Multivariate logistic regression analysis of the clinical factors influencing locally advanced prostate cancer.

Background: Locally advanced prostate cancer (PCa) carries a high risk of recurrence and metastasis after surgery, and the prognosis is poor. We explored the risk factors for locally advanced PCa among clinical factors (neutrophil: lymphocyte ratio, lymphocyte: monocyte ratio) and indicators of systemic inflammation [prostate-specific antigen (PSA) level, Gleason score, body mass index (BMI)] through retrospective evaluation of patients with PCa diagnosed at our center. The pathologic T stage was a key indicator of locally advanced PCa. Methods: Data from patients with pathologically confirmed PCa at our center from 1 January 2015 to 1 May 2020 were collected in strict accordance with inclusion and exclusion criteria. Clinical data were collected and the relationship between the indicators and the pathologic T stage was explored. First, Spearman rank correlation analysis was used to find the correlates of the pathologic T stage. Then, logistic ordered multiple regression analysis was used to identify independent risk factors. Finally, receiver operating characteristic (ROC) curves were used to assess the diagnostic accuracy for the T stage of PCa. Results: After rigorous screening, the data of 177 patients were obtained. Spearman correlation analysis showed that BMI, the PSA level, Gleason score, hypertension, N stage, and M stage were significantly correlated with the T stage (P<0.05), suggesting that these factors may be involved in locally advanced PCa. Analyses of ROC curves showed that the PSA level [area under the ROC curve (AUC) =0.802] had greater value than BMI (0.675) for the diagnosis of the pathologic T stage PCa, and that a combination of BMI and PSA (combined AUC =0.822) could improve locally advanced PCa diagnosis. Conclusions: BMI and PSA are independent risk factors for locally advanced PCa. They may play a key part in locally advanced PCa.

Efficient Recovery of Phosphate from Water Media by Iron-Magnesium Functionalized Lignite: Adsorption Evaluation, Mechanism Revelation and Potential Application Exploration.

Selective phosphorus removal from aquatic media has become an ideal strategy to mitigate eutrophication and meet increasingly stringent discharge requirements. To achieve phosphorus control and resource utilization of low-calorific-value lignite, iron and magnesium salts were used to functionalize lignite, and iron-magnesium functionalized lignite (called IM@BC) was prepared for phosphate recovery from water media. The adsorption properties of IM@BC were systematically evaluated, especially the influence of ambient pH and co-existing ions. The kinetic, isothermal, and thermodynamic adsorption behaviors of IM@BC were analyzed. The adsorption mechanism was revealed by microscopic characterization. The potential application of phosphate-containing IM@BC (P-IM@BC) was explored. The results show that IM@BC has a strong phosphate adsorption capacity, and the maximum adsorption capacity is 226.22 mgP/g at pH = 3. Co-existing CO32- inhibits phosphate adsorption, while coexisting Ca2+ and Mg2+ enhance the effect. At the initial adsorption stage, the amount of phosphate adsorbed by IM@BC continues to increase, and the adsorption equilibrium state is gradually reached after 24 h. The adsorption process conforms to the pseudo-second-order kinetic model (PSO) and Langmuir isothermal adsorption model, and the adsorption process is mainly chemical adsorption. The phosphate absorption capacity is positively correlated with temperature (283.15 K~313.15 K), and the adsorption process is spontaneous, endothermic, and entropy-increasing. Its adsorption mechanism includes electrostatic attraction, ion exchange, surface precipitation, and coordination exchange. IM@BC can efficiently recover phosphate from actual phosphorus-containing wastewater with a recovery efficiency of up to 90%. P-IM@BC slowly releases phosphate from pH 3 to 11. Plant growth experiments showed that P-IM@BC could be used as a slow-release fertilizer to promote the root growth of cowpeas. The novelty of this work lies in the development of a highly efficient phosphate recovery adsorbent, which provides a feasible method of phosphorus control in water media and resource utilization of lignite.

Updated review on analysis of long non-coding RNAs as emerging diagnostic and therapeutic targets in prostate cancers.

