MicroRNA-584-5p/RUNX family transcription factor 2 axis mediates hypoxia-induced osteogenic differentiation of periosteal stem cells.

BACKGROUND: The hypoxic environment during bone healing is important in regulating the differentiation of periosteal stem cells (PSCs) into osteoblasts or chondrocytes; however, the underlying mechanisms remain unclear. AIM: To determine the effect of hypoxia on PSCs, and the expression of microRNA-584-5p (miR-584-5p) and RUNX family transcription factor 2 (RUNX2) in PSCs was modulated to explore the impact of the miR-584-5p/RUNX2 axis on hypoxia-induced osteogenic differentiation of PSCs. METHODS: In this study, we isolated primary mouse PSCs and stimulated them with hypoxia, and the characteristics and functional genes related to PSC osteogenic differentiation were assessed. Constructs expressing miR-584-5p and RUNX2 were established to determine PSC osteogenic differentiation. RESULTS: Hypoxic stimulation induced PSC osteogenic differentiation and significantly increased calcified nodules, intracellular calcium ion levels, and alkaline phosphatase (ALP) activity in PSCs. Osteogenic differentiation-related factors such as RUNX2, bone morphogenetic protein 2, hypoxia-inducible factor 1-alpha, and ALP were upregulated; in contrast, miR-584-5p was downregulated in these cells. Furthermore, upregulation of miR-584-5p significantly inhibited RUNX2 expression and hypoxia-induced PSC osteogenic differentiation. RUNX2 was the target gene of miR-584-5p, antagonizing miR-584-5p inhibition in hypoxia-induced PSC osteogenic differentiation. CONCLUSION: Our study showed that the interaction of miR-584-5p and RUNX2 could mediate PSC osteogenic differentiation induced by hypoxia.

Deep convolutional neural network based on CT images of pulmonary nodules in the lungs of adolescent and young adult patients with osteosarcoma.

The aim of the present study was to explore the diagnostic value of a deep convolutional neural network (DCNN) model for the diagnosis of pulmonary nodules in adolescent and young adult patients with osteosarcoma. For the present study, 675 chest CT images were retrospectively collected from 109 patients with clinically confirmed osteosarcoma who underwent chest CT examination at Hangzhou Third People's Hospital (Hangzhou, China) from March 2011 to February 2022. CT images were then evaluated using the DCNN and manual models. Subsequently, pulmonary nodules of osteosarcoma were divided into calcified nodules, solid nodules, partially solid nodules and ground glass nodules using the DCNN model. Those patients with osteosarcoma who were diagnosed and treated were followed up to observe dynamic changes in the pulmonary nodules. A total of 3,087 nodules were detected, while 278 nodules were missed compared with those determined using the reference standard given by the consensus of three Experienced radiologists., which was analyzed by two diagnostic radiologists. In the manual model group, 2,442 nodules were detected, while 657 nodules were missed. The DCNN model showed significantly higher sensitivity and specificity compared with the manual model (sensitivity, 0.923 vs. 0.908; specificity, 0.552 vs. 0.351; P<0.05). In addition, the DCNN model yielded an area under the curve (AUC) value of 0.795 [95% confidence interval (CI), 0.743-0.846], outperforming that of the manual model (AUC, 0.687; 95% CI, 0.629-0.732; P<0.05). The film reading time of the DCNN model was also significantly shorter compared with that of the manual model [mean ± standard deviation (SD); 173.25±24.10 vs. 328.32±22.72 sec; P<0.05)]. The AUC of calcified nodules, solid nodules, partially solid nodules and ground glass nodules was calculated to be 0.766, 0.771, 0.761 and 0.796, respectively, using the DCNN model. Using this model, the majority of the pulmonary nodules were detected in patients with osteosarcoma at the initial diagnosis (69/109, 62.3%), and the majority of these were found with multiple pulmonary nodules instead of a single nodule (71/109, 65.1% vs. 38/109, 34.9%). These data suggest that, compared with the manual model, the DCNN model proved to be beneficial for the detection of pulmonary nodules in adolescent and young adult patients with osteosarcoma, which may reduce the time of artificial radiograph reading. In conclusion, the proposed DCNN model, developed using data from 675 chest CT images retrospectively collected from 109 patients with clinically confirmed osteosarcoma, may be used as an effective tool to evaluate pulmonary nodules in patients with osteosarcoma.

A novel hybrid scan approach enabling the ion-mobility separation and the alternate data-dependent and data-independent acquisitions (HDDIDDA): Its combination with off-line two-dimensional liquid chromatography for comprehensively characterizing the multicomponents from Compound Danshen Dripping Pill.

