OBJECTIVES: The aim of this study is to determine the effect of cannabinoid receptor (CB) 2 inhibitor on desmoglein 3 (DSG3) expression in HaCaT cells co-cultured with pemphigus serum. METHODS: Immunohistochemical staining was used to compare CB expression in pemphigus patients and normal individuals. Enzyme-linked immunosorbent assay (ELISA) was employed to quantify the concentration of CB2 in the serum of pemphigus patients and normal individuals. A correlation analysis was performed to examine the relationship between the serum CB2 and DSG of pemphigus patients. The CCK-8 assay was used to evaluate the inhibitory effect of AM630 on HaCaT cells, and the half-maximal inhibitory concentration (IC₅₀) value was utilized to determine the experimental concentration. Serum from normal individuals (negative control group) and pemphigus patients (pemphigus group) was co-cultured with HaCaT cells at a 1∶1 ratio. HaCaT cells cultured in complete medium were used as the control group. HaCaT cells in the pemphigus group treated with AM630 were employed as the AM630 group. Real-time polymerase chain reaction (PCR) and Western blot were conducted to assess the expression levels of CB2, DSG3, and β-catenin. Cell dissociation experiments were conducted to evaluate the effect of AM630 on the adhesion of HaCaT cells. RESULTS: Immunohistochemistry revealed significant differences in CB2 expression between pemphigus and normal mucosa (P<0.000 1), but no difference was found in CB1 expression. ELISA analysis revealed a statistically significant difference in the expression levels of CB2 in the serum between normal individuals and pemphigus patients (P<0.001). The expression of CB2 in the serum of pemphigus patients exhibited a significant positive correlation with that of DSG3 (r=0.831, P=0.003). The CCK-8 assay indicated that the IC₅₀ of AM630 on HaCaT cells was 0.55 μmol/L. Real-time PCR and Western blot showed that the expression levels of CB2 and DSG3 increased in the pemphigus group, while the expression level of β-catenin decreased compared with that in the AM630 groups (P<0.05). CONCLUSIONS CB2 is highly expressed in oral mucosal pemphigus. AM630 inhibits overexpression of CB2 and DSG3 and underexpression of β-catenin levels, which can provide new therapeutic targets for pemphigus.
Humans ; Pemphigus/pathology* ; Receptor, Cannabinoid, CB2/metabolism* ; Desmoglein 3/metabolism* ; Acantholysis/metabolism* ; Mouth Mucosa/pathology* ; HaCaT Cells ; Coculture Techniques ; beta Catenin/metabolism*

Objective To analyze the main epidemiological characteristics of fungal infection in this hospital in the past 7 years,and to provide reference for clinical treatment and prevention and control strategies of fun-gal infection.Methods The fungal data and clinical data of related patients isolated from clinical samples in Peking Union Medical College Hospital from early January 2018 to the end of May 2024 were selected,and the main epidemiological characteristics of fungal infection in this hospital were identified and described through multi-angle statistical analysis.Results A total of 4 479 patients with filamentous fungal infection were en-rolled.The proportion of male patients[57.5％(2 576/4 479)]was higher than that of female patients[42.5％(1 903/4 143)],mainly distributed in internal medicine,Intensive Care Unit(ICU)and emergency de-partment,among which internal medicine accounted for the highest proportion[50.0％(2 241/4 479)].About 90.0％of the specimens were from the lower respiratory tract,in addition to specimens from skin and soft tis-sue,tissue,ear and blood culture.In terms of seasonal distribution,there are more patients in winter.The fun-gi were mainly composed of Aspergillus,Mucor,Cerdosporium,Fusarium and Penicillium,among which As-pergillus was the most abundant,accounting for 74.6％of the total.Aspergillus fumigatus was the most a-bundant Aspergillus,accounting for 42.5％of the total Aspergillus(1 418/3 340).Among the related infec-tions caused by mold,Aspergillus was the most common in the lower respiratory tract,accounting for 76.8％.Among them,Aspergillus fumigatus accounted for the highest proportion(33.6％).98.6％of the molds infected the ear were Aspergillus,of which Aspergillus niger and Aspergillus terreus were the most common.Skin infections are mainly caused by Sporothrix schenckii,Trichophyton rubrum,Microsporum ca-nis.The results of in vitro drug sensitivity test showed that the four common Aspergillus isolated in this hos-pital were sensitive to voriconazole,and amphotericin B had better antifungal activity against Mucorales in vitro.Conclusion Based on the main epidemiological characteristics of fungal infections in this hospital,it is recommended that special attention be paid to the admission of patients in the respiratory department during the peak infection period in autumn and winter.In the treatment of fungal infections in different regions and on different body parts,attention should be paid to the differences in the distribution of bacterial species.

