Clinical evaluation of photodynamic therapy for oral leukoplakia: a retrospective study of 50 patients.

BACKGROUND: Topical photodynamic therapy (PDT) has demonstrated encouraging results in the treatment of oral leukoplakia (OLK). However, data on the clinical efficacy of PDT in Chinese patients with OLK are still limited. METHODS: Fifty patients diagnosed with OLK were enrolled, including patients with various dysplastic tissues. All patients received topical PDT with 5-aminolevulinic acid (5-ALA) as a photosensitizer. Clinical efficacy was evaluated 4 weeks after treatment. Follow-up was performed every 3 months during the first year and every 6 months during the second year. RESULTS: The overall response rate was 68% (34/50): 12% (n = 6) complete and 56% (n = 28) partial responses. Aneuploidy was reduced in the patients with dysplastic lesions. Oral pain and local ulcers developed in 52% of the patients (n = 26). Patients with a long history of OLK including hyperplasia and dysplastic lesions, as well as those with non-homogenous lesions, were more likely to develop pain and ulcer. During follow-up, the recurrence rate of hyperplasia and dysplastic lesions was 32% (n = 16) and the malignant transformation rate of dysplastic lesions was 4% (n = 2). Lesions on the buccal mucosa were associated with recurrence (P = 0.044; OR: 0.108, 95% CI: 0.013-0.915). CONCLUSION: Topical 5-ALA-mediated PDT is an effective treatment for OLK, particularly for homogenous leukoplakia, with few side effects. The buccal mucosa may be a protective factor that can reduce recurrence.

Clinical relevance of serum lipids in the carcinogenesis of oral squamous cell carcinoma.

BACKGROUND: Dyslipidaemia is associated with cancers. However, the specific expression of serum lipids in oral potentially malignant disorders (OPMD) and oral squamous cell carcinoma (OSCC) remains unclear, and it remains unknown whether serum lipids are associated with the development of OPMD and OSCC. This study investigated the serum lipid profiles of OPMD and OSCC patients, and the association of serum lipids with the occurrence of OPMD and OSCC. METHODS: A total of 532 patients were recruited from the Affiliated Hospital of Stomatology, Nanjing Medical University. Serum lipid parameters including total cholesterol (TC), triglycerides (TGs), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), apolipoprotein A (Apo-A), apolipoprotein B (Apo-B), and lipoprotein (a) (Lpa) were analysed, and clinicopathological data were collected for further analysis. Furthermore, a regression model was used to evaluate the relationship between serum lipids and the occurrence of OSCC and OPMD. RESULTS: After adjusting for age and sex, no significant differences were observed in serum lipid or body mass index (BMI) between OSCC patients and controls (P > 0.05). HDL-C, Apo-A, and Apo-B levels were lower in OSCC patients than in OPMD patients (P < 0.05); HDL-C and Apo-A levels were higher in OPMD patients than in controls (P < 0.05). Furthermore, female OSCC patients had higher Apo-A and BMI values than males. The HDL-C level was lower in patients under 60 years of age than in elders (P < 0.05); and age was related to a higher risk of developing OSCC. Female patients with OPMD had higher TC, HDL-C, and Apo-A levels than males (P < 0.05); OPMD patients over 60 years of age had higher HDL-C than youngers (P < 0.05), whereas the LDL-C level was lower in elders (P < 0.05). The HDL-C and BMI values of the patients with oral leukoplakia (OLK) with dysplasia were more elevated than those of the oral lichen planus group, and the LDL-C, and Apo-A levels in patients with OLK with dysplasia were decreased (P < 0.05). Sex, high HDL-C and Apo-A values were associated with the development of OPMD. CONCLUSION: Serum lipids exhibited certain differences according to the occurrence and development of OSCC; high levels of HDL-C and Apo-A might be markers for predicting OPMD.

Endoplasmic reticulum stress promotes the release of exosomal PD-L1 from head and neck cancer cells and facilitates M2 macrophage polarization.