Data-dependent acquisition (DDA) and data-independent acquisition (DIA)-based MSn strategies are extensively applied in metabolites characterization. DDA gives accurate MSn information, but receives low coverage, while DIA covers the entire mass range, but the precursor-product ions matching often yields false positives. Currently available MS scan approaches rarely integrate DIA and DDA within a duty circle. Utilizing a Vion™ IM-QTOF (ion mobility-quadrupole time-of-flight) mass spectrometer, we report a novel hybrid scan approach, namely HDDIDDA, which involves three scan events: 1) IM-enabled full scan (MS1), 2) high-definition MSE (HDMSE) of all precursor ions (MS2); and 3) high-definition DDA (HDDDA) of top N precursors (MS2). As a proof-of-concept, the HDDIDDA approach combined with off-line two-dimensional liquid chromatography (2D-LC) was applied to characterize the multiple ingredients from a reputable Chinese patent medicine, Compound Danshen Dripping Pill (CDDP) used for treating the cardiovascular diseases. An off-line 2D-LC system by configuring an XBridge Amide column and an HSS T3 column showed a measurable orthogonality of 0.92 and enhanced the separation of co-eluting components. A fit-for-purpose HDDIDDA methodology was developed in the negative mode to characterize saponins and salvianolic acids, while tanshinones in the positive mode. Computational workflows to efficiently process the acquired HDMSE and HDDDA data were established, and the searching of an in-house CDDP library (recording 712 compounds) eventually characterized 403 components from CDDP, indicating approximate 12-fold improvement compared with the previous report. The HDDIDDA approach can measure collision cross section of each component, and merges the merits of DIA and DDA in MS2 data acquisition.

A novel neutral loss/product ion scan-incorporated integral approach for the untargeted characterization and comparison of the carboxyl-free ginsenosides from Panax ginseng, Panax quinquefolius, and Panax notoginseng.

Differentiated composition in precursor ions for different subclasses of ginsenosides in the negative electrospray-ionization mode has been reported, which lays a foundation for the sorted and untargeted identification of ginsenosides. Carboxyl-free ginsenosides simultaneously from Panax ginseng, P. quinquefolius, and P. notoginseng, were comprehensively characterized and statistically compared. A neutral loss/product ion scan (NL-PIS) incorporated untargeted profiling approach, coupled to ultra-high performance liquid chromatography, was developed on a linear ion-trap/Orbitrap mass spectrometer for characterizing carboxyl-free ginsenosides. It incorporated in-source fragmentation (ISF) full scan-MS1, mass tag-MS2, and product ion scan-MS3. Sixty batches of ginseng samples were analyzed by metabolomics workflows for the discovery of ginsenoside markers. Using formic acid (FA) as the additive, carboxyl-free ginsenosides (protopanaxadiol-type, protopanaxatriol-type, and octillol-type) gave predominant FA-adducts, while rich deprotonated molecules were observed for carboxyl-containing ginsenosides (oleanolic acid-type and malonylated) when source-induced dissociation (SID) was set at 0 V. Based on the NL transition [M+FA‒H]- > [M-H]- and the characteristic sapogenin product ions, a NL-PIS approach was established. It took advantage of the efficient full-information acquisition of ISF-MS1 (SID: 50 V), the high specificity of mass tag (NL: 46.0055 Da)-induced MS2 fragmentation, and the substructure fragmentation of product ion scan-MS3. We could characterize 216 carboxyl-free ginsenosides, and 21 thereof were potentially diagnostic for the species differentiation. Conclusively, sorted and untargeted characterization of the carboxyl-free ginsenosides was achieved by the established NL-PIS approach. In contrast to the conventional NL or PIS-based survey scan strategies, the high-accuracy MSn data obtained can enable more reliable identification of ginsenosides.

Three novel indole alkaloids from Kopsia officinalis.

Three new indole alkaloids, named kopsioffines A-C (1-3), possessing relatively novel ten-membered lactam ring, and one known compound (11,12-demethoxy-16-deoxypauciflorine, 4) were isolated from the leaves and stems of Kopsia officinalis. Their structures were elucidated by means of spectroscopic methods. The absolute configuration of 1 was determined by calculated electronic circular dichroism data. The hypothetical biosynthetic pathway of 1 was postulated. All these isolates were evaluated for their inhibitory effects on α-glucosidase. None of them showed activity with IC50 far beyond 50µM.