Despite advancements, prostate cancers (PCa) pose a significant global health challenge due to delayed diagnosis and therapeutic resistance. This review delves into the complex landscape of prostate cancer, with a focus on long-noncoding RNAs (lncRNAs). Also explores the influence of aberrant lncRNAs expression in progressive PCa stages, impacting traits like proliferation, invasion, metastasis and therapeutic resistance. The study elucidates how lncRNAs modulate crucial molecular effectors, including transcription factors and microRNAs, affecting signaling pathways such as androgen receptor signaling. Besides, this manuscript sheds light on novel concepts and mechanisms driving PCa progression through lncRNAs, providing a critical analysis of their impact on the disease's diverse characteristics. Besides, it discusses the potential of lncRNAs as diagnostics and therapeutic targets in PCa. Collectively, this work highlights state of art mechanistic comprehension and rigorous scientific approaches to advance our understanding of PCa and depict innovations in this evolving field of research.

Recovery of ammonia nitrogen and phosphate from livestock farm wastewater by iron-magnesium oxide coupled lignite and its potential for resource utilization.

A new adsorbent called iron-magnesium oxide coupled lignite (CIMBC) was developed to address the challenges of recovering high concentrations of ammonia nitrogen and phosphate in livestock farm wastewater and improving the inefficient use of lignite (BC) with low calorific value. CIMBC was synthesized using the modified ferromagnesium salt double-coating method. The experiments demonstrated that Fe2O3 and MgO could be effectively loaded onto the surface of BC at a Fe/Mg molar ratio of 1:2 and pyrolysis temperature of 500 °C. The optimal conditions for adsorption were determined to be an N/P concentration ratio of 2:1, adsorbent dosage of 1 g/L, and pH of 7. The presence of coexisting cations (Ca2+ and Mg2+) inhibited the removal of ammonia nitrogen but enhanced the removal of phosphate. Likewise, the presence of coexisting anions (CO32- and SO42-) hindered the removal of both ammonia nitrogen and phosphate. The adsorption behavior followed the pseudo-second-order model and the Langmuir model, with a maximum adsorption capacity of 95.69 mg N/g for ammonia nitrogen and 101.32 mg P/g for phosphate. The adsorption process was a spontaneous endothermic process controlled by multiple levels. The main mechanisms of adsorption involved electrostatic attraction, intra-particle diffusion, ion exchange, chemical precipitation, and coordination exchange. After 5 times of adsorption-desorption, the recovery rate of CIMBC is less than 50%, and the removal rate of phosphate is less than 40%. Although the RCIMBC exhibited low reusability, but also it showed potential in removing heavy metals (Pb) from wastewater and for use as a slow-release fertilizer. CIMBC is a promising new adsorbent, which can realize resource utilization of lignite with low calorific value while removing nitrogen and phosphorus.

Experimental study on the treatment of AMD by SRB immobilized particles containing "active iron" system.

The inhibition and toxicity of high acidity and heavy metals on sulfate-reducing bacteria in acid mine drainage (AMD) were targeted. Highly active SRB immobilized particles were prepared using SRB, warm sticker wastes (iron powders), corncobs, and Maifan stones as the main matrix materials, employing microbial immobilization technology. The repair ability and reusability of highly active immobilized particles for AMD were explored. The results indicate that the adaptability of immobilized particles to AMD varied under different initial conditions, such as pH, Mn2+, and SO42-. The adsorption process of immobilized particles on Mn2+ follows the quasi-second-order kinetic model, suggesting that it involves both physical and chemical adsorption. The maximum adsorption capacity of immobilized particles for Mn2+ is 3.878 mg/g at a concentration of 2.0 mg/L and pH 6. On the other hand, the reduction process of immobilized particles on SO42- adheres to the first-order reaction kinetics, indicating that the reduction of SO42- is primarily driven by the dissimilation reduction of SRB. The maximum reduction rate of SO42- by immobilized particles is 94.23% at a concentration of 800 mg/L and pH 6. A layered structure with a flocculent appearance formed on the surface of the immobilized particles. The structure's characteristics were found to be consistent with sulfate green rust (FeII4FeIII2(OH)12SO4·8H2O). The chemisorption, ion exchange, dissimilation reduction, and surface complexation occurring between the matrices in the immobilized particles can enhance the alkalinity of AMD and decrease the concentration of heavy metals and sulfates. These results are expected to offer novel insights and materials for the treatment of AMD using biological immobilization technology, as well as improve our understanding of the mechanisms behind biological and abiotic enhanced synergistic decontamination.

Association of BMI with erectile dysfunction: A cross-sectional study of men from an andrology clinic.

