Application of mendelian randomization in ocular diseases: a review.

Ocular disorders can significantly lower patients' quality of life and impose an economic burden on families and society. However, for the majority of these diseases, their prevalence and mechanisms are yet unknown, making prevention, management, and therapy challenging. Although connections between exposure factors and diseases can be drawn through observational research, it is challenging to rule out the interference of confounding variables and reverse causation. Mendelian Randomization (MR), a method of research that combines genetics and epidemiology, has its advantage to solve this problem and thus has been extensively utilized in the etiological study of ophthalmic diseases. This paper reviews the implementation of MR in the research of ocular diseases and provides approaches for the investigation of related mechanisms as well as the intervention strategies.

Toxoplasma gondii Causes Adverse Pregnancy Outcomes by Damaging Uterine Tissue-Resident NK Cells That Secrete Growth-Promoting Factors.

Vertical transmission of the intracellular parasite, Toxoplasma gondii can lead to adverse pregnancy outcomes especially when infection occurs in early pregnancy. Decidual natural killer (dNK) cells accumulate at the maternal-fetal interface in large numbers during early pregnancy. Their nutritional roles during infection with T. gondii remain poorly defined. In the present study, we demonstrated that a functional deficiency of the uterine tissue-resident NK (trNK) cells, a subset of dNK cells, contributes to the adverse pregnancy outcomes induced by T. gondii in early pregnancy. Adverse pregnancy outcomes could be ameliorated by adoptive transfer of trNK cells. Moreover, fetal growth restriction could be improved after supplementation of growth-promoting factors. In addition to the widely recognized disturbance of the immune balance at the interface between the mother and the fetus, our study reveals a novel mechanism in T. gondii that contributes to the adverse pregnancy outcomes.

Quaternary Ammonium Salts: Insights into Synthesis and New Directions in Antibacterial Applications.

The overuse of antibiotics has led to the emergence of a large number of antibiotic-resistant genes in bacteria, and increasing evidence indicates that a fungicide with an antibacterial mechanism different from that of antibiotics is needed. Quaternary ammonium salts (QASs) are a biparental substance with good antibacterial properties that kills bacteria through simple electrostatic adsorption and insertion into cell membranes/altering of cell membrane permeability. Therefore, the probability of bacteria developing drug resistance is greatly reduced. In this review, we focus on the synthesis and application of single-chain QASs, double-chain QASs, heterocyclic QASs, and gemini QASs (GQASs). Some possible structure-function relationships of QASs are also summarized. As such, we hope this review will provide insight for researchers to explore more applications of QASs in the field of antimicrobials with the aim of developing systems for clinical applications.

An Improved RAPID Imaging Method of Defects in Composite Plate Based on Feature Identification by Machine Learning.

The RAPID (reconstruction algorithm for probabilistic inspection of defect) method based on Lamb wave detection is an effective method to give the position information of a defect in composite plate. In this paper, an improved RAPID imaging method based on machine learning (ML) is proposed to precisely visualize the location and features of defects in composite plate. First, the specific feature information of the defect, such as type, size and direction, can be identified by analyzing the detection signals through multiple machine learning models. Then, according to the obtained defect features, the scaling parameter ß of the RAPID method which controls the size of the elliptical area is revised, and weights are set to the important detection paths which are related to defect features to realize precise defect imaging. The simulation results show that the proposed method can intuitively characterize the location and related feature information of the defect, and effectively improve the accuracy of defect imaging.

An Intelligence Method for Recognizing Multiple Defects in Rail.

Ultrasonic guided waves are sensitive to many different types of defects and have been studied for defect recognition in rail. However, most fault recognition algorithms need to extract features from the time domain, frequency domain, or time-frequency domain based on experience or professional knowledge. This paper proposes a new method for identifying many different types of rail defects. The segment principal components analysis (S-PCA) is developed to extract characteristics from signals collected by sensors located at different positions. Then, the Support Vector Machine (SVM) model is used to identify different defects depending on the features extracted. Combining simulations and experiments of the rails with different kinds of defects are established to verify the effectiveness of the proposed defect identification techniques, such as crack, corrosion, and transverse crack under the shelling. There are nine channels of the excitation-reception to acquire guided wave detection signals. The results show that the defect classification accuracy rates are 96.29% and 96.15% for combining multiple signals, such as the method of single-point excitation and multi-point reception, or the method of multi-point excitation and reception at a single point.