Malonylginsenosides with Potential Antidiabetic Activities from the Flower Buds of Panax ginseng.

LC-MS-guided phytochemical isolation of malonylginsenosides, featuring neutral elimination of CO2 and C3H2O3 by the negative mode collision-induced dissociation, from the flower buds of Panax ginseng led to the isolation of 19 malonyl-substituted triterpenoid saponins. They include 15 new malonylginsenosides, malonylfloralginsenosides-Re1-Re3 (1-3), -Rb1 and -Rb2 (4, 5), -Rd1-Rd6 (6-11), and -Rc1-Rc4 (12-15), and the known m-Rb1, m-Rc, m-Rb2, and m-Rd (16-19). Compound 11 represents the first dimalonyl saponin isolated from the Panax genus, while 2-4, 9, and 10 are five ginsenosides with single malonylation at the C-20 sugar chain. The antidiabetic activities of nine of these malonyl-substituted ginsenosides (1, 3, 4, 8, 13, and 16-19) and five of the corresponding non-malonyl ginsenosides (Re, Rb1, Rb2, Rc, and Rd) were evaluated by L6 myotubes' glucose consumption and AMPKα2ß1γ1 activation. Ginsenoside Rb2, 1, and 18 promoted glucose consumption of differentiated L6 myotubes, while ginsenosides Rb1, Rb2, and Rd and the malonylginsenosides 4, 8, 13, 16, 17, and 19 activated AMPKα2ß1γ1 (EC50: 0.0168-2.8 µM, fold: 1.7-4.7).

A Strategy Combining Higher Energy C-Trap Dissociation with Neutral Loss- and Product Ion-Based MSn Acquisition for Global Profiling and Structure Annotation of Fatty Acids Conjugates.

Fatty acids conjugates (FACs) are ubiquitous but found in trace amounts in the natural world. They are composed of multiple unknown substructures and side chains. Thus, FACs are difficult to be analyzed by traditional mass spectrometric methods. In this study, an integrated strategy was developed to global profiling and targeted structure annotation of FACs in complex matrix by LTQ Orbitrap. Dicarboxylic acid conjugated bufotoxins (DACBs) in Venenum bufonis (VB) were used as model compounds. The new strategy (abbreviated as HPNA) combined higher-energy C-trap dissociation (HCD) with product ion- (PI), neutral loss- (NL) based MSn (n ≥ 3) acquisition in both positive-ion mode and negative-ion mode. Several advantages are presented. First, various side chains were found under HCD in negative-ion mode, which included both known and unknown side chains. Second, DACBs with multiple side chains were simultaneously detected in one run. Compared with traditional quadrupole-based mass method, it greatly increased analysis throughput. Third, the fragment ions of side chain and steroids substructure could be obtained by PI- and NL-based MSn acquisition, respectively, which greatly increased the accuracy of the structure annotation of DACBs. In all, 78 DACBs have been discovered, of which 68 were new compounds; 25 types of substructure formulas and seven dicarboxylic acid side chains were found, especially five new side chains, including two saturated dicarboxylic acids [(azelaic acid (C9) and sebacic acid (C10)] and three unsaturated dicarboxylic acids (u-C8, u-C9, and u-C10). All these results greatly enriched the structures of DACBs in VB. Graphical Abstract ᅟ.

New monoterpenoid oxindole alkaloid derivatives from the stems of Uncaria hirsuta Havil. and their cytotoxicity and tandem mass spectrometric fragmentation.

Four new alkaloids, comprising three 3-oxo-3,7-seco-oxindole alkaloids (hirsutanine D-F, 1-3) and one oxindole alkaloid N-oxide (uncarine B N-oxide, 4), together with four known heteroyohimbine-type oxindole alkaloids, were isolated from the stems of Uncaria hirsuta Havil. Structures of 1-4 were elucidated by extensive NMR and HR-ESIMS data analyses. Compound 3 is the first 3-oxo-3,7-seco-oxindole alkaloid with ring B opened and degraded isolated from the Uncaria genus. Compounds 1-3 exhibited slight inhibition effect on the proliferation of the breast cancer cell MDA-MB-231. The positive mode collision-induced dissociation of the 3-oxo-3,7-seco-oxindole alkaloids (1-3) was featured by the ß-cleavage and α-cleavage of the amido bond, while the N-oxide (4) showed characteristic neutral eliminations of ·OH and H2O.

