[Objective] To explore the expression of Nectin-4 in invasive bladder urothelial carcinoma (BUC) tissue and its clinical significance, so as to provide reference for clinical diagnosis and treatment of BUC. [Methods] Nectin-4 expression in 60 cases of invasive BUC and 40 cases of chronic inflammation of bladder mucosa was detected with immunohistochemical staining (IHC) and RNAscope.The results of the two methods were analyzed and compared, and the relationship between the two methods and the clinicopathological characteristics of invasive BUC was discussed.The correlation between the protein expression of Nectin-4 in BUC tissues, human epidermal growth factor receptor 2 (Her-2) and programmed death factor ligand 1 (PD-L1) was analyzed. [Results] The positive protein expression rates of Nectin-4 detected by IHC were 78.33%(47/60) and 17.50% (7/40) in the invasive BUC group and inflammatory group, respectively, while the positive mRNA expression rates of Nectin-4 detected by RNAscope were 83.33% (50/60) and 12.50% (5/40), respectively.The Kappa values of Nectin-4 in the invasive BUC group and inflammatory group were 0.732 and 0.610, respectively, with general consistency.The protein expression of Nectin-4 in invasive BUC was correlated with muscular invasion, histological grade, vascular thrombus, lymph node metastasis and clinical stage (P<0.05). The mRNA expression of Nectin-4 in invasive BUC was correlated with max tumor diameter, muscular invasion, histological grade, vascular thrombus, lymph node metastasis and clinical stage (P<0.05). The high expression of Nectin-4 in invasive BUC was positively correlated with the expression of Her-2 (P=0.002), but not with the expression of PD-L1 (P>0.05). [Conclusion] Nectin-4 is highly expressed in invasive BUC, and is usually associated with the pathological parameters of poor prognosis.Detection of Nectin-4 expression will help to guide clinical diagnosis and treatment.

Objective:To explore the clinical manifestations and genetic basis for a rare case of Generalized arterial calcification of infancy (GACI).Methods:A 44-day-old female infant who was treated at Baoding Hospital of Beijing Children′s Hospital Affiliated to Capital Medical University on August 26, 2022 was selected as the study subject. Clinical data of the child was collected, and Trio-whole exome sequencing (Trio-WES), whole genome copy number variation sequencing (CNV-seq) and minigene splicing assay were carried out to analyze the pathogenicity of the variants.Results:The child had presented with fever and high inflammatory indicators, for which treatment with various antibiotics was ineffective. Ultrasound had revealed extensive arterial calcification and arterial wall thickening. The child was suspected for GACI with arteritis related to the primary disease. Her fever was relieved by treatment with glucocorticoid and biological agents. Trio-WES revealed that she has harbored compound heterozygous variants of the ABCC6 gene, namely c. 4404-1G>A and c. 4041+ 5G>T, for which the latter was unreported previously. Based on the guidelines from the American College of Medical Genetics and Genomics, the variants were classified as likely pathogenic (PVS1+ PM2_Supporting) and variant of unknown significance (PM2_Supporting+ PM3+ PP3), respectively. The result of CNV-seq was negative. And the minigene splicing assay has further verified that both variants can result in alternative splicing. Conclusion:For pyrexia with unknown causes and refractory to conventional treatment, it is necessary to recommend early genetic testing to avoid missed diagnosis of GACI.

Objective:To construct and prepare recombinant virus strains chimeric with human metapneumovirus (HMPV) antigenic epitopes.Methods:Recombinant influenza virus vectors which chimeric with different HMPV antigenic epitopes were rescued by reverse genetics using eight-plasmid system. The recombinant influenza virus strain used the internal genes of A/PR/8/34 (PB1, PB2, PA, NP, NS, M, HA, and NA) as a backbone, with concomitant genetic modifications to insert the B-cell epitopes of HMPV into the HA gene, and the CTL+ Th cell epitopes of HMPV into the NA gene. Preparation of recombinant influenza virus strains using reverse genetics in a " 7+ 1" model. The recombinant virus strains were evaluated by measuring hemagglutinin (HA) titers, half tissue culture infectious dose (TCID 50) and growth curves. Sequencing analysis was conducted to verify whether the rescued viruses carried the chimeric HMPV epitopes. Results:The epitopes of HMPV were inserted into the influenza virus genome and two recombinant influenza virus strains were rescued successfully, named as FLU/HMPV/B and FLU/HMPV/CTL+ Th. HA titers of the recombinant strains were both 2 7, their TCID 50 were 10 5.2/ml and 10 5.0/ml, respectively. After cultured for three passages in chick embryo, these two recombinant strains could proliferate steadily. Whole genome sequencing verified that the FLU/HMPV/B carried the B-cell epitopes of HMPV, the FLU/HMPV/CTL+ Th carried the CTL and Th cell epitopes of HMPV. Growth curve tests also verified that the recombinant strains could proliferate steadily in eggs. Conclusions:Two recombinant influenza virus vector strains carrying the B cell, CTL and Th epitopes of HMPV were rescued successfully. The result of the recombinant virus strains in terms of growth characteristics as well as genetic stability indicate that they meet the requirements for proceeding to the next step of animal experiments. The immunogenicity and immunoprotective effect will be further evaluated by mouse experiments. Ultimately new ideas for the realization of " one vaccine for two uses" or " one vaccine formultiple uses", as well as a new strategy for the development of HMPV vaccine will be proposed.