Exploring gut microbiota and metabolite alterations in patients with thyroid-associated ophthalmopathy using high-throughput sequencing and untargeted metabolomics.

Introduction: Thyroid-associated ophthalmopathy (TAO) is an autoimmune-driven orbital inflammatory disease. Despite research efforts, its exact pathogenesis remains unclear. This study aimed to characterize the intestinal flora and metabolic changes in patients with TAO to identify the flora and metabolites associated with disease development. Methods: Thirty patients with TAO and 29 healthy controls were included in the study. The intestinal flora and metabolites were analyzed using high-throughput sequencing of the 16S rRNA gene and non-targeted metabolomics technology, respectively. Fresh fecal samples were collected from both populations for analysis. Results: Reduced gut richness and diversity were observed in patients with TAO. Compared to healthy controls, significant differences in relative abundance were observed in patients with TAO at the order level Clostridiales, family level Staphylococcaceae, genus level Staphylococcus, Fournierella, Eubacterium siraeum, CAG-56, Ruminococcus gnavus, Intestinibacter, Actinomyces, and Erysipelotrichaceae UCG-003 (logFC>1 and P<0.05). Veillonella and Megamonas were closely associated with clinical symptoms in patients with TAO. Among the 184 significantly different metabolites, 63 were upregulated, and 121 were downregulated in patients with TAO compared to healthy controls. The biosynthesis of unsaturated fatty acids was the significantly enriched metabolic pathway. Correlation analysis revealed Actinomyces was positively correlated with NAGlySer 15:0/16:0, FAHFA 3:0/20:0, and Lignoceric Acid, while Ruminococcus gnavu was positively correlated with Cer 18:0;2O/16:0; (3OH) and ST 24:1;O4/18:2. Conclusion: Specific intestinal flora and metabolites are closely associated with TAO development. Further investigation into the functional associations between these flora and metabolites will enhance our understanding of TAO pathogenesis.

Quaternary ammonium salts for water treatment with balanced rate of sterilization and degradation.

The growing number of infections caused by drug-resistant bacteria which arise from the overuse of antibiotics has severely affected the normal operation of human society. The high antibacterial activity of QAS makes it promising as an alternative to antibiotics, but it suffers from secondary pollution due to its non-degradation. Here we have synthesized a class of gemini quaternary ammonium salts (GQAS) with different carbon chain lengths containing ester groups by using facile methylation reaction. Quaternary ammonium groups contribute to insert negatively charged bacterial membranes, resulting in membrane damage and bacteria death. Compared with conventional single-chain QAS, except for the more efficient antibacterial efficiency attribute to the presence of the second carbon chain, GQAS with alterable antibacterial properties can minimize the possibility of bacterial resistance and reduce the accumulation of GQAS in the environment through the introduction of degradable ester groups. GQAS is completely superior to the commercial bactericide benzalkonium chloride (BAC) in both antibacterial activity and degrade performance, which can be used as a more environmentally friendly bactericide.

Relationship between bullying victimization and murderous behaviors and impulsivity's mediating role: Evidence from Chinese early adolescents.

