KDF1 Promoted Proliferation, Migration and Invasion of Lung Adenocarcinoma Cells through Activating STAT3 and AKT Pathway.

KDF1 has been reported to be correlated with carcinogenesis. However, its role and mechanism are far from clear. To explore the possible role and underlying mechanism of KDF1 in lung adenocarcinoma (LUAD), we investigated KDF1 expression in LUAD tissues and the influence of KDF1 in the phenotype of LUAD cells (A549 and PC-9) as well as the underlying mechanism. Compared to non-tumor lung epithelial cells, KDF1 was upregulated in the cancer cells of the majority of LUAD patients, and its expression was correlated with tumor size. Patients with enhanced KDF1 in cancer cells (compared with paired adjacent non-neoplastic lung epithelial cells) had shorter overall survival than patients with no increased KDF1 in cancer cells. Knockdown of KDF1 inhibited the migration, proliferation and invasion of LUAD cells in vitro. And overexpression of KDF1 increased the growth of the subcutaneous tumors in mice. In terms of molecular mechanisms, overexpression of KDF1 induced the expression of AKT, p-AKT and p-STAT3. In KDF1-overexpressing A549 cells, inhibition of the STAT3 pathway decreased the level of AKT and p-AKT, whereas inhibition of the AKT pathway had no effect on the activation of STAT3. Inhibition of STAT3 or AKT pathways reversed the promoting effects of KDF1 overexpression on the LUAD cell phenotype and STAT3 inhibition appeared to have a better effect. Finally, in the cancer cells of LUAD tumor samples, the KDF1 level was observed to correlate positively with the level of p-STAT3. All these findings suggest that KDF1, which activates STAT3 and the downstream AKT pathway in LUAD, acts as a tumor-promoting factor and may represent a therapeutic target.

Adhesion-regulating molecule 1 (ADRM1) can be a potential biomarker and target for bladder cancer.

Adhesion-regulating molecule 1 (ADRM1) has been implicated in tumor development, yet its specific role in bladder cancer (BC) remains undefined. This study aimed to elucidate the function of ADRM1 in BC through a combination of bioinformatics analysis and immunohistochemical analysis (IHC). Utilizing R version 3.6.3 and relevant packages, we analyzed online database data. Validation was conducted through IHC data, approved by the Institutional Ethics Committee (Approval No. K20220830). In both paired and unpaired comparisons, ADRM1 expression was significantly elevated in BC tissues compared to adjacent tissues, as evidenced by the results of TCGA dataset and IHC data. Patients with high ADRM1 expression had statistically worse overall survival than those with low ADRM1 expression in TCGA dataset, GSE32548 dataset, GSE32894 dataset, and IHC data. Functional analysis unveiled enrichment in immune-related pathways, and a robust positive correlation emerged between ADRM1 expression and pivotal immune checkpoints, including CD274, PDCD1, and PDCD1LG2. In tumor microenvironment, samples with the high ADRM1 expression contained statistical higher proportion of CD8 + T cells and Macrophage infiltration. Meanwhile, these high ADRM1-expressing samples displayed elevated tumor mutation burden scores and stemness indices, implying potential benefits from immunotherapy. Patients with low ADRM1 expression were sensitive to cisplatin, docetaxel, vinblastine, mitomycin C, and methotrexate. According to the findings from bioinformatics and IHC analyses, ADRM1 demonstrates prognostic significance for BC patients and holds predictive potential for both immunotherapy and chemotherapy responses. This underscores its role as a biomarker and therapeutic target in BC.

Tubastatin A potently inhibits GPX4 activity to potentiate cancer radiotherapy through boosting ferroptosis.

