[Malfunction of autophagy in tibial growth plate chondrocytes causes increased apoptosis rate in chronic renal insufficiency rats].

Objective: To observe the effect of autophagy of tibial growth plate chondrocyte on apoptosis in chronic renal insufficiency (CRI) rats. Methods: Male 4-week-old SD rats were randomly divided into two groups: (1) Sham group: only the left ureter was exposed (n=10); (2) CRI group: the left ureter was ligated to cause CRI (n=10). The urine from all the rats was collected 6 weeks after the operation and the total protein content was measured. Then all the rats were sacrificed and the concentrations of creatinine and urea nitrogen in intracardiac blood were detected. The proximal tibia were fixed and decalcified to prepare histological sections, and the number of chondrocytes of column cells in the proliferative area of tibia growth plate was observed by saffron O staining. The expression rate of protein Light Chain-3, an autophagy marker of chondrocytes, was detected by immunofluorescence. The apoptosis rate of chondrocytes was detected by the method of TUNEL assay. The level of glycogenin-1, a glycogen formation marker of chondrocyte was detected by immunohistochemistry in chondrocytes. Results: The 24 h urine total protein was higher in CRI group [(163.5±11.3) mg vs (38.6±9.8) mg, t=25.620, P<0.001]. The levels of blood creatinine [(67.3±16.2) µmol/L vs (28.4±11.5) µmol/L, t=5.974, P<0.001] and urea nitrogen [(16.4±6.4) mmol/L vs (4.8±2.0) mmol/L, t=5.198, P<0.001] were higher in CRI group. The number of chondrocytes of column cells in the proliferating area of tibia growth plate was lower in CRI group (4.2±2.1 vs 9.1±3.8, t=3.109, P=0.006). The expression rate of LC-3 protein in chondrocytes of CRI group was lower [(27.2±12.6)% vs (51.4±18.2)%, t=3.457, P=0.003]. The level of glycogenin-1 of chondrocytes in CRI group increased significantly (6.1±2.5 vs 3.5±1.8, t=2.669, P=0.016). The apoptosis rate of chondrocytes in CRI group also increased [(17.2±4.8)% vs (5.1±3.4)%, t=6.505, P<0.001]. Conclusion: Malfunction of autophagy in tibial growth plate chondrocytes causes increased apoptosis rate in CRI rats, which might be caused by the failure of glycogen degradation in chondrocytes.

[Effect of primary cilia expression rate on Wnt/ß signaling pathway in tibial growth plate chondrocytes from chronic renal insufficiency young rats].

Objective: To observe the effect of primary cilia expression rate on Wnt/ß signaling pathway in tibial growth plate chondrocytes from chronic renal insufficiency (CRI) young rats. Methods: Male 2-week-old SD rats were randomly divided into two groups: (1) Sham group: only the left ureter was exposed (n=6); (2) CRI group: the left ureter was ligated (n=6). Rats were sacrificed 2 weeks after the operation and the primary cilia expression rate of growth plate chondrocytes and key protein ß-catenin in Wnt/ß signaling pathway were observed in histological section of tibia specimen. Chondrocytes isolated from growth plate in two groups were cultured in vitro to P3 generation. The primary cilia expression rate and the level of ß-catenin were measured. The primary cilia expression rate was detected by agonists and antagonists Wnt/ß signaling pathway in chondrocytes of CRI group. The level of ß-catenin was detected by using serum-free culture and chloral hydrate to intervene chondrocytes in CRI group. Results: The primary cilia expression rate of growth plate chondrocytes in histological section of tibia specimen in CRI group was higher than that in Sham group [(17.5±7.7)% vs (8.7±3.6)%, t=3.237, P=0.005], and the level of ß-catenin was higher in CRI group (5.1±0.7 vs 1.9±0.8, t=6.731, P<0.001). The primary cilia expression rate of growth plate chondrocytes cultured in vitro in CRI group was higher than that in Sham group [(20.9±8.1)% vs (11.8±4.7)%, t=3.073, P=0.007], and the level of ß-catenin was higher in CRI group (0.49±0.12 vs 0.25±0.11, t=3.297, P=0.011). There was no significant change in primary cilia expression rate after intervention by using Wnt/ß signaling agonists and antagonists to change the level of ß-catenin [agonists group: (21.3±7.6)%, control: (20.6±6.8)%, antagonists group: (22.4±6.2)%, F=0.173, P=0.842]. The level of ß-catenin was significantly changed after intervention by using serum-free culture, chloral hydrate to alter the primary cilia expression rate (serum-free culture group: 0.61±0.23, control: 0.39±0.24, chloral hydrate group: 0.15±0.11, F=6.476, P=0.012). There was a positive correlation between the level of ß-catenin and primary cilia expression rate. Conclusion: The primary cilia expression rate and the level of Wnt/ß signaling pathway were higher in tibial growth plate chondrocytes in CRI rats model, and primary cilia might have positive regulatory effects on the Wnt/ß signaling pathway.