BACKGROUND: Evidence suggests a robust relationship between experiencing bullying victimization (BV) and engaging in murderous behaviors among adolescents. However, the potential mediating effect of impulsivity on the relationship between BV and murderous behaviors in early adolescents remains underexplored. METHODS: A total of 5724 adolescents, with a mean age of 13.5 years, were enrolled from three middle schools in Anhui Province, China. Participants completed self-report questionnaires detailing their experiences with bullying, impulsiveness, and murderous behaviors. To assess the relationship between BV and murderous behaviors, multivariate logistic regression and Poisson regression analyses were conducted. Mediation analysis was performed using structural equation modeling. RESULTS: After controlling for confounding factors, a positive association was found between experiencing BV and engaging in murderous behaviors (p < 0.05). Mediation analysis revealed a significant indirect effect of BV on the occurrence of murderous behaviors through impulsivity (indirect effect = 0.027, 95 % CI: 0.021, 0.033). BV appears to heighten levels of impulsivity, which in turn increases the likelihood of murderous behaviors. Additionally, sex-specific analysis indicated that impulsivity played a greater mediating role in the link between verbal and relational BV and murderous behaviors in females, while physical and cyber BV were more significant in males. CONCLUSIONS: Our findings underscore the necessity of early targeted interventions for adolescents experiencing BV and exhibiting high levels of impulsivity to mitigate their risk of engaging in murderous behaviors.

Predictive value of the domain specific PLA2R antibodies for clinical remission in patients with primary membranous nephropathy: A retrospective study.

BACKGROUND: M-type phospholipase A2 receptor (PLA2R) is a major auto-antigen of primary membranous nephropathy(PMN). Anti-PLA2R antibody levels are closely associated with disease severity and therapeutic effectiveness. Analysis of PLA2R antigen epitope reactivity may have a greater predictive value for remission compared with total PLA2R-antibody level. This study aims to elucidate the relationship between domain-specific antibody levels and clinical outcomes of PMN. METHODS: This retrospective analysis included 87 patients with PLA2R-associated PMN. Among them, 40 and 47 were treated with rituximab (RTX) and cyclophosphamide (CTX) regimen, respectively. The quantitative detection of -immunoglobulin G (IgG)/-IgG4 targeting PLA2R and its epitope levels in the serum of patients with PMN were obtained through time-resolved fluorescence immunoassays and served as biomarkers in evaluating the treatment effectiveness. A predictive PMN remission possibility nomogram was developed using multivariate logistic regression analysis. Discrimination in the prediction model was assessed using the area under the receiver operating characteristic curve (AUC-ROC).Bootstrap ROC was used to evaluate the performance of the prediction model. RESULTS: After a 6-month treatment period, the remission rates of proteinuria, including complete remission and partial remission in the RTX and CTX groups, were 70% and 70.21% (P = 0.983), respectively. However, there was a significant difference in immunological remission in the PLA2R-IgG4 between the RTX and CTX groups (21.43% vs. 61.90%, P = 0.019). Furthermore, we found differences in PLA2R-CysR-IgG4(P = 0.030), PLA2R-CTLD1-IgG4(P = 0.005), PLA2R-CTLD678-IgG4(P = 0.003), and epitope spreading (P = 0.023) between responders and non-responders in the CTX group. Multivariate logistic analysis showed that higher levels of urinary protein (odds ratio [OR], 0.49; 95% confidence interval [CI], 0.26-0.95; P = 0.035) and higher levels of PLA2R-CTLD1-IgG4 (OR, 0.79; 95%CI,0.62-0.99; P = 0.041) were independent risk factors for early remission. A multivariate model for estimating the possibility of early remission in patients with PMN is presented as a nomogram. The AUC-ROC of our model was 0.721 (95%CI, 0.601-0.840), in consistency with the results obtained with internal validation, for which the AUC-ROC was 0.711 (95%CI, 0.587-0.824), thus, demonstrating robustness. CONCLUSIONS: Cyclophosphamide can induce immunological remission earlier than rituximab at the span of 6 months. The PLA2R-CTLD1-IgG4 has a better predict value than total PLA2R-IgG for remission of proteinuria at the 6th month.

XAF1 promotes colorectal cancer metastasis via VCP-RNF114-JUP axis.

