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1.  Accuracy evaluation of a 3D-printed individual template for needle guidance in head and neck brachytherapy 
Journal of Radiation Research  2016;57(6):662-667.
To transfer the preplan for the head and neck brachytherapy to the clinical implantation procedure, a preplan-based 3D-printed individual template for needle insertion guidance had previously been designed and used. The accuracy of needle insertion using this kind template was assessed in vivo. In the study, 25 patients with head and neck tumors were implanted with 125I radioactive seeds under the guidance of the 3D-printed individual template. Patients were divided into four groups based on the site of needle insertion: the parotid and masseter region group (nine patients); the maxillary and paranasal region group (eight patients); the submandibular and upper neck area group (five patients); and the retromandibular region group (six patients). The distance and angular deviations between the preplanned and placed needles were compared, and the complications and time required for needle insertion were assessed. The mean entrance point distance deviation for all 619 needles was 1.18 ± 0.81 mm, varying from 0.857 ± 0.545 to 1.930 ± 0.843 mm at different sites. The mean angular deviation was 2.08 ± 1.07 degrees, varying from 1.85 ± 0.93 to 2.73 ± 1.18 degrees at different sites. All needles were manually inserted to their preplanned positions in a single attempt, and the mean time to insert one needle was 7.5 s. No anatomical complications related to inaccurately placed implants were observed. Using the 3D-printed individual template for the implantation of 125I radioactive seeds in the head and neck region can accurately transfer a CT-based preplan to the brachytherapy needle insertion procedure. Moreover, the addition of individual template guidance can reduce the time required for implantation and minimize the damage to normal tissues.
doi:10.1093/jrr/rrw033
PMCID: PMC5137284  PMID: 27422928
3D-printed; individual template; brachytherapy; head and neck
2.  High-Dose Fluoride Impairs the Properties of Human Embryonic Stem Cells via JNK Signaling 
PLoS ONE  2016;11(2):e0148819.
Fluoride is a ubiquitous natural substance that is often used in dental products to prevent dental caries. The biphasic actions of fluoride imply that excessive systemic exposure to fluoride can cause harmful effects on embryonic development in both animal models and humans. However, insufficient information is available on the effects of fluoride on human embryonic stem cells (hESCs), which is a novel in vitro humanized model for analyzing the embryotoxicities of chemical compounds. Therefore, we investigated the effects of sodium fluoride (NaF) on the proliferation, differentiation and viability of H9 hESCs. For the first time, we showed that 1 mM NaF did not significantly affect the proliferation of hESCs but did disturb the gene expression patterns of hESCs during embryoid body (EB) differentiation. Higher doses of NaF (2 mM and above) markedly decreased the viability and proliferation of hESCs. The mode and underlying mechanism of high-dose NaF-induced cell death were further investigated by assessing the sub-cellular morphology, mitochondrial membrane potential (MMP), caspase activities, cellular reactive oxygen species (ROS) levels and activation of mitogen-activated protein kinases (MAPKs). High-dose NaF caused the death of hESCs via apoptosis in a caspase-mediated but ROS-independent pathway, coupled with an increase in the phospho-c-Jun N-terminal kinase (p-JNK) levels. Pretreatment with a p-JNK-specific inhibitor (SP600125) could effectively protect hESCs from NaF-induced cell death in a concentration- and time-dependent manner. These findings suggest that NaF might interfere with early human embryogenesis by disturbing the specification of the three germ layers as well as osteogenic lineage commitment and that high-dose NaF could cause apoptosis through a JNK-dependent pathway in hESCs.
doi:10.1371/journal.pone.0148819
PMCID: PMC4747557  PMID: 26859149
3.  The Value of SPECT/CT in Monitoring Prefabricated Tissue-Engineered Bone and Orthotopic rhBMP-2 Implants for Mandibular Reconstruction 
PLoS ONE  2015;10(9):e0137167.
