Immunotherapy has emerged as a revolutionary approach to treat immune-related diseases. Dendritic cells (DCs) play a pivotal role in orchestrating immune responses, making them an attractive target for immunotherapeutic interventions. Modulation of gene expression in DCs using genome editing techniques, such as the CRISPR-Cas system, is important for regulating DC functions. However, the precise delivery of CRISPR-based therapies to DCs has posed a significant challenge. While lipid nanoparticles (LNPs) have been extensively studied for gene editing in tumor cells, their potential application in DCs has remained relatively unexplored. This study investigates the important role of cholesterol in regulating the efficiency of BAMEA-O16B lipid-assisted nanoparticles (BLANs) as carriers of CRISPR/Cas9 for gene editing in DCs. Remarkably, BLANs with low cholesterol density exhibit exceptional mRNA uptake, improved endosomal escape, and efficient single-guide RNA release capabilities. Administration of BLAN_mCas9/gPD-L1 results in substantial PD-L1 gene knockout in conventional dendritic cells (cDCs), accompanied by heightened cDC1 activation, T cell stimulation, and significant suppression of tumor growth. The study underscores the pivotal role of cholesterol density within LNPs, revealing potent influence on gene editing efficacy within DCs. This strategy holds immense promise for the field of cancer immunotherapy, offering a novel avenue for treating immune-related diseases.

Purpose To explore the value of iodine concentration(IC)and effective atomic number(Zeff)derived from dual-layer spectral detector CT in diagnosing the regional lymph nodes of gastric cancer.Materials and Methods From May 2021 to September 2022,108 patients pathologically confirmed with gastric cancer(92 non-metastatic lymph nodes and 116 metastatic lymph nodes),who underwent dual-phase enhanced dual-layer spectral detector CT scanning preoperatively in the Second Affiliated Hospital of Guangzhou University of Chinese Medicine,were included in the study.The imaging and clinical data of all cases were collected,retrospectively.According to preoperative CT images,the short diameter,margin and enhancement homogeneity of the lymph nodes were evaluated.The IC and Zeff were obtained from the delineated region of interest of the solid portion of the maximum layer of the target lymph nodes,and the normalization IC(nIC)and normalization Zeff(nZeff)were calculated.The difference of parameters between two groups were compared.The efficiency of differential diagnosis and combined diagnosis of each parameter was analyzed.The area under the receiver characteristic curve(AUC)for each parameter were compared.Results There were statistical differences in short diameter,margin and enhancement homogeneity between the two groups(t/x2=-7.575,30.971,20.913,all P＜0.001).In the benign lymph nodes group,all spectral parameters in the arterial phase(AP)were lower than those in malignant lymph nodes,while the parameters in the venous phase(VP)were higher(t=-10.723-12.610,all P＜0.001).Among the three conventional features,the AUC of short diameter was the highest(0.749).Compared with the AUC values of ICAP and ZeffVP,the AUC values of short diameter and quantitative parameters had no statistical difference(P＞0.05),but were lower than those of other spectral parameters.Among the spectral parameters,the AUC value of nICVP was the highest(0.925).The sensitivity and specificity of the spectral parameters were higher than those of conventional features.The diagnostic efficacy of the multi-parameter combined model was superior to that of the univariate model.The AUC value of the combined model with conventional features was 0.815.Among the combined model based on CT spectral parameters,the diagnostic efficacy of model combined with the parameters of arteriovenous phase was the highest(AUC=0.993).Conclusion The quantitative parameters of dual-phase enhancement derived from dual-layer spectral detector CT can effectively evaluate the characteristics of gastric cancer regional lymph nodes,and the diagnostic efficiency is higher than the conventional characteristics of lymph nodes.The multi-parameter combined model can improve the diagnostic efficiency.