Objective:To investigate the clinical features and therapeutic efficacy of patients with hereditary leiomyomatosis and renal cell carcinoma(RCC) syndrome-associated RCC (HLRCC-RCC) in China.Methods:The clinical data of 119 HLRCC-RCC patients with fumarate hydratase (FH) germline mutation confirmed by genetic diagnosis from 15 medical centers nationwide from January 2008 to December 2021 were retrospectively analyzed. Among them, 73 were male and 46 were female. The median age was 38(13, 74) years. The median tumor diameter was 6.5 (1.0, 20.5) cm. There were 38 cases (31.9%) in stage Ⅰ-Ⅱand 81 cases (68.1%) in stage Ⅲ-Ⅳ. In this group, only 11 of 119 HLRCC-RCC patients presented with skin smooth muscle tumors, and 44 of 46 female HLRCC-RCC patients had a history of uterine fibroids. The pathological characteristics, treatment methods, prognosis and survival of the patients were summarized.Results:A total of 86 patients underwent surgical treatment, including 70 cases of radical nephrectomy, 5 cases of partial nephrectomy, and 11 cases of reductive nephrectomy. The other 33 patients with newly diagnosed metastasis underwent renal puncture biopsy. The results of genetic testing showed that 94 patients had FH gene point mutation, 18 had FH gene insertion/deletion mutation, 4 had FH gene splicing mutation, 2 had FH gene large fragment deletion and 1 had FH gene copy number mutation. Immunohistochemical staining showed strong 2-succinocysteine (2-SC) positive and FH negative in 113 patients. A total of 102 patients received systematic treatment, including 44 newly diagnosed patients with metastasis and 58 patients with postoperative metastasis. Among them, 33 patients were treated with tyrosine kinase inhibitor (TKI) combined with immune checkpoint inhibitor (ICI), 8 patients were treated with bevacizumab combined with erlotinib, and 61 patients were treated with TKI monotherapy. Survival analysis showed that the median progression-free survival (PFS) of TKI combined with ICI was 18 (5, 38) months, and the median overall survival (OS) was not reached. The median PFS and OS were 12 (5, 14) months and 30 (10, 32) months in the bevacizumab combined with erlotinib treatment group, respectively. The median PFS and OS were 10 (3, 64) months and 44 (10, 74) months in the TKI monotherapy group, respectively. PFS ( P=0.009) and OS ( P=0.006) in TKI combined with ICI group were better than those in bevacizumab combined with erlotinib group. The median PFS ( P=0.003) and median OS ( P=0.028) in TKI combined with ICI group were better than those in TKI monotherapy group. Conclusions:HLRCC-RCC is rare but has a high degree of malignancy, poor prognosis and familial genetic characteristics. Immunohistochemical staining with strong positive 2-SC and negative FH can provide an important basis for clinical diagnosis. Genetic detection of FH gene germ line mutation can confirm the diagnosis. The preliminary study results confirmed that TKI combined with ICI had a good clinical effect, but it needs to be confirmed by the results of a large sample multi-center randomized controlled clinical study.

The 2024 American Society of Clinical Oncology Genitourinary (ASCO-GU) Cancers Symposium presented the latest research advancements in dose escalation and hypofractionation of radical radiotherapy (RT), the challenges associated with post-operative hypofractionated or ultrahypofractionated RT, and the technical developments in RT for prostate cancer. This article aims to interpret and review these significant studies, providing a comprehensive understanding of RT technical advancements, particularly focusing on the application of stereotactic body radiation therapy (SBRT) in localized prostate cancer.