Objective: To investigate whether fibroblast growth factor 18(FGF18) can induce human gingival fibroblasts(HGFs) isolatedin vitroto differentiate into osteoblast-like cells, and to explore the mechanism of osteogenesis. Methods : HGFs were isolated, cultured and identified by tissue block method. The third generation of HGFs were divided into experimental group and control group. FGF18 and L-DMEM was added to the experimental group while L-DMEM was added to the control group.The effects of different concentrations of FGF18(0, 0.01, 0.02, 0.04, 0.06 mg/L) on proliferation of HGFs were detected by Methylthiazolyldiphenyl-tetrazolium bromide(MTT) assay. Alkaline phosphatase(ALP) and alizarin red staining were used to detect the osteogenesis and mineralization ability of the cells after induction. RT-PCR, immunocytochemistry staining, and Western blot were used to detect the expression of genes and proteins related to osteogenesis and BMP2 in the BMP signaling pathway. Results: Compared with the control group, the experimental group could promote the proliferation of HGFs at 3, 5, 7, 9, and 11days(P<0.05),ALP activity and mineral salt deposition increased after induction at 14 and 21 days(P<0.05), and the expressions of ALP, OPN, OCN mRNA and BMP2 mRNA in BMP signaling pathway significantly increased(P<0.01). The expressions of OPN, OCN and BMP2 protein at 21 days were significantly higher than those at 14 days(P<0.01). Conclusion FGF18 can promote the proliferation of HGFs, and induce the differentiation of HGFs into functional osteoblasts. The osteogenic mechanism is related to the upregulation of BMP2.

This article reported a case of autosomal dominant intellectual developmental disorder type 22 caused by a heterozygous mutation in the ZBTB18 gene. At 24 +4 weeks of gestation, prenatal ultrasound indicated a short outer diameter of the fetal corpus callosum and bilateral ventricular dilatation. Whole-genome copy number variation analysis of the fetus showed no abnormalities. Whole exome sequencing (WES) and Sanger sequencing validation of the family revealed the fetus carried a c.1374_1383del(p.S459*) heterozygous mutation in the ZBTB18 gene (NM_205768.3), which was neither phenotypically present nor genotypically detected in the parents, suggesting a de novo mutation. Based on the clinical manifestations, the fetus was diagnosed with autosomal dominant intellectual developmental disorder type 22. After genetic counseling, the pregnant woman opted for termination of the pregnancy. This case highlights the correlation between prenatal ultrasonic detection of callosal dysgenesis and lateral ventricular enlargement and intellectual developmental disorders caused by gene mutations. Furthermore, it expands the mutation spectrum of the ZBTB18 gene, thereby facilitating prenatal diagnosis and genetic counseling.