[Effect of primary cilia on growth plate chondrocyte proliferation of young rats with chronic renal insufficiency].

Objective: To observe the effect of primary cilia on growth plate chondrocyte proliferation of young rats with chronic renal insufficiency (CRI). Methods: Male 2-week-old Sprague-Dawley rats were randomly divided into two groups (with 10 in each group): Sham group (only left ureter was exposed) and CRI group (left ureter was ligated). Rats were sacrificed 2 weeks after operation and the total length of tibia was measured. Histological sections of tibia were taken to observe the chondrocytes of growth plate proliferative region and the expression rate of primary cilia. Chondrocytes from growth plate in two groups were isolated and cultured in vitro to P3 generation and the chondrocyte proliferation rate at 24 h were detected. The primary cilia expression rate and cilia length of chondrocytes were measured. Western blot was used to detect the expression of intraflagellar transport 88 (IFT88) protein and the gray scale was analyzed. Results: The total length of tibia was shorter in CRI group [(35.84±4.56) mm vs (42.33±3.44) mm, P=0.002]. The results of tibial histological section showed that chondrocytes of growth plate proliferative region were unorganized and the number of chondrocyte with columnar structure was less in CRI group (2.71±1.10 vs 7.68±1.32, P<0.001). The primary cilia expression rate of chondrocytes was higher in CRI group [(35.53±7.41)% vs (18.31±5.12)%, P<0.001]. The chondrocyte proliferation rate at 24 h was lower in CRI group [(11.38±6.10)% vs (24.35±8.46)%, P=0.001]. The primary cilia expression rate of chondrocytes was higher in CRI group [(60.12±7.86)% vs (32.17±8.97)%, P<0.001], and the primary cilia length of chondrocytes was longer in CRI group [(3.54±1.61) µm vs (1.96±0.82) µm, P=0.012]. The protein IFT88 was more highly expressed in CRI group (0.47±0.23 vs 0.17±0.10, P=0.001). Conclusion: The primary cilia expression rate of growth plate chondrocytes was higher in the rats with CRI, resulting in decreased chondrocyte proliferation rate and growth retardation of tibial growth plate.

[Effect of low expression of PTHrp receptor on proliferation of tibial growth plate chondrocytes in chronic renal insufficiency young rats].

