Expression patterns of cancer stem cell markers ALDH1 and CD133 correlate with a high risk of malignant transformation of oral leukoplakia.

Molecular markers for predicting oral cancer development in premalignant oral leukoplakia (OL) are urgently needed. The objective of this study was to examine the expression patterns of cancer stem cell markers ALDH1 and CD133 in samples from patients with OL, and determine their prognostic values for subsequent development of oral cancer. Immunohistochemistry for ALDH1 and CD133 was performed in samples from a cohort of 141 patients with biopsy-proven OL who received a mean follow-up of 5.5 years. Patient clinicopathologic and follow-up data were analyzed. Expression of ALDH1 and CD133 was observed in 54 (38.3%) and 32 (22.7%) of 141 patients with OL, respectively. Kaplan-Meier analysis showed that 48.1% patients with ALDH1-positivity developed oral cancer compared with 12.6% those with ALDH1-negativity (p < 0.001). Meanwhile, 59.4% patients with CD133-positivity developed oral cancer compared with 16.5% those with CD133-negativity (p < 0.001). Multivariate analysis revealed that ALDH1 and CD133 expression was associated with 4.17-fold [95% confidence interval (CI), 1.96-8.90; p < 0.001] and 2.86-fold (95% CI, 1.48-5.55; p = 0.002) increased risk of OL transformation, respectively. Collectively, these data demonstrated for the first time that the expression of ALDH1 and CD133 correlated with malignant transformation in a large series of patients with OL who received a long-term follow-up, which suggests that they may serve as predictors to identify OL with a high risk of oral cancer development.

Expression of cancer stem cell markers ALDH1 and Bmi1 in oral erythroplakia and the risk of oral cancer.

BACKGROUND: Oral erythroplakia (OE) is a notoriously aggressive oral pre-malignant lesion with a high tendency to oral cancer development, but its biological behavior is largely unknown. The objective of this study was to determine the expression of cancer stem cell markers ALDH1 and Bmi1 in OE and their correlation with malignant transformation of OE. METHODS: In a retrospective case-control study, expression patterns of ALDH1 and Bmi1 were determined using immunohistochemistry in samples from 34 patients with OE, including patients with untransformed lesions (n=17) and patients with malignant transformed lesions (n=17). RESULTS: ALDH1 and Bmi1 expression was observed in 19 (55.9%) and 20 (58.8%) of 34 patients with OE, respectively. Multivariate analysis revealed that ALDH1 expression was significantly associated with increased risk of transformation (P<0.05), but Bmi1 expression was not a significant marker (P > 0.05). Notably, the coexpression of both ALDH1 and Bmi1 was a strong indicator associated with 8.56-fold (95% confidence interval [CI], 1.74-42.17; P<0.01) for malignant transformation. Point prevalence analysis revealed that 78.6% (95% CI, 54.0-100) of the patient with coexpression of both ALDH1 and Bmi1 developed oral cancer. CONCLUSION: Our data indicated that the expression patterns of ALDH1 and Bmi1 in OE were associated with malignant transformation, suggesting that they may be valuable predictors for evaluating the risk of oral cancer.

[Double-pulley combined with suture bridge technique for fixation of comminuted fractures of distal patella pole].

OBJECTIVE: To investigate the clinical effect of double pulley combined with suture bridge in the treatment of comminuted fracture of the lower pole of the patella. METHODS: From January 2018 to June 2020, 15 patients with comminuted fracture of the lower pole of the patella were treated with double pulley and suture bridge technology, including 9 males and 6 females, aged 28 to 68 years old with an average of (42.4±9.6) years old. All patients had obvious knee joint pain and limited movement after injury. All knee joints were examined by X-ray and CT, which confirmed that they were all comminuted fractures at the lower level of the patella. After operation, X-ray films of the knee joint were taken regularly to understand the fracture healing, the Insall Salvati index was measure, the range of motion of the joint was recorded, and the function of the knee joint was evaluateed by the Bostman scoring system. RESULTS: All the 15 patients were followed up for 7 to 24 months with an average of (11.4±4.2) months, and there was no obvious anterior knee pain. At the last follow-up, the knee joint range of motion of the affected limb was 105° to 140° with an average of (128.5±12.8) °, and the Insall Salvati index was 0.79 to 1.12 with an average of (0.92±0.18). The X-ray film showed that the patella was bone healing, and no anchor fell off, broken, or displaced fracture block was found. Bostman patellar fracture function score was 27.85±2.06, 13 cases were excellent, 2 cases were good. CONCLUSION: Double pulley technique combined with suture bridge technique is reliable for reduction and fixation of comminuted fracture of the lower pole of patella, and patients can start functional exercise early after operation.

