Purpose To investigate the clinicopathological features and cytogenetic characteristics in the fumarate hydratase(FH)-deficient uterine smooth muscle tumors.Methods A total of 334 paraffin-embedded specimen of be-nign,borderline,and malignant uterine smooth muscle tumors were collected.FH-deficient uterine smooth muscle tumors were screened using the immunohistochemical detection with FH and S-(2-succino)-cysteine(2SC)antibodies.A retrospective analysis of the clinicopathological features and FH mutations was conducted.Results Compared with the 310 non-FH-deficient uterine leiomyomas(non-FH-dUL),14 cases(93.3％)out of the 15 FH-dUL presented with staghorn blood vessels(x2=52.86,P＜0.000 1),8 cases(53.3％)exhibited pulmonary edema-like stroma(x2=26.41,P＜0.000 1),8 cases(53.3％)had cytoplasmic eosinophilic globules(x2=41.85,P＜0.000 1),5 cases(33.3％)showed multifocal bizarre nuclei(x2=72.54,P＜0.000 1),5 cases(33.3％)had large eosino-philic nucleoli with viral inclusion-like halos(x2=38.85,P＜0.000 1),and 5 cases(33.3％)demonstrated a sig-nificant increase in cell density(x2=8.782,P=0.003).The above morphological features were also observed in a case of FH-d uSTUMP and FH-d uLMS.Among the 14 cases with follow-up,except for one death in the FH-d uLMS,none reported the renal cell carcinoma and cutaneous leiomyoma.FH mutations were identified in 11 cases and the hotspot mutations were found in exon 1,2,4,5,6 and 7.Among these variations,c.125C＞T,c.151C＞T and c.704A＞C were found to be the hotspot gene variations.Furthemore,7 gene variations(c.197A＞G,c.426G＞T,c.505C＞T,c.704A＞C,c.812A＞C,c.847T＞C,c.1006A＞G)were not included in COSMIC and ClinVar database and were supposed to be novel.Conclusion The combination of FH/2SC immunohistochemical detection and FH exon mutation analysis are reliable tools for the early identification of FH-deficient uterine smooth muscle tumors or hereditary leiomyomatosis and renal cell carcinoma(HLRCC)syndrome.

Objective To investigate the expression of N-acetyltransferase 10(NAT10),serine hydroxymethyltransferase 2(SHMT2)and YTH domain family protein 1(YTHDF1)in lung cancer tissues and their correlation with clinicopathological characteristics and prognosis.Methods A total of 98 lung cancer patients admitted to our hospital from April 2020 to April 2021 were regarded as the observation subjects.The cancerous tissues and adjacent tissues of the patient during surgery were collected.Immunohistochemistry was used to detect the expression levels of NAT10,SHMT2 and YTHDF1.The patients were followed up for 3 years and divided into the survival group and the death group according to their prognosis.The data of general clinicopathological characteristics were collected and analyzed.The relationship between NAT10,SHMT2 and YTHDF1 with the prognosis of patients were analyzed.Multivariate Cox regression was used to analyze the influencing factors of lung cancer patients.Results The high expression ratios of NAT10,SHMT2 and YTHDF1 in cancer tissues were obviously higher than those in adjacent tissues(P＜0.05).The expression of NAT10 and YTHDF1 was related to clinical stage,degree of differentiation and lymph node metastasis(P＜0.05),and the expression of SHMT2 was correlated with clinical stage,degree of differentiation,tumor diameter,and lymph node metastasis(P＜0.05).There were statistically significant differences in tumor diameter,clinical stage,degree of differentiation,lymph node metastasis,and expressions of NAT10,SHMT2,and YTHDF1 between the survival group and the death group(P＜0.05).NAT10,SHMT2,YTHDF1 patient survival rates significantly below the low of high expression patients(x2=6.354,P=0.012,x2=8.512,P=0.004,x2=4.791,P=0.029).Lymph node metastasis,high expression of NAT10,SHMT2 and YTHDF1 are all risk factors affecting the prognosis of patients(P＜0.05).Conclusion NAT10,SHMT2,and YTHDF1 are all highly expressed in the tissues of lung cancer,and have a certain correlation with clinical pathological characteristics and prognosis.They may serve as relevant evaluation indicators for the prognosis of lung cancer patients.