Objective: To observe the effect of parathyroid hormone-related protein (PTHrp) receptor on the proliferation of tibial growth plate chondrocytes in chronic renal insufficiency (CRI) young rats. Methods: Two-week-old male SD rats were randomly divided into two groups: (1) Sham group (n=6), only left ureter was exposed; (2) CRI group(n=6), left ureter was ligated to induce chronic renal insufficiency. Rats were sacrificed 2 weeks after operation and the blood concentration of PTHrp was detected by intracardiac blood sampling. Chondrocytes isolated from growth plate in two groups were cultured in vitro to P3 generation. The level of PTHrp receptor in chondrocytes was observed by immunohistochemistry and quantitative analysis was completed by Western blot. The proliferation rate of chondrocytes from two groups at 24 h was detected by using 5-ethynyl-2'-deoxyuridine (EDU) technique. Three types of PTHrp receptor mRNA plasmids (overexpressed, empty vector and knockdown) were used to treat the chondrocytes from CRI group. The mRNA and protein levels of PTHrp receptor were detected after 24 h and 48 h intervention, respectively. The chondrocyte proliferation rate at 24 h was detected by EDU. Results: Blood concentration of PTHrp in CRI group was higher than that in Sham group [(1.36±0.42) ng/L vs (0.77±0.21) ng/L, t=3.913, P=0.001]. The results of Western blot showed that the level of PTHrp receptor in growth plate chondrocytes from CRI group decreased (0.15±0.07 vs 0.41±0.13, t=5.569, P<0.001). Chondrocyte proliferation rate of CRI group was lower than that in Sham group at 24 h [(11.3±3.1)% vs (24.6±5.7)%, t=6.482, P<0.001]. The mRNA and protein levels of PTHrp receptor increased in chondrocytes of CRI group after intervention with overexpressed plasmid. The chondrocyte proliferation rate increased at 24 h. On the contrary, the mRNA and protein levels of PTHrp receptor decreased afer intervention with knockdown plasmid, and the chondrocyte proliferation rate also decreased [overexpression: (22.8±6.5)%, empty carrier: (10.2±4.3)%, knockdown: (5.6±2.1)%, F=29.840, P<0.001]. Conclusion: Increased PTHrp concentration in the blood of CRI young rats leads to decreased PTHrp receptors in growth plate chondrocytes, which results in decreasing PTHrp activity and proliferation rate of chondrocyte.

Greater serum carotenoid concentration associated with higher bone mineral density in Chinese adults.

UNLABELLED: This cross-sectional study has been performed to investigate the relationship between serum carotenoids and bone mineral density (BMD) in Chinese population. We found that women with higher serum ß-cryptoxanthin, lycopene, or α-carotene exhibited higher BMD at various bone sites. Similar association was observed between α-carotene and BMD in men. INTRODUCTION: Carotenoids may positively regulate bone metabolism through their antioxidant properties; however, few studies have examined the relation between serum carotenoids and bone health. We aimed to determine the associations between the serum concentration of several carotenoid subclasses and BMD in a Chinese population. METHODS: This study was a community-based cross-sectional study. We measured 1898 women and 933 men aged 59.6 years who completed serum ß-cryptoxanthin, zeaxanthin + lutein, lycopene, and α-carotene concentration analyses and BMD assessments. Serum individual carotenoids were assessed by the methods of reverse-phase high-performance liquid chromatography. Dual-energy X-ray absorptiometry was applied to determine BMD at whole body, lumbar spine, total hip, femur neck, and trochanter. ANCOVA was used to examine the correlations between categorized individual carotenoids and BMD at measured sites. RESULTS: After adjusting for potential covariates, a monotonic dose-response positive correlation between circulating levels of ß-cryptoxanthin, lycopene, and α-carotene and BMD at various skeletal sites was observed in women. Women in the top (vs. bottom) quartiles of serum ß-cryptoxanthin, lycopene, or α-carotene exhibited 1.8-2.3, 1.5-2.0, or 1.3-2.7 % higher BMD at the bone sites with significant results (P-trend <0.05), respectively. For men, the corresponding values were 2.6-4.0 % for α-carotene at the whole body and hip regions (P-trend <0.001-0.023). CONCLUSION: These results suggest that serum carotenoids have a favorable association with bone health in the study population, especially in women.

Obatoclax induces Beclin 1- and ATG5-dependent apoptosis and autophagy in adenoid cystic carcinoma cells.