Abnormal body mass index (BMI) is associated with an increased risk of erectile dysfunction (ED). However, the relationship between different BMI categories and the levels of ED severity remains unclear. In the current study, 878 men from the andrology clinic in Central China were recruited. Erectile function was assessed by the International Index of Erectile Function (IIEF) scores. Questionnaires included questions about demographic characteristics (age, height, weight, educational status), lifestyle habits (drinking, smoking, sleep time), and medical history. Logistic regression was used to examine the association between ED risk and BMI. The incidence of ED was 53.1%. BMI was significantly higher in men from the ED group than in those from the non-ED group (P = 0.01). Compared with the normal weight group, obese men had a higher risk of ED (OR = 1.97, 95% CI = 1.25-3.14, P = 0.004), even after adjustment for potential confounders (OR = 1.78, 95% CI = 1.10-2.90, P = 0.02). Moreover, the positive correlation between obesity and moderate/severe ED severity was confirmed by logistic regression analysis (moderate/severe ED, OR = 2.71, 95% CI = 1.44-5.04, P = 0.002), even after adjusting for potential confounders (OR = 2.51 95% CI = 1.24-5.09, P = 0.01). Collectively, our findings indicate a positive correlation between obesity and the risk of moderate/severe ED. Clinicians could pay more attention to moderate/severe ED patients to maintain a healthy body weight to improve erectile function.

CD123 Antagonistic Peptides Assembled with Nanomicelles Act as Monotherapeutics to Combat Refractory Acute Myeloid Leukemia.

Acute myeloid leukemia (AML) is the most common type of acute leukemia in adults. Due to the development of drug resistance to traditional chemotherapies and high relapse rate, AML still has a low survival rate and there is in an urgent need for better treatment strategies. CD123 is widely expressed by AML cells, also associated with the poor prognosis of AML. In this study, we fabricated nanomicelles loaded with a lab-designed CD123 antagonistic peptide, which were referred to as mPO-6. The antagonistic and therapeutic effects were investigated with CD123+ AML cell lines and a refractory AML mouse (AE and CKITD816V) model. Results show that mPO-6 can specifically bind to the CD123+ AML cells and inhibit the cell viability effectively. Intravenous administration of mPO-6 significantly reduces the percentage of AML cells' infiltration and prolongs the median survival of AML mice. Further, the efficiency of mPO-6 is demonstrated to interfere with the axis of CD123/IL-3 via regulating the activation of STAT5, PI3K/AKT, and NF-κB signaling pathways related to cell proliferation or apoptosis at the level of mRNA and protein in vivo and in vitro. In conclusion, the novel CD123 antagonistic peptide micelle formulation mPO-6 can significantly enhance apoptosis and prolong the survival of AML mice by effectively interfering with the axis of CD123/IL-3 and therefore is a promising therapeutic candidate for the treatment of refractory AML.

Magnetic Nanofibrous Scaffolds Accelerate the Regeneration of Muscle Tissue in Combination with Extra Magnetic Fields.

The reversal of loss of the critical size of skeletal muscle is urgently required using biomaterial scaffolds to guide tissue regeneration. In this work, coaxial electrospun magnetic nanofibrous scaffolds were fabricated, with gelatin (Gel) as the shell of the fiber and polyurethane (PU) as the core. Iron oxide nanoparticles (Mag) of 10 nm diameter were added to the shell and core layer. Myoblast cells (C2C12) were cultured on the magnetic scaffolds and exposed to the applied magnetic fields. A mouse model of skeletal muscle injury was used to evaluate the repair guided by the scaffolds under the magnetic fields. It was shown that VEGF secretion and MyoG expression for the myoblast cells grown on the magnetic scaffolds under the magnetic fields were significantly increased, while, the gene expression of Myh4 was up-regulated. Results from an in vivo study indicated that the process of skeletal muscle regeneration in the mouse muscle injury model was accelerated by using the magnetic actuated strategy, which was verified by histochemical analysis, immunofluorescence staining of CD31, electrophysiological measurement and ultrasound imaging. In conclusion, the integration of a magnetic scaffold combined with the extra magnetic fields enhanced myoblast differentiation and VEGF secretion and accelerated the defect repair of skeletal muscle in situ.

Prognostic significance of CKAP2L expression in patients with clear cell renal cell carcinoma.