BACKGROUND: Endoplasmic reticulum (ER) stress has been found to foster the escape of cancer cells from immune surveillance and upregulate PD-L1 expression. However, the underlying mechanisms are unknown. METHODS: While analyzing the protein levels using immunofluorescence and Western blotting, the RNA levels were measured using qRT-PCR. Ten injection of exosomes into six-week-old nude mice was made through the tail vein once every other day in total. RESULTS: The expression of certain ER stress markers such as PERK (PKR-like endoplasmic reticulum kinase), ATF6 (activating transcription factor 6), and GRP78 (glucose-regulated protein 78), was found to be upregulated in the oral squamous cell carcinoma (OSCC) tissues and related to poor overall survival. There is a positive relationship between the extent of ER stress-related proteins and a cluster of PD-L1 expression and macrophage infiltration among the OSCC tissues. Further, incubation with exosomes derived from ER-stressed HN4 cells (Exo-ER) was found to upregulate PD-L1 extents in macrophages in vitro and in vivo, and macrophage polarization toward the M2 subtype was promoted by upregulating PD-L1. CONCLUSIONS: ER stress causes OSCC cells to secrete exosomal PD-L1 and upregulates PD-L1 expression in macrophages to drive M2 macrophage polarization. The delineation of a new exosome-modulated mechanism was made for OSCC-macrophage crosstalk driving tumor development and to be examined for its therapeutic use. Exosomal PD-L1 secreted by ER-stressed OSCC cells promoted M2 macrophage polarization. Video Abstract.

Efficacy and safety of miconazole muco-adhesive tablet versus itraconazole in oropharyngeal candidiasis: A randomized, multi-centered, double-blind, phase 3 trial.

Oropharyngeal candidiasis (OPC) is an opportunistic infection treated with anti-fungal agents. Herein, we evaluate the efficacy and safety of miconazole buccal tablets (MBT) and itraconazole capsules in the localized treatment of patients with OPC. In this multi-centered, double-blinded, phase III trial (CTR20130414), both males and non-pregnant females (≥18 years) with OPC were randomized (1:1) to MBT plus placebo (experimental group) or itraconazole capsules plus placebo (control group). The primary endpoint was clinical cure at the end-of-treatment period [visit 4 (V4)] while secondary endpoints were clinical remission rates, partial remission rates, mycological cure, clinical relapse, and adverse events (AEs). All endpoints were statistically analyzed in both the full analysis set (FAS) and per-protocol (PP) set. A total of 431 (experimental: 216; control: 215) subjects were included. At V4, in the FAS set, the clinical cure was achieved in 68% and 59% patients in experimental and control groups, respectively with a treatment difference of 9% [95% confidence interval (CI): -1,19; P < .001] demonstrating non-inferiority of MBT over itraconazole. At V4, mycological cure rates were 68.2% and 42.0% in the experimental group and control groups (P < .001), respectively in FAS. The relapse rates were 5.4% and 6.6%, respectively, in the experimental and control groups. A total of 210 patients experienced AEs during treatment with 47.7% in the experimental group and 49.8% in the control group with no deaths. This study demonstrated that once-daily treatment with MBT was non-inferior to itraconazole with higher mycological cure rates and was tolerable with mild AE in patients with OPC.

Miconazole is an antifungal drug against certain types of fungus or yeast infections. In this study, we showed that treatment with once-daily miconazole buccal tablets was as effective as systemic itraconazole capsules in Chinese patients infected by oropharyngeal candidiasis with minimum side effects.

MiR-155-5p modulates inflammatory phenotype of activated oral lichen-planus-associated-fibroblasts by targeting SOCS1.

BACKGROUND: Oral lichen planus (OLP) is a chronic inflammatory oral mucosal disease. Cytokines are closely associated with OLP development. In addition to immune cells, fibroblasts have been reported to induce regional inflammation. MicroRNA(miR)-155-5p is reportedly increased significantly in OLP and is known to regulate inflammation. This study aimed to investigate the role of miR-155-5p in fibroblasts of OLP lesions. METHODS AND RESULTS: Normal mucosal fibroblasts (NFs) and OLP associated-fibroblasts (OLP AFs) were isolated from the oral mucosa of 15 healthy controls and 30 OLP patients. We detected the expression of miR-155-5p and fibroblast activation protein alpha (FAP-α) using quantitative RT-PCR and analyzed their correlation. Interleukin (IL)-6 and IL-8 levels were determined using ELISA. Expression of suppressor of cytokine signaling (SOCS) 1 was analyzed by western blotting. A dual-luciferase reporter assay was performed to investigate the interaction between miR-155-5p and SOCS1. MiR-155-5p and FAP-α were significantly increased and positively correlated in OLP AFs. Overexpression of miR-155-5p in OLP AFs augmented IL-6 and IL-8 release and decreased SOCS1 expression, whereas knockdown of miR-155-5p in OLP AFs decreased IL-6 and IL-8 release. The expression of SOCS1 was downregulated in OLP AFs, and SOCS1 silencing augmented IL-6 and IL-8 production in OLP AFs. Furthermore, miR-155-5p inhibited SOCS1 expression by directly targeting its 3'-UTR in OLP AFs. CONCLUSIONS: MiR-155-5p regulates the secretion of IL-6 and IL-8 by downregulating the expression of SOCS1 in activated OLP AFs. Our results provide novel insights into the pathogenesis of OLP and identify a potential new target for OLP therapy.