Objective To evaluate the effect and mechanism of terminal fucosylation inhibitor 2-deoxy-D-galactose (2-D-gal) on ciclosporin (CsA)-induced renal epithelial-mesenchymal transition (EMT). Methods Fifteen male C57BL/6 mice aged 8-10 weeks were randomly and evenly divided into the control group (Ctrl group), CsA group and CsA+2-D-gal group (n=5). The expression levels of fucosyltransferase 1 (FUT1), EMT-associated proteins including E-cadherin, Vimentin, α-smooth muscle actin (α-SMA) in the kidney tissues of the Ctrl and CsA groups were detected by Western blot. The expression levels of terminal fucose in the kidney tissues of Ctrl and CsA groups were determined by immunofluorescence. The renal fibrosis of mice in each group was evaluated by Masson staining. The blood urea nitrogen and serum creatinine levels of mice in each group were detected. The in vitro EMT model of renal tubular epithelial cell HK2 was induced by CsA. HK2 cells were stimulated with 0, 2.5, 5.0 and 10.0 μmol/L CsA for 24 h, respectively. In addition, HK2 cells were divided into the Ctrl, 2-D-gal, CsA and CsA+2-D-gal groups. The morphology of HK2 cells after stimulation with different concentrations of CsA and in each group was observed. The expression levels of FUT1, E-cadherin, Vimentin and α-SMA in HK2 cells after stimulation with different concentrations of CsA and in each group were detected by Western blot. The expression level of terminal fucose in HK2 cells of the Ctrl and CsA groups was measured by immunofluorescence. Results Compared with the Ctrl group, the relative expression of E-cadherin protein was down-regulated, those of FUT1, Vimentin and α-SMA proteins were up-regulated (all P < 0.05), and that of terminal fucose in the mouse kidney tissues was up-regulated in the CsA group. Compared with the Ctrl group, the blood urea nitrogen and serum creatinine levels in the CsA and CsA+2-D-gal groups were up-regulated (all P < 0.05). Compared with the CsA group, the blood urea nitrogen and serum creatinine levels in the CsA+2-D-gal group were down-regulated (both P < 0.05). Compared with the Ctrl group, the collagen fiber deposition was increased and the relative expression of α-SMA protein was up-regulated in the mouse kidney tissues of CsA and CsA+2-D-gal groups (all P < 0.05). Compared with the CsA group, the collagen fiber deposition was decreased and the relative expression of α-SMA protein in the mouse kidney tissues was down-regulated in the CsA+2-D-gal group (both P < 0.05). With the increase of CsA concentration, the morphology of HK2 cells gradually became longer and thinner from original normal cobblestone shape, the relative expression levels of FUT1, Vimentin and α-SMA protein in HK2 cells were up-regulated, and that of E-cadherin protein was down-regulated in a concentration-dependent manner. Compared with the Ctrl group, the expression level of terminal fucose of HK2 cells was up-regulated in the CsA group. After CsA treatment combined with 2-D-gal intervention, the morphology of HK2 cells in the CsA+2-D-gal group was restored to resemble that of normal HK2 cells. Compared with the CsA group, the relative expression of E-cadherin protein in HK2 cells was up-regulated, whereas those of Vimentin and α-SMA proteins were down-regulated in the CsA+2-D-gal group (all P < 0.05). Conclusions CsA may induce EMT both in vivo and in vitro, and the terminal fucosylation is increased. 2-D-gal may inhibit CsA-induced EMT by suppressing the terminal fucosylation.

Objective:To investigate and analyze the somatic symptom disorder, anxiety and depression in patients with myocardial bridge.Methods:A total of 276 patients with myocardial bridge diagnosed by coronary angiography (CAG) were enrolled in the department of cardiology of Renji hospital in Shanghai from June to December in 2016. There were 151 cases of simple myocardial bridge (no coronary stenosis or coronary artery stenosis <30%) and 125 cases of complex myocardial bridge (combined with coronary stenosis >30%). A total of 1067 patients with myocardial bridge without coronary angiography were collected at the same time. Self-rating somatic symptom scale (SSS), generalized anxiety disorder (GAD-7) and patient health questionnaire (PHQ -9) were given to these patients during hospitalization. At the same time, somatic symptoms disorder and anxiety and depression in the myocardial bridge group and non-myocardial bridge group were compared.Results:The prevalence of somatic symptom disorder in patients with myocardial bridge was higher than that in non-myocardial bridge patients (35.86% vs 28.30%, P<0.05). There was significant correlation between somatic symptom disorder and depression and anxiety, with correlation coefficients of 0.629 and 0.565, respectively. The prevalence of depression and anxiety in myocardial bridge patients was higher than that in non-myocardial bridge patients (depression: 23.91% vs 22.11%. P=0.467; anxiety: 17.02% vs 14.15%, P=0.22), but there was no statistical difference. For male patients or female patients, the prevalence of somatic symptom disorder, depression and anxiety in the simple myocardial bridge patients were higher than those in the complex myocardial bridge patients, but there was no statistical difference. The most common non-specific somatic symptoms disorder in patients with myocardial bridge were fatigue (64.5%), followed by sleep disorders (63.8%) and decreased attention (63.0%). Conclusion:The somatic symptom disorder in patients with myocardial bridge is significantly higher than that in non-myocardial bridge group. Especially for patients with myocardial bridge with non-specific somatic symptoms, early identification of somatic symptoms disorder of myocardial bridge patients will be beneficial to proper clinical invitation.

