Objective To investigate the regulatory role of ubiquitin-specific protease 10(USP10)on the protein expression of Krüppel-like factor 4(KLF4)and its impact on the proliferation and invasion ability of hepatocellular carcinoma(HCC)cells.Methods The protein expression differences of USP10 and KLF4 in normal liver cell line L02 and HCC cell lines,including HepG2,HUH7,HCCLM3 were detected by immunoblotting(Western blot)methods.HCCLM3 and HUH7 cells were selected,and lentiviral particles overexpressing or silencing USP10(oe-USP10 or sh-USP10)was transfected into the cells,and they were designated as the oe-USP10 group and oe-NC group,respectively.Immunoprecipitation(Co-IP)experiments were conducted to examine whether USP10 could di-rectly interact with KLF4 in HCCLM3 or HUH7 cells.The Co-IP assay was repeated in HCC cells transfected with oe-USP10 or sh-USP10,with the addition of the proteasome inhibitor MG132,which used to detect the ubiquitina-tion level of KLF4 protein in the transfected HCC cells.The pcDNA3.1 vector containing overexpressed KLF4 or its negative control plasmid(pc-KLF4 or pc-NC)was co-transfected into cells of the sh-USP10 group or sh-NC group.These cells were designated as the sh-NC+pc-NC group,sh-USP10+pc-NC group,sh-NC+pc-KLF4 group,and sh-USP10+pc-KLF4 group.The cell proliferation activity of each group was measured using the CCK-8 assay,and the cell invasion ability was assessed using the Transwell assay.Results Compared to L02 cells,the protein expres-sion of USP10 and KLF4 significantly decreased in HepG2,HUH7,HCCLM3,and other cells(P＜0.05).In HC-CLM3 and HUH7 cells,USP10 protein directly interacted with KLF4.Furthermore,treatment with MG132 resulted in a time-dependent increase in KLF4 protein expression in HCCLM3 and HUH7 cells.Silencing USP10 increased the ubiquitination of KLF4 in HCCLM3 or HUH7 cells,while overexpressing USP10 decreased the ubiquitination level of KLF4 in cells.Compared to the sh-NC+pc-NC group,both the proliferation activity and invasion ability of HCCLM3 and HUH7 cells significantly increased in the sh-USP10+pc-NC group(P＜0.01),while they signifi-cantly decreased in the sh-NC+pc-KLF4 group and sh-USP10+pc-KLF4 group(P＜0.05).Compared to the sh-USP10+pc-NC group,the proliferation activity and invasion ability of cells significantly decreased in the sh-USP10+pc-KLF4 group(P＜0.05).Conclusion USP10 can promote the stability of KLF4 protein through deubiquiti-nation in HCC cell lines,thereby inhibiting the proliferation and invasion of tumor cells.

Objective To explore the clinical value of laparoscopic radical cystectomy based on fascia anatomy for bladder cancer treatment.Methods The clinical data of 51 patients with bladder cancer who underwent 3D laparoscopic radical cystectomy during Jan.2015 and Jun.2022 were retrospectively analyzed.The surgery was performed based on membrane anatomy technology along four longitudinal and two transverse planes to complete the radical cystectomy.The pelvic plexus was preserved for patients with normal preoperative sexual function.Results All surgeries were completed without conversion to open operation.The mean operation time was(502.52±108.99)min,mean intraoperative blood loss was(275.96±155.18)mL,mean postoperative drainage time was(4.14±2.41)d,and the mean postoperative hospital stay was(16.37±4.85)d.The mean number of lymph nodes removed was(17.98±11.48).The mean postoperative follow-up was(30.27±19.39)months.At the last follow-up,no Clavien ≥grade 3 complications were observed.The estimated overall survival(OS),tumor-specific survival(TSS),and recurrence-free survival(RFS)were 82.4％,92.2％,and 88.2％,respectively.The lymph node positive patients had shorter OS and RFS(60.0％,60.0％)than the lymph node negative patients(84.8％,91.3％).Among the 19 male patients who underwent radical cystectomy with pre-exposure and preservation of pelvic plexus,daytime and nocturnal continence rate were 83.3％and 72.2％,respectively,and 17 patients recovered potency within 6 months postoperatively.Conclusion Laparoscopic radical cystectomy based on fascia anatomy is safe and effective in laparoscopic radical cystectomy,with standardized surgical procedure,satisfactory oncological outcomes,little hemorrhage,few complications and fast recovery.

