Association between leptin receptor polymorphisms and polycystic ovary syndrome risk: a meta-analysis based on 11 studies.

OBJECTIVE: Published evidence indicated that the leptin receptor (LEPR) gene polymorphisms are associated with polycystic ovary syndrome (PCOS) risk. However, studies on the association between the polymorphisms of LEPR gene are inconsistent or even controversial. MATERIAL AND METHODS: We conducted this meta-analysis to explore the more precise relationship between LEPR polymorphisms and PCOS risk. Relevant articles were searched with five online databases up to March 1 2023. Odds ratios (OR) with 95% confidence intervals (CI) were selected to examine the statistical strength of each genetic model. Moreover, RNA secondary structure and variant effects of these loci were examined with in silico analysis. RESULTS: Overall, 11 publications were analyzed, and the pooled results did not present any significant association between rs1137101 A/G polymorphism and PCOS risk in general population and some subgroup analysis. But the significant association were observed in Asian population (AG vs. AA: OR = 0.51, 95%CI = 0.32-0.81, p = .01, I2=0%; AG + GG vs. AA: OR = 0.41, 95%CI = 0.26-0.65, p < .01, I2=25.9%). Moreover, similar positive associations were also observed in rs1805096 polymorphism with PCOS risk. CONCLUSION: In summary, our meta-analysis suggested that the LEPR gene polymorphisms might be associated with PCOS susceptibility. Owing to the limited studies and small sample size in our meta-analysis, more well-designed studies from different races were needed to be conducted to verify the current results.

SDHB reduction promotes oral lichen planus by impairing mitochondrial respiratory function.

Background: Oral lichen planus (OLP) is a type of chronic inflammatory disorder, which represents a potential risk of malignant transformation. Understanding the mechanism of OLP-related malignant transformation could reduce the risk of cancer. Accumulating evidence indicates that the expression of succinate dehydrogenase enzyme B (SDHB) is associated with the carcinogenesis of oral squamous cell carcinoma (OSCC). However, the function and underlying mechanism of SDHB in OLP remains unknown. Methods: In this study, we examined the expression of SDHB in tissues from OLP patients and normal oral mucosa (NOM) through immunohistochemical (IHC) staining, quantitative reverse transcription polymerase chain reaction (qRT-PCR), and western blot (WB). Adenosine triphosphate (ATP) assay, reactive oxygen species (ROS) assay, mitochondrial membrane potential (MMP) assay, and glucose uptake assay were used to explore the function of SDHB in mitochondrial injury and bioenergetic changes in OLP cell model and SDHB-overexpressing cells. Results: In current study, we found that the messenger RNA (mRNA) and protein expression of SDHB was significantly decreased in OLP patients, accompanied by the accumulation of succinate. In the lipopolysaccharide (LPS) or CoCl2-stimulated OLP cell model, the expression of SDHB was decreased along with treatment time and concentration. Mechanistically, decreased SDHB enhanced hypoxia-inducible factor (HIF)-1α activity, induced mitochondrial injury, bioenergetic changes, and cytokine release. Overexpression of SDHB could reverse the above biological process and switch bioenergetic metabolism during OLP process. Conclusions: Our study suggests that SDHB reduction promotes OLP by impairing mitochondrial respiratory function.

The circ-AMOTL1/ENO1 Axis Implicated in the Tumorigenesis of OLP-Associated Oral Squamous Cell Carcinoma.

BACKGROUND: Oral squamous cell carcinoma (OSCC) may develop from a variety of oral potentially malignant disorders, but the mechanism of malignant transformation is still unknown. Among them, oral lichen planus (OLP) has a high prevalence. Previous studies have shown that α-enolase (ENO1) can promote cell proliferation and play an important role in tumorigenesis. In this study, we aim to explore the mechanism of ENO1 regulation in the process of OSCC tumorigenesis from OLP. METHODS: ENO1 expression in tissues was determined by real-time quantitative PCR and immunohistochemistry. ENO1 was knocked down in cal-27 to observe the change in cell proliferation. Then, RNA-seq and bioinformatics analyses were conducted between OLP and OSCC samples. The expression of circ-AMOTL1, miRNA-22-3p, and miRNA-1294 was assessed using the real-time quantitative PCR. With knockdown and overexpression of circ-AMOTL1 in vitro, the change of ENO1 in the mRNA level was also assessed. RESULTS: ENO1 was enhanced in the OSCC samples in comparison with OLP. Immunohistochemistry and real-time quantitative PCR results showed that ENO1 was significantly higher in OSCC tissue than in the OLP group, with a statistically significant difference (p<0.05). When ENO1 was knocked down in cal-27, cell proliferation was inhibited (p<0.05). The expression of miR-22-3p and miR-1294 was decreased in OSCC tissues, whereas ENO1 and circ-AMOTL1 increased. In an in vitro study, knockdown of circ-AMOTL1 resulted in a decrease of ENO1, while overexpression of circ-AMOTL1 led to an increase of ENO1 in the mRNA level. CONCLUSION: We confirmed that ENO1 expression was elevated in OSCC and increased cell proliferation. In an in vitro study, ENO1 expression was promoted by circ-AMOTL1. ENO1 may play a role as a tumor-promoting gene in OSCC through the circ-AMOTL1/miR-22-3p/miR-1294 network. These novel findings may shed further light on the pathogenesis from OLP to OSCC and the potential precursor markers.