OBJECTIVES: Adenoid cystic carcinoma (ACC) is one of the most common salivary gland cancers. The prognosis of adenoid cystic carcinoma is poor for its high frequency of distant metastases and insensitivity to chemotherapy or molecular therapies. This study investigated the effect of Obatoclax on adenoid cystic carcinoma cells and its cytotoxic mechanism. METHODS: Western blot, transmission electron microscopy, and pEGFP-LC3 plasmids transfection were carried out to detect autophagy in ACC cells treated with Obatoclax. 3-MA and RNA interference against Beclin 1 and ATG5 were used to inhibit autophagy. Then we used Western blot and Hochest 33342 staining for apoptosis assessment. Finally, cell viability was assessed by MTT assay. RESULTS: We found that Obatoclax induced cytoprotective autophagy which depended on ATG5 and partly on Beclin 1 in adenoid cystic carcinoma cells. Furthermore, pharmacologically inhibiting Obatoclax-induced autophagy promoted apoptosis. Downregulation of Beclin 1 or ATG5 attenuated the cytotoxicity of Obatoclax by suppressing both autophagy and apoptosis. Finally, when apoptosis was pharmacologically inhibited, autophagic cell death was initiated in adenoid cystic carcinoma cells treated with Obatoclax. CONCLUSION: In summary, Beclin 1 and ATG5 play important roles in regulating both Obatoclax-induced autophagy and apoptosis in adenoid cystic carcinoma.

Association between dietary intake of flavonoid and bone mineral density in middle aged and elderly Chinese women and men.

SUMMARY: This large cross-sectional study examined the associations of dietary intakes of total flavonoids and their subtypes with bone density in women and men. We found that greater flavonoid intake was associated with higher bone density in women but not in men. INTRODUCTION: Studies in vitro and in animal models suggest a potential effect of flavonoids on bone health. Few studies have examined the association between the habitual intake of flavonoids and bone mineral density (BMD) in humans. METHODS: The cross-sectional study recruited 2,239 women and 1,078 men. A semiquantitative food frequency questionnaire was administered in face-to-face interviews to assess habitual dietary flavonoid intake using food composition databases. BMD was measured over the whole body (WB) and in the femoral neck (FN) and lumbar spine (LS) by dual-energy X-ray absorptiometry (DXA). RESULTS: After adjusting for covariates, women who consumed higher total flavonoids, and the subtypes of flavonols, flavan-3-ols, flavones, and proanthocyanidins tended to have greater BMD at the WB, LS, and FN (all P-trend < 0.05). Women in the highest (vs. the lowest) quartile of total flavonoids intake had 0.020 (1.91 %), 0.021 (2.51 %), and 0.013 (1.99 %) g/cm(2) greater BMD at the whole body, LS, and FN, respectively. For the subtypes of flavonoids, the corresponding differences in BMD (in g/cm(2)) were 0.012-0.021 (flavan-3-ols), 0.013-0.020 (flavonols), 0.016-0.019 (flavones), and 0.014-0.016 (proanthocyanidins), respectively. A higher intake of flavonones was associated with a greater BMD at the whole body (P-trend 0.041) and the FN (P-trend 0.022). In men, there were no significant positive associations between the consumption of total flavonoids and the subclasses and BMD at any sites. CONCLUSION: Dietary flavonoids intake was positively associated with BMD in women. Further large studies are needed to clarify this issue in men.

Lipoapoptosis pathways in pancreatic ß-cells and the anti-apoptosis mechanisms of adiponectin.