Ferroptosis, an iron-dependent lipid peroxidation-driven programmed cell death, is closely related to cancer therapy. The development of druggable ferroptosis inducers and their rational application in cancer therapy are critical. Here, we identified Tubastatin A, an HDAC6 inhibitor as a novel druggable ferroptosis inducer through large-scale drug screening. Tubastatin A directly bonded to GPX4 and inhibited GPX4 enzymatic activity through biotin-linked Tubastatin A putdown and LC/MS analysis, which is independent of its inhibition of HDAC6. In addition, our results showed that radiotherapy not only activated Nrf2-mediated GPX4 transcription but also inhibited lysosome-mediated GPX4 degradation, subsequently inducing ferroptosis tolerance and radioresistance in cancer cells. Tubastatin A overcame ferroptosis resistance and radioresistance of cancer cells by inhibiting GPX4 enzymatic activity. More importantly, Tubastatin A has excellent bioavailability, as demonstrated by its ability to significantly promote radiotherapy-induced lipid peroxidation and tumour suppression in a mouse xenograft model. Our findings identify a novel druggable ferroptosis inducer, Tubastatin A, which enhances radiotherapy-mediated antitumor effects. This work provides a compelling rationale for the clinical evaluation of Tubastatin A, especially in combination with radiotherapy.

A novel endothelial-related prognostic index by integrating single-cell and bulk RNA sequencing data for patients with kidney renal clear cell carcinoma.

Background: Endothelial cells in the tumor microenvironment play an important role in the development of kidney renal clear cell carcinoma (KIRC). We wanted to further identify the function of endothelial cells in KIRC patients by integrating single-cell and bulk RNA sequencing data. Methods: Online databases provide the original data of this study. An endothelial-related prognostic index (ERPI) was constructed and validated by R version 3.6.3 and relative packages. Results: The ERPI consisted of three genes (CCND1, MALL, and VWF). Patients with high ERPI scores were significantly correlated with worse prognosis than those with low ERPI scores in the TCGA training group, TCGA test group, and GSE29609 group. A positive correlation was identified between the ERPI score and poor clinical features. The results of functional analysis indicated that ERPI was significantly associated with immune-related activities. We suggested that patients with high ERPI scores were more likely to benefit from immunotherapy based on the results of immune checkpoints, tumor microenvironment, stemness index, and TCIA, while patients with low ERPI scores were sensitive to gemcitabine, docetaxel, paclitaxel, axitinib, pazopanib, sorafenib, and temsirolimus according to the results of the "pRRophetic" algorithm. Therefore, this ERPI may help doctors choose the optimal treatment for patients with KIRC. Conclusion: By integrating single-cell and bulk RNA sequencing data from KIRC patients, we successfully identified the key genes from the perspective of endothelial cells in the tumor microenvironment and constructed ERPIs that had positive implications in precision medicine.

Nonlinear Inverse Analysis for Predicting the Tensile Properties of Strain-Softening and Strain-Hardening UHPFRC.

The tensile stress-strain response is considered to be the most important and fundamental mechanical property of ultra-high-performance fiber-reinforced concrete (UHPFRC). Nevertheless, it is still a challenging matter for researchers to determine the tensile properties of UHPFRC. As a simpler alternative to the direct tensile test, bending tests are widely performed to characterize the tensile behavior of UHPFRC, but require further consideration and a sophisticated inverse analysis procedure. In order to efficiently predict the tensile properties of UHPFRC, a nonlinear inverse method based on notched three-point bending tests (3PBT) was proposed in this paper. A total of fifteen UHPFRC beams were fabricated and tested to evaluate the sensitivity of the predicted tensile behavior to variations in fiber volume fraction. A segmented stress-strain model was used, which is capable of describing the various tensile properties of UHPFRC, including strain softening and strain hardening. A more approximate formulation was adopted to simulate the load-deflection response of UHPFRC beam specimens. The closed-form analytical solutions were validated by tensile test results and existing methods in literature. Finally, parametric studies were also conducted to investigate the robustness of the proposed method. The load-deflection responses obtained from notched 3PBT could be easily converted into tensile properties with this inverse method.

Optimal Niacin Requirement of Oriental River Prawn Macrobrachium nipponense as Determined by Growth, Energy Sensing, and Glycolipid Metabolism.