A clinicopathological study on verrucous hyperplasia and verrucous carcinoma of the oral mucosa.

BACKGROUND: Oral verrucous hyperplasia (VH) and verrucous carcinoma (VC) are two clinicopathologically distinctive oral verrucous lesions. The objective of this study was to investigate the clinicopathological features of the two verrucous lesions and estimate their relationship from China. METHODS: Retrospective review of two series of patients with histologically confirmed VH (n = 121) and VC (n = 56) between 1996 and 2009 in our hospital were conducted. RESULTS: The average age of VH was 58.5 years (ratio male:female = 1.37) with the tongue being the predominant site. The average age of VC was 64.3 years (ratio male:female = 1.15) with the lower lip being the predominant site. Multivariate analysis revealed that the elderly patient with verrucous lesion (≥60 years) was associated with 3.06-fold (P = 0.007) increased carcinoma risk compared with the non-elderly patient. The lesion located on lower lip was associated with 13.54-fold (P < 0.001) increased carcinoma risk compared with other sites. CONCLUSION: Clinicopathological features of VH and VC in China were elucidated. Elderly patient with oral verrucous lesion located on the lower lip correlates with higher risk of carcinoma.

Application of acellular dermal matrix in reconstruction of oral mucosal defects in 36 cases.

PURPOSE: Artificial grafts have been investigated for use in the repair of oral mucosal defects. The aim of this retrospective study was to present the outcomes of the use of acellular dermal matrix (ADM) grafts to repair oral mucosal defects. MATERIALS AND METHODS: Data from 36 patients with oral mucosal defects reconstructed with ADM grafts from 2003 through 2009 were reviewed. All patients were followed-up for at least 6 months to observe the graft repair, wound-healing time, contracture, color, infection, pain, immunologic reaction, texture of the graft, and clinical course. Graft success was defined as the ADM graft being replaced by new mucosa-like tissue and the oral mucosal defect being covered with the new mucosa-like tissue. Any evidence of incomplete graft re-epithelialization or graft sloughing was considered a graft failure (complete or incomplete). RESULTS: Of the 36 cases, 34 grafts (94.4%) were successfully replaced with new mucosa-like tissues and only 2 grafts (5.6%) failed. No complaints such as pain, immunologic reaction, or infection were observed during the follow-up. Mild graft contraction occurred in 7 patients with lip or buccal defects, especially at approximately 3 to 5 weeks after the reconstructive surgery. CONCLUSIONS: The ADM grafts for oral mucosal defects were safe and effective. The present data support the clinical application of ADM grafts in reconstructing oral mucosal defects caused by various oral diseases.

Sequential damage assessment of the posterolateral complex of the knee joint: a finite element study.

BACKGROUND: The posterolateral complex (PLC), which consists of the popliteus tendon (PT), lateral collateral ligament (LCL), and popliteofibular ligament (PFL), is an indispensable structure of the knee joint. The aim of this study was to explore the functionality of the PLC by determining the specific role of each component in maintaining posterolateral knee stability. METHODS: A finite element (FE) model was generated based on previous material property data and magnetic resonance imaging of a volunteer's knee joint. The injury order of the PLC was set as LCL, PFL, and PT. A combined compressive load of 1150 N and an anterior tibial load of 134 N was applied to the tibia to investigate tibial displacement (TD). Tibial external rotation (TER) and tibial varus angulation (TVA) were measured under bending motions of 5 and 10 Nm. The instantaneous axis of rotation (IAR) of the knee joint under different rotation motions was also recorded. RESULTS: The TD of the intact knee under a combined compressive load of 1150 N and an anterior tibial load of 134 N matched the values determined in previous studies. Our model showed consistent increases in TD, TVA, and TER after sequential damage of the PLC. In addition, sequential disruption caused the IAR to shift superiorly and laterally during varus rotation and medially and anteriorly during external rotation. In the dynamic damage of the PLC, LCL injury had the largest effect on TD, TVA, TER, and IAR. CONCLUSIONS: Sequential injury of the PLC caused considerable loss of stability of the knee joint according to an FE model. The most significant structure of the PLC was the LCL.

