuclear factor erythroid 2-related factor 2 (NRF2)-antioxidant response element (ARE) pathway. Also, honokiol improved both mitochondrial function and morphology via fine-tuning mitochondrial dynamics in SOD1-G93A cells. Importantly, honokiol extended the lifespan of the SOD1-G93A transgenic mice and improved the motor function. The improvement of antioxidant capacity and mitochondrial function was further confirmed in the spinal cord and gastrocnemius muscle in mice. Overall, honokiol showed promising preclinical potential as a multiple target drug for ALS treatment.

Epilepsy is a common chronic neurological disease, and its comorbidity has attracted more attention.The proportion of epileptic children with mental disorders is also increasing year by year.Among them, children with epilepsy have more depression and anxiety disorders.Repeated seizures can easily cause depression and anxiety, and depression and anxiety can also induce epilepsy, thus the two affect each other.The assessment, screening, diagnosis and intervention of comorbid depression and anxiety in children with epilepsy have become an important part of clinical practice.This review summarized the relationship between epilepsy and depression and anxiety disorders in children, and its research progress on pathogenesis, clinical diagnosis, evaluation and treatment.

Objective:To explore the risk factors for early trauma-induced coagulopathy (TIC) following severe trauma in the elderly patients.Methods:A case-control study was used to analyze the clinical data of 317 elderly patients with severe trauma admitted to Second Affiliated Hospital of Zhejiang University School of Medicine between February 2015 and November 2020. There were 212 males and 105 females, aged 65-96 years [(72.6±6.8)years]. The patients were divided into TIC group ( n=32) and non-TIC group ( n=285) using the international normalised ratio (INR)>1.5 as the reference standard. Sex, age, trauma sites, injury severity score (ISS), Glasgow coma scale (GCS), first body temperature on admission, shock index(SI), first laboratory results of arterial blood gas, routine blood and coagulation, blood transfusion, usage of blood product, hospitalization days and clinical outcomes were compared between the two groups. Univariate and multivariate Logistic regression analysis were used to identify the risk factors for early TIC in patients with severe trauma. Results:Differences in sex, age, injuries to the face, chest and abdomen, GCS, first body temperature and hospitalization days were not statistically significant between the two groups (all P>0.05). The two groups showed statistical differences in the ratio of injuries to head, neck and extremities, ISS, SI, pH value, base excess (BE), lactate, hemoglobin (Hb), platelet (PLT) count (first detection, lowest level), activated partial thromboplastin time (APTT), thrombin time (TT), plasma fibrinogen (FIB), blood transfusion and usage of blood product and clinical outcomes (all P<0.05). According to the univariate analysis, injuries to the head, neck and extremities, ISS, first body temperature, SI, pH value, BE, lactate, Hb, PLT, APTT, TT and FIB were correlated with the occurrence of early TIC (all P<0.05). Multiple Logistic regressions analysis showed that SI ( OR=1.54, 95% CI 1.10-2.17, P<0.05), PLT ( OR=0.67, 95% CI 0.49-0.91, P<0.05) and FIB ( OR=0.56, 95% CI 0.40-0.78, P<0.01) were significantly correlated with the occurrence of early TIC. Conclusion:For elderly patients with severe trauma, higher SI, lower PLT count and lower concentration of FIB are independent risk factors for the incidence of early TIC.

Objective:To investigate the value of hypocalcaemia for predicting trauma-induced coagulopathy (TIC) in elderly trauma patients.Methods:The clinical data of elderly trauma patients in emergency ICU of the Second Affiliated Hospital of Zhejiang University School of Medicine from January 2015 to September 2020 were retrospectively analyzed, including age, sex, site of injury, injury severity score (ISS), Glasgow coma scale (GCS), admission arterial blood gas analysis (Ca 2+, K +), venous blood biochemical electrolyte (Ca 2+, K +, Na +); international normalized ratio (INR), activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen (FIB), use of blood products, length of stay, length of stay in ICU, total cost, and clinical prognosis. Receiver operating characteristic (ROC) curves and multivariate logistic regression analysis were performed to estimate the contribution of hypocalcaemia to triggering TIC in elderly trauma patients. Results:Totally 371 elderly trauma patients were included with a mean age of (72.5±6.8) years, and 248 (66.8%) were male. ISS score of the TIC group was higher than that of the non-TIC group [25(20, 34) vs. 21(16, 29)]. Compared with the non-TIC group, the incidence of chest injury, abdominal injury and limb injury were significantly higher , while the incidence of head and neck injury were significantly lower in the TIC group (all P<0.05). The biochemical blood calcium in the TIC group was significantly lower than that in the non-TIC group [(1.97±0.19) mmol/L vs. (2.15±0.16) mmol/L, P<0.001], but there was no significant difference in blood gas calcium between the two groups. The APTT value of the TIC group [(47.6±21.8) s vs. (33.8±4.1) s], PT value [(18.0±3.9) s vs. (13.7±0.8) s] were significantly higher than that of the non-TIC group, and FIB level was significantly lower than that of the non-TIC group[(1.7±0.8) g/L vs. (2.8±0.9) g/L] (all P<0.01). The utilization rate of blood products and the total cost in the TIC group were higher than that in the non-TIC group, while the recovery rate in the TIC group was lower than that in the non-TIC group (69.8% vs. 86.4%, P<0.001). Multivariate logistic regression analysis showed that hypocalcaemia was an independent risk factor for TIC in elderly trauma patients ( OR=5.830, 95% CI: 3.295-10.314). The area under ROC curve of correlation between biochemical calcium and TIC was 0.779 (95% CI: 0.728-0.831). The optimal diagnostic cut-off value was 2.06 mmol/L. Conclusions:The decrease of biochemical serum calcium level is an independent risk factor for TIC in elderly trauma patients. Positive correction of TIC in elderly trauma patients contributes to continuous improvement of clinical prognosis.