Niacin is indispensable for the growth and development of aquatic animals. However, the correlations between dietary niacin supplementations and the intermediary metabolism of crustaceans are still poorly elucidated. This study explored the effects of different dietary niacin levels on the growth, feed utilization, energy sensing, and glycolipid metabolism of oriental river prawn Macrobrachium nipponense. Prawns were fed with different experimental diets containing graded niacin levels (15.75, 37.62, 56.62, 97.78, 176.32, and 339.28 mg/kg, respectively) for 8 weeks. Weight gain, protein efficiency, feed intake, and hepatopancreas niacin contents all maximized in the 176.32 mg/kg group with significance noted with the control group (P <0.05), whereas the opposite was true for feed conversion ratio. Hepatopancreas niacin concentrations increased significantly (P < 0.05) as dietary niacin levels increased, and peaked at the 339.28 mg/kg group. Hemolymph glucose, total cholesterol, and triglyceride concentrations all maximized in the 37.62 mg/kg group, while total protein concentration reached the highest value in the 176.32 mg/kg group. The hepatopancreas mRNA expression of AMP-activated protein kinase α and sirtuin 1 peaked at the 97.78 and 56.62 mg/kg group, respectively, and then both decreased as dietary niacin levels increased furtherly (P < 0.05). Hepatopancreas transcriptions of the genes related to glucose transportation, glycolysis, glycogenesis, and lipogenesis all increased with increasing niacin levels up to 176.32 mg/kg, but decreased significantly (P < 0.05) as dietary niacin levels increased furtherly. However, the transcriptions of the genes related to gluconeogenesis and fatty acid ß-oxidation all decreased significantly (P < 0.05) as dietary niacin levels increased. Collectively, the optimum dietary niacin demand of oriental river prawn is 168.01-169.08 mg/kg. In addition, appropriate doses of niacin promoted the energy-sensing capability and glycolipid metabolism of this species.

Cyclic Behavior of Reinforced High Strain-Hardening UHPC under Axial Tension.

The cyclic tensile behavior of steel-reinforced high strain-hardening ultrahigh-performance concrete (HSHUHPC) was investigated in this paper. In the experimental program, 12 HSHUHPC specimens concentrically placed in a single steel reinforcement under cyclic uniaxial tension were tested, accompanied by acoustic emission (AE) source locating technology, and 4 identical specimens under monotonic uniaxial tension were tested as references. The experimental variables mainly include the loading pattern, the diameter of the embedded steel rebar, and the level of target strain at each cycle. The tensile responses of the steel-reinforced HSHUHPC specimens were evaluated using multiple performance measures, including the failure pattern, load-strain response, residual strain, stiffness degradation, and the tension-stiffening behavior. The test results showed that the enhanced bond strength due to the inclusion of steel fibers transformed the failure pattern of the steel-reinforced HSHUHPC into a single, localized macro-crack in conjunction with a sprinkling of narrow and closely spaced micro-cracks, which intensified the strain concentration in the embedded steel rebar. Besides, it was observed that the larger the diameter of the embedded steel rebar, the smaller the maximum accumulative tensile strain under cyclic tension, which indicated that the larger the diameter of the embedded steel rebar, the greater the contribution to the tensile stiffness of steel-reinforced HSHUHPC specimens in the elastic-plastic stage. In addition, it was found that a larger embedded steel rebar appeared to reduce the tension-stiffening effect (peak tensile strength) of the HSHUHPC. Moreover, the residual strain and the stiffness of the steel-reinforced HSHUHPC were reduced by increasing the number of cycles and finally tended toward stability. Nevertheless, different target strain rates in each cycle resulted in different eventual cumulative tensile strain rates; hence the rules about failure pattern, residual strain, and loading stiffness were divergent. Finally, the relationship between the accumulative tensile strain and the loading stiffness degradation ratio under cyclic tension was proposed and the tension-stiffening effect was analyzed.

KDF1, a Novel Tumor Suppressor in Clear Cell Renal Cell Carcinoma.