[Effect of different fluence rates applied in curcumin-photodynamic therapy on Candida albicans biofilms].

PURPOSE: In this study, the inactivation effect of different fluence rates on Candida albicans biofilms during curcumin-photodynamic therapy was investigated in vitro. METHODS: The standard Candida albicans and clinical isolated Candida albicans were selected as model fungus and different fluence rates (12, 22, 42, 62, 82, 102 mW/cm2) during curcumin-photodynamic therapy were applied to inactivate Candida albicans biofilm. To evaluate the inactivation effect, XTT assay and Live/Dead kit were employed to quantify and visualize the activities of Candida albicans biofilms. The data were analyzed with SPSS 19.0 software package. RESULTS: When 40 µmol/L of curcumin was applied followed by 4 min illumination, both standard Candida albicans and clinical isolated Candida albicans biofilms were greatly inactivated along with the increase of fluence rates. When fluence rate increased to 102 mW/cm2, there was no significant difference between the experimental group and the previous experimental group(P>0.05). CONCLUSIONS: Fluence rate plays an important role in inactivation of Candida albicans biofilms during curcumin-photodynamic therapy, with optimized value of fluence rate of 82 mW/cm2 in this study.

Metabolic changes during malignant transformation in primary cells of oral lichen planus: Succinate accumulation and tumour suppression.

Oral squamous cell carcinoma (OSCC) is usually diagnosed at late stages, which leads to high morbidity. There are evidence that chronic inflammation (eg oral lichen planus [OLP]) was a risk factor of OSCC, but often misdiagnosed or ignored until invasion and metastasis. By applying precision medicine, the molecular microenvironment variations and relevant biomarkers for the malignant transformation from OLP to OSCC can be fully investigated. Several studies pointed out that the metabolic pathway were suppressed in OSCC. However, it remains unclear how the systemic profile of the metabolites change during the malignant transformation. In this study, we examined and compared the mucosa samples from 11 healthy individuals, 10 OLP patients and 21 OSCC patients. Based on the results, succinate, a key metabolite of the tricarboxylic acid cycle pathway, was accumulated in the primary cultured precancerous OLP keratinocytes and OSCC cells. Then, we found that succinate activated the hypoxia-inducible factor-1 alpha (HIF-1α) pathway and induced apoptosis, which could also be up-regulated by the tumour suppressor lncRNA MEG3. These results suggested the critical roles of succinate and MEG3 in the metabolic changes during malignant transformation from OLP to OSCC, which indicated that succinate, HIF1α and downstream proteins might serve as new biomarkers of precancerous OLP for early diagnosis and therapeutic monitoring. In addition, succinate or its prodrugs might become a potential therapy for the prevention or treatment of OSCC.

Candida albicans induces TLR2/MyD88/NF-κB signaling and inflammation in oral lichen planus-derived keratinocytes.