Objective:To design protein binders of human epidermal growth factor receptor-2(HER2,also known as ErbB2)using de novo protein design and to preliminarily evaluate their biological effects on tumor cells.Methods:Based on de novo protein design strategy,RosettaDesign and ProteinMPNN algorithms were used to design the binding proteins targeting ErbB2.Concurrently,AlphaFold was applied to assess structural accuracy of the protein binders and their interactions with ErbB2.Subsequently,yeast surface display technology combined with dual-fluorescence high-throughput flow cytometry were performed to screen for protein binders which were capable of specifically binding to ErbB2 specifically.Selected binding proteins were expressed and purified in Eschrichia coli system.Bio-layer interferometry(BLI)was used to validate the binding specificity of the purified protein binders to ErbB2.Finally,Western blotting and CCK-8 assay were used to evaluate the effects of the candidate binding protein on the downstream signaling of ErbB2 and cell proliferation capacity in ovarian cancer(SK-OV-3)and pancreatic cancer(BxPC-3)cell lines,respectively.Results:In this study,the protein binders targeting ErbB2 were successfully designed,a corresponding binding protein library was established,and a protein binder that specifically binds to ErbB2 was screened out from the library.After BLI verification,this protein binder could effectively bind to ErbB2.Further investigation revealed that in ErbB2-high-expressing SK-OV-3 and BxPC-3 cells,this protein binder could effectively inhibit cell proliferation and reduce the phosphorylation level of AKT,a signaling molecule downstream of ErbB2.Conclusion:Based on the strategy of de novo protein design,this study successfully constructed a protein binder that can effectively bind to ErbB2.The binder can effectively inhibit the activation of the downstream signaling pathway mediated by ErbB2 and the proliferation of tumor cells.This indicates the potential application value of de novo designed protein binders targeting ErbB2 in cancer therapy,providing a novel research approach for the field of targeted tumor therapy.

Objective:To evaluate the safety and effectiveness of fuzuloparib for the treatment of ovarian epithelial cancer patients in the real world setting.Methods:A retrospective analysis was conducted on the baseline data of 4 620 ovarian cancer patients who had received fuzuloparib monotherapy or combination therapy. Another 224 ovarian cancer patients who were willing to receive fuzuloparib monotherapy or combination therapy were prospectively enrolled, and their baseline characteristics, drug effectiveness, and safety data were analyzed.Results:(1) Among the 4 620 patients in the retrospective cohort, the median age of patients was 60 years; tumor types: 89.8% (4 149/4 620) had ovarian cancer. Among patients with clearly documented information, the vast majority had a histological type of serous carcinoma (82.9%, 3 770/4 546) and International Federation of Gynecology and Obstetrics (FIGO) staging of Ⅲ-Ⅳ (90.9%, 1 537/1 691). (2) Among the 224 patients in the prospective cohort, the median age of patients was 57 years; tumor types: 83.9% (188/224) had ovarian cancer. Among patients with clearly documented records, the predominant pathologic type was serous carcinoma (91.9%, 193/210), and FIGO stage was Ⅲ-Ⅳ in 79.9% (139/174). (3) Among the 224 prospective patients: 84 patients received first-line fluzoparib maintenance therapy, 92 patients received fluzoparib maintenance therapy after platinum-sensitive recurrence, 23 patients received direct fluzoparib treatment after platinum-sensitive recurrence, 19 patients received direct fluzoparib treatment after platinum-resistant recurrence. The median follow-up durations were 8.5, 8.7, 7.9, and 6.7 months, respectively. The median durations of fluzoparib treatment were 6.7, 4.8, 3.1, and 1.9 months, respectively. The median progression-free survival (PFS) times were not reached during follow-up, 12.6 months, not reached during follow-up, and 4.8 months, respectively. The 1-year PFS rates were 84.1%, 55.0%, 69.8%, and 45.5%, respectively. The remaining 6 patients received other fluzoparib regimens. (4) Among the 224 patients in the prospective dataset, 205 had safety data recorded. Of these, 127 patients (62.0%, 127/205) experienced treatment-related adverse events, with common events including anemia (24.4%, 50/205), thrombocytopenia (21.0%, 43/205), and leukopenia (19.5%, 40/205). Among the 205 patients, 43 (21.0%, 43/205) experienced grade 3 or higher treatment-related adverse events, with common events including anemia (8.3%, 17/205) and thrombocytopenia (8.3%, 17/205).Conclusions:The effectiveness of fuzuloparib in clinical application is generally consistent with other drugs in the same class, with good safety. This study provids new clinical evidence for the treatment of ovarian cancer with fuzuloparib.