KDF1 has been identified as a key regulator of epidermal proliferation and differentiation, but it is unknown whether KDF1 is involved in the pathogenesis of malignancy. No study has reported the expression and function of KDF1 in renal cancer. To explore the pathologic significance of KDF1 in clear cell renal cell carcinoma (ccRCC), the expression level of KDF1 protein in the tumor tissue of ccRCC patients was examined by immunohistochemistry and Western blot while the expression level of KDF1 mRNA was analyzed by using the data from TCGA database. In vitro cell experiments and allogeneic tumor transplantation tests were performed to determine the effects of altered KDF1 expression on the phenotype of ccRCC cells. Both the KDF1 mRNA and protein were found to be decreasingly expressed in the tumor tissue of ccRCC patients when compared with the adjacent non-tumor control tissue. The expression level of KDF1 in the tumor tissue was found to correlate negatively with the tumor grade. Patients with higher KDF1 in the tumor tissue were found to have longer overall survival and disease-specific survival time. KDF1 was shown to be an independent factor influencing the disease-specific survival of the ccRCC patients. Overexpression of KDF1 was found to inhibit the proliferation, migration and invasion of ccRCC cells, which could be reversed by decreasing the expression of KDF1 again. ccRCC cells with KDF1 overexpression were found to produce smaller transgrafted tumors. These results support the idea that KDF1 is involved in ccRCC and may function as a tumor suppressor.

[Related factors of atrophic glossitis in 124 consecutive cases].

PURPOSE: To study the relationship between atrophic glossitis and anemia, anemia types and other related factors(oral candida infection, xerostomia) in 124 consecutive cases. METHODS: One hundred and twenty-four cases with atrophic glossitis and 53 healthy controls were collected from Qingdao local population. The main indexes including general status, oral examination findings, hemoglobin (Hb), mean red blood cell volume (MCV), vitamin B12, ferritin, folic acid, anemia and anemia type, xerostomia and candida infection were statistically analyzed using SPSS 20.0 software package for Student's t test. RESULTS: Among 124 cases of glossitis group, 48.39% were found with anemia, 41.94% with xerostomia, 79.03% with Candida infection, 29.03% with Vitamin B12 deficiency, 22.58% with ferritin deficiency, 11.29% with folic acid deficiency. The contents of hemoglobin, ferritin and vitamin B12 in glossitis group were significantly lower than those in the control group(P<0.05), and the number of glossitis patients with anemia, xerostomia and candida infection were significantly higher than those in the control group (P<0.05). There was no significant difference in folic acid content between the two groups(P<0.05). CONCLUSIONS: Occurrence of atrophic glossitis is closely related to anemia, vitamin B12 deficiency, ferritin deficiency, xerostomia, oral candida infection. There is no correlation with folic acid deficiency. Patients with atrophic glossitis accompanied by anemia have a higher proportion of macrocytic anemia.

Post-traumatic High-flow Priapism: Multidetector Computed Tomography Demonstration.

High-flow priapism due to pseudoaneurysm is a relatively rare urologic condition. Clear anatomic delineation of the number and origin of feeding vessels facilitates pre-embolization planning. Computed tomographic angiography can afford a three-dimensional display of the feeding vessels. We present a 26-year-old man with post-traumatic high-flow priapism, which is the first case studied with computed tomographic angiography.

[Characterization of the beta-actin gene of the rice field eel and its phylogeny in fish].

Beta-actin is a member of the actin family of genes,which play important roles in maintaining cytoskeletal structure, cell motility, cell division, intracellular movements and contractile processes. We report here the identification of a beta-actin cDNA of the rice field eel,a teleost fish with a characteristic of natural sex reversal. The cDNA sequence of this gene was 1860 bp in length,encoding a 375 amino acid protein. Amino acid identities of the beta-actin between the rice field eel and other vertebrates including human, chicken, and other fishes, were more than 98%. RT-PCR showed expression of the rice field eel beta-actin in testis, ovotestis, ovary, heart, liver, spleen and brain,suggesting a ubiquitous expression pattern. Phylogenetic tree including all beta-actin cds of teleost fishes was constructed,which suggested consistently that teleost beta-actin can be classified into four types,but no fish has been found contains all the four types of beta-actin gene in its genome. The results that suggest lineage-specific beta-actin loss might happen in the radical evolution of teleost fishes.

[Ribosomal protein genes highly expressed in swamp eel gonads].