Background: Cytoskeleton-associated protein 2-like protein (CKAP2L) is thought to promote the progression of glioma, breast cancer, and ovarian cancer. However, the role of cytoskeleton-associated protein 2-like protein in clear cell renal cell carcinoma (ccRCC) is still unclear. The study aimed to investigate the roles and mechanisms of cytoskeleton-associated protein 2-like protein in clear cell renal cell carcinoma. Methods: The level of cytoskeleton-associated protein 2-like protein in tumors was explored by using UALCAN and Oncomine databases. Gene expression datasets of clear cell renal cell carcinoma from The Cancer Genome Atlas and Gene Expression Omnibus (GEO) were also used to validate the cytoskeleton-associated protein 2-like protein level in clear cell renal cell carcinoma. Survival analysis was performed to investigate the relationship between cytoskeleton-associated protein 2-like protein level and prognosis of clear cell renal cell carcinoma patients. Cox regression analysis was used for identifying the independent prognostic factors. Gene set enrichment analysis (GSEA), gene set variation analysis (GSVA), protein-protein interaction analysis, co-expression analysis, and immune infiltration analysis were used to explore the potential mechanisms of cytoskeleton-associated protein 2-like protein in clear cell renal cell carcinoma. Moreover, the levels of cytoskeleton-associated protein 2-like protein in clinical clear cell renal cell carcinoma tissues were also measured using RT-PCR, immunohistochemical analysis, and Western blotting. M1 macrophages and CD4+ T cells were also detected by immunohistochemistry between tumor and normal tissues. Results: The level of cytoskeleton-associated protein 2-like protein was upregulated in clear cell renal cell carcinoma according to multiple databases and experimental verification. Upregulated cytoskeleton-associated protein 2-like protein is an independent prognostic factor, which might activate the JAK-STAT signaling pathway, the P53 signaling pathway, the TGF-ß signaling pathway, the WNT signaling pathway, etc., in clear cell renal cell carcinoma. Protein-protein interaction analysis and co-expression analysis suggest that cytoskeleton-associated protein 2-like protein might interact with some proliferation proteins. Immune infiltration analysis indicates that cytoskeleton-associated protein 2-like protein may affect the level of activated CD4+ memory T cells, M1 macrophages, CD8+ T cells, and neutrophils in clear cell renal cell carcinoma. More M1 macrophage infiltrations in tumor tissues with higher cytoskeleton-associated protein 2-like protein were validated by clear cell renal cell carcinoma tumor tissues. Conclusion: Cytoskeleton-associated protein 2-like protein is upregulated in clear cell renal cell carcinoma tissues, which may promote progression of the disease. Cytoskeleton-associated protein 2-like protein is a potential target for prognostic markers and a potential treatment target in clear cell renal cell carcinoma.

[Correlation of the platelet count in the peripheral blood with the prostate volume].

OBJECTIVE: To investigate the relationship of the prostate volume with the count of inflammatory cells in the peripheral blood and clarify the pathogenesis of BPH. METHODS: From 2015 to 2019, we enrolled 104 men pathologically diagnosed with BPH. Using univariate and multivariate linear regression analysis models, we analyzed the correlation of the prostate volume with the neutrophil count, platelet count, neutrophil-lymphocyte ratio (NLR), platelet-WBC ratio (PWR), and lymphocyte-monocyte ratio (LMR) in the peripheral blood of the patients. RESULTS: Both the platelet count (r = 0.401, P < 0.001) and PWR (r = 0.343, P < 0.001) in the peripheral blood were positively correlated with the prostate volume and serum PSA level, but not with IPSS. No evident relationship was found between the prostate volume and the systemic inflammatory markers NLR and LMR. CONCLUSIONS: The platelet count in the peripheral blood is an important predictor of BPH and may play an important role in the development and progression of BPH.

Expression of miR-410 in peripheral blood of patients with clear cell renal cell carcinoma and its effect on proliferation and invasion of Caki-2 cells.

PURPOSE: To explore the significance of miR-410 expression in clear cell renal cell carcinoma (CCRCC) and its biological function in CCRCC. METHODS: A total of 113 patients with CCRCC admitted to our hospital and 113 healthy individuals over the same period were enrolled. MiR-410 in the tissues and serum of patients with CCRCC was quantified, and the diagnostic value of miR-410 in CCRCC and the relationship between miR-410 and prognosis of patients with CCRCC were analyzed. In addition, miR-410 mimic and miR-410 inhibitor were adopted to regulate miR-410 in CCRCC cells (Caki-2), and then the changes in the proliferation, migration, invasion, and cell cycle of Caki-2 cells were determined. Moreover, tumorigenicity in nude mice was carried out to determine the effect of miR-410 on the tumor growth of CCRCC. RESULTS: MiR-410 was expressed at a high level in CCRCC patients, and had a high diagnostic accuracy [area under the curve (AUC) = 0.916]. In addition, miR-410 was an independent risk factor for the survival prognosis of patients with CCRCC, and its high expression indicated poor prognosis of the patients. Inhibiting miR-410 suppressed cell proliferation, cycle progression, migration, invasion and tumor growth in vivo and promoted cell apoptosis. CONCLUSION: MiR-410 is a possible biological indicator for the diagnosis and prognosis of CCRCC, and is also an independent risk factor for the survival prognosis of CCRCC patients. In addition, miR-410 plays a role as an oncogene in CCRCC and promotes the malignant progression of CCRCC.