Malignant transformation of oral epithelial dysplasia: clinicopathological risk factors and outcome analysis in a retrospective cohort of 138 cases.

AIMS: To explore the usefulness of a new binary system of grading dysplasia proposed by the World Health Organization and to identify significant risk factors for malignant transformation in a long-term follow-up cohort of patients with oral epithelial dysplasia. METHODS AND RESULTS: A total of 138 patients with histologically confirmed oral dysplasia between 1978 and 2008 were reviewed retrospectively in our department. The mean follow-up period was 5.1 years. Of these dysplasias, 37 (26.8%) developed into cancer, with a mean duration of 4.6 years. Cox regression analysis revealed that high-grade dysplasia was an independent risk factor for transition, but age, gender, lesion site, diet habit, smoking and alcohol intake were not risk factors. High-grade dysplasia was associated with a 2.78-fold (95% confidence interval 1.44-5.38; P = 0.002) increased risk of transition, as compared with low-grade dysplasia. Consistently, high-grade dysplasia had a significantly higher incidence of malignancy than low-grade dysplasia by Kaplan-Meier analysis (log-rank test, P = 0.001). CONCLUSIONS: The utilization of high-grade dysplasia as a significant indicator for evaluating malignant transformation risk in patients with potentially malignant lesions is suggested; this may be helpful to guide treatment selection in clinical practice.

Transcriptome array screening and verification of oral leukoplakia carcinogenesis-related hypoxia-responsive gene and microRNA.

OBJECTIVES: To study the hypoxia response gene and microRNA (miRNA) expression profiles in the pathogenesis and progression of oral leukoplakia (OLK). METHODS: Affymetrix GeneChip human transcriptome array 2.0 was used to detect the transcriptome of normal mucosa, low-risk OLK, high-risk OLK, and early squamous cell carcinoma (SCC). Gene ontology function analysis was used to screen genes and key miRNAs whose biological role is hypoxia response. Quantitative reverse transcription polymerase ch-ain reaction (qRT-PCR) was used to verify the expression of hypoxia response genes and miRNAs. RESULTS: A total of 7 different genes of hypoxia response between normal mucosa and low-risk OLK, 10 genes between low-risk and high-risk OLK, and 21 genes between high-risk OLK and SCC were identified. The results of qRT-PCR showed that the expression of hypoxia-inducible factor 1α, chemokine cc-motif ligand 2, and matrix metalloproteinase 3 mRNA and miR-21 in normal mucosa, OLK, and SCC increased in a stepwise manner. The expression difference between OLK and SCC was statistically significant and consistent with the results of transcriptome array. CONCLUSIONS: The hypoxia response gene and related miRNA play roles in the development and progression of OLK.

Structural validity and internal consistency of Picture My Participation: A measure for children with disability.

BACKGROUND: Picture My Participation (PMP) intended to measure participation, defined as attendance and involvement in everyday situations, of children with disabilities, particularly in low- and middle-income settings. OBJECTIVES: To explore structural validity of PMP by identifying possible subcomponents in the attendance scale and examining internal consistency of the total score and each subcomponent. METHOD: A picture-supported interview was conducted with 182 children, 7-18 years, with and without intellectual disability (ID). Frequency of attendance in 20 activities was rated on a four-point Likert scale (never, seldom, sometimes and always). RESULTS: An exploratory principal component analysis extracted four subcomponents: (1) organised activities, (2) social activities and taking care of others, (3) family life activities and 4) personal care and development activities. Internal consistency for the total scale (alpha = 0.85) and the first two subcomponents (alpha = 0.72 and 0.75) was acceptable. The two last subcomponents alpha values were 0.57 and 0.49. CONCLUSION: The four possible subcomponents of PMP can be used to provide information about possible domains in which participation and participation restrictions exist. This study provided further psychometric evidence about PMP as a measure of participation. The stability and the utility of these subcomponents needed further exploration.