Objective:To investigate the effect of Gexia Zhuyu decoction on Toll-like receptor 4 (TLR4)/Nuclear factor-kappaB (NF-κB) signaling pathway on ulcerative colitis and intestinal mucosal barrier.Methods:A total of 86 cases of ulcerative colitis admitted to Xi′an Traditional Chinese Medicine (TCM) Hospital from March 2019 to July 2023 were prospectively included and divided into observation group and control group according to random number table method, with 43 cases in each group. The control group was treated with mesalazine enteric-coated tablets; The observation group was treated with mesalazine enteric-coated tablets combined with Gexia Zhuyu decoction for removing stasis. The clinical efficacy of the two groups was compared in terms of TCM syndrome score, intestinal mucosal barrier function index [diamine oxidase (DAO), D-lactic acid (D-LA)], TLR4/NF-κB signaling pathway expression and inflammatory factor index [interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α)] before and after treatment. The occurrence of adverse reactions was also compared.Results:The total effective rate of observation group was higher than that of control group (χ 2=4.074, P=0.044). Before treatment, there was no significant difference in TCM syndrome scores between the two groups (all P>0.05). After treatment, the TCM syndrome scores of both groups were lower than those before treatment (all P<0.05), and the TCM syndrome scores of the observation group were lower than those of the control group (all P<0.05). Before treatment, there were no significant differences in D-LA and DAO levels between the two groups (all P>0.05). After treatment, the levels of D-LA and DAO in both groups were lower than before treatment (all P<0.05), and the levels of D-LA and DAO in the observation group were lower than those in the control group (all P<0.05). After treatment, the mRNA expressions of TLR4 and NF-κB in both groups were decreased compared with those before treatment, and the mRNA expressions of TLR4 and NF-κB in the observation group were lower than those in the control group (all P<0.05). After treatment, the levels of IL-6, IL-8 and TNF-α in both groups were decreased compared with those before treatment (all P<0.05), and the levels of IL-6, IL-8 and TNF-α in the observation group were lower than those in the control group (all P<0.05). There was no significant difference in the incidence of adverse reactions between the two groups (χ 2=2.346, P=0.126). Conclusions:The application of Gexia Zhuyu decoction in the treatment of ulcerative colitis can not only effectively improve the clinical symptoms, signs and intestinal mucosal barrier function, but also improve the inflammatory response by mediating TLR4/NF-κB signaling pathway.

Purpose To investigate the clinicopathological features and cytogenetic characteristics in the fumarate hydratase(FH)-deficient uterine smooth muscle tumors.Methods A total of 334 paraffin-embedded specimen of be-nign,borderline,and malignant uterine smooth muscle tumors were collected.FH-deficient uterine smooth muscle tumors were screened using the immunohistochemical detection with FH and S-(2-succino)-cysteine(2SC)antibodies.A retrospective analysis of the clinicopathological features and FH mutations was conducted.Results Compared with the 310 non-FH-deficient uterine leiomyomas(non-FH-dUL),14 cases(93.3％)out of the 15 FH-dUL presented with staghorn blood vessels(x2=52.86,P＜0.000 1),8 cases(53.3％)exhibited pulmonary edema-like stroma(x2=26.41,P＜0.000 1),8 cases(53.3％)had cytoplasmic eosinophilic globules(x2=41.85,P＜0.000 1),5 cases(33.3％)showed multifocal bizarre nuclei(x2=72.54,P＜0.000 1),5 cases(33.3％)had large eosino-philic nucleoli with viral inclusion-like halos(x2=38.85,P＜0.000 1),and 5 cases(33.3％)demonstrated a sig-nificant increase in cell density(x2=8.782,P=0.003).The above morphological features were also observed in a case of FH-d uSTUMP and FH-d uLMS.Among the 14 cases with follow-up,except for one death in the FH-d uLMS,none reported the renal cell carcinoma and cutaneous leiomyoma.FH mutations were identified in 11 cases and the hotspot mutations were found in exon 1,2,4,5,6 and 7.Among these variations,c.125C＞T,c.151C＞T and c.704A＞C were found to be the hotspot gene variations.Furthemore,7 gene variations(c.197A＞G,c.426G＞T,c.505C＞T,c.704A＞C,c.812A＞C,c.847T＞C,c.1006A＞G)were not included in COSMIC and ClinVar database and were supposed to be novel.Conclusion The combination of FH/2SC immunohistochemical detection and FH exon mutation analysis are reliable tools for the early identification of FH-deficient uterine smooth muscle tumors or hereditary leiomyomatosis and renal cell carcinoma(HLRCC)syndrome.