Rosiglitazone ameliorates palmitic acid-induced cytotoxicity in TM4 Sertoli cells.

The Sertoli cell is the only somatic cell within the seminiferous tubules, and is vital for testis development and spermatogenesis. Rosiglitazone (RSG) is a member of the thiazolidinedione family and is a peroxisome proliferator-activated receptor-γ (PPARγ) agonist. It has been reported that RSG protects various types of cells from fatty acid-induced damage. However, whether RSG serves a protective role in Sertoli cells against palmitic acid (PA)-induced toxicity remains to be elucidated. Therefore, the aim of the present study was to investigate the effect of RSG on PA-induced cytotoxicity in Sertoli cells. MTT assay and Oil Red O staining revealed that RSG ameliorated the PA-induced decrease in TM4 cell viability, which was accompanied by an alleviation of PA-induced lipid accumulation in cells. In primary mouse Sertoli cells, RSG also showed similar protective effects against PA-induced lipotoxicity. Knockdown of PPARγ verified that RSG exerted its protective role in TM4 cells through a PPARγ-dependent pathway. To evaluate the mechanism underlying the protective role of RSG on PA-induced lipotoxicity, the present study analyzed the effects of RSG on PA uptake, and the expression of genes associated with both fatty acid oxidation and triglyceride synthesis. The results demonstrated that although RSG did not affect the endocytosis of PA, it significantly elevated the expression of carnitine palmitoyltransferase (CPT)-1A, a key enzyme involved in fatty acid oxidation, which indicated that the protective effect of RSG may have an important role in fatty acid oxidation. On the other hand, the expression of CPT1B was not affected by RSG. Moreover, the expression levels of diacylglycerol O-acyltransferase (DGAT)-1 and DGAT2, both of which encode enzymes catalyzing the synthesis of triglycerides, were not suppressed by RSG. The results indicated that RSG reduced PA-induced lipid accumulation by promoting fatty acid oxidation mediated by CPT1A. The effect of RSG in protecting cells from lipotoxicity was also found to be specific to Sertoli cells and hepatocytes, and not to other cell types that do not store excess lipid in large quantities, such as human umbilical vein endothelial cells. These findings provide insights into the cytoprotective effects of RSG on Sertoli cells and suggest that PPARγ activation may be a useful therapeutic method for the treatment of Sertoli cell dysfunction caused by dyslipidemia.

Spermatogenesis improved by suppressing the high level of endogenous gonadotropins in idiopathic non-obstructive azoospermia: a case control pilot study.

BACKGROUND: Elevated plasma gonadotropins were associated with desensitization of Sertoli and Leydig cells in the male testis. Testis spermatogenesis ability would be improved via inhibiting high endogenous gonadotropin in patients with severe oligozoospermia. Whether it would be beneficial for non-obstructive azoospermia (NOA) patients was still unclear. METHODS: Goserelin, a gonadotropin releasing hormone agonist (GnRHα) was used to suppress endogenous gonadotropin levels (gonadotropin reset) in the NOA patients, improving the sensitization of the Sertoli and Leydig cells. Then human menopausal gonadotropin (hMG) and human chorionic gonadotropin (hCG) were injected to stimulate them to ameliorate the ability of testicular spermatogenesis. The main outcome measure was the existence of spermatozoa in the semen or by testicular sperm extraction (TESE). Elevation of inhibin B and/or ameliorative expression pattern of ZO-1 was the secondary objective. RESULTS: A total of 35 NOA men who failed to retrieve sperm via TESE were enrolled. Among these, 10 patients without treatment were selected as control group and secondary TESE was performed 6 months later. Of the 25 treated men, inhibin B was elevated in 11 patients in the first 4 weeks (Response group), while only 5 patients had constant increase in the following 20 weeks (Response group 2). Of the 5 men, 2 men acquired sperm (Response group 2B), while 3 failed (Response group 2A). Immunofluorescence of mouse vasa homologue (MVH) and ZO-1 showed that both positive MVH signals and ZO-1 expression were significantly increased in the Response group 2, but only Response group 2B showed ameliorative ZO-1 distribution. CONCLUSIONS: Gonadotropin reset, a new therapeutic protocol with GnRHα, was able to improve the ability of testicular spermatogenesis in the NOA patients through restoring the sensitivity of Sertoli and Leydig cells, which were reflected by elevated inhibin B and ameliorative ZO-1 expression and distribution. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT02544191 .