Objective To analyze the efficacy of entecavir (ETV) combined with Peg IFNα-2b in chronic hepatitis B ( CHB) patients with low levels HBsAg following initial ETV treatment.Methods Sixty-nine CHB outpatients achieving serum HBsAg ＜2 000 IU/mL and HBV DNA＜100 IU/mL following initial ETV treatment in Pujiang People's Hospital and the First Affiliated Hospital of Zhejiang University School of Medicine from January 2014 to January 2016 were enrolled.Patients were randomly assigned in two groups: 39 patients in combination group received ETV (0.5 mg/d ) and Peg IFNα-2b (1.5 μg· kg－1· week －1, hypodermic injection), and 30 patients in ETV group received ETV (0.5 mg/d) alone.Serum HBsAg quantification, negative conversion rate of HBsAg and HBeAg , and levels of aminotransferase (ALT) were measured at baseline , 12th, 24th, 48th, 72th and 96th week after treatment.Results The levels of HBsAg in the combination group decreased gradually with the prolongation of therapy , which were lower than those in ETV group 24 week after treatment (Z＝－2.566,P＜0.05),and at 48th, 72th and 96th week (Z＝－3.499,－3.825 and －3.864,P＜0.01).Clearance of HBsAg appeared in the combination group at 24th week,the clearance rates were 7.70%(3/39) and 28.20%(11/39) at 24th and 96th week, respectively;while the clearance of HBsAg occurred in ETV group at 96th week, the clearance rate was only 3.30%(1/30).The negative conversion rates of HBsAg in combination group were higher than those in ETV group at 48th,72th and 96th week (P＜0.05 or＜0.01).In the combination group, there were 11 cases of clinical cure , 11 cases of clinical efficacy and 17 cases of clinical effectiveness , while there were 1, 1 and 28 cases in ETV group,respectively.The treatment effect of the combination group was better than that of ETV group(χ2＝18.496,P＜0.01).Serological conversion rates of HBeAg were 30.00%(6/20) and 65.00%(13/20) in combination group at 12th and 96th week, while those were 11.11%(2/18) and 22.22%(4/18) in ETV group at 48th and 96th week.There were significant differences in the HBeAg serological conversion rates at 12th, 24th, 72th and 96th week between two groups (P＜0.05 or ＜0.01). The levels of ALT in combination group increased at 12th and 24th week, which had significant difference compared with ETV group (Z＝－1.236 and －2.658,P＜0.05), and the ALT levels gradually declined 48 week after treatment in combination group and there were no statistical differences between two groups at other time points.The ETV combined with Peg IFNα-2b and low baseline HBeAg levels were associated with the clearance rate of HBsAg (both P＜0.01).Conclusions CHB patients with low HBsAg levels following initial ETV monotherapy can achieve high negative conversion rate of HBeAg and HBsAg with the combination treatment of ETV and Peg IFN α-2b.

Objective To observe the effect of short-term treatment with metfbrmin on psychological distress and metabolic feature in patients with polycystic ovary syndrome(PCOS).Methods Ninety women were diagnosed as cases of PCOS based on the 2003 Rotterdam criteria.These patients were divided into three groups:group A,consisted of 26 subjects treated with mefformin only; group B,36 cases treated with metformin plus Dane-35 ; and group C,28 cases treated with placebo and Dane-35.The treatment was carried out for 3 months.Clinical and metabolic parameters were observed.The psychological distress was evaluated by the hospital anxiety and depression scale(HAD scale).Results Compared with group C,patients in group A and B showed significantly lower body mass index(BMI),waist circumference (WC),waist-hip ratio (WHR),serum fasting insulin,homeostasis model assessment for insulin resistance (HOMA-IR),total cholesterol (TC),triglyceride (TG),low density lipoproteincholesterol(LDL-C) levels as well as anxiety and depression scores,but higher high density lipoprotein-cholesterol (HDL-C) levels (all P＜0.05).Compared with group A,patients in Group B had significantly higher WC,WHR,and LDL-C levels(all P＜O.05).No significant differences in BMI,TG,HDL-C,anxiety and depression scores were found between group A and B (all P＞0.05).In group A,a positive correlation was found between the changes of metabolic feature (BMI,WHR,and HOMA-IR) and the changes of anxiety and depression scores (all P ＜0.05).Conclusion Short-term treatment with metformin will benefit patients with polycystic ovary syndrome,regarding psychological distress and metabolic features.