Objective:To investigate the effect of Gexia Zhuyu decoction on Toll-like receptor 4 (TLR4)/Nuclear factor-kappaB (NF-κB) signaling pathway on ulcerative colitis and intestinal mucosal barrier.Methods:A total of 86 cases of ulcerative colitis admitted to Xi′an Traditional Chinese Medicine (TCM) Hospital from March 2019 to July 2023 were prospectively included and divided into observation group and control group according to random number table method, with 43 cases in each group. The control group was treated with mesalazine enteric-coated tablets; The observation group was treated with mesalazine enteric-coated tablets combined with Gexia Zhuyu decoction for removing stasis. The clinical efficacy of the two groups was compared in terms of TCM syndrome score, intestinal mucosal barrier function index [diamine oxidase (DAO), D-lactic acid (D-LA)], TLR4/NF-κB signaling pathway expression and inflammatory factor index [interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α)] before and after treatment. The occurrence of adverse reactions was also compared.Results:The total effective rate of observation group was higher than that of control group (χ 2=4.074, P=0.044). Before treatment, there was no significant difference in TCM syndrome scores between the two groups (all P>0.05). After treatment, the TCM syndrome scores of both groups were lower than those before treatment (all P<0.05), and the TCM syndrome scores of the observation group were lower than those of the control group (all P<0.05). Before treatment, there were no significant differences in D-LA and DAO levels between the two groups (all P>0.05). After treatment, the levels of D-LA and DAO in both groups were lower than before treatment (all P<0.05), and the levels of D-LA and DAO in the observation group were lower than those in the control group (all P<0.05). After treatment, the mRNA expressions of TLR4 and NF-κB in both groups were decreased compared with those before treatment, and the mRNA expressions of TLR4 and NF-κB in the observation group were lower than those in the control group (all P<0.05). After treatment, the levels of IL-6, IL-8 and TNF-α in both groups were decreased compared with those before treatment (all P<0.05), and the levels of IL-6, IL-8 and TNF-α in the observation group were lower than those in the control group (all P<0.05). There was no significant difference in the incidence of adverse reactions between the two groups (χ 2=2.346, P=0.126). Conclusions:The application of Gexia Zhuyu decoction in the treatment of ulcerative colitis can not only effectively improve the clinical symptoms, signs and intestinal mucosal barrier function, but also improve the inflammatory response by mediating TLR4/NF-κB signaling pathway.

