Regulator of Ribosome Synthesis 1 (RRS1) Stabilizes GRP78 and Promotes Breast Cancer Progression.

Regulator of ribosome synthesis 1 (RRS1), a crucial regulatory factor in ribosome biogenesis, exerts a remarkable impact on the progression of breast cancer (BC). However, the exact mechanisms and pathways have not yet been fully elucidated. To investigate the impact of RRS1 on BC growth and metastasis, along with its underlying mechanisms. We discovered that RRS1 is overexpressed in BC tissues and cell lines. This study aims to regulate the level of RRS1 through lentiviral transfection technology to explore its potential function in BC cells. Knockdown of RRS1 resulted in the inhibition of cell proliferation, invasion, and migration, whereas overexpression had the opposite effects. We firstly identified the interaction between RRS1 and Glucose-Regulated Protein 78 (GRP78) using Co-immunoprecipitation (Co-IP) combined with mass spectrometry analysis, providing evidences of co-localization and positive regulation between RRS1 and GRP78. We observed that RRS1 inhibited the degradation of GRP78 through the ubiquitin-proteasome pathway, resulting in the stabilization of GRP78. In addition, our findings suggested that RRS1 promoted BC progression by activating the GRP78-mediated phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway. In conclusion, this newly discovered RRS1/GRP78 signaling axis provides a molecular and theoretical basis for further exploring the mechanisms of breast cancer invasion and metastasis.

Clinical significance of LAG-3 on microvessel density in primary hepatocellular carcinoma.

Aims: We aimed to determine whether lymphocyte activation gene 3 (LAG-3), also known as CD223, is associated with microvessel density (MVD) in primary hepatocellular carcinoma (HCC), as well as their clinical significance in predicting survival. Materials and methods: One hundred and twenty-seven patients were enrolled in the study. Samples were obtained on resection at the Department of Hepatobiliary Surgery of the Qingdao Municipal Hospital from June 2014 to June 2016. Immunohistochemistry was used to determine vessel density and LAG-3 abundance. Statistical analyses were performed to test for correlation of LAG-3 density and other clinicopathological variables with overall survival (OS). Results: High LAG-3 abundance was significantly correlated with increased MVD in primary HCC (P < 0.05). The χ2 test revealed a significant association of LAG-3 with preoperative AFP level, tumor diameter, N stage, and the presence of HBV infection (P < 0.05). Patients with high LAG-3 expression had shorter OS compared to those with low LAG-3 expression (P < 0.05). The Cox proportional hazards model showed that both higher LAG-3 and MVD density, age, the number of tumors, preoperative AFP level, tissue differentiation, Child-Pugh grade, and lymph node metastasis correlated with survival. Conclusions: High expression of LAG-3 is associated with angiogenesis and poor prognosis in HCC patients. With the deepening of research, LAG-3 is likely to become a novel biomarker for clinical diagnosis and prognosis and can even be a therapeutic target of HCC.

LAG-3 is a promising inhibitory immune checkpoint for antitumor immunotherapy.

INTRODUCTION: Lymphocyte activation gene-3 (LAG-3) is a member of the immunoglobulin superfamily. Engagement of LAG-3 by its ligands to trigger downstream signaling can inhibit immune responses and regulate the pathogenesis of many diseases, including cancer and inflammatory diseases. AREAS COVERED: We used keywords to search for relevant publications in PubMed and information on websites. After systematic analysis, we discuss the biological characteristics of LAG-3 and its ligands, LAG-3 related signaling, its roles in the pathogenesis of tumors, and its blockages for the treatment of cancers, as well as current challenges and future directions of research. EXPERT OPINION: Although the mechanisms underlying the action of LAG-3/ligand-related signaling in tumor development are not fully understood, advances in scientific research and LAG-3-based immunotherapies are promising. Further studies to explore its biological roles and molecular mechanisms may aid in developing new LAG-3- and ligand-based therapeutic drugs to benefit patients with different types of cancers.

Correlation Analysis Among the Level of IL-35, Microvessel Density, Lymphatic Vessel Density, and Prognosis in Non-Small Cell Lung Cancer.