Objective To develop the value of machine learning(ML)models based on contrast-transthoracic echocardiography(cTTE)and transesophageal echocardiography(TEE)combined with clinical and laboratory indicators for predicting patent foramen ovale-associated stroke(PFO-AS).Methods Totally 313 patients with PFO diagnosed with cTTE and TEE were retrospectively enrolled.Among them,65 cases were found complicated with ischemic stroke and confirmed as PFO-AS(PFO-AS group),and the rest 248 cases without ischemic stroke were classified as non-PFO-AS group.The patients were divided into training set(n=219,including 48 cases of PFO-AS and 171 cases of non-PFO-AS)and test set(n=94,including 17 cases of PFO-AS and 77 cases of non-PFO-AS)at the ratio of 7∶3.Univariable and multivariable logistic regression(LR)were used to analyze clinical and laboratory indicators as well as cTTE and TEE parameters in training set to screen independent predictive factors of PFO-AS.ML models,including LR,K-nearest neighbor(KNN),support vector machine(SVM),random forest(RF),decision tree(DT),back propagation neural network(BPNN)and gradient boosting machine(GBM)were constructed,and the predictive efficacy of the models for predicting PFO-AS was evaluated,then the optimal model was selected.Results Patient's age＞49-69 years,with smoking history,plasma albumin≥43.8 g/L,significant right-to-left shunt at rest shown on cTTE and complicated atrial septal aneurysm shown on TEE were all independent predictors of PFO-AS,which were used to construct ML models.The area under the curve(AUC)of LR,KNN,SVM,RF,DT,BPNN and GBM models in training set was 0.779-0.853,while in test set was 0.730-0.877.RF model had relatively high and comparable sensitivity,specificity and AUC in both training and test sets,also higher precision and smaller Brier score in test set,hence was regarded as the optimal ML model.Conclusion RF model based on cTTE and TEE combined with clinical and laboratory indicators could be used to effectively predict PFO-AS.

Objective:To investigate the clinical characteristics of encephalocraniocutaneous lipomatosis (ECCL) in pediatric patients.Methods:A retrospective analysis was conducted on the clinical data of 2 ECCL cases admitted to Children′s Hospital Affiliated to Zhengzhou University between January 2024 and December 2024. Additionally, a review of relevant literature was performed to summarize the clinical features of this condition.Results:Case 1 is a male patient aged 2 years and 10 months, while case 2 is a female patient aged 8 months. Both patients presented with seizures and exhibited nevus psiloliparus on the scalp, non-scarring alopecia, nodular skin tags around the eyes, and ocular choristomas. Brain magnetic resonance imaging revealed leptomeningeal angiomatosis in both cases, with case 1 also demonstrating an intracranial lipoma and case 2 showing localized cerebral atrophy and an arachnoid cyst. Whole-exome sequencing of peripheral blood and copy number variation analysis in both cases did not identify any pathogenic variants. Additionally, no relevant pathogenic variants were detected in the scalp lesion tissue of case 2. A review of the literature revealed that, to date, there have been 5 reported domestic cases, 132 reported foreign cases in pediatric populations, totally 139 cases including 2 cases described in this article. Among these patients, 86 are male, 49 are female, and the gender of 4 cases remains unspecified. Clinical manifestations observed included seizures in 79.0% (64/81) of cases and developmental delay in 64.7% (57/88). Cutaneous lesions were characterized by non-scarring alopecia in 100% (97/97) of cases,non-hair-bearing fatty tissue nevi in 98.3% (58/59), nodular skin tags in 96.5% (56/58), and subcutaneous lipomas in 94.8% (73/77). Ocular lesions predominantly involved choristomas, occurring in 91.8% (90/98) of cases. Central nervous system abnormalities were identified as ventricular dilatation or hydrocephalus in 85.0% (68/80) of cases, intracranial lipomas in 82.1% (69/84), localized cerebral atrophy in 80.9% (34/42), intracranial vascular anomalies in 74.1% (23/31), and spinal lipomas in 66.6% (30/45).Conclusions:ECCL is an uncommon neurocutaneous disorder with the potential to impact various organ systems, notably the integumentary, ocular, and central nervous systems. Pediatric patients may exhibit symptoms such as seizures, developmental delays, and additional clinical manifestations, necessitating vigilant monitoring and management.