Objectives:To investigate the effect of the expression of low-density lipoprotein receptor associated protein (LDLR) on the vascular abnormalities in hepatocellular carcinoma (HCC) and its mechanisms.Methods:Based on the information of Oncomine Cancer GeneChip database, we analyzed the correlation between the expression level of LDLR and the expression level of carcinoembryonic antigen (CEA) and CD31 in hepatocellular carcinoma tissues. Lentiviral transfection of short hairpin RNA target genes was used to construct LDLR-knockdown MHCC-97H and HLE hepatocellular carcinoma cells. The differential genes and their expression level changes in LDLR-knockdown hepatocellular carcinoma cells were detected by transcriptome sequencing, real-time fluorescence quantitative polymerase chain reaction, and protein immunoblotting. The gene-related signaling pathways that involve LDLR were clarified by enrichment analysis. The effect of LDLR on CEA was assessed by the detection of CEA content in conditioned medium of hepatocellular carcinoma cells. Angiogenesis assay was used to detect the effect of LDLR on the angiogenic capacity of human umbilical vein endothelial cells, as well as the role of CEA in the regulation of angiogenesis by LDLR. Immunohistochemical staining was used to detect the expression levels of LDLR in 176 hepatocellular carcinoma tissues, and CEA and CD31 in 146 hepatocellular carcinoma tissues, and analyze the correlations between the expression levels of LDLR, CEA, and CD31 in the tissues, serum CEA, and alanine transaminase (ALT).Results:Oncomine database analysis showed that the expressions of LDLR and CEA in the tissues of hepatocellular carcinoma patients with portal vein metastasis were negatively correlated ( r=-0.64, P=0.001), whereas the expressions of CEA and CD31 in these tissues were positively correlated ( r=0.46, P=0.010). The transcriptome sequencing results showed that there were a total of 1 032 differentially expressed genes in the LDLR-knockdown group and the control group of MHCC-97H cells, of which 517 genes were up-regulated and 515 genes were down-regulated. The transcript expression level of CEACAM5 was significantly up-regulated in the cells of the LDLR-knockdown group. The Gene Ontology (GO) function enrichment analysis showed that the differential genes were most obviously enriched in the angiogenesis function. The Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment analysis showed that the relevant pathways involved mainly included the cellular adhesion patch, the extracellular matrix receptor interactions, and the interactions with the extracellular matrix receptors. The CEA content in the conditioned medium of the LDLR-knockdown group was 43.75±8.43, which was higher than that of the control group (1.15±0.14, P＜0.001). The results of angiogenesis experiments showed that at 5 h, the number of main junctions, the number of main segments, and the total area of the lattice formed by HUVEC cells cultured with the conditioned medium of MHCC-97H cells in the LDLR-knockdown group were 295.3±26.4, 552.5±63.8, and 2 239 781.0±13 8211.9 square pixels, which were higher than those of the control group (113.3±23.5, 194.8±36.5, and 660 621.0±280 328.3 square pixels, respectively, all P＜0.01).The number of vascular major junctions, the number of major segments, and the total area of the lattice formed by HUVEC cells cultured in conditioned medium with HLE cells in the LDLR-knockdown group were 245.3±42.4, 257.5±20.4, and 2 535 754.5±249 094.2 square pixels, respectively, which were all higher than those of the control group (113.3±23.5, 114.3±12.2, and 1 565 456.5±219 259.7 square pixels, respectively, all P＜0.01). In the conditioned medium for the control group of MHCC-97H cells,the number of main junctions, the number of main segments, and the total area of the lattice formed by the addition of CEA to cultured HUVEC cells were 178.9±12.0, 286.9±12.3, and 1 966 990.0±126 249.5 spixels, which were higher than those in the control group (119.7±22.1, 202.7±33.7, and 1 421 191.0±189 837.8 square pixels, respectively). The expression of LDLR in hepatocellular carcinoma tissues was not correlated with the expression of CEA, but was negatively correlated with the expression of CD31 ( r=-0.167, P=0.044), the level of serum CEA ( r=-0.061, P=0.032), and the level of serum ALT (r=-0.147, P=0.05). The expression of CEA in hepatocellular carcinoma tissues was positively correlated with the expression of CD31 ( r=0.192, P=0.020). The level of serum CEA was positively correlated with the level of serum ALT ( r=0.164, P=0.029). Conclusion:Knocking down LDLR can promote vascular abnormalities in HCC by releasing CEA.