Bone tissue engineering shows good prospects for mandibular reconstruction. In recent studies, prefabricated tissue-engineered bone (PTEB) by recombinant human bone morphogenetic proteins (rhBMPs) applied in vivo has found to be an effective alternative for autologous bone grafts. However, the optimal time to transfer PTEB for mandibular reconstruction is still not elucidated. Thus, here in an animal experiment of rhesus monkey, the suitable transferring time for PTEB to reconstruct mandibular defects was evaluated by 99mTc-MDP SPECT/CT, and its value in monitoring orthotopic rhBMP-2 implants for mandibular reconstruction was also evaluated. The result of SPECT/CT showed higher 99mTc-MDP uptake, indicating osteoinductivity, in rhBMP-2 incorporated demineralized freeze-dried bone allograft (DFDBA) and coralline hydroxyapatite (CHA) implants than those without BMP stimulation. 99mTc-MDP uptake of rhBMP-2 implant peaked at 8 weeks following implantation while CT showed the density of these implants increased after 13 weeks’ prefabrication. Histology confirmed that mandibular defects were repaired successfully with PTEB or orthotopically rhBMP-2 incorporated CHA implants, in accordance with SPECT/CT findings. Collectively, data shows 99mTc-MDP SPECT/CT is a sensitive and noninvasive tool to monitor osteoinductivity and bone regeneration of PTEB and orthotopic implants. The PTEB achieved peak osteoinductivity and bone density at 8 to 13 weeks following ectopic implantation, which would serve as a recommendable time frame for its transfer to mandibular reconstruction.
doi:10.1371/journal.pone.0137167
PMCID: PMC4560383  PMID: 26340447
4.  Clinicopathological characteristics of immunoglobulin G4-related sialadenitis 
Introduction
Immunoglobulin G4-related disease (IgG4-RD) is a newly recognized fibro-inflammatory condition. Forty-two cases with immunoglobulin G4-related sialadenitis (IgG4-RS) confirmed by histopathological and immunohistochemical assessment were studied to clarify the clinicopathologic characteristics of the salivary glands involved in IgG4-RS, especially the relationship between the histopathologic features and function of salivary glands or serum levels of IgG4.
Methods
Clinical, serologic, imaging and histopathological data of these cases were analyzed. CT volumes of submandibular, parotid, and lacrimal glands were calculated. The saliva flow rate was measured. Scintigraphy with 99mTc-pertechnetate was undertaken in 31 cases, and the concentration index (CI) and secretion index (SI) was calculated. Relationships between fibrosis severity and salivary gland function or serum IgG4 levels were analyzed.
Results
The first symptom was swelling of bilateral submandibular or lacrimal glands. Physical examination showed multiple bilateral major salivary glands (including sublingual and accessory parotid glands) and lacrimal glands were enlarged in IgG4 RS. Multiple enlarged cervical lymph nodes were noted in 30 patients. Saliva flow at rest was lower than normal in 34 cases; stimulated saliva flow was lower than normal in 15 cases. Secretory function was reduced more severely in the submandibular glands than in the parotid glands. Serum levels of IgG4 were elevated in 95.2% of cases and 78.6% patients had increased IgE levels. Serum IgG4 level was higher and saliva secretion lower as glandular fibrosis increased.
Conclusions
Prominent changes in the morphology, histology, immunohistochemistry and secretion of the major salivary glands of IgG4-RS patients were accompanied by involvement of the lacrimal glands and cervical lymph nodes. Elevated IgE, allergic history, eosinophil infiltration suggest allergic reactions as a potential pathogenesis of IgG4-RS. Severity of glandular fibrosis correlated with salivary function and serum levels of IgG4.
doi:10.1186/s13075-015-0698-y
PMCID: PMC4508811  PMID: 26194097
5.  Microvascular autologous submandibular gland transplantation in severe cases of keratoconjunctivitis sicca 
Dry eye syndrome is a relatively common disease of the tears and ocular surfaces that results in discomfort, visual disturbance, and tear film instability with possible damage to the ocular surfaces. Microvascular submandibular gland (SMG) transfer offers a surgical alternative for a permanent autologous substitution of tears using the basal secretion of a transplanted SMG. Long-term follow-up reveals that this technique is a lasting and effective solution for patients with severe dry eye syndrome. The uncomfortable symptoms were relieved, and the frequency of use of pharmaceutical tear substitutes was reduced. Objective examination showed significant improvement in tear film and some features of ocular surface such as breakup time of tear film and corneal staining. Patients may suffer from obstruction of Wharton's duct or epiphora after surgery. Activation of secretion-related receptors could improve the early hypofunction of the denervated SMG and prevent the duct obstruction. Reduction surgery, partial SMG transplantation, uses of atropine gel or Botulinum toxin A could be the choices of treatment for epiphora.
doi:10.1186/s40902-015-0006-4
PMCID: PMC4317526  PMID: 25664315
Obstructive sialadenitis; Keratoconjunctivitis sicca; Submandibular gland transplantation
6.  miR-34a Inhibits Migration and Invasion of Tongue Squamous Cell Carcinoma via Targeting MMP9 and MMP14 
PLoS ONE  2014;9(9):e108435.