OBJECTIVE: To explore the genetic etiology of a Chinese pedigree with recurrent Joubert syndrome with negative results by whole-exome sequencing in the prior proband. METHODS: Chinese pedigree which opted elective abortion at the Women and Children's Hospital Affiliated to Chongqing Medical University in December 2024 was selected as the study subject. Whole-genome sequencing was carried out on fetal tissue after termination of pregnancy. Candidate variants were validated by Sanger sequencing and interpreted, while non-coding variant was analyzed using in silico prediction tools. RNA sequencing and cDNA sequencing were conducted on fetal brain tissue. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.2024YL045-02). RESULTS: Both the fetus and the affected child were found to harbor compound heterozygous variants of the CEP290 gene, namely c.7341dup (p.Leu2448fs*8) (pathogenic, maternally inherited) and c.1523-408G>A (likely pathogenic, paternally inherited). Both in silico analysis and fetal brain RNA sequencing confirmed aberrant RNA splicing caused by the intronic variant. CONCLUSION This case has highlighted the value of combining whole-genome sequencing with RNA functional validation. Above results not only enriched the spectrum of CEP290 gene mutations but also underscored its diagnostic value in resolving complex prenatal cases, providing critical clues for the prenatal diagnosis and recurrence risk assessment in genetic counseling.
Female ; Humans ; Pregnancy ; Abnormalities, Multiple/genetics* ; Antigens, Neoplasm/genetics* ; Cell Cycle Proteins/genetics* ; Cerebellum/abnormalities* ; Cytoskeletal Proteins/genetics* ; Eye Abnormalities/genetics* ; Introns/genetics* ; Kidney Diseases, Cystic/diagnosis* ; Pedigree ; Retina/abnormalities* ; Sequence Analysis, RNA/methods* ; Whole Genome Sequencing/methods* ; Child

Objective:To explore the influence of management mode of Deming cycle(plan,do,check,action,PDCA)combined with 6S management method on management effect of medical equipment in orthopedic department,and to improve the management quality for medical equipment in orthopedic department.Methods:In accordance with the combination of four stages of PDCA management and the six links(organization,tidying,cleaning,cleanliness,literacy and safety)of 6S management method,the medical equipment of orthopedic department was managed as stage by stage and link by link.Twelve using medical equipment in the orthopedic department of the Second Affiliated Hospital of Air Force Military Medical University were selected between 2022 and 2023.The management level between conventional management and management mode after the management mode of PDCA combined with 6S management mothed was implemented was compared,and the equipment fault and adverse event between within 87361 h before the management mode was used and within 87054 h after the management mode was used also were compared.The scores of satisfaction ratings of 15 medical staffs who involved in using equipment were investigated by using self-made questionnaire.Results:The scores of training assessment,regular maintenance,daily inspection,regular quality control and standardized use,and the total scores after used management mode of PDCA combined with 6S management method were respectively(18.87±0.66),(18.25±0.73),(18.74±0.69),(18.36±0.64),(18.89±0.54)and(92.11±4.20),all of which were higher than those before the management mode was implemented,and the differences of them were statistically significant(t=5.047,3.301,3.396,3.191,3.107,6.170,P<0.05).The standby time due to equipment fault and fault rate within 87054 h of using equipment after the management mode was implemented were respectively 262h and 0.30%,all of which were lower than those before the management mode was implemented,and the differences of them were statistically significant(x2=52.876,P<0.05).The incidence of adverse events of 12 equipment was 0.00%after the management mode was implemented,which was lower than that before it was implemented,and the difference was statistically significant(x2=4.800,P<0.05).The scores of 15 medical staffs on the performance,cleanliness,maintenance,consuming and technical support of equipment,and the total scores were respectively(18.41±1.11),(18.35±1.05),(18.58±0.87),(18.26±1.00),(18.77±0.89)and(92.21±4.12)scores after the management mode was implemented,which were higher than those before it was implemented,and the differences were statistically significant(t=5.257,4.939,5.429,3.191,4.705,9.858,P<0.05).Conclusion:The application of the management mode of PDCA combined with 6S management method in medical equipment management of orthopedic department can improve the management level for equipment,and reduce the incidences of equipment fault and adverse events in using equipment,and improve the satisfaction of medical staffs in using equipment.