The aim of this study was to determine the expression of IL-35 and the lymphatic vessel density (LVD) and microvessel density (MVD) in the pathological tissues from patients with non-small cell lung cancer (NSCLC) and to analyze their correlation with other common clinical prognostic factors, as well as patients' overall survival and progression-free survival. We analyzed the pathological characteristics of 130 patients with NSCLC and determined the IL-35 expression, MVD, and LVD changes in the pathological tissues by immunohistochemistry. The results showed that IL-35 expression was significantly correlated with tumor differentiation, lymph node metastasis, T staging, LVD, and MVD (P < 0.05) but was not associated with age, sex, smoking, and other factors. Univariate analysis of risk models showed that age, lymph node metastasis, T stage, and high IL-35 expression, LVD, and MVD were significantly associated with NSCLC prognosis (P < 0.05), whereas sex, smoking, and high differentiation were not correlated with prognosis. Multivariate analysis of the proportional risk model showed that the IL-35 expression, lymph node metastasis, high LVD, and high MVD were significantly correlated with NSCLC prognosis (P < 0.05). In conclusion, IL-35, MVD, and LVD may be independent prognostic markers. In addition, IL-35 might represent a promising clinical drug target for the treatment of NSCLC.

[Treatment of distal humerus fracture with unexposed ulnar nerve medial elbow incision and anatomical locking compression plate].

OBJECTIVE: To investigate the feasibility and effectiveness of unexposed ulnar nerve medial elbow incision, open reduction and internal fixation of anatomical locking compression plate (LCP) for distal humerus fractures. METHODS: Fourteen patients with distal humerus fracture were treated between January 2014 and June 2017. There were 5 males and 9 females, aged 18-85 years (mean, 65.5 years). The causes of injury included falling from height in 12 cases and traffic accident in 2 cases, all were closed fractures. Fractures were classified according to the AO/Association for the Study of Internal Fixation (AO/ASIF): 3 cases of type A2, 2 cases of type A3, 4 cases of type B2, 2 cases of type C1, 2 cases of type C2, and 1 case of type C3; without ulnar nerve damage. The time from injury to operation was 4-15 days, with an average of 7 days. The type B2 fractures were treated with unexposed ulnar nerve elbow medial incision and anatomic LCP internal fixation, the rest patients were all treated with unexposed ulnar nerve medial plus conventional lateral approach and bilateral LCP internal fixation. RESULTS: The operation time was 50-140 minutes (mean, 80 minutes), and the intraoperative blood loss was 20-200 mL (mean, 70 mL). There was no blood vessels or nerve damage during operation. All incisions healed by first intension, and no incision infection occurred. All the 14 cases were followed up 9-24 months (mean, 13 months). X-ray films showed that all fractures healed within 4 months without complications such as nonunion and osteomyelitis. No ulnar nerve injury, cubitus varus deformity, and ossifying myositis occurred during follow-up. At last follow-up, the elbow function was assessed by Mayo Elbow Performance score (MEPS), the results were excellent in 8 cases, good in 4 cases, fair in 1 case, and poor in 1 case (type C3 fracture), with the excellent and good rate of 85.7%. CONCLUSION: The unexposed ulnar nerve medial elbow incision can be used effectively to reduct the fracture, and it is not prone to ulnar nerve injury. Combined with the lateral approach to treat the distal humerus fracture, which has the advantages of short operation time, few trauma, little bleeding, and reliable effectiveness.

Clinical significance of cancer-associated fibroblasts and their correlation with microvessel and lymphatic vessel density in lung adenocarcinoma.