Lipoapoptosis is the main form of pancreatic ß-cell death in diabetes. Adiponectin is an adipocyte-derived hormone, which has anti-apoptosis effect in numerous cells. The present study was designed to explore the role of the 'extrinsic' (death receptor-induced) and the 'intrinsic' (mitochondrial) pathways in pancreatic ß-cell lipoapoptosis and the anti-apoptosis effect of adiponectin on pancreatic ß-cells. Palmitate (0.4 mmol/l) or oleate (0.4 mmol/l) was used to induce the apoptosis of Min 6 cells for 24 h. Z-LETD-FMK or Z-IEHD-FMK (40 µM) was used to inhibit the activity of caspase-8 or -9. When adiponectin was used, Min 6 cultures were pretreated in the absence or presence of fAd (5 µg/ml) for 2 h and then subjected to palmitate for 24 h. Apoptosis was evaluated using Annexin V-Cy3 kit. The expression levels of cleaved caspase-3, -8, -9, B-cell lymphoma 2 (BCL-2), and Bax were examined by Western blotting. Palmitate-induced pancreatic ß-cell apoptosis was accompanied by the activation of caspase-8, -9, and -3. Blockade of caspase-9 rather than caspase-8, showed an inhibitory effect on caspase-3 activation. Moreover, adiponectin treatment prevented palmitate-induced apoptosis by inhibition of caspase-9 activation, but not of caspase-8, and induced an upregulation of BCL-2 and a downregulation of Bax in protein level. Both extrinsic and intrinsic pathways are activated in pancreatic ß-cell lipoapoptosis, and the intrinsic apoptosis pathway is the major one. Adiponectin prevents pancreatic ß-cells from apoptosis by inhibition of intrinsic apoptosis pathway via regulation of the BCL2 family. Therefore, protection of intrinsic apoptosis pathway is a potential therapeutic strategy for diabetes.

Suppression of mesangial cell proliferation and extracellular matrix production in streptozotocin-induced diabetic mice by adiponectin in vitro and in vivo.

Renal growth, particularly hypertrophy, is a feature of diabetic nephropathy (DN). Adiponectin, an adipocyte-derived hormone, is an important regulator of cell proliferation. Recent studies have suggested that adiponectin has a protective effect in the kidney. The purpose of this study was to investigate the therapeutic effects and the underlying mechanisms of adiponectin in early DN. Mouse mesangial cells (MMCs) were cultured in media containing different concentrations of platelet-derived growth factor-BB (PDGF-BB) with or without adiponectin. MMC proliferation and expression of type IV collagen, laminin, and fibronectin were investigated. Streptozotocin-induced diabetic mice were injected intravenously with recombinant lentivirus encoding the mouse adiponectin gene (Lenti-Acdc-IRES-EGFP). Urinary microalbumin, serum adiponectin level, and expression of proliferating cell nuclear antigen, type IV collagen, laminin, and fibronectin were determined. Adiponectin inhibited the increases in MMC proliferation and expression of type IV collagen, laminin, and fibronectin induced by PDGF-BB. Adiponectin also effectively reduced renal cell proliferation and expression of type IV collagen, laminin, and fibronectin when it was introduced in vivo by lentivirus-mediated gene transfer. These findings suggest that adiponectin exerts renoprotective effects by inhibiting renal cell proliferation and reducing synthesis of extracellular matrix proteins, thus suppressing the development and progression of DN.

Adiponectin-induced inhibition of intrinsic and extrinsic apoptotic pathways protects pancreatic ß-cells against apoptosis.

Apoptosis is the main form of ß-cell death in type 2 diabetes mellitus. There are 2 major pathways leading to apoptosis, 'intrinsic and extrinsic pathways'. Adiponectin is an adipocyte-derived hormone, which plays an important role in glucose and lipid metabolism. The main aims of this study were to investigate related apoptotic pathways in diabetes and the anti-apoptosis effects of adiponectin on pancreatic ß-cell and the underlying mechanisms. Diabetic mice were generated by intraperitoneal injection of streptozotocin (STZ, 50 mg/kg/d for 5 days) and high-fat diet. Adiponectin overexpressing mice were developed by injecting lentivirus expressing mouse full length adiponectin (plenti-acdc-EGFP) through tail vein. Fourteen days after plenti-acdc-EGFP lentivirus injection, plasma adiponectin protein levels were increased 2-fold. Plasma triglyceride and glucose levels, but not total cholesterol, were significantly reduced in plenti-acdc-EGFP-treated mice. Pancreatic ß-cell apoptotic numbers and the expression of caspase-8, -9, -3 in islet increased in diabetic mice, which was reversed by elevated adiponectin in plenti-acdc-EGFP-treated mice. These results suggest that both intrinsic and extrinsic apoptotic pathways have an important role in diabetic ß-cell apoptosis. Adiponectin has antidiabetic and anti-apoptotic effects by regulating glucose and lipid metabolisms and inhibiting intrinsic and extrinsic apoptotic pathway in pancreatic ß-cells.

