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 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.

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 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.

Objective:To investigate whether IL-7-secreting oncolytic herpes simplex virus(HSV)could activate CD8+T cells and inhibit the growth of hepatocellular carcinoma.Methods:The expression of IL-7 was detected by Western blot.The in vitro cleavage of tumor cells by tumor oncolytic virus HSV and HSV-IL-7 were detected by crystal violet staining.The tumor inhibition ability of HSV-IL-7 and HSV were detected in subcutaneous transplanted tumor model.Levels of IL-7,IFN-γ and TNF-α in serum and tumor tissues were determined by ELISA.The infiltration of CD8+T cells in tumor tissues was detected by immunohistochemistry.Flow cytometry was used to detect Granzyme B secretion in CD8+T cells infiltrated by tumor.Results:Tumor cells infected with HSV-IL-7 expressed high level of IL-7.Both HSV and HSV-IL-7 can effectively lyse B16-F10,CT-26 and H22 tumor cell lines in a dose-dependent manner in vitro.HSV-IL-7 could significantly inhibit the growth of H22 hepatoma cells in vivo(P<0.01)and prolong the survival time of tumor-bearing mice(P<0.001).HSV-IL-7 could significantly increase the IL-7 content in tumor sites(P<0.000 1),and effectively increase the number of tumor infiltrating CD8+T cells(P<0.001).HSV-IL-7 significantly enhanced Granzyme B secretion of tumor-infiltrating CD8+T cells and IFN-γ and TNF-α in tumor tissues(P<0.000 1).Conclusion:HSV-IL-7 has well tumor inhibition activity in vivo and in vitro.It also can activate the anti-tumor activity of CD8+T cells in vivo by secreting IL-7,inhibit tumor growth and prolong the survival time of tumor-bearing mice.

Objective To detect the expressions of intercellular adhesion molecule-1(ICAM-1)and nu-clear factor(NF)-κB in hepatic tissues of the patients with chronic hepatitis B,and to analyze their correlation with the hepatic inflammatory activity and fibrosis degree.Methods The liver biopsy specimens from 66 pa-tients with hepatitis B and 10 non-hepatopathic controls were selected,and immunohistochemistry and in situ hybridization were used to detect ICAM-1 and NF-κB expression levels in different liver tissues.Results The positive rate of ICAM-1 and NF-κB expression in liver tissues of the patients with chronic hepatitis B was higher than that in normal liver tissues,and the difference was statistically significant(P＜0.05).The expres-sion of ICAM-1 and NF-κB in the patients with hepatitis B was positively correlated with the inflammatory ac-tivity and fibrosis degree(r=0.493,0.496,P＜0.01;r=0.580,0.519,P＜0.01).Conclusion ICAM-1 and NF-κB in the patients with chronic hepatitis B are highly expressed,which is useful in judging the hepatic in-flammatory activity and fibrosis degree.

Objective:To design a novel electromagnetic ejection device for endoscopic suturing to achieve continuous deployment of suture nails.Methods:An electromagnetic ejection device and its accompanying suture nail structure were designed and a prototype was fabricated based on electromagnetic ejection principles. A finite element model of the electromagnetic ejection device was constructed to study the effects of armature-coil center distance and different driving voltages on suture nail ejection speed. An experimental platform for testing electromagnetic ejection velocity was constructed, and a high-speed camera was used to detect the ejection velocity. A platform for the suture embedding experiment was built to measure the effects of different voltages on the inserting speed of suture into the gastric wall tissue. A platform for a suture extraction force experiment was built to evaluate the extraction force of sutures embedded in tissues under different driving voltages.Results:A suture nail structure and electromagnetic ejection device were designed, and a prototype was fabricated. The ejection velocity increased and then decreased with the increase of the armature-coil center distance, and the maximum ejection velocity was 15.81 m/s at the center distance of 18 mm. At this distance, the voltage was linearly related to the ejection velocity, and the experimental values of the staple basically coincided with the simulated values. When the driving voltage was in the range of 150 to 180 V, the suture nails could successfully insert in the tissues, and the 180 V voltage group had a greater insertion depth. The extraction force of the suture nails at 120, 150, 180, and 210 V voltages were (0.49 ± 0.19), (1.14 ± 0.19), (1.23 ± 0.15), and (1.85 ± 0.31) N, respectively.Conclusions:A novel electromagnetic ejection device for endoscopic suturing is proposed that is capable of continuous firing of suture nails. This device provides a new long-distance driving method for intelligent, minimally invasive surgical instruments.

Objective To investigate the efficacy and safety of Tubridge flow diverter(TFD)in the treatment of ruptured intracranial aneurysms.Methods The clinical data of 13 patients with aneurysmal subarachnoid hemorrhage,who received TFD treatment at the First Affiliated Hospital of Zhengzhou University between March 2019 and Jul 2022,were retrospectively collected.The perioperative materials and follow-up results were summarized and analyzed.Results Successful operation was accomplished in all the 13 patients(13 aneurysms in total).TFD and coil embolization were simultaneously performed in 10 patients(simultaneous treatment),spring coil filling followed by selective staged TFD placement was adopted in 2 patients(staged treatment),and pure TFD placement was employed in one patient.The incidence of perioperative complications was 15.4%(2/13),including asymptomatic ischemic event in one patient and extra-ventricular drainage-related postoperative bleeding in another patient,which caused death of the patient.The median follow-up time was 6.5 months,and 83.3%of patients(10/12)completed cerebral angiography reexamination with DSA.OKM grade D(complete occlusion of the aneurysm)was obtained in 8 patients(80%),and OKM grade C(residual aneurysm neck)in 2 patients.Conclusion For ruptured intracranial aneurysms,TFD implantation is a clinically feasible treatment with favorable safety and efficacy.