MicroRNA-134 inhibits tumor stem cell migration and invasion in oral squamous cell carcinomas via downregulation of PI3K-Akt signaling pathway by inhibiting LAMC2 expression.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is the most common malignant neoplasm of the mouth. Some studies have found that multiple microRNAs (miRs) participate in OSCC physiological and pathological processes. METHODS: We explored the mechanism of action of miR-134 in OSCC involving the PI3K-Akt signaling pathway. Different bioinformatics methods were used to analyze the potential genes and their related miRs in OSCC. Tumor stem cells were separated from OSCCs through magnetic cell sorting. Regulatory pattern between miR-134 and LAMC2 in OSCC was evaluated by ectopic expression, knockdown and reporter assay experiments. The expression of miR-134, LAMC2, genes in PI3K-Akt signaling pathway, and apoptosis-related genes was detected. Cell proliferation was assessed by MTT assay, cell invasion by scratch test, cell migration by Transwell assay, cell cycle and apoptosis by flow cytometry, and cell growth and migration by xenograft tumor in nude mice. LAMC2 was predicted as the crucial factor related to OSCC using different chip data, and miR-134 was predicted to specifically bind LAMC2 in all five databases. RESULTS: Overexpressed miR-134 or silenced LAMC2 was observed to inhibit cell proliferation, migration, invasion of OSCC cells, growth of subcutaneous xenograft in nude mice, as well as promote OSCC cell apoptosis. LAMC2, a target gene of miR-134, decreased following miR-134 promotion, while the PI3K-Akt signaling pathway was inactivated following LAMC2 knockdown. Furthermore, we also observed that the effect of overexpressed miR-134 was enhanced when LAMC2 was knocked down. CONCLUSIONS: Taken together, these findings suggest that miR-134-mediated direct downregulation of LAMC2 inhibits migration and invasion of tumor stem cells in OSCC by suppressing the PI3K-Akt signaling pathway.

Human papillomavirus genotypes and p16 expression in oral leukoplakia and squamous cell carcinoma.

Several studies have shown a broad variation in the prevalence of human papillomavirus (HPV) in oral leukoplakia (OLK) and oral squamous cell carcinoma (OSCC), whereas the relationship is less well-defined and specific HPV genotypes lack examination in OLK. In the present study, the role of HPV and surrogate p16 expression was investigated to explore the correlation and pathogenesis in OLK and OSCC. Polymerase chain reaction (PCR) and flow-through hybridization technology were utilized to detect HPV genotypes in oral exfoliated cells from 30 healthy volunteers, 103 OLK and 30 OSCC patients. Expression of p16 was assessed by immunohistochemistry (IHC) in biopsies from these OLK and OSCC, in addition to 15 normal oral mucosal tissues as the control group. The healthy controls showed 3.3% (1/30) HPV presence; In OLK and OSCC, the detection rate was 4.9% (5/103), 3.3% (1/30), respectively. No significant relationship between HPV and OLK or OSCC was observed when compared with the control group (P>0.05). All 6 HPV-positive OLK and OSCC cases had p16 overexpression. But the sensitivity of p16 IHC was poor, because 88.4% (38/43) of p16 over-expressed OLK were HPV negative. There was no statistical significance between HPV and the sex, age, site, alcohol consumption, or smoking. These findings suggested HPV had a low prevalence in OLK and OSCC. This suggests the detection of HPV genotypes by PCR in exfoliated cells combined with p16 IHC may be more accurate to represent HPV infection.

[Research progress of quantitative DNA analysis in oral carcinoma and precancerous lesions].

Normal individual cells had 23 pairs of chromosome and stable DNA content. DNA content was varied during malignant transformation, which was specific feature of tumor. Quantitative DNA analysis can reflect cellular physiological or pathological condition by nuclear DNA content, which had significant role in early diagnosis, predication of prognosis and treatment selection. This article summarized the research progress of quantitative DNA analysis in oral carcinoma and precancerous lesions in recent years.