Association between single nucleotide polymorphisms of the osteoprotegerin gene and postmenopausal osteoporosis in Chinese women.

Osteoporosis is an important and common complex health problem, particularly in postmenopausal women. It is characterized by a reduction in bone mineral density (BMD) and a deterioration of bone microarchitecture with a consequent increase of fracture risk. The osteoprotegerin (OPG) gene is considered to play an important role in the pathogenesis of osteoporosis. We analyzed SNPs of the OPG gene and associations between these polymorphisms and BMD in 399 Chinese postmenopausal women. BMD was quantified at the lumbar spine (L2-4), femoral neck, and total hip. The g.2264T>C and g.27676A>C SNPs were detected by PCR-RFLP and DNA sequencing methods. A significant association with spine BMD was found for g.27676A>C. The spine BMD value for subjects with genotype AA was significantly higher than those with genotypes GA and AA. No significant association was detected between any of the SNP marker genotypes and the other traits. We conclude that g.27676A>C in the OPG gene affects spine BMD and that the C allele is associated with increased risk for osteoporosis in Chinese postmenopausal women.

Sialendoscopic removal of metastatic adenocarcinoma in a mucous plug from Stensen's duct.

Sialendoscopy is a minimally invasive technique used mainly in the diagnosis and treatment of obstructive salivary gland disorders. There has yet to be a report on its use in the diagnosis of metastatic disease. While metastatic cancer has been described in numerous head and neck anatomic subsites, it has not been reported to be found in a mucous plug in Stensen's duct. Sialendoscopy was performed in a 68-year-old female patient who presented with symptoms of ductal obstruction. Basket removal of a mucous plug was done and histopathological analysis of this specimen found adenocarcinoma. The overall clinical picture, imaging, and final histopathological results suggested that this patient had metastatic breast carcinoma to a mucous plug in Stensen's duct, the diagnosis of which was made with the aid of interventional sialendoscopy. This is the first report in which metastatic cancer was identified in a mucous plug in Stensen's duct. Sialendoscopy can be a useful tool to aid in the diagnosis of metastatic diseases in rare and unusual clinical situations.

Optimization of total tongue functional reconstruction with the sushi roll technique and its application in pectoralis major myocutaneous flaps.

A protuberant shape and sufficient volume are the most important parameters for total tongue reconstruction. The conventional pectoralis major myocutaneous (PMMC) flap undergoes collapse due to atrophy of the denervated muscle. In a new technique, this flap was rolled up like sushi to reshape the neotongue. This study explored the feasibility and effect of the 'sushi roll' technique for precise total functional reconstruction of the tongue using a PMMC flap. Thirty patients scheduled for total glossectomy and PMMC flap reconstruction were recruited. The sushi roll technique was performed in 15 patients and the conventional repair in 15 patients. Outcomes were compared between the two groups. The flap survived in all 30 patients. The sushi roll group showed superior results to the conventional group in terms of time to oral alimentation (P = 0.012) and decannulation (P = 0.041), as well as swallowing function (P = 0.032), speech intelligibility (P < 0.001), shape (P < 0.001), and quality of life score (P < 0.001) at 12 months. The innovative sushi roll technique uses a folding method that utilizes the length rather than the thickness and width of the flap to maintain the volume and protuberance of the neotongue, which results in acceptable function and improved quality of life.