Expression and biological functions of the CCL5-CCR5 axis in oral lichen planus.

Oral lichen planus (OLP) is a chronic inflammatory oral mucosal disorder with T cell-mediated immunological pathogenesis. C-C motif chemokine ligand 5 (CCL5) and its receptor C-C motif chemokine receptor 5 (CCR5) play important roles in the activation and recruitment of T cells. This study sought to explore the expression and biological functions of the CCL5-CCR5 axis in OLP. We examined the expression of the CCL5-CCR5 axis in the peripheral blood and oral tissue of healthy controls and patients with OLP using quantitative real-time PCR, Simple Western assays, enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry. In addition, we investigated the effects of the CCL5-CCR5 axis on the proliferation, apoptosis and migration of OLP T cells using Cell Counting Kit-8 (CCK8) assay, flow cytometry and transwell assay. We found that the expression of the CCL5-CCR5 axis was significantly elevated both in the peripheral blood and oral tissue of patients with OLP compared with healthy controls. CCL5 not only promoted OLP T-cell proliferation and migration but also inhibited OLP T-cell apoptosis. Moreover, CCR5 inhibition suppressed OLP T-cell proliferation and migration, whereas OLP T-cell apoptosis was promoted. In conclusion, our data suggest that the CCL5-CCR5 axis may be closely related to the inflammatory infiltration of T cells in OLP.

Discovery and preclinical validation of proteomic biomarkers in saliva for early detection of oral squamous cell carcinomas.

OBJECTIVES: Our aim was to identify and prevalidate a set of salivary proteins that can distinguish oral squamous cell carcinomas (OSCC) patients from healthy individuals and patients with oral potentially malignant disorders (OPMD). SUBJECTS AND METHODS: Proteomes of 60 saliva samples from healthy individuals, OPMD patients, and OSCC patients were assayed using the isobaric tags for relative and absolute quantitation (iTRAQ) method. Enzyme-linked immunosorbent assay (ELISA) was used to prevalidate the candidate biomarkers in an independent sample set (n = 90). RESULTS: In total, 246 differentially expressed proteins were identified by comparing each two groups, and 21 proteins were differentially expressed when OSCC was compared with both OPMD and Control. Three proteins, namely, solute carrier family 3 member 2 (SLC3A2), S100 calcium-binding protein A2 (S100A2), and interleukin-1 receptor antagonist protein (IL1RN), were selected as candidate biomarkers. Comparing the OSCC group with the healthy group, the area under curve (AUC) of the three combined biomarkers was 0.89, with a sensitivity of 83.33% and a specificity of 83.33%. Comparing the OSCC group with the OPMD group, the AUC value was 0.87, with a sensitivity of 93.33% and a specificity of 70.00%. CONCLUSION: Our study indicates that salivary proteomics is promising for the discovery of OSCC biomarkers.

Macrophage-Derived Exosomal miR-31-5p Promotes Oral Squamous Cell Carcinoma Tumourigenesis Through the Large Tumor Suppressor 2-Mediated Hippo Signalling Pathway.

Tumour-associated macrophages (TAMs) are thought to contribute to oral squamous cell carcinoma (OSCC) initiation and progression. However, the underlying mechanism through which TAMs foster OSCC progression is still unclear. This study intended to determine whether there are exclusively exosomal miRNAs-derived macrophages that are functionally necessary for OSCC progression. The phenotype of TAM recruitment in OSCC tissue samples was assessed, subsequently identifying the influence of M2 macrophages and exosomes derived from M2 macrophages on OSCC proliferation and tumorigenesis in vitro and in vivo. CD68 and CD163, the specific markers of M2 type macrophages, were upregulated in TAMs presented in intra-cancer tissues. M2 macrophages and M2 macrophage-derived exosomes (M2 exos) both can promote OSCC growth and tumorigenicity. An exosomal RNA-seq analysis was conducted to predict regulatory exosomal miRNAs related to OSCC growth, which determined miR-31-5p and LATS2 for subsequent experiments. Mechanistically, miR-31-5p was delivered to recipient OSCC cells through M2 exos and complementary pairing with the large tumor suppressor 2 (LATS2) coding sequence, thus suppressing the expression of LATS2 and inactivation the Hippo signaling pathway to support OSCC growth. Collectively, our findings demonstrate that M2 macrophage-derived exosomal miR- 31-5p can make tumor suppressor LATS2 gene inhibited and facilitate the progression of OSCC via inhibiting the Hippo signaling pathway, which possibly provides new targets for the molecular therapy of OSCC.