Objective To investigate the expression of N-acetyltransferase 10(NAT10),serine hydroxymethyltransferase 2(SHMT2)and YTH domain family protein 1(YTHDF1)in lung cancer tissues and their correlation with clinicopathological characteristics and prognosis.Methods A total of 98 lung cancer patients admitted to our hospital from April 2020 to April 2021 were regarded as the observation subjects.The cancerous tissues and adjacent tissues of the patient during surgery were collected.Immunohistochemistry was used to detect the expression levels of NAT10,SHMT2 and YTHDF1.The patients were followed up for 3 years and divided into the survival group and the death group according to their prognosis.The data of general clinicopathological characteristics were collected and analyzed.The relationship between NAT10,SHMT2 and YTHDF1 with the prognosis of patients were analyzed.Multivariate Cox regression was used to analyze the influencing factors of lung cancer patients.Results The high expression ratios of NAT10,SHMT2 and YTHDF1 in cancer tissues were obviously higher than those in adjacent tissues(P＜0.05).The expression of NAT10 and YTHDF1 was related to clinical stage,degree of differentiation and lymph node metastasis(P＜0.05),and the expression of SHMT2 was correlated with clinical stage,degree of differentiation,tumor diameter,and lymph node metastasis(P＜0.05).There were statistically significant differences in tumor diameter,clinical stage,degree of differentiation,lymph node metastasis,and expressions of NAT10,SHMT2,and YTHDF1 between the survival group and the death group(P＜0.05).NAT10,SHMT2,YTHDF1 patient survival rates significantly below the low of high expression patients(x2=6.354,P=0.012,x2=8.512,P=0.004,x2=4.791,P=0.029).Lymph node metastasis,high expression of NAT10,SHMT2 and YTHDF1 are all risk factors affecting the prognosis of patients(P＜0.05).Conclusion NAT10,SHMT2,and YTHDF1 are all highly expressed in the tissues of lung cancer,and have a certain correlation with clinical pathological characteristics and prognosis.They may serve as relevant evaluation indicators for the prognosis of lung cancer patients.

Objective This study aimed to explore the regulatory role of autophagy in acute kidney injury(AKI)-induced acute liver injury(ALI).Methods Forty-eight male Sprague-Dawley rats were randomly divided into four groups:sham operation group,IRI group,3-MA group and RA group.Except for the sham operation group,a rat model of AKI induced by IRI was established in all groups.The AKI model was established by removing the right kidney,separating the left renal artery,and clamping the left renal artery,followed by reper-fusion for 12,24,48,or 72 h.The 3-MA and RA groups were intraperitoneally injected with 3-MA(15 mg/kg,1 mL)or RA(2 mg/kg,1 mL)12 h before and after IRI treatment.The structure and function of the rat lung and kidney tissues were evaluated,and the expression levels of autophagy-related proteins,oxidative stress,and apoptosis were measured.Results Renal IRI led to ALI after AKI,and the levels of blood urea nitrogen,creatinine,tumor necrosis factor-α,and interleukin-1βwere all significantly increased.In addition,compared to the IRI group,the expression levels of P62 and caspase-3 significantly decreased in the RA group,whereas the expression levels of LC3-Ⅱ/LC3-Ⅰ,Beclin-1,Bcl-2,and ULK1 increased.Autophagy reduced pathological damage to kidney and lung tissues by inhibiting inflammation and oxidative stress and effectively ameliorated AKI-induced ALI.Conclusion Autophagy plays an important role in the regulation of ALI induced by AKI and can be used as a new target for AKI treatment and to reduce complication-related mortality.

Objective To construct and validate a risk prediction model for poor inhalation in chronic obstructive pulmonary disease(COPD)patients receiving inhaler therapy,providing a decision support tool for personalized prevention of poor inhalation.Methods A cross-sectional study was conducted to collect data related to COPD patients receiving inhaler therapy,forming a dataset.The dataset was randomly divided into a training set and a test set in a ratio of 4∶1.Four different methods for missing value imputation,3 methods for variable feature selection,and 18 machine learning algorithms were employed to successfully construct 216 models on the training set.The monte carlo simulation method was used for resampling in the test set to validate the models,with the area under curve(AUC),accuracy,precision,recall,and F1 score used to evaluate model performance.The optimal model was selected to build the poor inhalation prediction platform.Results A study involving 308 patients with COPD found that 135(43.8%)were at risk of adverse inhalation.Using 33 predictor variables,216 risk prediction models were developed.Of these models,the ensemble learning algorithm yielded the highest average AUC of 0.844,with a standard deviation of 0.058[95%CI=(0.843,0.845)].The differences in predictive performance among the 216 models were statistically significant(P＜0.01).Under the ensemble learning algorithm,adherence to inhaler use(38.087 4%),inhaler satisfaction(25.680 1%),literacy(24.031 3%),number of inhalers(5.482 3%),age(4.204 5%)and number of acute exacerbations in the past year(2.184 7%)contributed most to the predictive model.The model exhibited superior performance,with an AUC of 0.869 3,an accuracy of 83.87%,a precision of 86.96%,a recall of 74.07%,and an F1 score of 0.8.Conclusion This study has developed a predictive model for poor inhalation risk in COPD inhaler therapy patients using machine learning algorithms,which exhibits strong predictive capabilities and holds potential clinical application value.

