OBJECTIVES: Uridine diphospho-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE) myopathy is a progressive neurodegenerative disease associated with homozygous or compound heterozygous missense mutations in the GNE gene. This study aims to explore the impact of the homozygous p.V543M mutation in on cell phenotype and to gain preliminary insights into the underlying mechanisms. METHODS: Human embryonic kidney 293T (HEK 293T) cells were used to construct wild-type (WT-GNE) and mutant (MUT-GNE) GNE overexpression models. Western blotting and immunofluorescence were used to assess GNE protein expression levels and subcellular localization. Cell adhesion, proliferation, apoptosis, and mitochondrial membrane potential were evaluated using the cell counting kit-8 (CCK-8) assay, crystal violet staining, flow cytometry, Hoechst 33342/propidium iodide (PI) staining, and tetramethylrhodamine ethyl ester (TMRE) staining. Sialic acid synthesis levels and GNE enzymatic activity were measured, and the mRNA expression of sialic acid biosynthesis-related enzymes was quantified by real-time PCR. RESULTS: Western blotting confirmed successful establishment of GNE overexpression models. Immunofluorescence showed significantly reduced co-localization of GNE protein with Golgin-97 in the MUT-GNE group compared to WT-GNE (Pearson's correlation coefficient: 0.65±0.08 vs 0.83±0.06, P<0.05). Compared with WT-GNE, cells in the MUT-GNE group exhibited increased adhesion, decreased proliferation, and reduced mitochondrial membrane potential (P<0.05). No significant differences in apoptosis were observed between groups. The MUT-GNE group showed reduced sialic acid production, significantly decreased kinase activity, and downregulated transcription of sialic acid biosynthesis-related enzymes compared to WT-GNE (P<0.001). CONCLUSIONS The p.V543M mutation in the GNE gene alters cellular phenotype by reducing GNE enzymatic activity and the transcription of sialic acid biosynthesis enzymes, ultimately impairing sialic acid production.
Humans ; Mutation, Missense ; HEK293 Cells ; Apoptosis/genetics* ; Phenotype ; Multienzyme Complexes/metabolism* ; Cell Proliferation ; Homozygote ; Cell Adhesion/genetics* ; Distal Myopathies/genetics*

Objective:To explore the clinical features, histopathological morphology, and differential diagnosis of lymphoepithelioma-like carcinoma with abnormal expression of follicular dendritic cell markers.Methods:From 2020 to 2021, 4 cases of lymphoepithelioma-like carcinoma with abnormal expression of follicular dendritic cell markers diagnosed in Fujian Cancer Hospital (2 cases) and the Second Affiliated Hospital of Fujian Medical University (2 cases) were collected. Different ancillary procedures such as HE, special stains, immunohistochemistry, and in situ hybridization techniques were used to assess the histopathological features and immunophenotypes. The clinical data were collected and literature was reviewed.Results:All 4 cases of lymphoepithelioma-like carcinoma with abnormal expression of follicular dendritic cell markers were male. They were 32, 45, 67 and 39 years old, respectively. The main clinical manifestations were bloody phlegm, abdominal pain, fatigue and anorexia. The clinical stages at diagnosis were stage Ⅳ (3 cases) and stage Ⅱ (1 case). Cases 2 and 3 had two pathological examinations at different sites, with a total of six pathological examinations. The histomorphology showed singly scattered or nests of tumor cells in a background of abundant small lymphocytes. The tumor cells were enlarged and pleomorphic, some appeared polygonal with inconspicuous cell borders, and they were arranged in a syncytial pattern. There were megakaryocytes, multinucleated tumor cells, and a few spindle-shaped cells seen. Atypical mitosis was commonly noted. By immunohistochemistry, the tumor cells were positive for CKpan(5/6), CK8/18(4/4), CAM5.2(2/5), CK-H(0/4), CK-L(3/4), EMA(4/5), CK5/6(3/6), p63(1/6), p40(1/6), E-cadherin (4/6), SSTR2(6/6), PD-L1(5/5), LCA(0/6), vimentin(5/6), CD2 (6/6), CD23(6/6), CD35(5/6), CXCL-13(4/5) and D2-40(1/5). The Ki-67 proliferative index was 60%-95%. In situ hybridization for EBER were all positive (6/6). Special stain for reticulin showed positive staining surrounding nests of tumor cells.Conclusions:The expression of follicular dendritic cell markers in lymphoepithelioma-like carcinoma is very rare, which may be related to EBV infection. Occasionally, it can overlap with follicular dendritic cell sarcoma by morphology and immunophenotype, which can lead to misdiagnosis. Only by combining clinical information, morphological characteristics and immunophenotype can an appropriate diagnosis be made.