Metastasis is the main cause of colorectal cancer (CRC)-related death, and the 5-year relative survival rate for CRC patients with distant metastasis is only 14%. X-linked inhibitor of apoptosis (XIAP)-associated factor 1 (XAF1) is a zinc-rich protein belonging to the interferon (IFN)-induced gene family. Here, we report a metastasis-promoting role of XAF1 in CRC by acting as a novel adaptor of valosin-containing protein (VCP). XAF1 facilitates VCP-mediated deubiquitination of the E3 ligase RING finger protein 114 (RNF114), which promotes K48-linked ubiquitination and subsequent degradation of junction plakoglobin (JUP). The XAF1-VCP-RNF114-JUP axis is critical for the migration and metastasis of CRC cells. Moreover, we observe correlations between the protein levels of XAF1, RNF114, and JUP in clinical samples. Collectively, our findings reveal an oncogenic function of XAF1 in mCRC and suggest that the XAF1-VCP-RNF114-JUP axis is a potential therapeutic target for CRC treatment.

PLA2R-IgG4 antibody as a predictive biomarker of treatment effectiveness and prognostic evaluation in patients with idiopathic membranous nephropathy: a retrospective study.

Background: The Kidney Disease Improving Global Outcomes (KDIGO) 2021 guidelines recommend Rituximab (RTX) as the first-line therapy and phospholipase A2 receptor (PLA2R) antibody as a biomarker for remission and prognosis in patients with idiopathic membranous nephropathy (IMN). Methods: This study was a retrospective analysis of 70 patients with IMN treated with either rituximab (RTX) or cyclophosphamide (CTX) and steroid. Quantitative detection of PLA2R-IgG and PLA2R-IgG4 antibodies at sixth month after treatment, determined using time-resolved fluoroimmunoassay (TRFIA), were used for treatment effectiveness analysis and prognostic evaluation in patients with IMN. Results: After 12 months of therapy, the remission rate of proteinuria, including complete remission (CR) and partial remission (PR) in the RTX group and the CTX group, were 74% versus 67.5% (P = 0.114), respectively. Both PLA2R-IgG and PLA2R-IgG4 levels were decreased in patients with remission of proteinuria after 6 months of therapy. Receiver operating characteristic curve (ROC) curve analysis exhibited that the AUC of PLA2R-IgG4 and the PLA2R-IgG as laboratory criteria for proteinuria remission were 0.970 versus 0.886 (P = 0.0516), respectively, after 6 months of treatment. The cut-off value of PLA2R-IgG4 was 7.67 RU/mL and the sensitivity and specificity of remission rate at 6th month were 90.9% and 100%, respectively. Furthermore, the AUC of the PLA2R-IgG4 and PLA2R-IgG to predict the outcome after 12 months of treatment were 0.922 versus 0.897 (P = 0.3270), respectively. With the cut-off value of PLA2R-IgG4 being 22.985 RU/mL, the sensitivity and specificity of remission rate at 12th month were 100% and 87.1%, respectively. Logistic regression analysis revealed that the PLA2R-IgG4 level (P = 0.023), the rate of decrease of PLA2R-IgG4 level (P = 0.034), and eGFR level (P = 0.012) were significantly associated with remission. Conclusions: We found that the patients in the RTX group and CTX group achieved effective remission of proteinuria after 12 months of treatment. PLA2R-IgG4 may be a more effective biomarker for treatment effectiveness analysis and prognostic assessment, compared with anti-PLA2R-IgG for PLA2R associated IMN.

Kernelized k-Local Hyperplane Distance Nearest-Neighbor Model for Predicting Cerebrovascular Disease in Patients With End-Stage Renal Disease.

Detecting and treating cerebrovascular diseases are essential for the survival of patients with chronic kidney disease (CKD). Machine learning algorithms can be used to effectively predict stroke risk in patients with end-stage renal disease (ESRD). An imbalance in the amount of collected data associated with different risk levels can influence the classification task. Therefore, we propose the use of a kernelized k-local hyperplane nearest-neighbor model (KHKNN) for the effective prediction of stroke risk in patients with ESRD. We compared our proposed method with other conventional machine learning methods, which revealed that our method could effectively perform the task of classifying stroke risk.