Background::Heterogeneity of tumor cells and the tumor microenvironment (TME) is significantly associated with clinical outcomes and treatment responses in patients with urothelial carcinoma (UC). Comprehensive profiling of the cellular diversity and interactions between malignant cells and TME may clarify the mechanisms underlying UC progression and guide the development of novel therapies. This study aimed to extend our understanding of intra-tumoral heterogeneity and the immunosuppressive TME in UC and provide basic support for the development of novel UC therapies.Methods::Seven patients with UC were included who underwent curative surgery at our hospital between July 2020 and October 2020. We performed single-cell RNA sequencing (scRNA-seq) analysis in seven tumors with six matched adjacent normal tissues and integrated the results with two public scRNA-seq datasets. The functional properties and intercellular interactions between single cells were characterized, and the results were validated using multiplex immunofluorescence staining, flow cytometry, and bulk transcriptomic datasets. All statistical analyses were performed using the R package with two-sided tests. Wilcoxon-rank test, log-rank test, one-way analysis of variance test, and Pearson correlation analysis were used properly.Results::Unsupervised t-distributed stochastic neighbor embedding clustering analysis identified ten main cellular subclusters in urothelial tissues. Of them, seven urothelial subtypes were noted, and malignant urothelial cells were characterized with enhanced cellular proliferation and reduced immunogenicity. CD8 + T cell subclusters exhibited enhanced cellular cytotoxicity activities along with increased exhaustion signature in UC tissues, and the recruitment of CD4 + T regulatory cells was also increased in tumor tissues. Regarding myeloid cells, coordinated reprogramming of infiltrated neutrophils, M2-type polarized macrophages, and LAMP3 + dendritic cells contribute to immunosuppressive TME in UC tissues. Tumor tissues demonstrated enhanced angiogenesis mediated by KDR + endothelial cells and RGS5 +/ACTA2 + pericytes. Through deconvolution analysis, we identified multiple cellular subtypes may influence the programmed death-ligand 1 (PD-L1) immunotherapy response in patients with UC. Conclusion::Our scRNA-seq analysis clarified intra-tumoral heterogeneity and delineated the pro-tumoral and immunosuppressive microenvironment in UC tissues, which may provide novel therapeutic targets.