Objective To estimate the disease burden of silicosis in China from 1990 to 2021 and analyze its changing trend using the data from the Global Burden of Disease Study 2021 (GBD 2021), and to provide evidence for the prevention and control of silicosis. Methods Data on the incidence, prevalence, morbidity and death, mortality and disability-adjusted life year (DALY) of silicosis in China were extracted from the GBD 2021 to analyze the disease burden and age distribution of silicosis. The average annual percentage change (AAPC) was calculated to reflect the temporal trend of various disease burden indicators from 1990 to 2021. The results were then compared with those of the global population. Results In China, prevalent cases of silicosis increased by 116.62% from 79 075 in 1990 to 171 291 in 2021; incident cases of silicosis increased by 75.75% from 13 315 in 1990 to 23 401 in 2021; deaths of silicosis increased by 30.76% from 4 837 in 1990 to 6 326 in 2021; DALYs of silicosis increased by 14.84% from 150,729.65 person-years in 1990 to 173 091.06 person-years in 2021. The age-standardized rate of prevalence, incidence, mortality, DALY, YLL, and YLD of silicosis in China all showed a downward trend. The AAPC (95% CI) was -0.42% (-0.56% ~ -0.29%), -1.02% (-1.16 %~ -0.88%), -2.16% (-2.49% ~ -1.83%), -2.24% (-2.63% ~ -1.84%), -2.45% (-2.76% ~ -2.14%), and -0.42% (-0.54% ~ -0.29%), respectively. From 1990 to 2021, the age-standardized indicators of silicosis in China were all higher than the global level, and the differences were statistically significant (all P <0.05). The proportion of silicosis incident cases in the total incident cases of pneumoconiosis in China increased from 68.49% in 1990 to 78.58% in 2021. Conclusion The age-standardized indicators of silicosis in China showed a downward trend from 1990 to 2021, but the number of prevalent cases, incident cases, and DALYs showed an increasing trend. The age-standardized incident rate of silicosis in China remains at a high level, suggesting that further efforts should be made to prevent and control silicosis.

Objectives:To investigate the effect of the expression of low-density lipoprotein receptor associated protein (LDLR) on the vascular abnormalities in hepatocellular carcinoma (HCC) and its mechanisms.Methods:Based on the information of Oncomine Cancer GeneChip database, we analyzed the correlation between the expression level of LDLR and the expression level of carcinoembryonic antigen (CEA) and CD31 in hepatocellular carcinoma tissues. Lentiviral transfection of short hairpin RNA target genes was used to construct LDLR-knockdown MHCC-97H and HLE hepatocellular carcinoma cells. The differential genes and their expression level changes in LDLR-knockdown hepatocellular carcinoma cells were detected by transcriptome sequencing, real-time fluorescence quantitative polymerase chain reaction, and protein immunoblotting. The gene-related signaling pathways that involve LDLR were clarified by enrichment analysis. The effect of LDLR on CEA was assessed by the detection of CEA content in conditioned medium of hepatocellular carcinoma cells. Angiogenesis assay was used to detect the effect of LDLR on the angiogenic capacity of human umbilical vein endothelial cells, as well as the role of CEA in the regulation of angiogenesis by LDLR. Immunohistochemical staining was used to detect the expression levels of LDLR in 176 hepatocellular carcinoma tissues, and CEA and CD31 in 146 hepatocellular carcinoma tissues, and analyze the correlations between the expression levels of LDLR, CEA, and CD31 in the tissues, serum CEA, and alanine transaminase (ALT).Results:Oncomine database analysis showed that the expressions of LDLR and CEA in the tissues of hepatocellular carcinoma patients with portal vein metastasis were negatively correlated ( r=-0.64, P=0.001), whereas the expressions of CEA and CD31 in these tissues were positively correlated ( r=0.46, P=0.010). The transcriptome sequencing results showed that there were a total of 1 032 differentially expressed genes in the LDLR-knockdown group and the control group of MHCC-97H cells, of which 517 genes were up-regulated and 515 genes were down-regulated. The transcript expression level of CEACAM5 was significantly up-regulated in the cells of the LDLR-knockdown group. The Gene Ontology (GO) function enrichment analysis showed that the differential genes were most obviously enriched in the angiogenesis