A PLA2R-IgG4 Antibody-Based Predictive Model for Assessing Risk Stratification of Idiopathic Membranous Nephropathy.

Background: Known as an autoimmune glomerular disease, idiopathic membranous nephropathy (IMN) is considered to be associated with phospholipase A2 receptor (PLA2R) in terms of the main pathogenesis. The quantitative detection of serum PLA2R-IgG and PLA2R-IgG4 antibodies by time-resolved fluoroimmunoassay (TRFIA) was determined, and the value of them, both in the clinical prediction of risk stratification in IMN, was observed in this study. Methods: 95 patients with IMN proved by renal biopsy were enrolled, who had tested positive for serum PLA2R antibodies by ELISA, and the quantitative detection of serum PLA2R-IgG and PLA2R-IgG4 antibodies was achieved by TRFIA. All the patients were divided into low-, medium-, and high-risk groups, respectively, which were set as dependent variables, according to proteinuria and renal function. Random forest (RF) was used to estimate the value of serum PLA2R-IgG and PLA2R-IgG4 in predicting the risk stratification of progression in IMN. Results: Out-of-bag estimates of variable importance in RF were employed to evaluate the impact of each input variable on the final classification accuracy. The variable of albumin, PLA2R-IgG, and PLA2R-IgG4 had high values (>0.3) of 0.3156, 0.3981, and 0.7682, respectively, which meant that these three were more important for the risk stratification of progression in IMN. In order to further assess the contribution of PLA2R-IgG and PLA2R-IgG4 to the model, we built four different models and found that PLA2R-IgG4 played an important role in improving the predictive ability of the model. Conclusions: In this study, we established a random forest model to evaluate the value of serum PLA2R-IgG4 antibodies in predicting risk stratification of IMN. Compared with PLA2R-IgG, PLA2R-IgG4 is a more efficient biomarker in predicting the risk of progression in IMN.

A Self-Representation-Based Fuzzy SVM Model for Predicting Vascular Calcification of Hemodialysis Patients.

In end-stage renal disease (ESRD), vascular calcification risk factors are essential for the survival of hemodialysis patients. To effectively assess the level of vascular calcification, the machine learning algorithm can be used to predict the vascular calcification risk in ESRD patients. As the amount of collected data is unbalanced under different risk levels, it has an influence on the classification task. So, an effective fuzzy support vector machine based on self-representation (FSVM-SR) is proposed to predict vascular calcification risk in this work. In addition, our method is also compared with other conventional machine learning methods, and the results show that our method can better complete the classification task of the vascular calcification risk.

The unique molecular mechanism of diabetic nephropathy: a bioinformatics analysis of over 250 microarray datasets.

BACKGROUND/AIMS: Diabetic nephropathy (DN) is one of the main causes of end-stage kidney disease worldwide. Emerging studies have suggested that its pathogenesis is distinct from nondiabetic renal diseases in many aspects. However, it still lacks a comprehensive understanding of the unique molecular mechanism of DN. METHODS: A total of 255 Affymetrix U133 microarray datasets (Affymetrix, Santa Calra, CA, USA) of human glomerular and tubulointerstitial tissues were collected. The 22 215 Affymetrix identifiers shared by the Human Genome U133 Plus 2.0 and U133A Array were extracted to facilitate dataset pooling. Next, a linear model was constructed and the empirical Bayes method was used to select the differentially expressed genes (DEGs) of each kidney disease. Based on these DEG sets, the unique DEGs of DN were identified and further analyzed using gene ontology and pathway enrichment analysis. Finally, the protein-protein interaction networks (PINs) were constructed and hub genes were selected to further refine the results. RESULTS: A total of 129 and 1251 unique DEGs were identified in the diabetic glomerulus (upregulated n = 83 and downregulated n = 203) and the diabetic tubulointerstitium (upregulated n = 399 and downregulated n = 874), respectively. Enrichment analysis revealed that the DEGs in the diabetic glomerulus were significantly associated with the extracellular matrix, cell growth, regulation of blood coagulation, cholesterol homeostasis, intrinsic apoptotic signaling pathway and renal filtration cell differentiation. In the diabetic tubulointerstitium, the significantly enriched biological processes and pathways included metabolism, the advanced glycation end products-receptor for advanced glycation end products signaling pathway in diabetic complications, the epidermal growth factor receptor (EGFR) signaling pathway, the FoxO signaling pathway, autophagy and ferroptosis. By constructing PINs, several nodes, such as AGR2, CSNK2A1, EGFR and HSPD1, were identified as hub genes, which might play key roles in regulating the development of DN. CONCLUSIONS: Our study not only reveals the unique molecular mechanism of DN but also provides a valuable resource for biomarker and therapeutic target discovery. Some of our findings are promising and should be explored in future work.