Background
miR-34a is an important tumor suppressor gene in various cancer types. But little is known about the dysregulation of miR-34a in tongue squamous cell carcinoma (TSCC). In this study, we investigate the expression and potential role of miR-34a in TSCC.
Methods
We evaluated miR-34a expression and its relationship with clinicopathological characters in 75 pairs of TSCC samples, and confirmed the role of miR-34a for predicting lymph node metastases from a further 15 pairs of paraffin-embedded TSCC specimens with stringent clinicopathological recruitment criteria using quantitative reverse transcription polymerase chain reaction (qRT-PCR). The effects of miR-34a on cell proliferation, migration and invasion were examined in TSCC cell lines using Cell Counting Kit-8 assay, wound healing assay and transwell assay, respectively. The effects of miR-34a on the expression of matrix metalloproteinase (MMP) 9 and 14 were detected by luciferase reporter assays and Western blot analysis. The expression of miR-34a, MMP9 and MMP14 were also confirmed in TSCC samples by in situ hybridization and immunohistochemistry.
Results
miR-34a expression in tumor tissues from TSCC patients with positive lymph node metastases was significantly lower than that with negative lymph node metastases. Overexpression of miR-34a significantly suppressed migration and invasion in TSCC cells and simultaneously inhibited the expression of MMP9 and MMP14 through targeting the coding region and the 3′untranslated region, respectively. Moreover, miR-34a expression in TSCC was inversely correlated with protein expression of MMP9 and MMP14 in the TSCC samples.
Conclusions
miR-34a plays an important role in lymph node metastases of TSCC through targeting MMP9 and MMP14 and may have potential applications in prognosis prediction and gene therapy for lymph node metastases of TSCC patients.
doi:10.1371/journal.pone.0108435
PMCID: PMC4182478  PMID: 25268950
7.  Botulinum toxin A inhibits salivary secretion of rabbit submandibular gland 
Botulinum toxin A (BTXA) has been used in several clinical trials to treat excessive glandular secretion; however, the precise mechanism of its action on the secretory function of salivary gland has not been fully elucidated. In this study, we aimed to investigate the effect of BTXA on secretion of submandibular gland in rabbits and to identify its mechanism of action on the secretory function of salivary gland. At 12 weeks after injection with 5 units of BTXA, we found a significant decrease in the saliva flow from submandibular glands, while the salivary amylase concentration increased. Morphological analysis revealed reduction in the size of acinar cells with intracellular accumulation of secretory granules that coalesced to form a large ovoid structure. Expression of M3-muscarinic acetylcholine receptor (M3 receptor) and aquaporin-5 (AQP5) mRNA decreased after BTXA treatment, and distribution of AQP5 in the apical membrane was reduced at 1, 2 and 4 weeks after BTXA injection. Furthermore, BTXA injection was found to induce apoptosis of acini. These results indicate that BTXA decreases the fluid secretion of submandibular glands and increases the concentration of amylase in saliva. Decreased expression of M3 receptor and AQP5, inhibition of AQP5 translocation, and cell apoptosis might involve in BTXA-reduced fluid secretion of submandibular glands.
doi:10.1038/ijos.2013.82
PMCID: PMC3967320  PMID: 24158141
aquaporin-5; apoptosis; botulinum toxin A; M3-muscarinic acetylcholine receptor; submandibular gland
8.  Oral and maxillofacial surgery: Current and future 
Annals of Maxillofacial Surgery  2013;3(2):111-112.
doi:10.4103/2231-0746.119209
PMCID: PMC3814657  PMID: 24205468
9.  Adiponectin Increases Secretion of Rat Submandibular Gland via Adiponectin Receptors-Mediated AMPK Signaling 
PLoS ONE  2013;8(5):e63878.
Adiponectin and adiponectin receptors (AdipoR1/2) are expressed in various tissues and are involved in the regulation of multiple functions such as energy metabolism and inflammatory responses. However, the effect of adiponectin and AdipoRs in submandibular glands has not been fully evaluated. In the present study, we found that mRNA and protein of both adiponectin and AdipoR1/2 were expressed in rat submandibular glands and in the SMG-C6 cell line, as evidenced by RT-PCR and Western blot analysis. Immunofluorescence staining showed that adiponectin was diffused in the cytoplasm, while AdipoR1/2 was concentrated in the membrane of acinar cells. Saliva flow was significantly increased by full length adiponectin (fAd) or globular adiponectin (gAd) perfusion in isolated rat submandibular glands. 5-Aminoimidazole-4-carboxamide-1-4-ribofuranoside (AICAR), an adenosine monophosphate activated protein kinase (AMPK) activator, also increased saliva secretion. fAd, gAd, and AICAR all increased the average width of apical tight junctions in perfused submandibular glands, and decreased transepithelial electrical resistance (TER) in SMG-C6 cells, suggesting that adiponectin promoted secretion by modulating paracellular permeability. fAd and gAd increased p-AMPK levels, while AraA, an AMPK antagonist, abolished fAd- and gAd-induced changes in secretion, tight junction ultrastructure, and TER. Moreover, both AdipoR1 and AdipoR2 were required for fAd- or gAd-induced p-AMPK and TER responses, suggesting from their inhibition following AdipoR1 or AdipoR2 knockdown, and co-knockdown of AdipoRs by RNA interference. Our results suggest that adiponectin functions as a promoter of salivary secretion in rat submandibular glands via activation of AdipoRs, AMPK, and paracellular permeability.