8 cDNA clones have been isolated from a cDNA library prepared from swamp eel testies by macroarray. DNA sequence analysis and database search showed that they encode 8 proteins which are highly homologous to 40S ribosomal proteins S4,S9,S16,S17,S20 and 60S riobosomal proteins L7, L18a,L29. Phylogenetic trees (ML) based on ribosomal protein genes from swamp eel and other organisms has been reconstructed, which showed that ribosomal protein genes were highly conserved during evolution. These results suggested that ribosomal protein genes as house keeping genes may play roles in developmental regulation such as sexual differentiation and can also be used as markers for the study of molecular evolution.

[Phylogenetic tree and synteny of DMRT genes family of vertebrates].

Vertebrates contain a family of genes related to the Drosophila doublesex and C. elegans mab-3 genes, which encode transcription factors including a DNA-binding motif, DM domain. Evolution and function of different DMRT genes of vertebrates have not been understood yet,although some DM proteins are involved in sex determination, sexual differentiation and early embryonic development among different phyla. By genomic analysis of zebrafish and rat DMRT genes, all protein sequences of the vertebrate DMRTs were searched from gene databases and aligned. Phylogenetic tree of all these DMRT genes was reconstructed and evaluated by Bootstrap method. These DMRT genes were clustered into seven subfamilies. Results from analysis of gene structure and cluster organization of DMRT genes showed that synteny of DMRT genes of vertebrates were highly conserved among human, mouse, rat, fugu, medaka and zebrafish, with two syntenic groups, DMRT 1 approximately 3 and DMRT 5 approximately 6.

[Cloning of four members of giant panda Dmrt genes].

Sex determining genes Mab-3 of C. elegans and Doublesex of Drosophila contain a common DNA binding motif called DM (Doublesex and Mab-3) domain, both of which regulate similar aspects of sexual development. Human Doublesex-related gene DMRT1 has been identified, which also contains the conserved DM-related DNA-binding domain and plays an essential role in gonadal differentiation. We amplified genomic DNA of the giant panda using the DM degenerate primers and detected two bands, approximately 140 bp and 250 bp. After cloned into T-easy vector and sequenced, four sequences showed high homology with the DM domain. Amino acid sequence of the first clone is 100% identical with the Dmrt1 of human, mouse and pig, hence we named it as pDmrt1. The second clone is 96% identical with human DMRTB1, and the third one 100% with the Dmrt3 of mouse and medaka, which were named as pDmrtb1 and pDmrt3 respectively. The last sequence contains an intron of 116 bp within the DM domain, which encodes an amino acid sequence 100% identical with human DMRTC2, accordingly we named it as pDmrtc2. Based on similarities of amino acid sequences of the DM domain, Dmrt protein sequences from human, mouse and giant panda were included in a phylogenetic tree. They revealed seven distinct subgroups: Dmrt1, Dmrt2, Dmrt3, Dmrt4 (DMRTA1), Dmrt5 (DMRTA2), Dmrt6 (DMRTB1) and Dmrt7 (DMRTC2). Our results further reveal the unexpected complexity and the evolutionary conservation of the DM domain gene family in both invertebrates and vertebrates.

The effect of hepatitis B vaccination on the incidence of childhood HBV-associated nephritis.

The aim of the present study was to investigate the effect of a vaccination program for hepatitis B virus (HBV) on the incidence of HBV-associated glomerulonephritis (HBV-GN). In total, 727 renal biopsies were carried out at our hospital from November 1979 through March 2002. Two groups were established. Group A included those biopsied from November 1979 through December 1991 (prior to the HBV vaccination program) and group B from January 1992 through March 2002. Group B was divided into five subgroups (B(1 )to B(5)), with an interval of 2 years between each subgroup. Patients were divided into those with or without a history of HBV vaccination. Of the 727 renal biopsies, 64 fulfilled the criteria of HBV-GN, There were 28 cases of the 211 cases in group A and 36 cases of the 516 cases in group B ( X(2)=7.397, P<0.01). The incidence in group A and group B(1 )through B(5 )was 13.27% (28/211), 13.04% (9/69), 7.32 (6/82), 6.25% (4/64), 4.88% (4/82), and 5.94% (13/219), respectively ( X(2)=9.627, P<0.01). Only 8 of the 231 vaccinated children had HBV-GN, while there were 48 HBV-GN cases of the 381 non-vaccinated children ( X(2)=14.44, P<0.001). There were only 6 cases of membranous nephropathy (MN) in the vaccinated group, while 40 cases of MN occurred in the non-vaccinated group ( X(2)=12.92, P<0.01). There were 8 children that developed HBV-GN with abnormal serum HBV markers despite HBV vaccination. Two mothers of these 8 children had evidence of HBV infection. The incidence of HBV-GN in children has been decreasing each year since the implementation of the nationwide HBV vaccination program in Shanghai, China. Furthermore, since childhood MN is associated with HBV, vaccination can also reduce the incidence of childhood MN.

