Objective: To reveal the molecular biological mechanism of a rare Dc-variant individual using PacBio third-generation sequencing technology. Methods: ABO and Rh blood type identification, DAT, unexpected antibody screening and D antigen enhancement test were conducted by serological testing. The absorption-elution test was used to detect the e antigen. RHCE gene typing was performed by PCR-SSP, and the 1-10 exons of RHCE were sequenced by Sanger sequencing. The full-length sequences of RHCE, RHD and RHAG were detected by PacBio third-generation sequencing technology. Results: Serological findings: Blood type O, Dc-phenotype, DAT negative, unexpected antibody screening negative; enhanced D antigen expression; no detection of e antigen in the absorption-elution test. PCR-SSP genotyping indicated the presence of only the RHCE c allele. Sanger sequencing results: Exons 5-9 of RHCE were deleted, exon 1 had a heterozygous mutation at c. 48G/C, and exon 2 had five heterozygous mutations at c. 150C/T, c. 178C/A, c. 201A/G, c. 203A/G and c. 307C/T. Third-generation sequencing results: RHCE genotype was RHCE 02N. 08/RHCE-D(5-9)-CE; RHD genotype was RHD 01/RHD 01; RHAG genotype was RHAG 01/RHAG 01 (c. 808G>A and c. 861G>A). Conclusion: This Dc-individual carries the allele RHCE 02N. 08 and the novel allele RHCE-D(5-9)-CE. The findings of this study provide data support and a theoretical basis for elucidating the molecular mechanisms underlying RhCE deficiency phenotypes.

Acute lung injury (ALI) is a severe disease caused by viral infection that triggers an uncontrolled inflammatory response. This study investigated the capacity of jasurolignoside (JO), a natural compound, to bind to Toll-like receptor 4 (TLR4) and treat ALI. The anti-inflammatory properties of JO were evaluated in vitro through Western blotting, enzyme-linked immunosorbent assay (ELISA), immunofluorescence staining, and co-immunoprecipitation. The investigation utilized a lipopolysaccharide (LPS)-induced ALI animal model to examine the therapeutic efficacy and mechanism of JO in vivo. JO attenuated inflammatory symptoms in infected cells and tissues by modulating the NOD-like receptor family pyrin domain containing protein 3 (NLRP3) inflammasome and the nuclear factor κB (NF-κB)/mitogen-activated protein kinase (MAPK) pathway. Molecular docking simulations revealed JO binding to TLR4 active sites, confirmed by cellular thermal shift assay. Surface plasmon resonance (SPR) demonstrated direct interaction between JO and TLR4 with a Kd value of 35.1 μmol·L^-1. Moreover, JO inhibited tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and IL-6 secretion and reduced leukocyte, neutrophil, lymphocyte, and macrophage infiltration in ALI-affected mice. JO also enhanced lung function and reduced ALI-related mortality. Immunohistochemical staining demonstrated JO's ability to suppress TLR4 expression in ALI-affected mouse lung tissue. This study establishes that JO can bind to TLR4 and effectively treat ALI, indicating its potential as a therapeutic agent for clinical applications.
Toll-Like Receptor 4/chemistry* ; Animals ; Acute Lung Injury/chemically induced* ; Mice ; Humans ; Ilex/chemistry* ; Molecular Docking Simulation ; Male ; NF-kappa B/immunology* ; Mice, Inbred C57BL ; NLR Family, Pyrin Domain-Containing 3 Protein/immunology* ; Tumor Necrosis Factor-alpha/genetics* ; Interleukin-1beta/genetics* ; RAW 264.7 Cells ; Disease Models, Animal

Objective T o explore the correlation between adult chest CT quantitative parameters and traditional Chinese medicine(TCM)constitution classification in adult persons with different body mass index(BMI).Methods The basic information of 205 adult subjects in the hospital was collected,including gender,age,height,weight,BMI,etc.They were divided into the low body mass group(BMI＜18.5 kg/mi2),normal body mass group(BMI 18.5-＜24.0 kg/m2)and overweight/obese group(BMI≥24.0 kg/m2)according to BMI.All subjects underwent the TCM constitution classification.The distribution of TCM constitutions a-mong the three groups with different BMI was analyzed.The quantitative chest CT parameters were collected,including fat content,muscle content,area of the middle slice of the transverse section image,transverse diam-eter,anteroposterior diameter,scan length,etc.The differences were compared among different TCM constitu-tions,individual basic information and chest CT quantitative parameters.Results In the low BMI group,the Qi-stagnation constitution was the highest;in the BMI normal group,the gentler constitution was the highest,while in the overweight/obese group,the phlegm-damp constitution was the highest.The gender,age,height,weight and BMI had significant correlation with the TCM constitution(P＜0.05).The fat content,fat vol-ume,muscle volume,area of transverse section ROI(AROI),transverse diameter,anteroposterior diameter and scan length/transverse diameter had significant correlation with the TCM constitution(P＜0.05).The differ-ences in BMI and fat content had statistical difference among the phlegm-damp constitution,Qi-stagnation constitution and the other Chinese medicine constitutions(P＜0.01).The fat content,fat volume and muscle volume had significant correlation with BMI(P＜0.01).The muscle content had no significant correlation with BMI(P＞0.05).The muscle content,scan length and transverse diameter/anteroposterior diameter had no significant correlation with the TCM constitution(P＞0.05).Conclusion The distribution difference of TCM constitution in adult persons with different BMI has the statistical significance.The chest CT quantita-tive parameters are closely correlated with TCM constitution classification.