Adiponectin inhibits PDGF-induced mesangial cell proliferation: regulation of mammalian target of rapamycin-mediated survival pathway by adenosine 5-monophosphate-activated protein kinase.

An aberrant proliferation of mesangial cells (MCs) is one of the more important features of diabetic nephropathy (DN). Adiponectin, an adipocyte-derived hormone, has been associated with type 2 diabetes, a known cause of DN. Recent studies have suggested that adiponectin has a protective effect on the kidney. To elucidate the potential protective mechanism of adiponectin on kidney, we investigated the effects of adiponectin on platelet-derived growth factor (PDGF)-induced cell proliferation and intracellular signaling pathways in cultured Human MCs (HMCs). PDGF-induced HMC proliferation was significantly inhibited by the co-treatment of adiponectin. Adiponectin alone had no effect on HMC proliferation. The mammalian target of rapamycin (mTOR) and 40 S ribosomal S6 kinase 1 (S6K1) were activated by PDGF stimulation in HMCs. PDGF-induced mTOR and S6K1 phosphorylations were significantly attenuated by the co-treatment of adiponectin in HMC. Adiponectin alone had no effects on PDGF-receptor autophosphorylation by PDGF. We also confirmed that the inhibitory effect of adiponectin on PDGF-induced HMC proliferation was significantly suppressed by compound C, an adenosine 5'-monophosphate-activated protein kinase (AMPK) inhibitor. From these findings, it is implied that adiponectin could attenuate renal dysfunction associated with MC disorders through AMPK-mTOR signal pathway.

Myeloid-derived suppressor cells contribute to oral cancer progression in 4NQO-treated mice.

OBJECTIVE: Abnormal myelopoiesis especially the expansion of myeloid-derived suppressor cells (MDSCs) is increasingly recognized as an important reason for the escape of tumor from immune surveillance. This study aims to investigate the role of this specific population of cells in oral cancer progression. MATERIALS AND METHODS: 4-Nitroquinoline 1-oxide (4NQO) was used to induce oral cancer in C57BL/6 mice. The tongue mucosa was examined by hematoxylin and eosin staining. The distribution of MDSCs in the spleen and peripheral blood and T cell subsets in the spleen was analyzed by flow cytometry. The expression of MDSCs in the tongue tissues was investigated by immunohistochemical staining, and the expression of arginase-1 (ARG-1) and NOS-2 in the tongue tissues was detected by real-time PCR. RESULTS: We found that during tumor progression, significantly increased frequency of MDSCs was observed in the spleens and peripheral blood of 4NQO-treated mice, and the frequency of MDSCs in the spleens was positively correlated with systemic CD3(+) CD8(+) T cells. Moreover, 4NQO-treated mice showed significantly higher MDSCs infiltration and ARG-1 mRNA level in the tumor site. CONCLUSIONS: Myeloid-derived suppressor cells contribute to oral tumor progression and represent a potential target for immunotherapy of oral cancer.

Prediction of ameloblastoma recurrence using random forest-a machine learning algorithm.

The purpose of this study was to investigate whether ameloblastoma with a high likelihood of recurrence can be predicted using random forest model, a machine learning algorithm. Data were collected from patients treated for ameloblastoma between 1999 and 2019 at the University of Hong Kong. Fourteen clinical parameters were used to grow the decision trees to classify patients with or without ameloblastoma recurrence in the follow-up period. The random forest algorithm was computed 100 times in the training cohort (n = 100) and verified in the testing cohort (n = 50). The receiver operating characteristic curve (ROC) and area under the curve (AUC) were used as the performance measurement of separability. One hundred and fifty patients (76 female, 74 male) were recruited, with a mean follow-up time of 103 months. Recurrence occurred in a total of 25 cases (16.7%) over the 20-year period. The AUC were calculated for the median and mean ROC curves; these were 0.777 and 0.825, respectively. The results showed that random forest model was able to predict recurrence of ameloblastoma with reliable accuracy. The four most important variables influencing ameloblastoma recurrence were the time elapsed from treatment, initial surgical treatment, tumour size, and radiographic presentation. This study provides insights into the detection of high-risk patient groups to monitor recurrence. Further application of random forest to other diseases could greatly benefit clinical decisions.