MicroRNA-155 in oral squamous cell carcinoma: Overexpression, localization, and prognostic potential.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is a common malignancy with poor prognosis. MicroRNAs (miRNAs) play an important role in cancer, but their role in OSCC is not clarified. METHODS: We performed miRNA microarray, quantitative reverse transcription polymerase chain reaction (qRT-PCR), and fluorescence in situ hybridization (FISH) to examine miRNA expression in OSCC and paired adjacent cancer-free mucosal (ACF) tissues. RESULTS: Thirteen miRNAs, including miRNA-155, were upregulated (>2-fold) in OSCC against ACF. MiRNA-155 was confirmed to have significantly higher expression in OSCC against ACF (paired-samples t test; p = .041) and it was localized in the cancer nest, inflammatory area, and vascular endothelium of OSCC. High expression of miRNA-155 in ACF tissue was an independent prognostic indicator for OSCC survival. CONCLUSION: MiRNA-155 was overexpressed in OSCC and it was located in the cancer nest, inflammatory area, and vascular endothelium of OSCC. High miRNA-155 expression level in ACF may predict poor prognosis in patients with OSCC.

[Locking plate percutaneous external fixation for the treatment of pediatric tibial fractures].

OBJECTIVE: To explore treatment effect of the locking plate percutaneous external fixation to tibial fractures. METHODS: From July 2010 to February 2013, 8 cases with pediatric tibial fractures were treated by using unilateral locking plate percutaneous external fixation,including 6 males and 2 females with an average age of 7 years old ranging from 4 to 10. Among them, 5 cases were open fractures involving 1 case of Gustilo-Anderson type II, 3 cases of type III A, 1 case of type III B; and the other 3 cases were closed fractures involving 2 cases of AO type A3, 1 cases of type B2. The postoperative bone healing and gait impact were observed and the function was evaluated by Johner-Wruhs scores. RESULTS: All fractures healed successfully without infection. The fracture healing time was from 3 to 6 months with an average of 3.9 months. The locking plate removal time was from 4 to 7 months with an average of 4.3 months. Among them, 7 cases were visually normal after walking with stand, 1 case of anterior tibial tendon defect affected gait. The results of Johner-Wruhs assessment were excellent in 7 cases, good in 1 case. No rub contralateral medial calf skin wounds occurenced. CONCLUSION: The method is simple, stable and reliable. The fixation strength is suitable for children using locking plate percutaneous external fixation. The postoperative functional recovery was excellent and the walking gait was less affected. But the point of LCP pedicle screw should be carefully selected before installation with good skin coverage.

Upregulation of miR-31* is negatively associated with recurrent/newly formed oral leukoplakia.

BACKGROUND: Oral leukoplakia (OLK) is a potentially malignant disorder of the oral cavity. However, the underlying mechanism of OLK is still unclear. In this study, we explore possible miRNAs involved in OLK. METHODOLOGY/PRINCIPAL FINDINGS: Using miRNA microarrays, we profiled miRNA expression in OLK and malignantly transformed OLK (mtOLK) tissue samples. The upregulation of miR-31*, miR-142-5p, miR-33a, miR-1259, miR-146b-5p, miR-886-3p, miR-886-5p, miR-519d, and miR-301a along with the downregulation of miR-572, miR-611, miR-602, miR-675, miR-585, miR-623, miR-637, and miR-1184 in mtOLK were new observations. Fluorescence in situ hybridization (FISH) analyses confirmed that miR-31* is highly expressed in mtOLK. There was a significant difference between the FISH score (p<0.05) in patients with or without recurrent/newly formed OLK. Functional analyses demonstrated that a miR-31* inhibitor decreased apoptosis in the Leuk-1, which is an immortalized oral epithelial cell line spontaneously derived from an oral leukoplakia lesion. miR-31* regulated apoptosis, cell proliferation, migration, and invasion in the HOIEC, which is a HPV E6/E7-immortalized oral epithelial cell line. Furthermore, miR-31* modulated the biological functions of apoptosis, cell proliferation, cell cycle, migration, and invasion in the oral squamous cell carcinoma cell line, Cal-27. Using bioinformatic analyses and dual luciferase reporter assays, we determined that the 3' untranslated region of fibroblast growth factor 3 (FGF3) is the target of miR-31*. Expression of FGF3 was downregulated or upregulated in the presence of a miR-31* mimic or inhibitor, respectively. CONCLUSIONS/SIGNIFICANCE: Upregulation of miR-31* is negatively associated with recurrent/newly formed OLK. MiR-31* may exert similar but distinguishable effects on biological function in oral cells with different malignant potential. FGF3 is the target of miR-31*. miR-31* may play an important role during OLK progression through regulating FGF3. MiRNA* strands may also have prominent roles in oral carcinogenesis.