INTRODUCTION: The risk of oral lichen planus (OLP), a chronic inflammatory oral mucosal disease, becoming malignant increases by 21-fold in patients with fungal infection. This study examined the impact of Candida albicans exposure on Toll-like receptor (TLR) signaling in primary keratinocyte cultures obtained from OLP patients. METHODOLOGY: Following co-culture of primary OLP keratinocyte cultures with C. albicans for 24 hours, inflammatory cytokine concentrations were determined by ELISA. TLR2, MyD88, and NF-κBp65 mRNA and protein expression were assessed using quantitative RT-PCR and Western blot analyses, respectively. Keratinocyte apoptosis was also determined by flow cytometry. RESULTS: IL-10, IL-8, IL-2, and TNF-ɑ levels were significantly higher following co-culture with C. albicans (all p ≤ 0.034). MyD88, NF-κB p65, and TLR2 mRNA (all p < 0.001) and protein (all p ≤ 0.004) expression levels were significantly higher in OLP keratinocytes following C. albicans exposure. Finally, the apoptosis rates of OLP keratinocytes were 21.2%, 29.4%, and 25.4% for the control cells and 3.9%, 5.6%, and 4.4% for those exposed to C. albicans, suggesting that co-culture with C. albicans inhibits the apoptosis of OLP keratinocytes. CONCLUSIONS: C. albicans activates the TLR2/MyD88/NF-κB signaling pathway in OLP keratinocytes, resulting in increased cytokine expression and decreased keratinocyte apoptosis. Two key events in the pathogenesis of OLP and its progression to malignancy, namely increased inflammation and decreased apoptosis, were induced by exposure to C. albicans. Thus, targeting this signaling pathway may represent a novel therapeutic strategy to prevent OLP malignant transformation.

A genome-wide association scan of biological processes involved in oral lichen planus and oral squamous cell carcinoma.

BACKGROUND: In this study, the molecular mechanisms underlying malignant transformation from oral lichen planus (OLP) to oral squamous cell carcinoma (OSCC) were examined. METHODS: High-throughput sequencing of long noncoding RNAs (lncRNAs) and mRNAs of normal subjects and patients with OLP and OSCC was conducted. RNA-seq reads were mapped, lncRNA and mRNA transcripts were assembled, and expression levels were estimated. The targets of lncRNAs were predicted. Finally, Gene Ontology (GO) and pathway enrichment analyses of differentially expressed genes (DEGs) and lncRNA targets were performed. RESULTS: High-quality sequence data were generated and the mapping ratios for OSCC, normal, and OLP samples were high. In total, 820, 656, and 582 DEGs were obtained from OPL vs. normal, OSCC vs. normal, and OSCC vs. OPL, respectively. A total of 1721 known lncRNAs and 133 predicted lncRNAs and targets were obtained. Keratinization was significantly enriched by OSCC-related DEGs, but not OPL-related DEGs. The pathway of olfactory transduction was enriched by OPL- and OSCC-related DEGs. Defense response to virus and viral carcinogenesis were enriched by DEGs and lncRNA targets in all comparisons. GO term related to the metabolic process was enriched by lncRNA targets in the OPL vs normal comparison, and antigen processing and presentation via MHC class I was significantly enriched by lncRNA targets in the other 2 comparisons. CONCLUSION: Keratinization and MHC class I antigen processing and presentation were activated during the malignant transformation from OLP to OSCC. Additionally, the olfactory transduction pathway may be important for OSCC.

Identification of the key genes implicated in the transformation of OLP to OSCC using RNA-sequencing.

Oral lichen planus (OLP) is a chronic inflammatory disease that may transform to oral squamous cell carcinoma (OSCC), while its carcinogenesis mechanisms are not entirely clear. This study was designed to identify the important genes involved in the malignant transformation of OLP to OSCC. After RNA-sequencing, the differently expressed genes (DEGs) in OLP vs. normal and OSCC vs. normal groups, respectively, were identified by limma package in R language, and then clustering analysis were conducted by Pheatmap package in R language. Weighed gene co-expression network analysis (WGCNA) was performed for the DEGs to screen disease-associated modules. Using Cytoscape software, co-expression networks were constructed for the genes involved in the modules. Enrichment analysis was conducted for the genes involved in the co-expression networks using GOstat package in R language. Finally, quantitative real-time PCR (qRT-PCR) experiments were conducted to validate the key genes. There were, respectively, 223 and 548 DEGs in OLP vs. normal and OSCC vs. normal groups. WGCNA identified the blue modules for the DEGs in the two groups as disease-associated modules. Moreover, 19 common DEGs (including upregulated BCL9L, PER2 and TSPAN33, and downregulated GMPS and HES1) associated with both OLP and OSCC were identified. In the co-expression networks, BCL9L, HES1, PER2 and TSPAN33 might function in OLP via interactions (such as BCL9L-TSPAN33 and HES1-PER2). qRT-PCR analysis showed that BCL9L, PER2 and TSPAN33 were significantly upregulated, and GMP and HES1 were downregulated. These findings indicated that BCL9L, GMPS, HES1, PER2 and TSPAN33 affected the transformation of OLP to OSCC.