Objective To investigate the application of the Health Effective Data and Information System(HEDIS)-based phased health education in infertile patients treated with assisted reproductive technology(ART).Methods A total of 120 infertile patients who were admitted to The First Affiliated Hospital of Naval Medical University from March 2023 to September 2023 were consecutively selected and randomly assigned to observation group or control group at a ratio of 1:1 using a random number table.During the ART treatment period,the control group was given conventional nursing care,while the observation group was given HEDIS based phased health education for nursing intervention.The negative emotion score,shame score,self-management ability score,and nursing satisfaction were compared between the two groups.Results After intervention,the Hamilton Anxiety Scale(HAMA)score and Hamilton Depression Scale(HAMD)score,and self-rating scale(ISS)scores of the observation group were lower than those of the control group(P<0.05).The degree of nursing satisfaction of the observation group was higher than that of the control group(P<0.05).Conclusion The HEDIS-based phased health education can alleviate the negative emotions of anxiety and depression,reduce the sense of shame,and enhance nursing satisfaction in infertile patients during ART treatment.

Objective:To analyze the early warning value of laboratory examination on admission of patients with hemorrhagic fever with renal syndrome to critically ill patients.Meetods:In this study, a retrospective case-control study was used to analyze the clinical data and laboratory examination results of patients with hemorrhagic fever with renal syndrome admitted to the emergency department of Tangdu Hospital of Air Force Medical University from January 2021 to January 2022. According to the patient's laboratory indexes and clinical symptoms, the patients were divided into mild, moderate, severe and critical groups. The general data of the two groups were compared, and the independent risk factors of critically ill patients were screened by multi-factor logistic regression analysis, the predictive model of severe HFRS patients was constructed, and the ROC curve was drawn. .Results:Of the 164 patients with HFRS, 50 were in the severe group and 114 in the mild group. The serum levels of WBC, AST, ALT, Cr, BUN, DD and PCT in the severe group were higher than those in the mild group, while the levels of PLT, ALB and PTA in the severe group were lower than those in the mild group. Multiple logistic regression analysis showed that WBC, PLT and PCT were independent influencing factors for the progression of critically ill patients. The predictive model of severe HFRS was established as follows: logit (P) = -0.321 + 0.040 WBC (×10 9/L) -0.045 PLT (×10 9/L) + 0.086 PCT(ng/mL). The early warning ef?cacy of WBC, PLT, And PCT for severe HFRS was further analyzed. The area under the ROC curve (area under curve, AUC) was 0.779, 0.842, 0.862, and the optimal threshold was 10.435×109/L, 41.5 ×109/Land 2.97 ng/mL, respectively. The AUC of joint detection is 0.900, the sensitivity is 88.0%, and the speci?city is 82.5%, which is better than that of a single laboratory. . Conclusions:HFRS laboratory indexes have certain clinical signi?cance for the identi?cation of critically ill patients, in which serum WBC, PLT and PCT indexes are the risk factors of severe HFRS, which provides a theoretical basis for clinical diagnosis, treatment and prognosis of severe HFRS patients.

Objective To investigate the protective effect of Guanxinning(GXN)tablet on dilated cardiomyopathy(DCM),and to explore its effect and mechanism in pyroptosis of cardiomyocytes via the NLRP3/ASC/Caspase-1 pathway.Methods Rats were divided into GXN low-dose,GXN high-dose,digoxin,model control,and normal control groups.The DCM model was induced by multiple intraperitoneal injections of 17.5 mg/kg doxorubicin(DOX).The drug was administered at the same time as the model was established for 10 weeks.After the last administration,echocardiography was used to assess cardiac function indexes.After sacrificing the rats,serum was collected to measure IL-1β and IL-18 levels.RT-PCR was used to detect mRNA expression of NLRP3,ASC,Caspase-1,NF-κB,TXNIP,IL-1β,and IL-18.Immunohistochemistry and immunofluorescence staining and Western Blot were used to assess NLRP3,ASC,Caspase-1,IL-1β,and IL-18,GSDMD and GSDMD-NT protein,and TUNEL staining result.Changes in the microstructure of cardiomyocytes were observed by transmission electron microscopy.Results Compared with the normal control group,IVSs,IVSd,LVPWs,FS,SV,EF,and HR of the model control group were significantly reduced,LVIDs,ESV,and serum IL-1β and IL-18 were significantly increased,NLRP3,ASC,Caspase-1,NF-κB,TXNIP,IL-1β and IL-18 mRNA expression was significantly increased,and NLRP3,ASC,Caspase-1,IL-1β,IL-18 and GSDMD-NT protein expression and the TUNEL staining area were increased significantly,and the microstructure of cardiomyocytes changed significantly.Compared with the model control group,GXN significantly increased IVSs,SV,FS,EF,and HR,significantly reduced LVIDs,ESV,and the serum levels of IL-1β and IL-18,and reduced NLRP3,ASC,Caspase-1,NF-κB,TXNIP,IL-1β,and IL-18 mRNA expression,NLRP3,ASC,Caspase-1,IL-1β,IL-18 and GSDMD-NT protein expression,and the TUNEL staining area.Additionally,the microstructure was improved significantly.Conclusions GXN alleviates cardiomyocyte pyroptosis in rats with DCM by inhibiting the NLRP3/ASC/Caspase-1 pathway.

