Associations of Urinary Polycyclic Aromatic Hydrocarbons With Bone Mineral Density at Specific Body Sites in U.S. Adults, National Health and Nutrition Examination Survey 2001 to 2004.

OBJECTIVE: We aimed to analyze the association between certain types of urinary polycyclic aromatic hydrocarbons (PAHs) and bone mineral density (BMD) at specific sites of the body. METHODS: A total of 2978 eligible participants from the National Health and Nutrition Examination Survey 2001 to 2004 were included in this study. Data of 8 urinary PAHs and BMDs of 3 skeleton sites and the total body were analyzed. Univariate and multivariate linear regression analyses were performed to explore the association between urinary PAHs and BMDs. Subgroup analyses stratified by sex and body mass index were also performed. RESULTS: After adjustment for all confounders, elevated 3-fluorene (ß = 0.046; 95% confidence intervals [CIs], 0.007-0.084) and 2-fluorene (ß = 0.054; 95% CI, 0.007-0.100) levels were associated with greater left arm BMD, whereas no statistical differences were observed in the relationship between other PAHs and BMDs (all P > .05). Higher 3-fluorene and 2-fluorene levels were still associated with increased left arm BMD in men (P < .05), whereas the higher 2-phenanthrene level was related to decreased left arm BMD (ß = -0.062; 95% CI, -0.105 to -0.019), right arm BMD (ß = -0.059; 95% CI, -0.091 to -0.027), and total BMD (ß = -0.065; 95% CI, -0.119 to -0.012) in women. Similar results were also found in different body mass index populations (all P < .05). CONCLUSION: Certain urinary PAHs are associated with BMDs at specific body sites. Future studies are needed to illustrate the mechanisms behind the association to establish a causal relationship.

INHBA gene silencing inhibits gastric cancer cell migration and invasion by impeding activation of the TGF-ß signaling pathway.

Gastric cancer (GC) is the fourth largest cancer in the world, with a 5-year survival rate of <30%. Thus, this study intends to investigate the effects of inhibin ßA (INHBA) gene silencing on the migration and invasion of GC cells via the transforming growth factor-ß (TGF-ß) signaling pathway. Initially, this study determined the expression of INHBA and the TGF-ß signaling pathway-related genes in GC tissues. After that, to assess the effect of INHBA silencing on GC progression, GC cells were transfected with short hairpin RNAs that targeted INHBA in order to detect the expression of INHBA and the TGF-ß signaling pathway-related genes, as well as cell migration, invasion, and proliferation abilities. Finally, a tumor xenograft model in nude mice was constructed to verify the effect that the silencing of INHBA had on tumor growth. Highly expressed INHBA and activated TGF-ß signaling pathways were observed in GC tissues. In response to shINHBA-1 and shINHBA-2, the TGF-ß signaling pathway was inhibited in GC cells, whereas the GC cell migration, invasion, proliferation, and tumor growth were significantly dampened. On the basis of the observations and findings of this study, INHBA gene silencing inhibited the progression of GC by inactivating the TGF-ß signaling pathway, which provides a potential target in the treatment of GC.

The long non-coding RNA NEAT1 interacted with miR-101 modulates breast cancer growth by targeting EZH2.

Nuclear enriched abundant transcript 1 (NEAT1), an important cancer-associated long non-coding RNA (lncRNA), contributes to the development and progression of several cancers. An increased expression of NEAT1 was observed in cancers including bladder cancer, lung cancer and breast cancer (BC). However, the exact effect of NEAT1 in BC progression and the underlying molecular mechanisms are still unknown up to now. Here, we investigated the detailed role of NEAT1 in human BC cell lines and clinical tumor samples in order to validate the function of this molecule. In our research, lncRNA-NEAT1 was specifically upregulated in BC cell lines and promoted BC cell growth through targeting miR-101. Knockdown of NEAT1 inhibited the proliferation and DNA synthesis of human BC cell in vitro. In addition, the regulation of EZH2 by miR-101 was required in NEAT1 induced BC cell growth. These findings indicated that NEAT1 might suppress the tumor growth via miR-101 dependent EZH2 regulation. Taken together, our data indicated that NEAT1 might be an oncogenic lncRNA that promoted proliferation of BC and could be regarded as a therapeutic target in human BC.