Hypoxia-inducible factor-1α and glucose transporter 1 in the malignant transformation of oral lichen planus.

Hypoxia-inducible factor-1α (HIF-1α) and glucose transporter 1 (GLUT1) are key factors in numerous physiological and pathological processes. However, studies on their involvement in the pathogenesis of oral lichen planus (OLP) and its progression toward oral squamous cell carcinomas (OSCC) are scarce. In this study, we examined the protein and gene expressions of both HIF-1α and GLUT1 in normal mucosa, nonatrophic OLP (OLPI), atrophic OLP (OLPII), and OSCC resulting from OLP. Tissues were obtained from 60 cases of OLP patients (n=36 for OLPI, n=24 for OLPII), 20 cases of OSCC patients and 30 healthy control individuals. In addition, in order to investigate if the pathological changes are due to hypoxia, we cultured keratinocytes under hypoxia conditions and measured the expression of HIF-1α and GLUT1. The results indicated that the expressions of HIF-1α and GLUT1 were gradually amplified from normal mucosa to OLPI, OLPII, and OSCC. The expression of both HIF-1α and GLUT1 in OLPII was significantly greater than OLPII. Likewise, the HIF-1α and GLUT1 expressions in OSCC were markedly higher compared to both OLPI and OLPII. Similar trends were obtained in real time PCR and Western blot analyses. A progressive increased micro-vessel density (MVD) was also recorded from normal mucosa to OLPI, OLPII, and OSCC. Moreover, the correlation analysis revealed significant positive correlations between HIF-1α and GLUT1 which were both correlated with MVD in the OLP and OSCC groups. Culture of keratinocytes isolated from OLP tissues under hypoxic and normoxic conditions showed a time-dependent inhibition of keratinocyte proliferation and increased expression of HIF-1α and GLUT1 under hypoxia conditions. In summary, we provided new evidence that hypoxia markers HIF-1α and GLUT1 are upregulated in OLP and are potentially involved in pathological changes leading to malignant transformation of OLP. Further characterization of these factors will provide new ideas for the diagnosis and treatment of OLP.

Effects of icariin on reproductive functions in male rats.

The present study investigated the effects and potential mechanism(s) of action of icariin on the reproductive functions of male rats. Adult rats were treated orally with icariin at doses of 0 (control), 50, 100, or 200 mg/kg body weight for 35 consecutive days. The results show that icariin had virtually no effect on the body weight or organ coefficients of the testes or epididymides. However, 100 mg/kg icariin significantly increased epididymal sperm counts. In addition, 50 and 100 mg/kg icariin significantly increased testosterone levels. Real-time PCR suggests icariin may be involved in testosterone production via mRNA expression regulation of genes such as peripheral type benzodiazepine receptor (PBR) and steroidogenic acute regulatory protein (StAR). Furthermore, 100 mg/kg icariin treatment also affected follicle stimulating hormone receptor (FSHR) and claudin-11 mRNA expression in Sertoli cells. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels were measured in the testes; 50 and 100 mg/kg icariin treatment improved antioxidative capacity, while 200 mg/kg icariin treatment upregulated oxidative stress. These results collectively suggest that icariin within a certain dose range is beneficial to male reproductive functions; meanwhile, higher doses of icariin may damage reproductive functions by increasing oxidative stress in the testes.

Dermatopontin is a novel regulator of the CdCl2-induced decrease in claudin-11 expression.

Cadmium (Cd) is a ubiquitous environmental heavy metal, which may be harmful to the reproductive functions through injury to the blood-testis barrier (BTB). However, the underlying mechanism of this adverse effect on the BTB remains uncharacterized. A preliminary study revealed that dermatopontin (DPT) expression was significantly increased in Cd chloride (CdCl2)-treated Sertoli cells in vitro, which suggested that an increase in DPT expression is crucial for CdCl2-induced BTB damage. To explore this further, in the present study we initially determined that DPT is expressed in testis Sertoli cells. The treatment of cells with CdCl2 resulted in a significant increase in DPT expression and a parallel decrease in claudin-11 expression, both in vivo and in vitro. To confirm the relationship between DPT and claudin-11, a DPT-silenced 15P-1 Sertoli cell model was established. We determined that DPT silencing could partly reduce the CdCl2-induced decrease in claudin-11 expression. Additionally, western blot analyses demonstrated that the p38 signaling pathway is involved in the effect of CdCl2 on DPT expression. In conclusion, the present study provides the first evidence that DPT may be a novel effector of CdCl2, highlighting the significant role of DPT in the regulation of claudin-11 expression.