[Effect of hepatitis B vaccine immunization on HBV associated nephritis in children].

OBJECTIVE: Hepatitis B has been extensively prevalent in China and hepatitis B virus associated nephritis (HBV-GN) has been one of the common renal damages secondary to HBV infection in Chinese children. Regular vaccination against hepatitis B has been carried out nation-wide in China since January 1st, 1992. The present study was conducted to evaluate the effect of regular vaccination against hepatitis B virus on the incidence of childhood HBV-GN and membranous nephropathy (MN). METHODS: Retrospective analysis on the results of renal biopsy in 727 patients (from Nov. 1979 to March 2002) was carried out. The patients were first divided into two groups according to the date when the patients were seen. Group A patients were seen from Nov. 1979 through Dec. 1991; Group B patients were seen from Jan. 1992 through March 2002. Group B patients were further divided into 5 subgroups (Group B(1) to B(5)), with a 2-year interval after 1992. Secondly, each of these groups and subgroups were again divided into two groups, vaccinated and unvaccinated groups. RESULTS: In 727 renal biopsies, 64 cases (8.80%) met HBV-GN diagnostic criteria. Twenty-eight cases were diagnosed as HBV-GN in Group A (211 cases), accounting for 13.27%, while there were 36 cases with HBV-GN in 516 renal biopsies of Group B, accounting for 6.98% (chi(2) = 7.397 and P < 0.01). The frequency in Group B was significantly lower. Prevalence rate (from Group A to Group B(5)) was 13.3% (28/211), 13.0% (9/69), 7.3% (6/82), 6.3% (4/64), 4.9% (4/82), 5.9% (13/219), respectively, which showed a tendency of decline. Only 8 cases of HBV-GN occurred in vaccinated group (231 cases), accounting for 3.5%, while 48 cases of HBV-GN were seen in unvaccinated group (381 cases), accounting for 12.6% (chi(2) = 14.44 and P < 0.001), vaccination history was unknown in 115 of the 727 cases. In 727 renal biopsies, pathological type of 46 cases (6.3%) was membranous nephropathy and all of them had HBV-GN. Six cases of MN occurred in vaccinated group, accounting for 2.60%, while 40 cases with membranous nephropathy were found in unvaccinated group, accounting for 10.5% (chi(2) = 12.92 and P < 0.001). On the other hand, in vaccinated group there still were 8 cases of HBV-GN whose serum markers of HBV were positive. Two of their mothers had apparent evidence of hepatitis B virus infection. CONCLUSION: The frequency of HBV-GN has decreased significantly after vaccination against hepatitis B virus was routinely carried out since 1992; at the same time, childhood membranous nephropathy might be decreasing gradually, too. The cause of individual cases of HBV-GN who has be vaccinated was probably due to maternal-infant transmission and immunization failure. Attention should be paid to interruption of maternal-infant transmission and serological follow-up should be performed in high-risk newborns after vaccination to further lower the incidence of hepatitis B virus associated nephritis.

[Phylogenic analysis of the Sox gene family of vertebrate].

Sox genes of vertebrate are highly evolutionarily conserved, which encode different transcriptional factors involved in various developmental processes. Sox family is characterized by a sequence-specific DNA binding HMG-box containing about 79 amino acids. To realize the complexity of genes of the Sox family in the structures, functions and their evolutionary relationships, in the present study by utilizing all available complete nucleotide/protein sequence data of vertebrate Sox genes, we performed multi-sequence comparison and construction of phylogenic tree, and the grouping of Sox family members and the pattern of their molecular evolution was also analyzed.