Design and Characterization of a Novel eEF2K Degrader with Potent Therapeutic Efficacy Against Triple-Negative Breast Cancer.

Dysregulated eEF2K expression is implicated in the pathogenesis of many human cancers, including triple-negative breast cancer (TNBC), making it a plausible therapeutic target. However, specific eEF2K inhibitors with potent anti-cancer activity have not been available so far. Targeted protein degradation has emerged as a new strategy for drug discovery. In this study, a novel small molecule chemical is designed and synthesized, named as compound C1, which shows potent activity in degrading eEF2K. C1 selectively binds to F8, L10, R144, C146, E229, and Y236 of the eEF2K protein and promotes its proteasomal degradation by increasing the interaction between eEF2K and the ubiquitin E3 ligase ßTRCP in the form of molecular glue. C1 significantly inhibits the proliferation and metastasis of TNBC cells both in vitro and in vivo and in TNBC patient-derived organoids, and these antitumor effects are attributed to the degradation of eEF2K by C1. Additionally, combination treatment of C1 with paclitaxel, a commonly used chemotherapeutic drug, exhibits synergistic anti-tumor effects against TNBC. This study not only generates a powerful research tool to investigate the therapeutic potential of targeting eEF2K, but also provides a promising lead compound for developing novel drugs for the treatment of TNBC and other cancers.

Circular RNA circPOFUT1 enhances malignant phenotypes and autophagy-associated chemoresistance via sequestrating miR-488-3p to activate the PLAG1-ATG12 axis in gastric cancer.

Circular RNAs are key regulators in regulating the progression and chemoresistance of gastric cancer (GC), suggesting circular RNAs as potential therapeutic targets for GC. The roles of a novel circular RNA circPOFUT1 in GC are unknown. Here, we found that circPOFUT1 was upregulated in GC tissues and cells, and increased circPOFUT1 expression indicated poor prognosis. Overexpression of circPOFUT1 enhanced cell proliferation, migration, invasion and autophagy-associated chemoresistance in GC, which were suppressed by miR-488-3p overexpression. CircPOFUT1 reduced miR-488-3p expression via sponging miR-488-3p in GC cells. PLAG1 interacted with ATG12 and promoted its expression. MiR-488-3p bound to PLAG1 and suppressed the expression of PLAG1 and ATG12 in GC cells. Overexpression of circPOFUT1 enhanced autophagy-associated chemoresistance of GC cells in vivo, but it was inhibited by overexpression of miR-488-3p. Collectively, circPOFUT1 directly sponged miR-488-3p to activate the expression of PLAG1 and ATG12, thus enhancing malignant phenotypes and autophagy-associated chemoresistance in GC. Our findings show the potential of circPOFUT1 as biomarkers and targeting circPOFUT1 as a therapeutic strategy for GC.

Targeting lncRNA DDIT4-AS1 Sensitizes Triple Negative Breast Cancer to Chemotherapy via Suppressing of Autophagy.

In this study, it is found that the lncRNA, DNA damage inducible transcript 4 antisense RNA1 (DDIT4-AS1), is highly expressed in triple-negative breast cancer (TNBC) cell lines and tissues due to H3K27 acetylation in the promoter region, and promotes the proliferation, migration, and invasion of TNBC cells via activating autophagy. Mechanistically, it is shown that DDIT4-AS1 induces autophagy by stabilizing DDIT4 mRNA via recruiting the RNA binding protein AUF1 and promoting the interaction between DDIT4 mRNA and AUF1, thereby inhibiting mTOR signaling pathway. Furthermore, silencing of DDIT4-AS1 enhances the sensitivity of TNBC cells to chemotherapeutic agents such as paclitaxel both in vitro and in vivo. Using a self-activatable siRNA/drug core-shell nanoparticle system, which effectively deliver both DDIT4-AS1 siRNA and paclitaxel to the tumor-bearing mice, a significantly enhanced antitumor activity is achieved. Importantly, the codelivery nanoparticles exert a stronger antitumor effect on breast cancer patient-derived organoids. These findings indicate that lncRNA DDIT4-AS1-mediated activation of autophagy promotes progression and chemoresistance of TNBC, and targeting of DDIT4-AS1 may be exploited as a new therapeutic approach to enhancing the efficacy of chemotherapy against TNBC.