Objective:To investigate the expressions of miR-20a-5p and lysine (K) demethylase 6B (KDM6B) in osteosarcoma tissues and the effects of miR-20a-5p targeting KDM6B on the proliferation, migration and invasion of osteosarcoma cells and tumor growth.Methods:The clinicopathological and paracancerous tissues of 20 patients with osteosarcoma admitted to the First Affiliated Hospital of Chinese Medical University from January 2017 to March 2019 were collected. Quantitative real-time PCR (qRT-PCR) was used to detect the expression levels of miR-20a-5p and KDM6B mRNA in tissues. The osteosarcoma MG63 cells were divided into control group, mimic NC group, miR-20a-5p mimic group, and NC+ empty vector group, miR-20a-5p+ empty vector group, miR-20a-5p+ KDM6B group. The expression levels of miR-20a-5p and KDM6B mRNA of all groups were detected by qRT-PCR. Western blotting was used to detect the expression level of KDM6B. CCK-8 assay, cell scratch test and Transwell test were used to detect cell proliferation, migration and invasion ability. According to the random number table method, nude mice were divided into NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group, with 5 mice in each group. Tumor growth ability was detected by tumor xenograft nude mouse models.Results:The relative expression level of miR-20a-5p mRNA in osteosarcoma tissues was 0.55±0.27, and that in paracancerous tissues was 1.22±0.28, with a statistically significant difference ( t=7.701, P<0.001). The relative expression level of KDM6B mRNA in osteosarcoma tissues was 1.66±0.19, and that in paracancerous tissues was 1.00±0.15, with a statistically significant difference ( t=12.219, P<0.001). After transfection of miR-20a-5p, KDM6B mRNA and protein expression levels decreased with the increase of miR-20a-5p expression level. After miR-20a-5p transfection for 48 h, the cell proliferation abilities of the blank control group, mimic NC group and miR-20a-5p mimic group were 0.83±0.04, 0.81±0.03 and 0.52±0.01 ( F=89.655, P<0.001), compared with the blank control group and mimic NC group, the cell proliferation ability was significantly inhibited in the miR-20a-5p mimic group (both P<0.001). The cell proliferation abilities of NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group were 0.83±0.05, 0.52±0.01 and 0.67±0.05 ( F=43.919, P<0.001), compared with the NC+ empty vector group, the cell proliferation ability was significantly inhibited in the miR-20a-5p+ empty vector group ( P<0.001); compared with the miR-20a-5p+ empty vector group, the cell proliferation ability of miR-20a-5p+ KDM6B group increased significantly ( P<0.001). The scratch healing rates of the blank control group, mimic NC group and miR-20a-5p mimic group were (32.51±2.73)%, (30.26±3.22)% and (13.52±1.77)% ( F=46.314, P<0.001), compared with the control group and the mimic NC group, the scratch healing rate of the miR-20a-5p mimic group was significantly decreased (both P<0.001). The scratch healing rates of NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group were (31.34±3.11)%, (12.15±1.64)% and (28.93±2.89)% ( F=47.511, P<0.001), compared with the NC+ empty vector group, the scratch healing rate of the miR-20a-5p+ empty vector group was significantly decreased ( P<0.001); compared with the miR-20a-5p+ empty vector group, the scratch healing rate of miR-20a-5p+ KDM6B group was significantly increased ( P=0.001). The numbers of transmembrane cells in the blank control group, mimic NC group and miR-20a-5p mimic group were 114±16, 108±11 and 42±6 ( F=36.282, P<0.001), compared with the control group and mimic NC group, the number of transmembrane cells of the miR-20a-5p mimic group was significantly decreased (both P<0.001). The numbers of transmembrane cells in the NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group was 143±11, 39±4 and 139±12 ( F=112.120, P<0.001), compared with the NC+ empty vector group, the number of transmembrane cells of the miR-20a-5p+ empty vector group was significantly decreased ( P<0.001); compared with the miR-20a-5p+ empty vector group, the number of transmembrane cells of the miR-20a-5p+ KDM6B group was increased significantly ( P<0.001). The tumor volumes of mice for 21 d in the NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group were (1 667.50±250.40) mm 3, (129.20±21.00) mm 3 and (775.41±77.51) mm 3 respectively, with a statistically significant difference ( F=77.651, P<0.001). The tumor weights of the 3 groups were (1.35±0.18) g, (0.12±0.01) g and (0.61±0.03) g respectively, with a statistically significant difference ( F=104.191, P<0.001). Conclusion:The expression of miR-20a-5p is significantly decreased in osteosarcoma tissues, and the expression of KDM6B is significantly increased in osteosarcoma tissues. Overexpression of miR-20a-5p may inhibit the proliferation, migration and invasion of osteosarcoma cells and tumor growth by targeting to reduce the expression of KDM6B.