New mutations of Nogo-C in hepatocellular carcinoma.

The neurite outgrowth inhibitor Nogo has attracted great interest, but its relevance with hepatocellular carcinoma has not been reported. This paper first found mutations of Nogo-C in HCC patients from Qidong in China: A172G (Thr58Ala), A340G (Arg114Gly), A571G (Ile191Val). In six examined patient cases from Qidong, the mutations occurred in five cases. The mutation Arg114Gly was predicted bioinformatically to affect Nogo-66 dimensional structure of Nogo-C. Our previous works also had indicated that mutant Nogo-C promoted liver cancer cell line apoptosis and resulted in molecular marker of HCC p53 gene transfer from nucleus to cytoplast. Above results revealed a new physiological role and clinical implications of Nogo-C on HCC.

Hsp70 inhibits lipopolysaccharide-induced NF-kappaB activation by interacting with TRAF6 and inhibiting its ubiquitination.

Inducible heat shock protein 70 (Hsp70) is one of the most important HSPs for maintenance of cell integrity during normal cellular growth as well as pathophysiological conditions. Tumor necrosis factor receptor-associated factor 6 (TRAF6) is a crucial signaling transducer that regulates a diverse array of physiological and pathological processes and is essential for activating NF-kappaB signaling pathway in response to bacterial lipopolysaccharide (LPS). Here we report a novel mechanism of Hsp70 for preventing LPS-induced NF-kappaB activation in RAW264.7 macrophage-like cells. Our results show that Hsp70 can associate with TRAF6 physically in the TRAF-C domain and prevent TRAF6 ubiquitination. The stimulation of LPS dissociates the binding of Hsp70 and TRAF6 in a time-dependent manner. Hsp70 inhibits LPS-induced NF-kappaB signaling cascade activation in heat-shock treated as well as Hsp70 stable transfected RAW264.7 cells and subsequently decreases iNOS and COX-2 expression. Two Hsp70 mutants, Hsp70DeltaC(1-428aa) with N-terminal ATPase domain and Hsp70C(428-642aa) with C-terminal domain, lack the ability to influence TRAF6 ubiquitination and TRAF6-triggered NF-kappaB activation. Taken together, these findings indicate that Hsp70 inhibits LPS-induced NF-kappaB activation by binding TRAF6 and preventing its ubiquitination, and results in inhibition of inflammatory mediator production, which provides a new insight for analyzing the effects of Hsp70 on LPS-triggered inflammatory signal transduction pathways.

[Inhibitory effect of iguratimod on TNFalpha production and NF-kappaB activity in LPS-stimulated rat alveolar macrophage cell line].