Long-term reproductive consequences of no-scalpel vasectomy in beagles.

The effects of vasectomy on the reproductive organs in various species are controversial. This study investigated the morphological change and apoptosis of the testis, epididymis, and vas deferens in beagle dogs 12 months after vasectomy. The male beagles were divided into two groups: vasectomized and sham-operated groups (n=5 in each). Histopathological, ultrastructural, and TUNEL evaluation of the changes in the testis, epididymis, and ductus deferens of each animal were conducted 12 months after surgery. The mean lumen diameter, cellular thickness, mean interstitial distance, and lumen area fraction of each seminiferous tubule and ductus epididymis were measured by stereological analysis. The results showed that, compared with the sham-operated group, the seminiferous tubular epithelial cells of the testes in the vasectomized group were disorderly arranged and scattered. Significant atrophy and apoptosis were found in the endothelial cells, and a range of ultrastructural variations were observed in the cells of testes, epididymis, and vas deferens in vasectomized group. It was concluded that complete obstruction of the vas deferens as a traditional contraception method is not absolutely safe in terms of the reversal of fertility in the long run. Techniques of relieving the inner pressure in the vas deferens while maintaining the efficacy of male contraception need to be explored.

Synthesis, characterization and thermal behavior of plasticized poly (vinyl chloride) doped with folic acid-modified titanium dioxide.

To inhibit the agglomeration of nanotitanium dioxide, a poly (vinyl chloride) (PVC) composite film doped with folic acid-modified titanium dioxide was synthesized and characterized using X-ray powder diffraction, X-ray photoelectron spectroscopy and thermogravimetric analysis coupled with Fourier transform infrared spectroscopy. The average grain size of the folic acid-modified titanium dioxide was found to decrease by 1.3 nm, indicating that the cohesiveness of the nanoparticles is decreased. The lowest temperature for 1.0% thermal decomposition of PVC was determined to be 230.0 °C. The decomposition rate at the peak temperature is found to be 39.6% lower than that of a control sample. The stability of the PVC is improved due to a lower number of surface chlorine atoms as well intermolecular attraction. A mechanism for folic acid modification of titanium dioxide-doped PVC is proposed. After doping, the ester groups in the plasticizer show a significant decrease in the vibration peak intensities observed at 1264 cm-1, 1736 cm-1 and 1106 cm-1. The doped PVC film suppresses the release of CO2, and the strongest vibration peak at 1264 cm-1 is found to be 17.2% lower than that for the blank sample, indicating that doping is beneficial for plasticizer recovery.

[Lateral decubitus position assisted plate internal fixation through a lateral incision to assist reduction combined with intramedullary nail in treatment of complicated subtrochanteric femoral fracture].

