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.

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.

BACKGROUND: Immune checkpoint inhibitors (ICIs) are increasingly used as first-line therapy for patients with advanced non-small cell lung cancer (NSCLC) harboring no actionable mutations; however, data on their efficacy among patients presenting with intracranial lesions are limited. This study aimed to explore the efficacy and safety of ICIs combined with chemotherapy in advanced NSCLC patients with measurable brain metastasis at initial diagnosis. METHODS: Our study retrospectively analyzed clinical data of a total of 211 patients diagnosed with driver gene mutation-negative advanced NSCLC with measurable, asymptomatic brain metastasis at baseline from Hunan Cancer Hospital between January 1, 2019 and September 30, 2021. The patients were stratified into two groups according to the first-line treatment regimen received: ICI combined with chemotherapy ( n = 102) or chemotherapy ( n = 109). Systemic and intracranial objective response rates (ORRs) and progression-free survival (PFS) were analyzed. Adverse events were also compared between the groups. RESULTS: Compared with the chemotherapy-based regimen, the ICI-containing regimen was associated with a significantly higher intracranial (44.1% [45/102] vs . 28.4% [31/109], χ2 = 5.620, P = 0.013) and systemic (49.0% [50/102] vs . 33.9% [37/109], χ2 = 4.942, P = 0.019) ORRs and longer intracranial (11.0 months vs . 7.0 months, P <0.001) and systemic (9.0 months vs . 5.0 months, P <0.001) PFS. Multivariable analysis consistently revealed an independent association between receiving ICI plus platinum-based chemotherapy as a first-line regimen and prolonged intracranial PFS (hazard ratio [HR] = 0.52, 95% confidence interval [CI]: 0.37-0.73, P <0.001) and systemic PFS (HR = 0.48, 95% CI: 0.35-0.66, P <0.001). No unexpected serious adverse effects were observed. CONCLUSION: Our study provides real-world clinical evidence that ICI combined with chemotherapy is a promising first-line treatment option for driver gene mutation-negative advanced NSCLC patients who present with brain metastasis at initial diagnosis. CLINICAL TRIAL REGISTRATION https://www.clinicaltrials.gov/ , OMESIA, NCT05129202.
Humans ; Carcinoma, Non-Small-Cell Lung/genetics* ; Lung Neoplasms/genetics* ; Immune Checkpoint Inhibitors/therapeutic use* ; Retrospective Studies ; Brain Neoplasms/genetics*

Objective:To study the multi-locus sequence typing (MLST) gene characteristics of Brucella isolates in Guizhou Province. Methods:Brucella strains, which were isolated from 2017 to 2021 in Guizhou Province (preserved in the Bacterial and Viral Seed Bank of Guizhou Center for Disease Control and Prevention) were identified Brucella and species/types by BCSP31-PCR and AMOS-PCR methods, respectively. MLST method was used for genotyping, and Biometrics 8.0 software was used for cluster analysis of the typing results. Results:A total of 32 strains of Brucella were isolated in Guizhou Province and identified as Brucella melitensis ( B.melitensis) by BCSP31-PCR and AMOS-PCR methods. These strains were classified into 2 ST types (ST8 and ST39) by MLST method, with 28 strains of ST8 type(87.5%) and 4 strains of ST39 type (12.5%). The 28 strains of ST8 type were distributed in 7 cities (prefectures) of Guizhou Province, while the 4 strains of ST39 type were only found in Qianxinan Buyi and Miao Autonomous Prefecture. The cluster analysis results showed that ST8 and ST39 types strains were clustered in a group with the reference strain of B.melitensis, and there was only one nucleotide site difference between ST39 and ST8 types in the glk gene, indicating a close genetic relationship. Conclusions:B.melitensis is the main pathogen of the brucellosis epidemic in Guizhou Province in recent years. ST8 is the dominant MLST genotype in Guizhou Province.