BACKGROUND: To determine whether cancer-associated fibroblasts (CAFs) are associated with microvessel density (MVD) and lymphatic vessel density (LVD) in lung adenocarcinoma (ADC) or are not prognostic. METHODS: Ninety-three lung adenocarcinoma patients without adjuvant therapy between January 2010 and June 2011 were enrolled. CAFs, MVD, and LVD were identified by α-smooth muscle actin (α-SMA), CD34 and D2-40 staining via immunohistochemistry. Staining intensities were assessed and quantified. For statistics, Pearson's chi-square test, logistic regression, Kaplan-Meier, and log-rank tests were applied. In addition, the Cox proportional hazards model was used for multifactor analysis to predict survival. RESULTS: CAFs abundance in lung adenocarcinoma is associated with higher MVD and LVD. In addition, a correlation was demonstrated between MVD and LVD (P < 0.05). CAFs, MVD, and LVD are significantly correlating with age, tumor size, differentiation grade, clinical stage, and lymph node metastasis (P < 0.05), but not influenced by gender, tumor location, and smoking history. Three-year overall survival in the CAFs-poor group is 64.5%, which is significant higher than that in the CAFs-rich cohort (41.9%). Further, we found that age, clinical stage, α-SMA, CD34, D2-40 positivity, tumor size, differentiation grade, and lymph node metastasis significantly correlate with overall survival of patients with lung adenocarcinoma. However, sex, smoking history, and tumor location have no association with 3-year survival. The clinical stage is an independent prognostic factor in overall survival (P < 0.05). CONCLUSIONS: The density of CAFs identified by α-SMA staining is associated with progression and metastasis of lung adenocarcinoma and affects the patient's disease outcome.

Clinical significance of FAP-α on microvessel and lymphatic vessel density in lung squamous cell carcinoma.

AIMS: We aimed to determine whether cancer-associated fibroblasts (CAFs) are associated with microvessel density (MVD) and lymphatic vessel density (LVD) in lung squamous cell carcinoma, as well as their clinical significance in predicting survival. METHODS: 122 patients were enrolled in the study. Samples were obtained on resection at the Department of Thoracic Surgery of the Qingdao Municipal Hospital between January 2011 and December 2014. Immunohistochemistry was used to determine vessel and lymphatic vessel density, and CAF abundance (fibroblast activation protein α (FAP-α) positivity). Statistical analyses were performed on 85 patients to test for correlation of CAF density and other clinicopathological variables with 3-year overall survival (OS) and disease-free survival (DFS). RESULTS: High stromal CAF abundance significantly correlated with increased MVD and LVD in lung squamous cell carcinoma (p<0.05). χ2 test revealed a significant association of CAF density with lymph node metastasis. Cox proportional hazards model showed that both higher CAF density and lymph node metastasis negatively correlate with survival. CAF density or lymph node status can be used as an independent prognostic factor to predict 3-year OS and DFS. CONCLUSIONS: CAF density, identified by FAP-α staining pattern, should be considered as a novel biomarker for disease prognosis in patients with lung squamous cell carcinoma.

IL-33 reflects dynamics of disease activity in patients with autoimmune hemolytic anemia by regulating autoantibody production.

BACKGROUND: Autoimmune hemolytic anemia (AIHA), a life-threatening anemia with rapid onset, is caused by autoantibody directed to self red blood cells (RBCs). Currently, mechanisms underlying AIHA pathogenesis are largely undefined. Here we explored the correlation of IL-33 with AIHA disease activity and evaluated IL-33 based therapeutics in AIHA treatment. METHODS: Thirty patients diagnosed with AIHA of warm-type autoantibodies without treatment were enrolled and followed up for 6 months. Levels of cytokines including IL-33, IL-4, IL-6 and IL-13 was determined with ELISA. AIHA disease activity was presented by levels of reticulocyte count, hemoglobin and lactate dehydrogenase. Serum RBC-bound IgG autoantibody was detected using anti-IgG antibody with flow cytometry. To evaluate the effect of IL-33 blockade on AIHA development, groups of B6 mice were immunized with rat RBCs plus recombinant IL-33 protein or IL-33 neutralizing antibody respectively and detected for levels of anti-RBC antibody, frequency of reticulocytes and destruction of transfused syngeneic mouse RBCs. RESULTS: Serum level of IL-33 was higher in AIHA patients compared with healthy individuals. Of interest, serum IL-33 was positively correlated with AIHA disease activity and sensitive to their changes in AIHA patients under clinical management. Mechanistically, IL-33 could promote the production of anti-RBC autoantibody. Serum IL-33 was closely associated with serum anti-RBC autoantibody and sensitive to their changes in AIHA patients. Accordingly, blockade of IL-33 interfered with AIHA incidence and ameliorated disease activity. Vice vasa, enforced IL-33 promoted AIHA incidence and disease activity. CONCLUSIONS: IL-33 was a potential biomarker for monitoring disease activity and therapeutic response in AIHA patients. Targeting IL-33 was a promising strategy for controlling autoantibody production in AIHA patients.