Objective: To investigate the effectiveness of lateral decubitus position assisted plate internal fixation through a lateral incision to assist reduction combined with intramedullary nail in the treatment of complicated subtrochanteric femoral fracture. Methods: The clinical data of 16 patients with complicated subtrochanteric femoral fractures (Seinsheimer type Ⅲ-Ⅴ) treated with lateral decubitus position assisted plate internal fixation through a lateral incision to assist reduction combined with intramedullary nail between September 2017 and August 2020 were retrospectively analyzed. There were 13 males and 3 females with an average age of 47 years (range, 26-85 years). There were 12 cases of high-energy injury and 4 cases of low-energy injury. According to Seinsheimer classification, there were 3 cases of type ⅢA, 2 cases of type ⅢB, 7 cases of type Ⅳ, and 4 cases of type Ⅴ. The time from injury to operation ranged from 2 to 6 days, with an average of 4.7 days. The operation time, intraoperative blood loss, postoperative drainage volume, hospitalization stay, surgical complications, fracture healing time, and collodiaphyseal angle of the affected and healthy sides before and after operation were recorded. Hip fracture Harris score was used to evaluate hip function. Results: The operation time was 90-180 minutes (mean, 135.9 minutes), the intraoperative blood loss was 200-400 mL (mean, 288.8 mL), the postoperative drainage volume was 120-220 mL (mean, 140.0 mL), and the hospitalization stay was 12-22 days (mean, 15.8 days). All the 16 patients were followed up 9-12 months (mean, 9.9 months). There was 1 case of incision superficial infection after operation, which healed after anti-infection treatment; no complication such as deep venous thrombosis of lower limbs, coxa vara deformity, re-fracture, or broken nails occurred. All the fractures healed successfully, the healing time ranged from 12 to 20 weeks, with an average of 17.5 weeks. At 6 months after operation, the Harris score was 87-96, with an average of 91.5; the results were excellent in 11 cases and good in 5 cases, with the excellent and good rate of 100%. The collodiaphyseal angle of the affected side was (124.0±5.7)°, while that of the healthy side was (132.0±2.1)°, showing significant difference between the two sides ( t=-7.376, P=0.001). At last follow-up, the collodiaphyseal angle of the affected side was (129.0±3.2)°, which significantly improved when compared with that before operation ( t=-6.175, P=0.002), and there was no significant difference between the affected side and the healthy side ( t=-2.648, P=0.181). Conclusion: Lateral decubitus position assisted plate internal fixation through a lateral incision to assist reduction combined with intramedullary nail is a reliable internal fixation method for the treatment of complicated subtrochanteric femoral fractures. The use of plate reduction is conducive to maintaining the force line of the femoral trochanter. The enlargement of the incision is conducive to the accurate implantation of intramedullary nails without affecting fracture healing.

Optical coherence tomography evaluation of retinal nerve fiber layer thickness in non-arteritic anterior ischemic optic neuropathy and primary open angle glaucoma: a systematic review and Meta-analysis.

AIM: To assess the differences in average and sectoral peripapillary retinal nerve fiber layer (pRNFL) thickness using spectral domain optical coherence tomography (SD-OCT) in patients with non-arteritic anterior ischemic neuropathy (NAION) compared with those with primary open angle glaucoma (POAG). METHODS: A comprehensive literature search of the PubMed, Cochrane Library, and Embase databases were performed prior to October, 2021. Studies that compared the pRNFL thickness in NAION eyes with that in POAG eyes with matched mean deviation of the visual fields were included. The weighted mean difference (WMD) with 95% confidence interval (CI) was used to pool continuous outcomes. RESULTS: Ten cross-sectional studies (11 datasets) comprising a total of 625 eyes (278 NAION eyes, 347 POAG eyes) were included in the qualitative and quantitative analyses. The pooled results demonstrated that the superior pRNFL was significantly thinner in NAION eyes than in POAG eyes (WMD=-6.40, 95%CI: -12.22 to -0.58, P=0.031), whereas the inferior pRNFL was significant thinner in POAG eyes than in NAION eyes (WMD=11.10, 95%CI: 7.06 to 15.14, P≤0.001). No difference was noted concerning the average, nasal, and temporal pRNFL thickness (average: WMD=1.45, 95%CI: -0.75 to 3.66, P=0.196; nasal: WMD=-2.12, 95%CI: -4.43 to 0.19, P=0.072; temporal: WMD=-1.24, 95%CI: -3.96 to 1.47, P=0.370). CONCLUSION: SD-OCT based evaluation of inferior and superior pRNFL thickness can be potentially utilized to differentiate NAION from POAG, and help to understand the different pathophysiological mechanisms between these two diseases. Further longitudinal studies and studies using eight-quadrant or clock-hour classification method are required to validate the obtained findings.