Objective:To explore the clinical features, histopathological morphology, and differential diagnosis of lymphoepithelioma-like carcinoma with abnormal expression of follicular dendritic cell markers.Methods:From 2020 to 2021, 4 cases of lymphoepithelioma-like carcinoma with abnormal expression of follicular dendritic cell markers diagnosed in Fujian Cancer Hospital (2 cases) and the Second Affiliated Hospital of Fujian Medical University (2 cases) were collected. Different ancillary procedures such as HE, special stains, immunohistochemistry, and in situ hybridization techniques were used to assess the histopathological features and immunophenotypes. The clinical data were collected and literature was reviewed.Results:All 4 cases of lymphoepithelioma-like carcinoma with abnormal expression of follicular dendritic cell markers were male. They were 32, 45, 67 and 39 years old, respectively. The main clinical manifestations were bloody phlegm, abdominal pain, fatigue and anorexia. The clinical stages at diagnosis were stage Ⅳ (3 cases) and stage Ⅱ (1 case). Cases 2 and 3 had two pathological examinations at different sites, with a total of six pathological examinations. The histomorphology showed singly scattered or nests of tumor cells in a background of abundant small lymphocytes. The tumor cells were enlarged and pleomorphic, some appeared polygonal with inconspicuous cell borders, and they were arranged in a syncytial pattern. There were megakaryocytes, multinucleated tumor cells, and a few spindle-shaped cells seen. Atypical mitosis was commonly noted. By immunohistochemistry, the tumor cells were positive for CKpan(5/6), CK8/18(4/4), CAM5.2(2/5), CK-H(0/4), CK-L(3/4), EMA(4/5), CK5/6(3/6), p63(1/6), p40(1/6), E-cadherin (4/6), SSTR2(6/6), PD-L1(5/5), LCA(0/6), vimentin(5/6), CD2 (6/6), CD23(6/6), CD35(5/6), CXCL-13(4/5) and D2-40(1/5). The Ki-67 proliferative index was 60%-95%. In situ hybridization for EBER were all positive (6/6). Special stain for reticulin showed positive staining surrounding nests of tumor cells.Conclusions:The expression of follicular dendritic cell markers in lymphoepithelioma-like carcinoma is very rare, which may be related to EBV infection. Occasionally, it can overlap with follicular dendritic cell sarcoma by morphology and immunophenotype, which can lead to misdiagnosis. Only by combining clinical information, morphological characteristics and immunophenotype can an appropriate diagnosis be made.

Objective:To explore the relationship between the life satisfaction of family caregivers and the de-gree of disability of disabled elderly people in Xinjiang Uygur and Kazak nationality,and the role of family mem-bers'participation in the relationship.Methods:A total of 431 elderly people with disabilities at home and their fam-ily caregivers(247 without family members and 184 with family members)were selected from Xinjiang Uygur and Kazak ethnic groups,and the survey was conducted with the Activity of Daily Living Scale(ADL)and Life Satis-faction Index B(LSIB).Results:The LSIB scores in family caregivers were negatively correlated with the ADL scores in the disabled elderly(r=-0.19,P＜0.01),and the family members'participation in care was positively correlated with the LSIB scores of family caregivers(r=0.52,P＜0.01).Family members'participation in care could moderate the negative effect of the ADL scores in the disabled elderly on the LSIB scores in family caregivers(β=0.08,P＜0.05).Conclusion:The involvement of family members in care has a moderating effect on the life satisfaction of Uyghur and Kazak family caregivers and the degree of disability of disabled elderly people.

Objective To investigate the relationship between regulatory T(Treg)cells and recurrence after treatment with mometasone furoate in children with adenoid hypertrophy complicated with allergic rhinitis.Methods A total of 104 children with adenoid hyperplasia complicated with allergic rhinitis admitted to Chan-gzhou Maternal and Child Health Care Hospital from January 2021 to January 2023 were selected as the study subjects.All the children were treated with mometasone furoate for 1 month.According to whether the drug relapsed after 3 months,the patients were divided into recurrence group and the non-recurrence group.The pe-ripheral blood Treg cells,intracellular cytokine interleukin-10(IL-10),transforming growth factor β1(TGF-β1),IgE,IgG4 levels,and eosinophilic count(EOS)were measured in both groups before and after treatment.The adenoid/nasopharynx(A/N)value and symptom scores of allergic rhinitis were measured 3 months after drug withdrawal in the two groups.Pearson analysis was used to evaluate the correlation between Treg cells and A/N value and symptom scores.The effectiveness of Treg cells,IL-10,TGF-β1,IgE,IgG4,and EOS in predicting the recurrence was evaluated by receiver operating characteristic(ROC)curve.Results The recur-rence rate of 104 children with adenoid hypertrophy complicated with allergic rhinitis was 41.35％(43/104).After treatment,Treg cells,IL-10,TGF-β1 and IgG4 in the recurrent group were lower than those in the non-recurrent group,while IgE,EOS,A/N value and symptom score were higher than those in the non-recurrent group,with statistical significance(P＜0.05).Pearson analysis showed that Treg cells were negatively corre-lated with A/N value(r=-0.470,P＜0.001)and rhinitis symptom scores(r=-0.872,P＜0.001).The accuracy,sensitivity and specificity of Treg cells in predicting the prognosis were 94.6％,95.1％and 93.0％respectively.Conclusion Treg cells are effective in predicting the prognosis of children with adenoid hypertro-phy complicated with allergic rhinitis.