Spatial deviations of the temporomandibular joint after oncological mandibular reconstruction.

Spatial deviations of the temporomandibular joint (TMJ) after oncological mandibular reconstruction are important to the aesthetic and functional rehabilitation. The aim of this study was to clarify whether and how three dimensionally (3D) printed patient-specific surgical plates, and the preservation of the condyle or ramus, affect spatial deviations of the TMJ. A total of 33 patients who underwent mandibular reconstruction via computer-assisted surgery were included. Regarding absolute deviations, patients in the 3D-printed plate group showed smaller TMJ deviations compared to those in the conventional plate group. There was no difference in absolute deviations of the TMJ regardless of whether the condyle or ramus was preserved. Regarding physiological deviations, the impact on the contralateral TMJ was smaller in the 3D-printed plate group. Patients with both the condyle and ramus removed had significantly higher deviations of the condyle and joint space. In summary, 3D-printed patient-specific surgical plates improved the spatial accuracy of the TMJ. Under physiological conditions, TMJ deviations on the operated side were mainly affected by the preservation of the condyle. Removal of both the condyle and ramus caused more severe spatial interference to the TMJ; this should be further confirmed.

Surgical morbidities of sagittal split ramus osteotomy versus intraoral vertical ramus osteotomy for the correction of mandibular prognathism: a randomized clinical trial.

The sagittal split ramus osteotomy (SSRO) and intraoral vertical ramus osteotomy (IVRO) are two common orthognathic procedures for the treatment of mandibular prognathism. This randomized clinical trial compared the surgical morbidities between SSRO and IVRO for patients with mandibular prognathism over the first 2 years postoperative. Ninety-eight patients (40 male, 58 female) with a mean age of 24.4±3.5 years underwent bilateral SSRO (98 sides) or IVRO (98 sides) as part or all of their orthognathic surgery. IVRO presented less short-term and long-term surgical morbidity in general. The SSRO group had a greater incidence of inferior alveolar nerve deficit at all follow-up time points (P< 0.01). There was more TMJ pain at 6 weeks (P= 0.047) and 3 months (P= 0.001) postoperative in the SSRO group. The SSRO group also presented more minor complications, which were related to titanium plate exposure and infection. There were no major complications for either technique in this study. Despite the need for intermaxillary fixation, IVRO appears to be associated with less surgical morbidity than SSRO when performed as a mandibular setback procedure to treat mandibular prognathism.

Reliable ductal access in sialoendoscopy.

During sialoendoscopy, insertion of the endoscope through the ductal orifice is usually a challenging procedure, especially for beginners. Based on our experience, we have found that using the Kolenda Salivary Access Introducer Set is the most reliable and easily learned ductal access approach. We report details of the technique and discuss the advantages of this approach.

Oral Epstein-Barr virus-positive mucocutaneous ulcer: gingival presentation of a benign lymphoproliferative lesion.

Epstein-Barr virus-positive mucocutaneous ulcer (EBVMCU) is a benign lymphoproliferative lesion related to iatrogenic or age-related immunosuppression in patients with prior Epstein-Barr virus (EBV) infection. Although the clinical presentation may resemble malignant disease, the course of EBVMCU is indolent, and regression is expected when immunosuppression is reduced. We present a case of EBVMCU in the gingiva of a 59-year-old male patient with long-standing pemphigus vulgaris. The initial presentation was suspicious for oral cavity cancer, which was ruled out by biopsy. After reduction of immunosuppression, the ulceration regressed and an area of exposed necrotic bone remained. Complete healing was achieved after sequestrectomy and primary closure with a local gingival flap.