The prognostic value of circular RNA regulatory genes in competitive endogenous RNA network in gastric cancer.

Accumulating evidence shows that circular RNA (circRNA) is an important regulator of many diseases, especially cancer. Gastric cancer (GC) is a malignant tumor of the digestive system. The regulatory role and potential mechanism of circRNAs in GC remain unknown. This study aims to explore the function and regulatory mechanism of circRNA-related competitive endogenous RNA (ceRNA) in GC. The circRNA expression profile was downloaded from the Gene Expression Omnibus (GEO) database. The RNA expression profile and clinical data were downloaded from The Cancer Genome Atlas (TCGA) database. Difference analysis was conducted after quality control. Based on CircInteractome, TargetScan, and miRDB databases, a circRNA-related ceRNA network was constructed. R package "clusterProfiler" was used for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Then, a univariate and multivariate Cox regression was used to construct a prognostic-related gene model to predict survival models. Finally, a gene set enrichment analysis (GSEA) analysis was performed to elucidate the function of genes related to prognosis. Altogether, 23 DEcircRNAs, 319 DEmiRNAs, and 14,541 DEmRNAs were identified. Based on ceRNA trends, the ceRNA network included 15 DEcircRNAs, 25 DEmiRNAs, and 1099 DEmRNAs in GC. Univariate and multivariate Cox proportional hazards regression analysis was used to establish a survival model with 11 prognosis-related genes and its AUC was 0.741, indicating good sensitivity and specificity in the prediction of GC prognosis. Finally, three prognostic-related genes were selected randomly to verify expression levels, which were consistent with the analysis result. The prognostic genes were significantly enriched in cancer-related biological processes, suggesting their roles in the onset and progression of GC. Our study constructs a prognostic model of GC, deepens our understanding of circRNA-related ceRNA networks in GC biology, and provided further implications for the diagnosis and treatment of GC.

microRNA-6785-5p-loaded human umbilical cord mesenchymal stem cells-derived exosomes suppress angiogenesis and metastasis in gastric cancer via INHBA.

OBJECTIVE: Exosomes (Exos) are known to transfer microRNAs (miRNAs) to participate in human diseases. We aim to identify the role of human umbilical cord mesenchymal stem cells (HUCMSCs)-derived Exos (HUCMSC-Exos) conveying miR-6785-5p in gastric cancer (GC). METHODS: MiR-6785-5p and inhibin subunit beta A (INHBA) expression in GC tissues and cells were determined. GC cells were transfected with the vectors that altered miR-6785-5p or INHBA expression. HUCMSCs were transfected with altered miR-6785-5p or INHBA vectors, and the HUCMSC-Exos were extracted. Then, HUCMSC-Exos were co-cultured with GC cells. The proliferation, migration, invasion, apoptosis and angiogenesis of GC cells were assessed. The binding relationship between miR-6785-5p and INHBA was verified. RESULTS: MiR-6785-5p was down-regulated and INHBA was up-regulated in GC tissues and cells. Elevation of miR-6785-5p or inhibition of INHBA restricted the malignant development of GC cells. HUCMSC-Exos suppressed malignant episodes of GC cells, which could be further enhanced by up-regulated miR-6785-5p or down-regulated INHBA. Elevated INHBA abolished the impacts of up-regulated miR-6785-5p in HUCMSC-Exos on GC cells. INHBA was confirmed as a target gene of miR-6785-5p. CONCLUSION: HUCMSC-Exos containing elevated miR-6785-5p suppress angiogenesis and metastasis in GC via inhibiting INHBA. This study may further the understanding on molecular mechanisms of GC.

HDAC3 increases HMGB3 expression to facilitate the immune escape of breast cancer cells via down-regulating microRNA-130a-3p.