Localization and expression patterns of prolactin-like protein J in mouse testis.

Prolactin (PRL)­like protein J (PLP­J) is a member of the prolactin family, mainly expressed in the placental decidua tissues of females, and is involved in gestation. To the best of our knowledge, it has not previously been shown to be expressed in males. Preliminary experiments of the present study indicated that PLP­J is expressed in the testis of male mice and is implicated in the regulation of testicular function. To definitively address whether PLP­J is expressed in the mouse testis, the expression pattern and cellular localization of PLP­J in mouse testes during postnatal development were characterized in the current study using molecular and immunological methods. Reverse transcription (RT)­polymerase chain reaction (PCR) was performed to amplify gene fragments from mouse testis specimens, which yielded sequences matching those of the PLP­J gene in Genbank. Subsequently, in situ hybridization showed that PLP­J was localized in interstitial tissue of the mouse testis. Immunofluorescence results indicated that PLP­J and 3ß­hydroxysteroid dehydrogenase 1 were colocalized in testis Leydig cells, confirming PLP­J expression in Leydig cells. In addition, PLP­J gene expression levels were examined at different stages of postnatal mouse development in male testis tissues using quantitative RT­PCR and western blotting. The results revealed that PLP­J expression levels were lowest in 18­day­old mice and highest in adults aged 4 months. Levels observed in 16­month­old individuals were lower than those observed in the 4­month­old mice, but remained significantly higher than the levels observed in 18­day­old mice. Furthermore, the roles of PLP­J in the murine testis TM3 Leydig cell line were studied. The results demonstrated that the upregulation of PLP­J expression in TM3 Leydig cells did not affect testosterone production or the cell cycle. In conclusion, this study demonstrated that PLP­J, a known member of the PRL family that was previously considered to be expressed solely in females, is also expressed in the testis of males with an age­dependent expression profile. Nevertheless, the physiological role of PLP­J in males remains unclear.

Proliferin-related protein overexpression in SGC­7901 gastric cancer cells inhibits in vitro cell growth and tumorigenesis in nude mice.

As reported in the literature, the worldwide 5-year overall survival rate for patients with gastric cancer receiving surgical treatment in the progressive stage is less than 25%. Therefore, there is an urgent need for the development of novel therapeutic strategies. Our preliminary studies demonstrated that proliferin-related protein (PRP) inhibits the proliferation of TM3 Leydig testicular cells. To evaluate whether PRP has antitumor effects in vitro and in vivo, we stably expressed PRP in SGC-7901 gastric carcinoma cells. PRP inhibited the proliferation and cell cycle progression of SCG-7901 cells, as determined by cell growth and cell cycle assays. Transwell experiments demonstrated that PRP inhibited the cell migration and invasion of SCG-7901 cells. Western blotting demonstrated that PRP-overexpressing cells had upregulated matrix metalloproteinase 9 (MMP-9) and downregulated tissue inhibitor of metalloproteinases-1 (TIMP-1). In a xenograft tumor formation assay using nude mice, tumors formed by PRP-overexpressing cells had significantly lower weights than those formed by control cells, and the tumor inhibitory rate reached 71.9%. We demonstrated for the first time that PRP inhibits gastric carcinoma cell proliferation, motility, and tumorigenicity in vivo, suggesting that PRP may become an important target for the development of gastric cancer gene therapy.

Study on adsorption of glyphosate (N-phosphonomethyl glycine) pesticide on MgAl-layered double hydroxides in aqueous solution.

MgAl-layered double hydroxides with different interlayer anions (nitrate, carbonate and chloride) were evaluated for their abilities to adsorb the organic pesticide glyphosate (N-phosphonomethyl glycine, Gly). The adsorption isotherms of Gly on layered double hydroxides (LDHs) nitrate were described by the Langmuir equation at lower equilibrium concentration of Gly (C(e)<1.0 mmol/L), and the Gly adsorption capacity on LDHs increased with the layer charge density, i.e. the structural Al3+/Mg2+ ratio. Gly adsorption on LDHs nitrate generally occurred through two processes, external surface adsorption and interlayer anion exchange. The adsorption amount on LDHs at C(e)=1.0 mmol/L decreased in the order of interlayer anions: Cl(-)>NO3(-)>CO3(2-).