An in-source multiple collision-neutral loss filtering based nontargeted metabolomics approach for the comprehensive analysis of malonyl-ginsenosides from Panax ginseng, P. quinquefolius, and P. notoginseng.

The simultaneous identification and quantification of target metabolites from herbal medicines are difficult to implement by the full-scan MS based nontargeted metabolomics approaches. Here an in-source multiple collision-neutral loss filtering (IMC-NLF) based nontargeted metabolomics approach is developed and applied to identify and quantify the variations of malonyl-ginsenosides, a common group of acyl saponins with potential anti-diabetic activity, among Panax ginseng, P. quinquefolius, and P. notoginseng. The key steps of the IMC-NLF strategy are the acquisition of specific high-resolution neutral loss data and the efficient filtering of target precursor ions from the full-scan spectra. Using a hybrid LTQ-Orbitrap mass spectrometer after UHPLC separation, abundant in-source product ions, [M-H-CO2]- (due to the vulnerability of the carboxyl group) and [M-H-Mal.]-, were generated at the energies of 70 V and 90 V, respectively. After spectral deconvolution, the generated peak list was screened by dual NLF using a Neutral Loss MS Finder software (NL of 43.9898 Da for CO2 and 86.0004 Da for the malonyl substituent). By combining the precursor ions list-triggered HCD-MS/MS and basic hydrolysis, a total of 101 malonyl-ginsenosides (including 69 from P. ginseng, 52 from P. quinquefolius, and 44 from P. notoginseng) were identified or tentatively characterized. The variations of 81 characterized malonyl-ginsenosides among 45 batches of Ginseng samples were statistically analyzed disclosing ten potential markers. It is the first systematic analysis of malonyl-ginsenosides. The IMC-NLF approach by a single analytical platform is promising in targeted analyses of modification-specific metabolites in metabolomics and drug metabolism.

Nontargeted metabolomic analysis and "commercial-homophyletic" comparison-induced biomarkers verification for the systematic chemical differentiation of five different parts of Panax ginseng.

A key segment in authentication of herbal medicines is the establishment of robust biomarkers that embody the intrinsic metabolites difference independent of the growing environment or processing technics. We present a strategy by nontargeted metabolomics and "Commercial-homophyletic" comparison-induced biomarkers verification with new bioinformatic vehicles, to improve the efficiency and reliability in authentication of herbal medicines. The chemical differentiation of five different parts (root, leaf, flower bud, berry, and seed) of Panax ginseng was illustrated as a case study. First, an optimized ultra-performance liquid chromatography/quadrupole time-of-flight-MS(E) (UPLC/QTOF-MS(E)) approach was established for global metabolites profiling. Second, UNIFI™ combined with search of an in-house library was employed to automatically characterize the metabolites. Third, pattern recognition multivariate statistical analysis of the MS(E) data of different parts of commercial and homophyletic samples were separately performed to explore potential biomarkers. Fourth, potential biomarkers deduced from commercial and homophyletic root and leaf samples were cross-compared to infer robust biomarkers. Fifth, discriminating models by artificial neutral network (ANN) were established to identify different parts of P. ginseng. Consequently, 164 compounds were characterized, and 11 robust biomarkers enabling the differentiation among root, leaf, flower bud, and berry, were discovered by removing those structurally unstable and possibly processing-related ones. The ANN models using the robust biomarkers managed to exactly discriminate four different parts and root adulterant with leaf as well. Conclusively, biomarkers verification using homophyletic samples conduces to the discovery of robust biomarkers. The integrated strategy facilitates authentication of herbal medicines in a more efficient and more intelligent manner.

Venenum Bufonis induces rat neuroinflammation by activiating NF-κB pathway and attenuation of BDNF.