OBJECTIVE: Histone deacetylase 3 (HDAC3) has been reported to repress the expression of various genes by eliminating acetyl group from histone. The objective of this study was to discuss the effect of HDAC3/microRNA-130a-3p (miR-130a-3p)/high-mobility group box 3 (HMGB3) on immune escape of breast cancer. METHODS: HDAC3, miR-130a-3p and HMGB3 expression in breast cancer tissues and cells were tested, and the correlation between HDAC3, miR-130a-3p and HMGB3 was analyzed. CD8, CD69 and programmed cell death protein 1 (PD-1) expression was detected. MDA-MB-231 cells were treated with relative plasmid of HDAC3 or miR-130a-3p to test cell viability, migration, epithelial-mesenchymal transition (EMT) and apoptosis in MDA-MB-231 cells. The cytotoxicity of CD8+/CD69+/PD-1+T cells in MDA-MB-231 cells was tested, and CD8+/CD69+/PD-1+T cell proliferation and apoptosis before and after co-culture with MDA-MB-231 cells were detected. RESULTS: HDAC3 and HMGB3 expression were raised and miR-130a-3p expression was diminished in breast cancer tissues and cells. HDAC3 was negatively correlated with miR-130a-3p while miR-130a-3p was negatively correlated with HMGB3. Down-regulating HDAC3 or up-regulating miR-130a-3p restrained cell viability, migration, EMT and anti-CD8+/CD69+/PD-1+T cytotoxicity and facilitated apoptosis of breast cancer cells. HDAC3 regulated HMGB3 by mediating miR-130a-3p expression. Down-regulating miR-130a-3p reversed the role of HDAC3 reduction on breast cancer cells. HDAC3 regulated CD8+/CD69+/PD-1+T cell proliferation and apoptosis by mediating miR-130a-3p. CONCLUSION: This study provides evidence that HDAC3 increases HMGB3 expression to promote the immune escape of breast cancer cells via down-regulating miR-130a-3p.

Long non-coding RNA LINC00426 contributes to doxorubicin resistance by sponging miR-4319 in osteosarcoma.

BACKGROUND: LINC00426 is a newly identified long non-coding RNA (lncRNA) with unacknowledged biological roles. Here we set out to characterize the expression status of LINC00426 in osteosarcoma and understand its mechanistic involvement in incidence of doxorubicin (Dox) resistance. METHODS: The relative expression of LINC00426 and miR-4319 was determined by real-time PCR. Cell viability and proliferation in response to LINC00426 silencing or miR-4319 over-expression was measured with CCK-8 kit and colony formation assay, respectively. The direct association between LINC00426 and miR-4319 was analyzed by pulldown assay with biotin-labelled probes. RESULTS: LINC00426 was significantly up-regulated in Dox-resistant osteosarcoma (OS) both in vitro and in vivo, which intimately associated with unfavorable prognosis. SiRNA-mediated knockdown of LINC00426 remarkably compromised cell viability and proliferation in Dox-resistant OS cells, which accompanied with decrease of IC50 and activation of caspase-3. We further predicted and validated the regulatory effects of miR-4319 on LINC00426 expression. Simultaneously, we provided evidences in support of direct binding between LINC00426 and miR-4319 by pulldown assay. Reciprocally negative regulation was observed between LINC00426 and miR-4319 each other. CONCLUSION: Ectopic introduction of miR-4319 significantly surmounted the Dox resistance in OS cells, while miR-4319 inhibition in LINC00426-deficient cells greatly restore this phenotype. We uncovered the important contribution of LINC00426/miR-4319 to Dox resistance in osteosarcoma. REVIEWERS: This article was reviewed by Bo Liang and Sinan Zhu.

Expression of receptor protein tyrosine phosphatase ζ is a risk factor for triple negative breast cancer relapse.