Objective To assess the reliability and validity of the Somatic Self-rating Scale(SSS). Methods The sample consisted of 589 outpatients and 64 inpatients, which all the patients completed Zung's Scales at the same time to test criterion validity of the SSS. And 24 inpatients were selected randomly to take a retest after two weeks without antidepressant treatment. Result The test-retest reliability of the SSS was 0.96 and the Cronbach' s αcoefficients was 0.89. The correlation coefficients of the four factors with the total scale score ranged from 0.76 to 0.88, the correlation coefficients of the four factors ranged from 0. 56 to 0.70. The confirmatory factor analysis showed that the fit index for GFI, NFI, NNFI, CFI, IFI were all approaching 0.9, REMEA = 0. 064,x2/df = 3.67. The factor loadings ranged from 0. 427 to 0.732. Correlations with Zung's Scales were in range of 0.74to 0. 80 while the four factors were in range of 0.55 to 0.74. Drawing the ROC curve and the area under the curve was 0.841. Conclusion The SSS has a high reliability and a good validity and can be used in general hospital.

Objective To evaluate the influence of right ventricular outflow tract septal ( RVS) pacing with right ventricular apical ( RVA) pacing on left ventricular remodeling and brain natriuretic peptide ( BNP). Methods Sixty patients with indication of pacemaker implantation were randomized into two groups, RVA group and RVS group. BNP was measured with ELISA, and echocardiography was performed to measure the left ventricular end diastolic volume ( LVEDD), left ventricular end systolic volume ( LVEDV) and left ventricular ejection fraction (LVEF) at pre-operation,and after 6,12,24 months pacing. The difference of cardiac remodeling and BNP in the two groups was observed. Results Compared to BNP at pre-operation (( 60. 2 ± 15. 7 ) ng/L) , BNP increased significantly in the RVA group at the 6th,12th and 24th month after operation( ( 108. 2 ±29. 8) , ( 190. 3 ±46. 7) ,(308. 2 ±56. 5)ng/L,respectively) (P <0. 05). In the RVS group,BNP increased only at 24 months after pacing ( (75. 2 ± 15. 8) ng/L vs. (63. 9 ± 15. 1 ) ng/L) (P < 0. 05). There was significant difference on BNP between the two groups. LVEDD,LVEDV increased,LVEF declined at 12 months after pacing in the RVA group,which were not observed in the RVS group. There was significant difference on LVEDD,LVEDV and LVEF in the RVA group (P< 0. 05) between the 12th month and pre-operation,and there were no significant difference in the RVS group (P > 0. 05). Conclusions Compared to RVA pacing,RVS pacing was more beneficial to improve heart function,prevent cardiac remodeling and decline the activation of nerve-endocrine.

OBJECTIVETo determine the economic effects on the prevention of dental fear (DF) by pre-operation-education or local anesthesia method during the process of tooth filling.

METHODSOne hundred and fifty school children of age 7 to 12, participating in this study, suffered from occlusal caries on their mandibular first molars. They were divided into 3 groups, with 25 boys and 25 girls each. Group A (pre-operation-education) was showed video tape on caries to, familiarized with clinic environment, including dental chairs and instruments; Group B (local anesthesia) received injection of 1.8 ml of 2% lidocaine for local anesthesia about 15 minutes before treatment; Group C (blank) received no intervention for DF. Thereafter, all subjects received same filling treatment. During that process, each DF case was evaluated blindly based on Venham's clinical ratings of anxiety and cooperation. Cost-minimization analysis was then studied.

RESULTSSignificant difference was found between groups, but not between group A and B in DF rate (5/50, 9/50 and 21/50, chi(2) = 15.503 1, P = 0.000 4). The costs of each case in 3 groups were 83.99 Yuan, 87.09 Yuan and 87.76 Yuan respectively.

CONCLUSIONBetween pre-operation-education and local anesthesia methods, the former was worth promoted for it's better clinical effect in DF prevention with lower cost.

Child ; Dental Anxiety ; economics ; epidemiology ; prevention & control ; Dental Caries ; therapy ; Dental Cavity Preparation ; economics ; methods ; Female ; Humans ; Incidence ; Male

ObjectiveTo establish and evaluate the methods which can quantify the ocular hemodynamics by combined scanning laser ophthalmoscopy video fluorescein angiography with computerized digital image processing system．MethodsAfter acquiring serial digital images by video capture from some angiography videotapes and measuring the gray values of some fixed areas over time in these serial images，the gray value curves and some parameters were obtained．The inter- and intra-observer reproducibility surveys were carried out．ResultsThe hemodynamic parameters were obtained，which included sectional choroidal filling time and rate，sectional papillary filling time and rate，retinal arterial and venous filling time and rate，arterovenous passing time and diameter of artery and vein．The intra- and inter-observer reproducibility were fairly good．Conclusioncomputerized digital image processing system is useful for understanding retinal and choroidal circulation．