function. The Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment analysis showed that the relevant pathways involved mainly included the cellular adhesion patch, the extracellular matrix receptor interactions, and the interactions with the extracellular matrix receptors. The CEA content in the conditioned medium of the LDLR-knockdown group was 43.75±8.43, which was higher than that of the control group (1.15±0.14, P＜0.001). The results of angiogenesis experiments showed that at 5 h, the number of main junctions, the number of main segments, and the total area of the lattice formed by HUVEC cells cultured with the conditioned medium of MHCC-97H cells in the LDLR-knockdown group were 295.3±26.4, 552.5±63.8, and 2 239 781.0±13 8211.9 square pixels, which were higher than those of the control group (113.3±23.5, 194.8±36.5, and 660 621.0±280 328.3 square pixels, respectively, all P＜0.01).The number of vascular major junctions, the number of major segments, and the total area of the lattice formed by HUVEC cells cultured in conditioned medium with HLE cells in the LDLR-knockdown group were 245.3±42.4, 257.5±20.4, and 2 535 754.5±249 094.2 square pixels, respectively, which were all higher than those of the control group (113.3±23.5, 114.3±12.2, and 1 565 456.5±219 259.7 square pixels, respectively, all P＜0.01). In the conditioned medium for the control group of MHCC-97H cells,the number of main junctions, the number of main segments, and the total area of the lattice formed by the addition of CEA to cultured HUVEC cells were 178.9±12.0, 286.9±12.3, and 1 966 990.0±126 249.5 spixels, which were higher than those in the control group (119.7±22.1, 202.7±33.7, and 1 421 191.0±189 837.8 square pixels, respectively). The expression of LDLR in hepatocellular carcinoma tissues was not correlated with the expression of CEA, but was negatively correlated with the expression of CD31 ( r=-0.167, P=0.044), the level of serum CEA ( r=-0.061, P=0.032), and the level of serum ALT (r=-0.147, P=0.05). The expression of CEA in hepatocellular carcinoma tissues was positively correlated with the expression of CD31 ( r=0.192, P=0.020). The level of serum CEA was positively correlated with the level of serum ALT ( r=0.164, P=0.029). Conclusion:Knocking down LDLR can promote vascular abnormalities in HCC by releasing CEA.

Objective:To investigate the effects of microRNA (miR) -125b on the proliferation and phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway of hepatocellular carcinoma (HCC) cell by targeting Polo like kinases (PLK4).Methods:The tumor tissues and adjacent tissues of 65 patients with HCC were collected from March 2022 to March 2023 in Tianjin Medical University Cancer Hospital, including 33 males and 32 females, aged (60.1±5.6) years. The expressions of miR-125a and miR-125b in liver cancer, adjacent tissues and liver cancer cells were detected by fluorescence quantitative polymerase chain reaction. Low expression liver cancer cell was selected to transfect negative control (NC) sequences of miR, miR-125a and miR-125b. Subsequently, miR-NC and NC plasmid, miR-125b sequence and NC plasmid, and miR-125b sequence and PLK4 plasmid were co-transfected. Cell proliferation was detected by cell counting assay, the expression of PLK4, phosphorylated PI3K (p-PI3K) and phosphorylated Akt (p-Akt) was detected by Western blot, and miR-125b-targeting PLK4 were detected by bioinformatics analysis and dual luciferase reporter gene.Results:The relative expressions of miR-125a and miR-125b in HCC patients were (0.62±0.08) and (0.58±0.07), respectively, lower than those in adjacent tissues (1.00±0.12) and (1.00±0.13), and the differences were statistically significant ( t=21.24, 22.93, P=0.005, P<0.001). HepG2 cells with low expression of miR-125a and miR-125b and miR-125b targeting PI3K/Akt were selected for transfection. Bioinformatic analysis and dual luciferase reporter gene assay confirmed that miR-125b binds to PLK4. Overexpression of miR-125b could inhibit the proliferation of HepG2 cells and the expression of p-PI3K and p-Akt, while overexpression of PLK4 could partially reverse the proliferation inhibition caused by miR-125b and the expression of p-PI3K and p-Akt, (0.91±0.07) vs(0.41±0.04), (0.97±0.08) vs (0.32±0.03)( t=13.87, 17.01, both P<0.001). Conclusion:The inhibitory effect of miR-125b on HepG2 cell proliferation and PI3K/Akt signaling pathway is partly mediated by targeted inhibition of PLK4.