doi:10.1371/journal.pone.0063878
PMCID: PMC3646765  PMID: 23667684
10.  Prognostic Implications of MicoRNA miR-195 Expression in Human Tongue Squamous Cell Carcinoma 
PLoS ONE  2013;8(2):e56634.
Background
miR-195 is aberrantly expressed in multiple types of disease. But little is known about the dysregulation of miR-195 in tongue squamous cell carcinoma (TSCC). In this study, we investigated the roles of miR-195 in the development and progression of TSCC.
Methods
Using quantitative reverse transcription-polymerase chain reaction (qRT-PCR), we evaluated miR-195 expression in TSCC samples from 81 patients. Overall survival of these patients was examined using Kaplan–Meier curves with log-rank tests and the Cox proportional hazards model. The expression of two known miR-195 target genes, Cyclin D1 and Bcl-2, was also examined in the TSCC samples by immunohistochemistry. The effects of miR-195 overexpression on cell cycle progression and apoptosis and its effects on the expression of Cyclin D1 and Bcl-2 were examined in transfected TSCC cell lines (SCC-15 and Cal27) using fluorescence-activated cell sorting assays, luciferase reporter assays, and Western blots.
Results
Reduced miR-195 expression was associated with tumor size and the clinical stage of TSCC tumors. Kaplan–Meier survival analysis indicated that the TSCC patients with reduced expression of miR-195 had poor overall survival and in multivariable analyses low levels of miR-195 emerged as an independent prognostic factor for this clinical outcome. Levels of miR-195 expression were inversely correlated with the expression of Cyclin D1 and Bcl-2. Overexpression of miR-195 inhibited cell cycle progression, promoted apoptosis, and reduced Cyclin D1 and Bcl-2 expression in two TSCC cell lines.
Conclusions
miR-195 may have potential applications as a prognostic factor for TSCC patients.
doi:10.1371/journal.pone.0056634
PMCID: PMC3579853  PMID: 23451060
11.  A digital model individual template and CT-guided 125I seed implants for malignant tumors of the head and neck 
Journal of Radiation Research  2012;53(6):973-977.
To enhance the accuracy of radioactive seed implants in the head and neck, a digital model individual template, containing information simultaneously on needle pathway and facial features, was designed to guide implantation with CT imaging. Thirty-one patients with recurrent and local advanced malignant tumors of head and neck after prior surgery and radiotherapy were involved in this study. Before 125I implants, patients received CT scans based on 0.75mm thickness. And the brachytherapy treatment planning system (BTPS) software was used to make the implantation plan based on the CT images. Mimics software and Geomagic software were used to read the data containing CT images and implantation plan, and to design the individual template. Then the individual template containing the information of needle pathway and face features simultaneously was made through rapid prototyping (RP) technique. All patients received 125I seeds interstitial implantation under the guide of the individual template and CT. The individual templates were positioned easily and accurately, and were stable. After implants, treatment quality evaluation was made by CT and TPS. The seeds and dosages distribution (D90,V100,V150) were well meet the treatment requirement. Clinical practice confirms that this approach can facilitate easier and more accurate implantation.
doi:10.1093/jrr/rrs046
PMCID: PMC3483848  PMID: 22859564
Digital model; individual template; radioactive seeds; head and neck
12.  7-Diethyl­amino-2-propyl­sulfan­yl-3-(1H-1,2,4-triazol-1-yl)-4H-thio­chromen-4-one 
In the title compound, C18H22N4OS2, the six-membered rings are almost coplanar, showing a dihedral angle between the mean planes of 9.0 (4)°, while the triazol ring is nearly perpendicular to the thio­chromen-4-one unit, making an angle of 89.8 (4)°. In the crystal, C—H⋯N hydrogen bonds link the mol­ecules in a stacked arrangement along the c axis.