OBJECTIVE: To explore the genetic characteristics of a child with Focal segmental glomerulosclerosis and neurodevelopmental syndrome (FSGSNEDS). METHODS: A child with FSGSNEDS who had visited Shengli Oilfield Central Hospital on September 15, 2019 was selected as the study subject. Clinical data of the child was collected, and trio-whole exome sequencing (trio-WES), Sanger sequencing, chromosomal karyotyping analysis, and copy number variation sequencing (CNV-seq) were used to analyze the child and his parents. RESULTS: The child, a 3-year-old boy, had manifested developmental delay, nephrotic syndrome, and epilepsy. Trio-WES and Sanger sequencing showed that he has carried a heterozygous c.1375C＞T (p.Q459*) variant of the TRIM8 gene, for which both his parents were of the wild type. Based on guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was predicted to be pathogenic. No abnormality was found in the chromosomal karyotyping and CNV-seq results of the child and his parents. CONCLUSION The child was diagnosed with FSGSNEDS, for which the c.1375C＞T variant of the TRIM8 gene may be accountable.
Male ; Humans ; Child ; Child, Preschool ; DNA Copy Number Variations ; Glomerulosclerosis, Focal Segmental/genetics* ; Genomics ; Heterozygote ; Karyotyping ; Carrier Proteins ; Nerve Tissue Proteins

Osteoporosis seriously threatens the living quality of people, especially the elderly, and causes a huge economic burden to society. In the past, bisphosphonates, denosumab and other first-line drugs were used in the treatment of osteoporosis. However, these drugs can only inhibit bone resorption, but can not promote bone formation. Studies have shown that mesenchymal stem cells (MSCs) can be used to treat osteoporosis, however, it has some defects and deficiencies, such as genetic instability, limited cell survival and increased risk of cancer. However, mesenchymal stem cell-derived exosomes (MSCs-Exos) can regulate the differentiation and proliferation of osteoblasts by mediating wingless and int-1 (Wnt)/β-catenin and mitogen-activated protein kinase (MAPK) signaling pathways, promote bone regeneration, and thus has an impact on osteoporosis. In this paper, preclinical studies on MSCs and MSCs-Exos in the treatment of osteoporosis in recent years were reviewed, in order to provide a new idea for the treatment of osteoporosis.

Objective: To investigate the effects of antimicrobial peptides merecidin on the biological functions of human lung adenocarcinomaA549 cells and the potential signaling pathways and targets that involved in promoting apoptosis, by studying the changes of phosphorylation levels of proteins in A549 cells after merecidin treatment. Methods: The antibacterial peptide mericidin (9 μmol/L) was applied to treat A549 cells for 6 h, and the total protein was collected and extracted. The peptide was digested by trypsin and labeled with TMT, and then fractionated by HPLC. The phosphorylated peptides were enriched with IMAC-Fe, and finally subjected to mass spectrometry analysis. Library identification and quantification of phosphorylated peptides obtained by mass spectrometry were processed using MaxQuant software, to further analyze the functions and pathways of differentially expressed phosphorylated proteins by combining with bioinformatic analysis. Results: Through IPA analysis of phosphorylated proteins in the normal control group and the antibacterial peptide merecidin treatment group, 753 differentially phosphorylated proteins in mericidin treatment group were screened out under the conditions of |Fold Change|≥2 and P<0.05, including 229 significantly up-regulated genes and 417 down-regulated genes. Among them, the differentially expressed proteins related to apoptosis included RB1, MAPK1, ARAF, PTK2, FOXO, MARCKSandsoon.Theresultsofbiologicalprocessanalysisshowedthatdifferentiallyexpressed phosphorylated proteins were mainly concentrated in cell signal transduction, degradation and transport of nucleic acid, and cellular energy metabolism, protein translation and synthesis, and cytoskeleton formation etc. The enrichment results showed that the differentially expressed phosphorylated proteins were mainly involved in apoptosis-related MAPK, ErbB, PI3K-Akt, and Ras signaling pathways. Protein-protein interaction analysis indicated the associations among apoptosis-related proteins PTK2, PRKCA, MA2PK2, MAPK1, and LMNA. Conclusion: The antibacterial peptide merecidin may induce apoptosis and alteration of other cell functions by affecting a variety of genes and signaling pathways such as RB1, MAPK1,ARAF, PTK2, FOXO and MARCKS etc.