Microbial Community Analysis of Saliva and Biopsies in Patients With Oral Lichen Planus.

The specific etiology and pathogenesis of oral lichen planus (OLP) remain elusive, and microbial dysbiosis may play an important role in OLP. We evaluated the saliva and tissue bacterial community of patients with OLP and identified the colonization of bacteria in OLP tissues. The saliva (n = 60) and tissue (n = 24) samples from OLP patients and the healthy controls were characterized by 16S rDNA gene sequencing and the bacterial signals in OLP tissues were detected by fluorescence in situ hybridization (FISH) targeting the bacterial 16S rDNA gene. Results indicate that the OLP tissue microbiome was different from the microbiota of OLP saliva. Compared with the healthy controls, Capnocytophaga and Gemella were higher in OLP saliva, while Escherichia-Shigella and Megasphaera were higher in OLP tissues, whereas seven taxa, including Carnobacteriaceae, Flavobacteriaceae, and Megasphaera, were enriched in both saliva and tissues of OLP patients. Furthermore, FISH found that the average optical density (AOD) of bacteria in the lamina propria of OLP tissues was higher than that of the healthy controls, and the AOD of bacteria in OLP epithelium and lamina propria was positively correlated. These data provide a different perspective for future investigation on the OLP microbiome.

Corrigendum: Microbial Community Analysis of Saliva and Biopsies in Patients With Oral Lichen Planus.

[This corrects the article on p. 629 in vol. 11, PMID: 32435231.].

Photobiomodulation therapy for repeated closed head injury in rats.

Repeated traumatic brain injury, leads to cumulative neuronal injury and neurological impairments. There are currently no effective treatments to prevent these consequences. Growing interest is building in the use of transcranial photobiomodulation (PBM) therapy to treat traumatic brain injury. Here, we examined PBM in a repeated closed head injury (rCHI) rat model. Rats were administered a total of three closed head injuries, with each injury separated by 5 days. PBM treatment was initiated 2 hours after the first injury and administered daily for a total of 15 days. We found that PBM-treated rCHI rats had a significant reduction in motor ability, anxiety and cognitive deficits compared to CHI group. PBM group showed an increase of synaptic proteins and surviving neurons, along with a reduction in reactive gliosis and neuronal injury. These findings highlight the complexity of gliosis and neuronal injury following rCHI and suggest that PBM may be a viable treatment option to mitigate these effects and their detrimental consequences.

Increased granulysin in the peripheral blood and tissues of patients with oral lichen planus.

Oral lichen planus (OLP) is a chronic inflammatory disease of unclear etiology and pathogenesis. Granulysin (GNLY) participates in various immune responses and mediates various skin diseases. However, its expression in OLP has not been reported. This study was to investigate whether there was an abnormal expression of GNLY in the peripheral blood and tissues of patients with OLP. Twenty patients with non-erosive OLP, twenty patients with erosive OLP, and twenty healthy controls were enrolled. The mRNA expression of GNLY in the peripheral blood and tissues was detected using RT-qPCR. The protein expression of GNLY in the peripheral blood plasma was measured using ELISA. The GNLY in tissues was investigated using immunohistochemistry. The mRNA and protein expression of GNLY in non-erosive and erosive OLP patients, when compared with the controls, were upregulated both in the peripheral blood and tissue. Also, in non-erosive and erosive OLP lesion tissues, there was a weak positive expression of GNLY in the lymphocyte membrane of the lamina propria layer and was less expressed in the epithelial layer. In normal mucosa, GNLY was hardly expressed. Furthermore, the immunohistochemical levels of GNLY in non-erosive and erosive OLP lesion were significantly higher than that in the normal oral mucosa. In conclusions, the expression of GNLY in the peripheral blood and tissues of OLP patients were significantly higher than controls, suggesting that there exists an abnormality in the expression of GNLY in OLP which might be involved in the pathogenesis of OLP.

Changes of gene expression in T lymphocytes following Golli­MBP gene RNA interference.