Objective:To investigate the epidemiological and genetic characteristics of human metapneumovirus (hMPV) in Tianjin during two influenza epidemic seasons from October 2020 to March 2021 and from October 2021 to March 2022, and enrich the whole genome database of hMPV in China.Methods:A total of 1 040 pharyngeal swab samples collected from patients with influenza-like illness (ILI) were analyzed using microfluidic chip fluorescence quantitative PCR. RT-PCR was used to amplify the whole genome in hMPV-positive samples, and the second-generation sequencing was performed for complete genome sequencing. Bioinformatics software including CLC, DNAStar, and MEGA was used for sequence assembly, nucleotide and amino acid homology analysis, and phylogenetic tree mapping.Results:Among the 1 040 samples, 25 were positive for hMPV with a positive rate of 2.40%. The highest positive rate was observed in the age group of 3 to 5 years, reaching 3.71% (16/431). During the influenza epidemic seasons, the detection rate of hMPV peaked in December, reaching 6.67% (12/180). Twelve strains were successfully sequenced, and there were seven of type B2, four of type A2b, and one of type B1. More variations were detected in the G gene, with 111nt-dup sequence repeats observed in the G gene of three A2b strains.Conclusions:The prevalence of hMPV peaks in December during the influenza epidemic seasons in Tianjin, with Type B2 being the predominant type. Except for the G gene with more mutations, other genes remain stable.

Objective To investigate the differential expression of proteins in the serum of diabetic patients quantitative proteomics technologies, and to identify specific protein biomarkers. Methods Serum samples from three diabetic patients and three healthy controls in Shenzhen in 2023 were collected. Immunoprecipitation and nano liquid chromatography-quadrupole-electrostatic orbitrap-linear ion trap mass spectrometry (easy-nanoLC-Q-orbitrap-fusion) were employed to perform fraction and no-fraction-based quantitative analysis of the serum proteome in diabetic patients and healthy controls. The mass spectrometry data were analyzed by MaxQuant1.6.1.0, and the differential proteins identified by the fraction-based method were subjected to gene ontology (GO) functional clustering and kyoto encyclopedia of genes and genomes (KEGG) pathway analysis. Results A total of 251 reliable proteins were identified by the no-fraction-based method, among which 17 differentially expressed proteins were selected. In contrast, 494 reliable proteins were identified by the fraction-based method, and 74 differentially expressed proteins were selected. Among them, seven proteins showed consistent changes in both methods. Five proteins were downregulated, including pregnancy zone protein (PZP), carbonic anhydrase 1 (CAH1), mannose-binding protein C (MBL2), fibrinogen alpha chain (FIBA), and haptoglobin (HPT). Two proteins were upregulated, including apolipoprotein(a) (LPA) and complement factor H-related protein 4 (CFH4). GO analysis revealed that the differentially expressed proteins were mainly involved in molecular functions such as glycosaminoglycan binding and heparin binding. KEGG pathway analysis showed that the differentially expressed proteins were mainly associated with complement and coagulation cascades, cell adhesion, PI3K-Akt signaling pathway, and other pathways. Conclusions Fraction-based no-fractional quantitative proteomics analysis provides an effective approach for analyzing differential protein expression in the serum of diabetic patients, and offers a valuable technique for screening specific protein biomarkers of serum samples in large-scale cohorts of disease.

Objective To study the expression of domain protein ubiquitin 5(OTUD5)and ring finger protein 186(RNF186)in non-muscle invasive bladder cancer(NMIBC)and their relationship with clinicopathological characteristics and prognostic value.Methods A total of 106 NMIBC patients admitted to Tangshan Traditional Chinese Medicine Hospital from January 2019 to January 2020 were collected.OTUD5 and RNF186 expression in NMIBC cancer tissue and para-cancer tissue were detected by immunochemistry.The differences between OTUD5 and RNF186 expressions in NMIBC cancer tissues with different clinical and pathological characteristics were compared.Kaplan-Meier curve analysis(Log Rank test)was used to analyze the impact of OTUD5 and RNF186 expression on progression free survival of NMIBC.COX regression model analysis was used to analyze factors affecting progression free survival in NMIBC patients.Results The positive rates of OTUD5(62.26％)and RNF186(66.04％)in NMIBC cancer tissue were higher than those in adjacent tissues(11.32％,8.49％),with significant differences(x2=59.146,75.079,all P＜0.05).There was a significant positive correlation between OTUD5 and RNF186 expressions in NMIBC cancer(r=0.659,P＜0.05).The positive rates of OTUD5(72.97％,84.21％)and RNF186(77.03％,86.84％)in T1 stage and high-grade NMIBC cancer tissues were higher than those in Ta/Tis stage(37.50％,50.00％)and low-grade cancer tissues(40.63％,54.41％),and the differences were significant(x2=11.964,13.199;12.143,11.431,all P＜0.05).The 3-year cumulative progression free survival rate of patients in the OTUD5 positive group(40.91％)and RNF186 positive group(45.70％)were lower than that in the OTUD5 negative group(90.00％)and RNF186 negative group(86.11％)(Log Rank test x2=24.710,14.586,all P＜0.05).OTUD5 positive(OR=1.446,95％CI:1.086～1.925),RNF186 positive(OR=1.579,95％CI:1.132～2.024),Tumor TNM stage T1(OR=1.582,95％CI:1.219～2.054)and high pathological grade(OR=1.570,95％CI:1.188～2.074)were independent risk factors affecting progression free survival in NMIBC patients(all P＜0.001).Conclusion The increased expression of OTUD5 and RNF186 in cancer tissue of NMIBC patients are related to tumor TNM staging and pathological grading and are independent risk factors affecting the prognosis of NMIBC patients.