Oral cancer development in patients with leukoplakia--clinicopathological factors affecting outcome.

BACKGROUND: Oral leukoplakia (OL) is the best-known potentially malignant disorder. The objective of the current study was to evaluate the clinicopathological factors predictive of outcome in a large cohort of patients with OL, and report our experience in the early detection of malignant events. METHODS: A total of 320 patients with biopsy-proven OL were retrospectively reviewed from the study institution who had a mean follow-up of 5.1 years. Data on patient and lesion at initial diagnosis and patient underwent sequential biopsies were reviewed. Multiple biopsies indicates > = 3 times sequential biopsies. Oral cancer-free survival rate (OCFS) was determined by the Kaplan-Meier method and significant factors were identified by Cox regression analysis. RESULTS: The 3-year and 5-year OCFS was 86.6% and 82.0%, respectively. A new binary system of grading oral dysplasia was performed and Kaplan-Meier analysis indicated that high-grade dysplasia had significantly higher malignant incidence than low-grade dysplasia (5-year OCFS, 90.5% vs 59.0%; P<0.001), especially during the first 2-3 years of follow-up. Multivariate analysis revealed that the 4 factors including patient aged >60 years, lesion located at lateral/ventral tongue, non-homogenous lesion, high-grade dysplasia were independent significant indicators for OL malignant transformation. In addition, significant positive correlation between the multiple biopsies and these 4 factors and malignant outcome was established. CONCLUSIONS: Elderly patients with OL located at lateral/ventral tongue and who had non-homogenous lesion with high-grade dysplasia correlated much higher risk of transformation. This high-risk subpopulation was suggested to undergo sequential biopsies and histologic examination contributing to early detection of malignant event.

Expression of podoplanin and ABCG2 in oral erythroplakia correlate with oral cancer development.

Oral erythroplakia (OE) is a notoriously aggressive oral premalignant lesion with a high tendency to oral cancer development, but it's biological behavior is largely unknown. The objective of the current study was to determine podoplanin and ABCG2 immunoexpression in OE and both correlation to malignant transformation of OE. In a retrospective follow-up study, the expression patterns of podoplanin and ABCG2 were determined using immunohistochemistry in samples from 34 patients with OE, including patients with untransformed lesions (n=17) and patients with malignant transformed lesions (n=17). Podoplanin and ABCG2 expression was observed in 15 (44.1%) and 21 (61.8%) of 34 patients, respectively. Multivariate analysis revealed that podoplanin and ABCG2 expression was associated with 6.31-fold (95% confidence interval [CI], 1.02-38.92; P=0.047) and 14.39-fold (95% CI, 2.02-102.29; P=0.008) increased the risk of transformation, respectively. Point prevalence analysis revealed that 90.9% (95% CI, 70.7-100) of the patient with both podoplanin and ABCG2 positivity developed oral cancer. Collectively, our data indicated that the expression patterns of podoplanin and ABCG2 in OE were associated with oral cancer development, suggesting that podoplanin and ABCG2 may be valuable predictors for evaluating oral cancer risk.

[In vitro activity of matrine against Candida albicans biofilms].

PURPOSE: To establish a model of Candida albicans biofilms and to examine the effect of matrine on C.albicans biofilms and ultrastructure. METHODS: C. albicans collection strain ATCC76615 was obtained and propagated. Biofilms were formed in 96-well microtiter plates. Antifungal susceptibility testing of C. albicans biofilms were assessed with the tetrazolium salt (XTT) reduction assay. Confocal laser scanning microscopy (CLSM) and dead/live fluorescent staining technique were combined to detect the effects of Matrine on preformed C. albican biofilms' composition and ultrastructure. RESULTS: Matrine was active against different growth stages (early,middle,mature) of biofilms; The bioactivity and drug-resistance of C. albican biofilm increased with culturing time. CLSM showed that C. albicans biofilms were inhibited and growth were predominantly composed of yeast cells and pseudohyphae. CONCLUSIONS: This study demonstrates that Matrine has potent activity against C.albicans biofilms in vitro and potential therapeutic implication for biofilm-associated candidal infections.