Electrospun hierarchical TiO2 nanorods with high porosity for efficient dye-sensitized solar cells.

Ultraporous anatase TiO2 nanorods with a composite structure of mesopores and macropores fabricated via a simple microemulsion electrospinning approach were first used as photoanode materials for high-efficiency dye-sensitized solar cells (DSSCs). The special multiscale porous structure was formed by using low-cost paraffin oil microemulsion droplets as the soft template, which can not only provide enhanced adsorption sites for dye molecules but also facilitate the electrolyte diffusion. The morphology, porosity, and photovoltaic and electron dynamic characteristics of the porous TiO2 nanorod based DSSCs were investigated in detail by scanning electron microscopy (SEM), N2 sorption measurements, current density-voltage (J-V) curves, UV-vis diffuse reflectance spectra, electrochemical impedance spectroscopy (EIS), intensity modulated photocurrent/photovoltage spectroscopy (IMPS/IMVS), and open-circuit voltage decay (OCVD) measurements. The results revealed that, although fewer amounts of dyes were anchored on the porous TiO2 nanorod films, they exhibited stronger light scattering ability, fast electrolyte diffusion, and extended electron lifetime compared to the commercial P25 nanoparticles. A power conversion efficiency of 6.07% was obtained for the porous TiO2 nanorod based DSSCs. Moreover, this value can be further improved to 8.53% when bilayer structured photoanode with porous TiO2 nanorods acting as the light scattering layer was constructed. This study demonstrated that the porous TiO2 nanorods can work as promising photoanode materials for DSSCs.

Critical role of Th17 cells in development of autoimmune hemolytic anemia.

Autoimmune hemolytic anemia (AIHA) is defined as the increased destruction of red blood cells (RBCs) in the presence of anti-RBC autoantibodies with or without complement activation. However, the underlying mechanism for the development of AIHA remains largely unclear. In this study, we carefully evaluated the potential role of Th17 cells in the development of AIHA. We found an elevated frequency of Th17 cells in patients with AIHA, which were closely correlated with their disease activity, including the level of anti-RBC IgG antibodies, hemoglobin, serum C3, and lactate dehydrogenase activity. Furthermore, we observed that interleukin (IL)-17 was also closely correlated with the disease activity in AIHA patients. To further elucidate the potential role of Th17 cells in induction of AIHA, we used the Marshall-Clarke and Playfair model of murine AIHA. Notably, we found that Th17 cells affected development of AIHA by enhancing the adaptive humoral responses. Specifically, we found that adoptive transfer of Th17 cells heightened the initial anti-rat RBC antibody responses and concomitantly increased the onset of AIHA. In addition, in vivo neutralization of IL-17 abrogated the development of AIHA, while initiation of anti-rat RBC IgG responses and induction of AIHA in IL-17(-/-) mice were impaired. Our findings suggest that Th17 cells contribute to the development of AIHA, which could facilitate our better understanding of AIHA pathogenesis and provide clues to developing novel forms of immunotherapy against AIHA.

[Expression of miR-21 in multiple myeloma and its clinical significance].

This study was aimed to explore the expression of microRNA-21 (miR-21) in multiple myeloma (MM), and its correlation with plasma ß2-microglobulin (ß2-MG) and staging of MM by Durie-Salmon (D-S) classification. The expression level of miR-21 in bone marrow mono-nuclear cells (BMMNC) of 43 patients with MM and 20 healthy individuals was examined by real-time polymerase chain reaction (real-time PCR), and the correlations among the expression level of miR-21 and related clinical pathologic features, plasma ß2-MG and staging of MM by D-S classification were analyzed. The results showed that the expression of miR-21 in BMMNC of MM patients was obviously higher than that in normal controls (P < 0.05). The expression of miR-21 in BMMNC of relapsed/refractory MM patients was obviously higher than that in newly diagnosed MM patients. The expression of miR-21 in MM patients decreased after chemical therapy, especially in effective group (P < 0.05), there was no significant change of miR-21 expression level in ineffective/progressive group before and after chemical therapy (P > 0.05). The expression of miR-21 was related with staging of MM and plasma ß2-MG level. It is concluded that the expression levels of miR-21 in BMMNC of MM patients are significantly higher than in normal bone marrow, these data indicated that miR-21 may play an important role in the development of MM. Super expression of miR-21 is positively correlated with level of plasma ß2-MG and staging of MM by D-S classification.