The major cell types expressing Golli in the immune system are the T­lineage cells. The aim of the current study was to investigate the changes of gene expression in T lymphocytes subsequent to downregulation of the Golli­myelin basic protein (MBP) gene. RNA interference technology was used to suppress the expression of Golli­MBP in Jurkat cells and DNA microarray techniques were applied to investigate the alterations of gene expression profiles. The results indicated that there were 387 differentially expressed genes. In the Golli­MBP knockdown Jurkat cells, the expression of 108 genes was enhanced, 279 genes were suppressed. Gene ontology analysis identified differentially expressed genes involved in several biological progresses, including cell adhesion and immune responses. Pathway analysis demonstrated that the majority of the differentially expressed genes (23.3%) were involved in cytokine­cytokine receptor interaction. Subsequent to Golli­MBP knockdown, the mechanisms that changed the biological characteristics of Jurkat cells were complex, involving numerous types of functional proteins, and metabolic and signaling pathways. However, further experiments are required to confirm these results.

Intracellular Ca2+ and related proteins in patients with oral lichen planus.

Oral lichen planus (OLP) is suggested to be a T cell-mediated chronic inflammatory oral mucosal disease. Gene expressed in the oligodendrocyte lineage-myelin basic proteins (Golli-MBP) and stromal interaction molecule 1 (STIM1) are important in the activation and function of T lymphocytes. This study aimed to analyze and compare the expression of Golli-MBP and STIM1 between OLP patients and healthy controls and to analyze the level of intracellular Ca(2+), which is involved in lymphocyte activation. The Ca(2+) fluorescent probe, Fluo-3/AM, was used to test the level of intracellular Ca(2+) in patients with OLP and healthy controls peripheral blood lymphocytes. Golli-MBP and STIM1 mRNA and protein levels were analyzed using quantitative real-time PCR and Western blot, respectively. Following lymphocyte activation, the intracellular Ca(2+) in OLP patients was markedly lower than that in the control group (P < 0.001). In OLP patients, the expression of Golli-MBP mRNA and protein was significantly upregulated compared to those of the control group (P < 0.001). Similarly, OLP patients showed markedly upregulated levels of STIM1 mRNA expression (P < 0.01) and protein compared to healthy controls. The intracellular Ca(2+) of OLP patients was markedly lower than that of healthy controls. This evidence may indicate that Ca(2+) signaling pathways in OLP patients are abnormal. The overexpression of Golli-MBP and STIM1 may play a role in the pathogenesis of OLP.

[The progress in expression of genes of the oligodendrocyte lineage-myelin basic protein in oral lichen planus].

OBJECTIVE: The aim of this study was to determine the expression of genes of the oligodendrocyte lineage-myelin basic protein (Golli-MBP) in peripheral blood mononuclear cell (PBMC) in oral lichen planus (OLP) and to further understand the pathogenesis of OLP. METHODS: PBMC was obtained by density gradient centrifugation, and the expression of Golli-MBP in PBMC was investigated in erythematous/erosive OLP (20 cases), reticular OLP (16 cases) and normal controls (19 cases) using reverse transcription-polymerase chain reaction (RT-PCR) and Western blot methods. RESULTS: RT-PCR results showed that Golli-MBP mRNA was overexpressed in erythematous/erosive and reticular OLP as compared with normal control group (P < 0.01). Western blot assay indicated that erythematous/erosive and reticular OLP patients had a higher expression level of Golli-MBP protein in PBMC than normal controls (P < 0.01). However, there were no significant differences between erythematous/erosive and reticular groups in Golli-MBP mRNA and protein expression (P > 0.05). CONCLUSION: The data accumulated here strongly indicate that Golli-MBP was involved in the pathogenesis of OLP.

[Effect of Yupingfeng on the level of salivary epidermal growth factor in oral lichen planus].

OBJECTIVE: To examine the effect of mouthwash Yupingfeng on the level of salivary epidermal growth factor (sEGF) in oral lichen planus (OLP). METHODS: The level of sEGF was measured by radioimmunoassay with ligand 125I-EGF. The saliva samples were taken from the normal control group and the OLP patients before and after treatment with Yupingfeng. RESULTS: The levels of sEGF in OLP patients before treatment with Yupingfeng were (4.09 +/- 3.64) microg/L, which was significantly higher than that in normal control [(2.15 +/- 1.62) microg/L, P = 0.013], and (2.57 +/- 1.19) microg/L after treatment,which was significantly lower than before treatment (P = 0.05). CONCLUSIONS: Yupingfeng can modulate the level of sEGF in OLP patients.