Pulmonary fibrosis(PF)is a chronic progressive end-stage lung disease.However,the mechanisms un-derlying the progression of this disease remain elusive.Presently,clinically employed drugs are scarce for the treatment of PF.Hence,there is an urgent need for developing novel drugs to address such diseases.Our study found for the first time that a natural source of Prismatomeris connata Y.Z.Ruan(Huang Gen,HG)ethyl acetate extract(HG-2)had a significant anti-PF effect by inhibiting the expression of the transforming growth factor beta 1/suppressor of mothers against decapentaplegic(TGF-β1/Smad)pathway.Network pharmacological analysis suggested that HG-2 had effects on tyrosine kinase phosphorylation,cellular response to reactive oxygen species,and extracellular matrix(ECM)disassembly.Moreover,mass spec-trometry imaging(MSI)was used to visualize the heterogeneous distribution of endogenous metabolites in lung tissue and reveal the anti-PF metabolic mechanism of HG-2,which was related to arginine biosyn-thesis and alanine,asparate and glutamate metabolism,the downregulation of arachidonic acid meta-bolism,and the upregulation of glycerophospholipid metabolism.In conclusion,we elaborated on the relationship between metabolite distribution and the progression of PF,constructed the regulatory metabolic network of HG-2,and discovered the multi-target therapeutic effect of HG-2,which might be conducive to the development of new drugs for PF.

Objective To investigate the classification,concomitant injuries,and their correlations of medial meniscus posterior root(MMPR)injuries through a large-sample analysis,to enhance the comprehensive understanding of MMPR and related injuries.Methods A total of 240 patients with MMPR injuries were divided into 5 types.The distance of the torn end separation and the value of meniscus protrusion of MMPR were measured,and the grading of cartilage injury in the medial tibiofemoral compartment was recorded.The relationships between MMPR injuries and meniscus tear location,tear type,meniscus protrusion,and grading of cartilage injury were analyzed.Results The incidence of MMPR injuries was 2.82％,with females being 3.14 times more affected than males.Medial meniscus tears in type 1 and type 4 MMPR injuries were predominantly located in the posterior horn and posterior root,while there were no statistical differences among types 2,3,and 5.Type 1 MMPR injuries were predominantly oblique tears,types 2,3,and 5 were predominantly radial and complex tears,and type 4 was predominantly complex tears.The incidence of meniscus protrusion was sig-nificantly higher in types 3 and 4 MMPR injuries compared to other types.The value of medial meniscus protrusion was greater in type 4 MMPR than in type 3.In type 3 MMPR injuries,a larger torn end separation distance correlated with a greater value of medial meniscus protrusion.The severity of MMPR injuries correlated positively with the grading of cartilage injury in the medial tibiofemo-ral compartment.Conclusion Females are more prone to MMPR injuries than males.The classification of MMPR injuries correlates with the location and type of medial meniscus tears,as well as medial meniscus protrusion.There is a positive correlation between the torn end separation distance and the value of meniscus protrusion in MMPR injuries.The severity of MMPR injuries correlates with the degree of cartilage injury in the medial tibiofemoral compartment.

ObjectiveTo explore the relationship between the occupational noise exposure and arteriosclerosis in mechanical manufacturing workers. Methods A total of 453 employees of a machinery manufacturing enterprise were selected as the study subjects using the judgment sampling method. The noise exposure levels in their workplaces were measured, and their cumulative noise exposure (CNE) was assessed based on the type of job-noise exposure matrix and occupational hazard exposure history. Pure-tone audiometry was performed on the research subjects, and their brachial-ankle pulse wave velocity (baPWV) was measured. Results The CNE was (91±11) dB(A) per year and the median baPWV was 1 278.0 cm/s in the research subjects. The results of the generalized linear regression model analysis showed that for every one dB(A) per year increase in CNE, the baPWV of the general population increased by 0.20% ［95% confidence interval (CI) 0.10%-0.30%, P<0.01］, with an increase of 0.17% in males (95%CI 0.06%-0.28%, P<0.01) and 0.28% in females (95%CI 0.07%-0.49%, P<0.01). Using the hearing loss as an outcome indicator for high intensity noise exposure, the results showed that baPWV increased by 7.04% (95%CI 2.42%-11.87%, P<0.01) in individuals with bilateral hearing loss, and by 9.84% and 6.53% (95%CI 3.07%-17.07% and 2.13%-11.11%, all P<0.01) in individuals with elevated high-frequency hearing thresholds in both ears and in either ear, respectively. There was no significant association in elevated speech-frequency hearing thresholds and arteriosclerosis (P>0.05). Conclusion Occupational noise exposure may increase the risk of arteriosclerosis.