Objective:To investigate the prognosis and risk factors of IgA vasculitis nephritis (IgAVN) in children.Methods:A retrospective cohort study was conducted. Clinical data were collected from 264 children who were pathologically diagnosed with IgAVN at Department of Pediatric Nephrology, Tongji Hospital, affiliated with Tongji Medical College, Huazhong University of Science and Technology, between January 2011 and December 2017. All patients had a follow-up period of more than 3 years. Clinical characteristics, renal pathology, 3-year and 5-year prognosis were analyzed. The patients were grouped based on gender, age of onset (≤6 years, >6-9 years, and >9 years), pathological classification (≤Ⅲ and>Ⅲ),whether the prognosis was complete remission at 3 and 5 years. Independent sample t-tests, ANOVA or chi-squared test were used for intergroup comparisons. Spearman correlation analysis was applied for ordinal data, and multivariate Logistic regression was used to analyze factors affecting the prognosis. Receiver operating characteristic (ROC) curve was utilized to evaluate the predictive value of these factors. Results:Of the 264 children with IgAVN, 153 were male and 111 were female, the age of onset was 8.3 (6.7, 10.3) years, 118 patients (45%) with onset age >6-9 years accounted for the highest proportion. All patients presented with skin purpura and renal involvement, primarily manifesting as hematuria and/or proteinuria. Microscopic hematuria was observed in 253 patients (95.8%), while 246 patients (93.2%) showed proteinuria. In 256 patients (97.0%), hematuria or proteinuria urinalysis was detected within 6 months of skin purpura onset, and 243 patients (92.0%) underwent renal biopsy within 6 months of renal involvement. The most common clinical subtype in 264 IgAVN children was hematuria and proteinuria (204 cases, 77.3%), with grade Ⅲ being the predominant pathological classification (181 cases, 68.6%). Among children ≤6 years old, the 3-year complete remission rate was higher in males than in females (83.9% (26/31) vs. 7/16, χ2=8.12, P=0.012). Factors independently associated with poor 5-year prognosis included time from hematuria or proteinuria urinalysis to renal biopsy >6 months, elevated serum cholesterol levels, and incomplete remission 3 years post-biopsy ( OR=5.41, 1.39, 6.02, 95% CI 1.40-20.86, 1.04-1.84, 2.61-13.88, all P<0.05). The serum cholesterol has a predictive value for 5-year prognosis ( P=0.020, AUC=0.62, 95% CI 0.52-0.71, Youden index=0.27, cutoff=4.37). Conclusions:For children with IgAVN aged≤6 years, the 3-year prognosis is better in males than in females. Time from hematuria or proteinuria urinalysis to renal biopsy >6 months, elevated serum cholesterol levels, and incomplete remission at 3 years post-biopsy may be independent risk factors for poor 5-year prognosis in children with IgAVN.

Objective:To study the association of frailty status with nutritional risk and the effect on clinical outcomes among elderly surgical inpatients.Methods:Elderly inpatients from the surgery department of Beijing Hospital were enrolled from January to June 2021. Frail scale and nutritional risk screening 2002 (NRS 2002) were used for frailty evaluation and nutrition risk screening. The influence of frailty and associated nutrition risk in elderly surgical inpatients was analyzed.Results:487 elderly surgical patients were included, of whom 131 cases were in the non-frailty group, 279 cases were in the pre-frailty group and 77 cases were in the frailty group, according to the Frail scale score. 146 cases were at nutritional risk, of whom 8 (6.1% of 131) were in the non-frailty group, 87 (31.2% of 279) in the pre-frailty group and 51 (66.2% of 77) were in the frailty group. According to univariate/multivariate logistic regression analysis of frailty in elderly surgical patients, a higher NRS 2002 score, older age, and the presence of multiple concurrent diseases (≥ 5) were significantly associated with frailty ( P < 0.001). The Frail scale score was positively correlated with NRS 2002 score ( r = 0.448, P < 0.01). Multiple comparisons showed that frailty had statistically significant effects on hospital stay and medical costs in elderly surgical patients ( P < 0.05). Conclusions:The prevalence of frailty is higher in elderly surgical patients, and the prevalence of nutritional risk increases with the progression of frailty. Frailty can lead to prolonged hospital stays and increased hospital costs in elderly surgical patients.