[Expression and significance of B cell-activating factor of TNF family (BAFF) and B cell lymphoma/leukemia-2 (BCL-2) in multiple myeloma].

This study was aimed to explore the expression of B cell-activating factor of TNF family (BAFF) and B cell lymphoma/leukemia-2 (BCL-2) in bone marrow mononuclear cells (BMMNC) of multiple myeloma (MM) and the significance of BAFF and BCL-2 for occurrence, development and prognosis of MM. The bone marrow of 40 cases of MM and 10 healthy persons was collected, the mononuclear cells (MNC) were isolated, the expression of BAFF and BCL-2 mRNA in BMMNC was detected by real-time PCR; the plasma was simultaneously collected and the ß2-MG level was determined; the clinical staging of MM patients was performed according to Durie-Salmon (D-S) staging criterion. The results indicated that the expression level of BAFF and BCL-2 mRNA in MM patients increased, as compared with normal controls, the difference was statistical significant (p < 0.05); the expression level of BAFF and BCL-2 mRNA in plateau stage after treatment obviously decreased. The expression level of BAFF and BCL-2 mRNA in relapsed/refractory MM patients was significantly higher than that in normal controls and patients reached plateau stage (p < 0.05), there was no statistically significant difference between newly diagnosed and relapsed/refractory MM patients (p > 0.05). The expression of BAFF and BCL-2 mRNA related with D-S staging and ß2-MG level. It is concluded that the expression levels of BAFF and BCL-2 mRNA increase, moreover the expression levels of BAFF and BCL-2 mRNA in newly diagnosed and relapsed/refractory MM patients are higher than those in patients reached plateau stage, which suggest the BAFF and BCL-2 may be involved in occurrence and development of MM; the relation of expression level of BAFF and BCL-2 mRNA to MM load is positive, which indicates the expression level of BAFF and BCL-2 mRNA may be a new indicator for evaluating the prognosis of MM patients.

[Mechanism underlying IL-2 inhibition on T cell specific immune response].

The study was aimed to investigate the effect of IL-2 on the acquired immune response of naive T cells, and to explore the influence of IL-2 on suppressor of cytokine signaling 3 (SOCS-3) expression of naive T cells, as well as to elucidate the role of SOCS-3 on antigen specific immune response in vitro. Naive DO11.10 T cells were pre-stimulated with IL-2 (50 U/ml), and stimulated with OVA(323 - 329) antigen after removing IL-2, then the proliferation of naive DO11.10 T cells was detected. Naive DO11.10 T cells were stimulated with IL-2 (50 U/ml), and SOCS-3 expression was detected by real-time PCR. Naive DO11.10 T cells were stimulated with OVA(323 - 329) antigen, and SOCS-3 expression was detected by means of (3)H-TdR. The results showed that after IL-2 pre-stimulation, the proliferation of naive DO11.10 T cells decreased significantly when stimulated with OVA(323 - 329) antigen; SOCS-3 expression of naive DO11.10 T cells was up-regulated significantly after IL-2 stimulation, the up-regulation began obviously at 4 hours and reached peak at 6 hours. SOCS-3 expression on naive DO11.10 T cells was down-regulated markedly after OVA(323 - 329) antigen stimulation, the expression level of SOCS-3 was lowest on day 2 and returned to normal on day 4 after stimulation. It is concluded that the antigen specific immune response of naive DO11.10 T cells is inhibited after pre-stimulation with IL-2, which may be mediated by SOCS-3.