Objective To study the effects and mechanisms of Smilax glabra flavonoids(SGF)on myocardial hypertrophy and cardiac inflammation induced by isoproterenol(ISO)in mice.Methods C57/6J mice were randomly divided into a normal group,ISO model group(1.25 mg/(kg·d)),SGF low-dose group(50 mg/(kg·d)),and SGF high-dose group(100 mg/(kg·d)).The SGF groups were given prophylactic SGF for 7 days before modeling,then subcutaneous ISO injection was given to each group,and echocardiography was performed after continuous injection for 7 days.Serum was separated from orbital blood,and the NT-proBNP and inflammatory factor(IL-1β,IL-6,and TNF-α)contents of the blood were detected.Myocardial specimens were collected,and pathological changes to myocardial tissue were observed.Myocardial ROS levels were detected by DHE staining.The mRNA levels of heart-related hypertrophy genes and changes in the expression of key proteins in the inflammatory pathway of NLRP3/Caspase-1/IL-18 in myocardial tissue were detected.Results SGF prophylactic administration decreased IVSd,IVSs,and EF in echocardiography and increased LVIDs,LVIDd,and LVESV.The IL-1 β,IL-6,TNF-α,and NT-proBNP contents of blood decreased,and the mRNA expression levels of the heart-related hypertrophy genes for ANP,BNP,and β-MHC were subdued.The increase in myocardial ROS levels was also inhibited.The protein expression of NLRP3,Caspase-1(p20),and IL-18,which are key factors in the NLRP3/caspase-l/IL-18 inflammatory pathway,was down-regulated in myocardial tissue.The hypertrophy and disordered arrangement of cardiomyocytes were improved,the increase in fibroblast numbers outside myocardial fibers was reduced,and the infiltration of inflammatory cells and fibrosis of myocardial tissue were alleviated.Conclusions SGF can improve ISO-induced myocardial hypertrophy and cardiac inflammation in mice and may act through the NLRP3/Caspase-1/IL-18 inflammatory pathway.

Objective:To investigate the utility of 68Ga-fibroblast activation protein (FAP) inhibitor (FAPI)-04 PET imaging in assessing the therapeutic response in pressure overload-induced heart failure. Methods:Rat models of pressure overload-induced heart failure were established by abdominal aortic constriction (AAC). Thirty rats were categorized into AAC group, 7 days reverse AAC (rAAC) group, and sham operation (sham) group ( n=10 per group) using completely random grouping method. All rats underwent 68Ga-FAPI-04 PET/CT imaging at 4 and 8 weeks after the ACC operation, while echocardiography, pathological examination, and immunohistochemical analysis were performed at 8 weeks postoperation. One-way analysis of variance, independent-sample t test and Pearson correlation analysis were used for data analysis. Results:68Ga-FAPI-04 PET/CT imaging showed that the target-to-background ratios of the heart and liver had significant differences among three groups both at 4 and 8 weeks postoperation ( F values: 2 547.12, 2 041.71, 462.65, 1 210.97, all P<0.001). Echocardiography revealed left ventricular ejection fraction (LVEF), left ventricular internal diameter at end-diastole (LVIDd), and left ventricular internal diameter at end-systole (LVIDs) in three groups at 8 weeks postoperation were significantly different ( F values: 118.92, 9.11, 10.63, all P<0.01). Rats were sacrificed at 8 weeks postoperation, and Masson staining showed that the fibrosis in the heart and liver of the rAAC group was significantly improved compared with that of the AAC group, and immunohistochemical analysis revealed significantly lower FAP levels in the heart and liver of the rAAC group compared with those of the AAC group ( t values: from -11.27 to -4.16, all P<0.01). FAPI uptake in the heart of the AAC group and rAAC group at 8 weeks postoperation were significantly positively correlated with FAPI uptake in the liver, LVIDd and LVIDs, FAPI uptake in the heart was significantly negatively correlated with LVEF, and FAPI uptake in the heart and liver were significantly positively correlated with fibrosis degree and FAP levels of corresponding organs ( r values: -0.89, -0.88, 0.72-0.97, all P<0.05). Conclusions:68Ga-FAPI-04 PET/CT can show the improvement process of cardio-liver fibrosis following the unloading of excessive pressure in heart failure. Myocardial FAPI uptake is closely related to the extent of heart failure improvement.