Patients with triple negative breast cancer (TNBC) have a higher rate of distant recurrence and a poorer prognosis than those with other breast cancer subtypes. Therefore, it is important to study the mechanism of TNBC relapse. A retrospective immunohistochemical analysis of the expression of receptor protein tyrosine phosphatase ζ (PTPRZ1) and pleiotrophin (PTN) was performed for 325 cases of breast cancer. These samples included 66 cases of luminal A breast cancer, 67 cases of luminal B breast cancer, 78 cases of Her-2-enriched breast cancer, 78 cases of TNBC and 36 cases of relapsed TNBC (RTNBC). In addition, 30 control specimens and 30 cases of metastasized lymph nodes were examined. PTPRZ1 and PTN were highly expressed in the RTNBC group. Compared with the RTNBC group, significant differences in the expression of PTPRZ1 were observed between the TNBC, BC and control groups. A significant difference was observed in the expression of PTN in the BC group (P<0.05) compared to RTNBC, and there were no significant differences in the expression of PTPRZ1 and PTN among the molecular subtypes. No significant correlation was observed between the expression of PTPRZ1, PTN, ER, PR, Her-2 and ALN and the tumor size or menopause status. No significant correlation was identified between the expression of PTPRZ1 and PTN and the expression of CD24 and CD44. In summary, high expression of PTPRZ1 may be an independent risk indicator for TNBC recurrence and metastasis.

Intratumoral CD8⁺ cytotoxic lymphocyte is a favorable prognostic marker in node-negative breast cancer.

BACKGROUND: The prognostic effect of tumor infiltrating CD8+ cytotoxic lymphocytes (CTLs) in breast cancer is controversial. We analyzed the association between CD8+ CTLs and survival of untreated node-negative breast cancer patients. MATERIAL AND METHODS: CD8+ CTLs infiltrate was evaluated by immunostaining in a cohort of 332 node-negative breast cancer patients with a median follow-up of 152 months. The prognostic significance of CD8+ CTLs for disease-free survival (DFS) and breast cancer-specific overall survival (OS) was evaluated with Kaplan-Meier survival analysis as well as univariate analysis and multivariate Cox analysis adjusted for age at diagnosis, pT stage, histological grade, estrogen receptor (ER) status, progesterone receptor (PR) status, Ki-67 expression and human epidermal growth factor receptor 2 (HER-2) status. RESULTS: 285 (85.8%) patients showed strong CD8+ CTLs infiltrate positive status. Univariate analysis showed that CD8+ CTLs had statistically significant association with DFS (P = 0.004, hazard ratio [HR] = 0.454, 95% confidence interval [CI] = 0.265-0.777) and OS (P = 0.014, HR = 0.430, 95% CI = 0.220-0.840) in the entire cohort. The significance of CD8+ CTLs was especially strong in ER negative, HER-2 negative and ER, PR, HER-2 triple-negative breast cancers. In Kaplan-Meier analysis, CD8+ CTLs had significant effect on prognosis of patients (Log-rank test: P = 0.003 for DFS and P = 0.011 for OS), independent of established clinical factors for DFS (P = 0.002, HR = 0.418, 95% CI = 0.242-0.724) as well as for OS (P = 0.009, HR = 0.401, 95% CI = 0.202-0.797).

Immunoglobulin kappa C predicts overall survival in node-negative breast cancer.

BACKGROUND: Biomarkers of the immune system are currently not used as prognostic factors in breast cancer. We analyzed the association of the B cell/plasma cell marker immunoglobulin kappa C (IGKC) and survival of untreated node-negative breast cancer patients. MATERIAL AND METHODS: IGKC expression was evaluated by immunostaining in a cohort of 335 node-negative breast cancer patients with a median follow-up of 152 months. The prognostic significance of IGKC for disease-free survival (DFS) and breast cancer-specific overall survival (OS) was evaluated with Kaplan-Meier survival analysis as well as univariate and multivariate Cox analysis adjusted for age at diagnosis, pT stage, histological grade, estrogen receptor (ER) status, progesterone receptor (PR) status, Ki-67 and human epidermal growth factor receptor 2 (HER-2) status. RESULTS: 160 patients (47.7%) showed strong expression of IGKC. Univariate analysis showed that IGKC was significantly associated with DFS (P = 0.017, hazard ratio [HR] = 0.570, 95% confidence interval [CI] = 0.360-0.903) and OS (P = 0.011, HR = 0.438, 95% CI = 0.233-0.822) in the entire cohort. The significance of IGKC was especially strong in ER negative and in luminal B carcinomas. In multivariate analysis IGKC retained its significance independent of established clinical factors for DFS (P = 0.004, HR = 0.504, 95% CI = 0.315-0.804) as well as for OS (P = 0.002, HR = 0.371, 95% CI = 0.196-0.705). CONCLUSION: Expression of IGKC has an independent protective impact on DFS and OS in node-negative breast cancer.