Identification and preliminary study of immunogens involved in autoimmune prostatitis in human males.

BACKGROUND: Experimental models have confirmed that autoimmunity is an important factor in the onset of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS); however, there is no conclusive evidence on whether autoimmune prostatitis exists in human males. METHODS: Rabbits were immunized with either human prostate tissue homogenates or normal saline and the antiserum was collected. Two-dimensional electrophoresis (2-DE) was performed on the homogenates and Western blotting was conducted on the sera. The identified human prostate tissue immunodominant antigens (HPTIAs) were detected by mass spectrometry. The serum immunoglobulin (Ig)G from the immunized rabbits was purified with protein A-agarose, and the purified IgG was linked with Sepharose to purify HPTIAs by affinity chromatography. Non-obese diabetic (NOD) mice were immunized with the purified HPTIAs, and the levels of serum antibodies, INF-γ, and histopathological changes in their prostate tissues were detected. The purified HPTIAs were coated into polystyrene pores and serum autoantibodies in CP/CPPS patients were detected by enzyme-linked immunosorbent assay (ELISA). Meanwhile, serum interleukin 2 (IL-2), interferon gamma (IFNγ), and tumor necrosis factor alpha (TNFα) levels in CP/CPPS patients were also determined by ELISA. RESULTS: Sixteen HPTIAs were identified. Among them, three types were reported to be associated with prostatic diseases. Prostatitis was induced in mice immunized with the 16-HPTIA complex, with positive serum autoantibody and increased prostatic IFN-γ levels. The positive rate of serum autoantibodies against HPTIAs was significantly higher in CP/CPPS patients (23.1%, 18/78) than in the control (2.7%, 2/75). But there was no significant difference in serum TNFα, IFNγ, and IL-2 levels between the CPPS patients with positive and negative autoantibodies against HPTIAs. CONCLUSIONS: Autoantibodies against HPTIAs exist in part in CP/CPPS patients, which implies that autoimmunity and the 16 HPTIAs are important factors in the onset of CP/CPPS. The detection of serum autoantibodies could be applied in clinical diagnoses of autoimmune prostatitis; treatment protocols might change. Additional studies are needed to determine which of the 16 HPTIAs is the most important.

Effects of saturated palmitic acid and omega-3 polyunsaturated fatty acids on Sertoli cell apoptosis.

Obesity is believed to negatively affect male semen quality and is accompanied by dysregulation of free fatty acid (FFA) metabolism in plasma. However, the implication of dysregulated FFA on semen quality and the involvement of Sertoli cells remain unclear. In the present study, we report obesity decreased Sertoli cell viability through dysregulated FFAs. We observed an increased rate of apoptosis in Sertoli cells, accompanied with elevated FFA levels, in the testes of obese mice that were provided a high-fat diet (HFD). Moreover, the levels of reactive oxygen species were elevated. Furthermore, we demonstrated by in vitro assays that saturated palmitic acid (PA), which is the most common saturated FFA in plasma, led to decreased cell viability of TM4 Sertoli cells in a time- and dose-dependent manner. A similar finding was noted in primary mouse Sertoli cells. In contrast to saturated FFA, omega-3 (ω-3) polyunsaturated fatty acids (PUFAs) protected Sertoli cells from PA-induced lipotoxicity at the physiologically relevant levels. These results indicated that the lipotoxicity of saturated fatty acids might be the cause of obesity-induced Sertoli cell apoptosis, which leads to decreased semen quality. In addition, ω-3 PUFAs could be classified as protective FFAs. ABBREVIATIONS: FFA: free fatty acid; HFD: high-fat diet; SD: standard diet; PA: palmitic acid; PUFA: polyunsaturated fatty acid; AI: apoptotic index; MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide; ROS: reactive oxygen species; HE: Hematoxylin and eosin; WT1: Wilm Tumor 1; NAFLD: non- alcoholic fatty liver disease; DCFH-DA: 2', 7' dichlorofluorescin diacetate; 36B4: acidic ribosomal phosphoprotein P0; SD: standard deviation; EPA: eicosapentaenoic acid; PI: propidium iodide; DHA: docosahexenoic acid.