doi:10.1107/S1600536812015954
PMCID: PMC3344555  PMID: 22590317
13.  (Z)-1-(2,4-Difluoro­phen­yl)-2-(1H-1,2,4-triazol-1-yl)ethanone oxime 
In the title compound, C10H8F2N4O, the dihedral angle between the rings is 65.4 (1)°. In the crystal, inter­molecular O—H⋯N and C—H⋯F hydrogen bonds link the mol­ecules in a stacked arrangement along the a and c axes, respectively.
doi:10.1107/S1600536811050008
PMCID: PMC3239095  PMID: 22199943
14.  1-(4-Fluoro­phen­yl)-2-(1H-imidazol-1-yl)ethanone 
In the title compound, C11H9FN2O, the dihedral angle between the rings is 87.50 (4)°. In the crystal, inter­molecular C—H⋯N hydrogen bonds link the mol­ecules in a stacked arrangement along the c axis.
doi:10.1107/S1600536811026432
PMCID: PMC3213485  PMID: 22091064
15.  7-Fluoro-2-(prop-2-en-1-ylsulfan­yl)-3-(1H-1,2,4-triazol-1-yl)-4H-thio­chromen-4-one 
The asymmetric unit of the title compound, C14H10FN3OS2, contains two independent mol­ecules which differ in the relative orientations of the triazole and allyl­sulfanyl groups with respect to the planar thio­chromen-4-one frameworks. The N—N—C—C torsion angles are 128.2 (5) and −120.9 (5)°, while the C—S—C—S torsion angles are −17.4 (4) and 16.4 (4)°. In the crystal, inter­molecular C—H⋯O and C—H⋯N hydrogen bonds link the mol­ecules in a stacked arrangement along the a axis.
doi:10.1107/S1600536811022665
PMCID: PMC3151981  PMID: 21837153
16.  2-Ethyl­sulfanyl-7-fluoro-3-(1H-1,2,4-triazol-1-yl)-4H-thio­chromen-4-one 
The asymmetric unit of the title compound, C13H10FN3OS2, contains two independent mol­ecules, which differ slightly in the relative orientations of the triazole and ethyl­sulfanyl groups with respect to the planar thio­chromen-4-one frameworks. The dihedral angles between the mean planes of the triazole groups and the corresponding six-membered C5OS rings are 56.8 (1) and 52.9 (1)°, while the S—C—S—C dihedral angles are −11.7 (2) and −16.3 (2)°. In the crystal structure, inter­molecular C—H⋯O and C—H⋯N hydrogen bonds link the mol­ecules in a stacked arrangement along the a axis. A weak intramolecular C—H⋯·O interaction results in the formation of a non-planar five-membered ring.
doi:10.1107/S1600536810030175
PMCID: PMC3007947  PMID: 21588584
17.  Ethyl 2-{[7-fluoro-4-oxo-3-(1H-1,2,4-triazol-1-yl)-4H-thio­chromen-2-yl]sulfan­yl}acetate 
In the title compound, C15H12FN3O3S2, the two six-membered rings are essentially coplanar, their mean plnes making a dihedral angle of 1.1 (2)°. The carbonyl C, the two attached non-fused C atoms and the S atom deviate from the plane of the benzene ring by −0.046 (5), −0.017 (5), 0.000 (6), 0.026 (4) Å, respectively. The angle between the mean planes of the triazole ring and the sulfur heterocycle is 53.3 (1)°. In the crystal, inter­molecular C—H⋯O hydrogen bonds link the mol­ecules in a stacked arrangement along the a axis.
doi:10.1107/S1600536810027467
PMCID: PMC3007513  PMID: 21588373
18.  Methyl 7-chloro-2-ethyl­sulfanyl-6-fluoro-4-oxo-4H-thio­chromene-3-carboxyl­ate 
In the title compound, C13H10ClFO3S2, the two-ring system is essentially planar, the mean plane of the benzene ring being inclined at 6.0 (2)° to the plane of the remaining four atoms. The ethyl­sulfanyl group is almost coplanar with the two rings [dihedral angle = 6.4 (2)°], while the carboxyl­ate group is almost perpendicular to it [dihedral angle = 72.4 (2)°]. In the crystal structure, inter­molecular C—H⋯O and C—H⋯F hydrogen bonds link the mol­ecules in a stacked arrangement along the a axis.
doi:10.1107/S1600536810002667
PMCID: PMC2983649  PMID: 21580435

Results 1-18 (18)