As research delves deeper into the mechanisms of tumor immune responses,studies reveal the importance of microbial com-munities within the tumor microenvironment in tumor progression and their interactions with the host immune system.Intratumoral micro-biota could influence the tumor microenvironment,thereby promoting or inhibiting tumor growth and development.Despite this import-ance,the specific role of intratumoral microbiota impacting cancer immunotherapeutic efficacy remains largely unexplored.A deeper under-standing of the characteristics and biological functions of tumor-specific microbiota heralds a potential revolutionary innovation in cancer treatment.In this review,we introduce the discovery and sources of intratumoral microbiota,also addressing its composition,and discuss tumor tissue characteristics.Moreover,we briefly review the history of cancer immunotherapy development with a particular focus on the research progress concerning the impact of intratumoral microbiota on cancer immunotherapy.Furthermore,we explore emerging strategies that combine targeting intratumoral microbiota with immunotherapy to enhance immune efficacy,inhibit tumor progression,and improve cure rates,anticipating that this approach could represent a new direction for enhancing treatment outcomes and prospects.

Objective:To explore the gene microarray of patients with ankylosing spondylitis in GEO database by using various bioinformatics methods, and to explore the possible targets and mechanisms of action.Methods:The GEO database was searched with "ankylosing spondylitis" the keyword, and the expression profile of genes related to AS was selected as the research object. Standard difference analysis, weighted co-expression analysis and gene set enrichment analysis were conducted to construct the disease set. GO and KEGG enrichment analysis were performed on the disease sets. The NCC algorithm identifies the first five key genes. THP-1 cells were implanted into RPMI-1640 culture medium containing 10% fetal bovine serum to multiply and construct the cell model of AS in vitro. The expression levels of 5 key genes were detected by qRT-PCR and Western blot. The experimental measurement data were expressed as mean± standard deviation, and the t test was used in comparison between the two groups. Results:One thousand six hundred and sixty seven disease genes were analyzed, functional annotation was mainly concentrated in 689 molecular components of cytoplasmic ribosomes, ribosomal subunits, ribosomes, cytoplasmic large ribosomal subunits, the structural composition of ribosomal REDOX enzyme activity, 1 002 molecular functions of NADH dehydrogenase activity, NADH dehydrogenase activity, and 5 764 molecular processes of mRNA catabolism and RNA catabolism The physical process involved 1 002 signaling pathways involved in Alzheimer′s disease, Prion disease, Parkinson′s disease, and the first 5 key genes were identified as RPS11, RPL4, RPL37A, RPS23, and RPS9. The experimental results were obtained by t test. The results showed that TNF-α mRNA ( t=5.59, P=0.001) and protein ( t=20.14, P<0.001) were significantly increased, indicating that LPS had induced inflammatory response in THP-1 cells, while RPL37AmRNA ( t=5.87, P=0.001), RPS11 mRNA ( t=3.88, P=0.008), RPS23 mRNA ( t=2.64, P=0.038), RPL37A protein ( t=3.18, P=0.030), RPS11 protein ( t=11.26, P<0.001), RPS23 protein ( t=5.64, P<0.001), increased, while RPS9 mRNA ( t=3.16, P=0.020), RPL4 mRNA ( t=2.54, P=0.044), RPS9 protein ( t=5.85, P<0.001) and RPL4 ( t=2.93, P=0.040) protein expressions decreased. RPL23 stimulated the joint synovial tissue to produce effect-T lymphocytes and release a large number of IL-2 and other inflammatory cytokines. RPS9 acts on the early stages of ribosomogenesis, and knocking down RPS9 reduced overall protein synthesis. RPL4 interacted with TTC22 protein to enhance the binding of WTAP mRNA to RPL4, which was associated with immune diseases. The nucleoprotein OGFOD1 catalyzed the hydroxylation of RPS23 and participated in the inflammatory process. The chromosome conformation confirmed the single nucleotide polymorphism function of IL23R genomic locus in AS disease. Conclusion:Ribosomal protein may be an important target for exploring the mechanism of AS inflammation.