AIM: To investigate the effect of iguratimod (T-614), a non-steroidal anti-inflammatory drug, on TNFalpha mRNA expression and TNFalpha production, and on the activity of nuclear factor-kappaB (NF-kappaB) in the rat alveolar macrophage cell line (NR8383) activated by LPS. METHODS: NR8383 cells were pretreated with T-614 (13.4, 26.7, 53.4 micromol x L(-1)), then were stimulated with LPS. The production of TNFalpha in the supernatant of NR8383 was assayed by enzyme-linked immunosorbent assay (ELISA). The TNFalpha mRNA level was determined by a semi-quantitative PCR assay. Assessment of the NF-kappaB DNA binding activity was performed by an ELISA kit. RESULTS: T-614 inhibited LPS-stimulated mRNA expression and production of TNFalpha in a concentration-dependent manner, as well as the activity of NF-kappaB. The IC50 value of effect of T-614 on TNFalpha level was 26.2 micromol x L(-1). CONCLUSION: The inhibitory effect of T-614 on the production of TNFalpha in LPS-stimulated NR8383 cells may be mediated by suppression of NF-kappaB activity.

Cloning of the nucleostemin gene and its function in transforming human embryonic bone marrow mesenchymal stem cells into F6 tumor cells.

We recently established a novel tumor cell line denominated as F6, which was derived from mutated human embryonic bone marrow mesenchymal stem cells (MSCs). The difference between gene expression of F6 cells and MSCs was distinguished by fluorescent differential display. Results showed that the expression of nucleostemin, a novel factor participating in the control of stem and cancer cell proliferation, was different in F6 cells and MSCs. To further understand its role in transforming human embryonic bone marrow mesenchymal stem cells into F6 tumor cells, the full-length nucleostemin gene (1650 bp) from an LTEP-a-2 cell line was cloned, and GST-nucleostemin protein was expressed in E. coli. The characteristics of nucleostemin expression in F6 cells and other human cancer cell lines were investigated by RT-PCR, Western blot analysis, immunocytochemical staining, fluorescent microscope and confocal laser scanning microscope. The levels of nucleostemin gene expression were detected by real-time PCR in F6 tumor tissue obtained from SCID nude mice at 4, 6 and 7 weeks after the injection of F6 cells, and from the lung tissue of five lung cancer patients. Results showed that nucleostemin gene expression increased significantly in F6 tumor tissue and lung cancer tissue. The results also showed that transfection with pcDNA3.1(+)-GFP-nucleostemin for 4-20 weeks promoted cell size augmentation and nuclei multiplication, and the cells were converted to giant cell-like cells. Western blot analysis revealed that expression levels of nucleostemin in the nuclei and cytoplasm of cancer cell lines, e.g. F6, LTEP-a-2, U937, SW480 and 95D, were higher than those in MSCs and COS-7 cells. Levels of nucleostemin in F6 cells were notably high and confirmed with immunohistochemical staining. These results implied that nucleostemin may play an important role in both tumorigenesis and transforming human embryonic bone marrow mesenchymal stem cells into F6 tumor cells.

RTN4-C gene expression in hepatocellular carcinoma and its influence on SMMC7721 cell growth and apoptosis.

RTN4-C gene is a member of RTNs. To investigate its expression in human hepatocellular carcinoma tissues and study its function on SMMC7721 cells, RT-PCR was conducted to detect its expressions in human hepatocellular carcinoma tissues. RTN4-C-pcDNA3. 1 plasmid was reconstructed and stably transfected into SMMC7721 cells by Lipofect AMINE. Growth curve of SMMC7721 measured by MTT and apoptosis indentified with acridine orange staining were examined to demonstrate the effect of RTN4-C on SMMC7721. Immunocytochemical analysis for mutant p53, c-Fos, Hsp70 proteins were conducted. The results showed that the transfected SMMC7721 cells grew more slowly than control and the average inhibition rate at the 1st, 2nd and 3rd day were 33.8% +/- 0.026, 56.2% +/- 0. 094, 34.8% +/- 0.077 respectively. In transfected SMMC7721 cell line, the apoptosis was strengthened,mutant p53 protein tranferred from nucleus to cytoplasm and c-Fos, Hsp70 proteins were decreased. Our data indicated RTN4-C gene was expressed differently in hepatocellular carcinoma and its paracancerous tissues. By tranferring mutant p53 protein from nucleus to cytoplasm and decreasing c-Fos, Hsp70 protein expression, RTN4-C inhibited SMMC7721 cells growth and promoted its apoptosis.