A comprehensive analysis of human gene expression profiles identifies stromal immunoglobulin κ C as a compatible prognostic marker in human solid tumors.

PURPOSE: Although the central role of the immune system for tumor prognosis is generally accepted, a single robust marker is not yet available. EXPERIMENTAL DESIGN: On the basis of receiver operating characteristic analyses, robust markers were identified from a 60-gene B cell-derived metagene and analyzed in gene expression profiles of 1,810 breast cancer; 1,056 non-small cell lung carcinoma (NSCLC); 513 colorectal; and 426 ovarian cancer patients. Protein and RNA levels were examined in paraffin-embedded tissue of 330 breast cancer patients. The cell types were identified with immunohistochemical costaining and confocal fluorescence microscopy. RESULTS: We identified immunoglobulin κ C (IGKC) which as a single marker is similarly predictive and prognostic as the entire B-cell metagene. IGKC was consistently associated with metastasis-free survival across different molecular subtypes in node-negative breast cancer (n = 965) and predicted response to anthracycline-based neoadjuvant chemotherapy (n = 845; P < 0.001). In addition, IGKC gene expression was prognostic in NSCLC and colorectal cancer. No association was observed in ovarian cancer. IGKC protein expression was significantly associated with survival in paraffin-embedded tissues of 330 breast cancer patients. Tumor-infiltrating plasma cells were identified as the source of IGKC expression. CONCLUSION: Our findings provide IGKC as a novel diagnostic marker for risk stratification in human cancer and support concepts to exploit the humoral immune response for anticancer therapy. It could be validated in several independent cohorts and carried out similarly well in RNA from fresh frozen as well as from paraffin tissue and on protein level by immunostaining.

Biological evaluation of a novel copper-containing composite for contraception.

OBJECTIVE: To investigate the biocompatibility of a novel copper-containing composite to provide preclinical data for clinical application of intrauterine device (IUD) or intra-vas device (IVD). DESIGN: Prospective experimental study. SETTING: Good laboratory practices laboratories. ANIMALS: Twenty healthy adult mice (SPF grade Kunming white mice, animal code SCXK 2003-0005). INTERVENTION(S): Cytotoxicity tests in vitro were conducted to evaluate the influence of the materials on the morphology, growth, and proliferation of cultured L929 mouse fibroblasts. Acute systemic toxicity tests were conducted to investigate the acute systemic toxic reaction with mice, and then the materials were implanted into the spinal muscle of rabbits (n = 15). The rabbits were sacrificed for pathologic examination at 1, 4, and 12 weeks after surgery. MAIN OUTCOME MEASURE(S): Evaluation of cytotoxicity by MTT assay, cytotoxicity test by direct contact assay, acute systemic toxicity test, and material implantation test. RESULT(S): The cytotoxicity grade of the copper-containing composite was 0-1, suggesting that the material was free of cytotoxicity; no acute systemic toxicity was found in any mice; mild inflammatory reaction was observed in the surrounding tissues of the implanted material in the early implantation stage, which was similar to that of the sham-operated sides. Twelve weeks after implantation, the inflammatory reaction was completely disappeared in the implanted tissue, similarly to the sham-operated sides. The fibrosis membrane surrounding the material became stable gradually over time. CONCLUSION: The copper-containing composite has excellent biocompatibility, which is feasible and safe for the clinical application as a novel contraceptive material.