Objective To analyze the scores of National Institutes of Health stroke scale(NIHSS),geriatric nutritional risk index(GNRI),motor function in dependence measure(MFIM)and the risk of stroke-related pneumonia(SAP).Methods Patients with stroke admitted from November,2021 to May,2022 were included.The scores of NIHSS,GNRI and MFIM at admission were collected,and they were divided into non-SAP group(n=232)and SAP group(n=86)according to whether pneumonia occurred within one week after stroke.According to the receiver operating characteristic(ROC)curve,the predictive ability of each score to SAP was evaluated,and each score was grouped according to the best cut-off point.logistic regression model and restricted cubic spline were used to analyze the relationship between each score and SAP.Results A total of 318 stroke patients were enrolled in the study including 86 in SAP group and 232 in non-SAP group.Logistic regression shows that,levels of NIHSS score(OR=32.783,95％CI:16.366～65.671,P<0.001),MFIM score(OR=0.052,95％CI:0.027～0.100,P<0.001)and GNRI score(OR=0.262,95％CI:0.144～0.476,P<0.001)were associated with SAP.Restricted cubic spline analysis shows that,there was a nonlinear dose-response relationship between SAP risk and NIHSS score(P-general trend<0.001,P-nonlinear=0.002),GNRI score(P-general trend<0.001,P-nonlinear<0.001).Conclusion NIHSS,MFIM,and GNRI scores are associated with the risk of SAP in stroke patients,and some of them have nonlinear relationships.

Objective:To screen high-risk population of chronic obstructive pulmonary disease (COPD) and to analyze the risk factors in Haicang District of Xiamen City.Methods:A questionnaire survey was conducted from February 2023 to May 2023 among residents who visited or underwent physical examinations at five community health service centers in Haicang District of Xiamen City selected by cluster sampling method. The self-designed general information questionnaire, COPD population screening questionnaire (COPD-PS) and COPD screening questionnaires (COPD-SQ) were applied in the survey. Individuals with COPD-PS scale>5 or COPD-SQ scale>16 were defined as COPD high-risk group. The association of COPD risk with gender, age, smoking, family history of COPD, history of tuberculosis, history of COVID-19 infection, and using coal/woodstove for cooking or heating was analyzed with chi-square test and binary logistic regression analysis.Results:A total of 4 260 questionnaires were distributed and 4 221 valid questionnaires were collected with a recovery rate of 99.6%. Among all respondents there were 1 904 males (45.11%) and 2 317 females (54.89%); and 217 individuals aged 40-<50 (5.14%), 434 aged 50-<60 (10.28%), 2 194 aged 60-<70 (51.98%), 1 302 aged 70-<80 (30.85%) and 74 aged≥80 (1.76%). The results showed that there were 269 respondents (6.4%) scored≥5 on the COPD-PS scale, 534 residents (12.7%) scored≥16 on the COPD-SQ scale, 646 (15.3%) scored≥5 on the COPD-PS scale or≥16 on the COPD-SQ scale. Male gender ( OR=2.592, 95% CI:2.135-3.146), second-hand smoke exposure ( OR=3.763, 95% CI:2.944-4.810), frequently catching cold before the age of 14 ( OR=3.804, 95% CI:2.927-4.944), history of tuberculosis ( OR=2.575, 95% CI:1.224-5.418), hypertension ( OR=1.547, 95% CI:1.277-1.875), and diabetes ( OR=1.791, 95% CI:1.027-3.121) were independently associated with the high-risk of COPD, while the history of COVID-19 ( OR=0.583, 95% CI:0.476-0.714) was a protective factor for COPD risk. Conclusion:Males, exposure to second-hand smoke, frequently catching cold before the age of 14, history of tuberculosis, hypertension, and diabetes will increase the risk of COPD, while the history of COVID-19 is a protective factor.

Genome-scale metabolomics analysis is increasingly used for pathway and function dis-covery in the post-genomics era.The great potential offered by developed mass spectrometry(MS)-based technologies has been hindered,since only a small portion of detected metabolites were iden-tifiable so far.To address the critical issue of low identification coverage in metabolomics,we adopted a deep metabolomics analysis strategy by integrating advanced algorithms and expanded reference databases.The experimental reference spectra and in silico reference spectra were adopted to facilitate the structural annotation.To further characterize the structure of metabolites,two approaches were incorporated into our strategy,i.e.,structural motif search combined with neutral loss scanning and metabolite association network.Untargeted metabolomics analysis was performed on 150 rice cultivars using ultra-performance liquid chromatography coupled with quadrupole-Orbitrap MS.Consequently,a total of 1939 out of 4491 metabolite features in the MS/MS spectral tag(MS2T)library were annotated,representing an extension of annotation coverage by an order of magnitude in rice.The differential accumulation patterns of flavonoids between indica and japon-ica cultivars were revealed,especially O-sulfated flavonoids.A series of closely-related flavonolignans were characterized,adding further evidence for the crucial role of tricin-oligolignols in lignification.Our study provides an important protocol for exploring phytochemical diversity in other plant species.