ETHNOPHARMACOLOGICAL RELEVANCE: Venenum Bufonis (VB), also called toad venom, has been widely used in clinic as a cardiotonic, anohyne and antineoplastic agents both in China and other Asian countries. However, its neurotoxicity and cardiotoxicity limit its wide clinical application. Compared with extensive attention attracted with cardiotoxicity, the toxic effect of VB on Central Nervous System (CNS) is much less studied. AIM OF THE RESEARCH: This study was performed to examine the neurotoxicity caused by VB on Sprague Dawley (SD) rats, then to clarify the mechanism in vivo by investigating its action on the neuroinflammation which possibly attributed to the activation of nuclear factor κB (NF-κB) pathway and the attenuation of brain-derived neurotrophic factor (BDNF). MATERIALS AND METHODS: Rats administrated with 0.5% carboxymethyl cellulose sodium salt (CMC-Na) aqueous solution and VB (100mg/kg, 200mg/kg and 400mg/kg) were sacrificed at 2h, 4h, 6h, 8h, 24h and 48h. The brain level of neurotransmitters and their corresponding receptors, pro-inflammatory cytokines, BDNF/TrkB and NF-κB pathway-related proteins were examined, respectively. RESULTS: VB administration induced severe neurologic damage and neuroinflammation, as indicated by the disordered 5-hydroxytryptamine (5-HT), dopamine (DA) and their corresponding receptors, together with the over production of inflammatory cytokines including interleukin-6 (IL-6), interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α). VB also notably promoted the expression of p-NF-κBp65, p-IκBα, p-IKKα and p-IKKß and down-regulated the expression of BDNF and TrkB. CONCLUSION: This study demonstrates that VB triggers neurotoxicity which probably is induced by neuroinflammation via activating of NF-κB pathway and attenuating the expression of BDNF.

TXNIP/TRX/NF-κB and MAPK/NF-κB pathways involved in the cardiotoxicity induced by Venenum Bufonis in rats.

Venenum Bufonis (VB) is a widely used traditional medicine with serious cardiotoxic effects. The inflammatory response has been studied to clarify the mechanism of the cardiotoxicity induced by VB for the first time. In the present study, Sprague Dawley (SD) rats, were administered VB (100, 200, and 400 mg/kg) intragastrically, experienced disturbed ECGs (lowered heart rate and elevated ST-segment), increased levels of serum indicators (creatine kinase (CK), creatine kinase isoenzyme-MB (CK-MB), alanine aminotransferase (ALT), aspartate aminotransferase (AST)) and serum interleukin (IL-6, IL-1ß, TNF-α) at 2 h, 4 h, 6 h, 8 h, 24 h, and 48 h, which reflected that an inflammatory response, together with cardiotoxicity, were involved in VB-treated rats. In addition, the elevated serum level of MDA and the down-regulated SOD, CAT, GSH, and GPx levels indicated the appearance of oxidative stress in the VB-treated group. Furthermore, based on the enhanced expression levels of TXNIP, p-NF-κBp65, p-IκBα, p-IKKα, p-IKKß, p-ERK, p-JNK, and p-P38 and the obvious myocardial degeneration, it is proposed that VB-induced cardiotoxicity may promote an inflammatory response through the TXNIP/TRX/NF-κB and MAPK/NF-κB pathways. The observed inflammatory mechanism induced by VB may provide a theoretical reference for the toxic effects and clinical application of VB.

A green protocol for efficient discovery of novel natural compounds: characterization of new ginsenosides from the stems and leaves of Panax ginseng as a case study.

Exploration of new natural compounds is of vital significance for drug discovery and development. The conventional approaches by systematic phytochemical isolation are low-efficiency and consume masses of organic solvent. This study presents an integrated strategy that combines offline comprehensive two-dimensional liquid chromatography, hybrid linear ion-trap/Orbitrap mass spectrometry, and NMR analysis (2D LC/LTQ-Orbitrap-MS/NMR), aimed to establish a green protocol for the efficient discovery of new natural molecules. A comprehensive chemical analysis of the total ginsenosides of stems and leaves of Panax ginseng (SLP), a cardiovascular disease medicine, was performed following this strategy. An offline 2D LC system was constructed with an orthogonality of 0.79 and a practical peak capacity of 11,000. The much greener UHPLC separation and LTQ-Orbitrap-MS detection by data-dependent high-energy C-trap dissociation (HCD)/dynamic exclusion were employed for separation and characterization of ginsenosides from thirteen fractionated SLP samples. Consequently, a total of 646 ginsenosides were characterized, and 427 have not been isolated from the genus of Panax L. The ginsenosides identified from SLP exhibited distinct sapogenin diversity and molecular isomerism. NMR analysis was finally employed to verify and offer complementary structural information to MS-oriented characterization. The established 2D LC/LTQ-Orbitrap-MS/NMR approach outperforms the conventional approaches in respect of significantly improved efficiency, much less use of drug materials and organic solvent. The integrated strategy enables a deep investigation on the therapeutic basis of an herbal medicine, and facilitates new compounds discovery in an efficient and environmentally friendly manner as well.

Comparison of distraction osteogenesis versus costochondral graft in reconstruction of temporomandibular joint condylectomy with disc preservation.

PURPOSE: This study compared transport distraction with costochondral graft in the reconstruction of temporomandibular joint (TMJ) with disc preservation. MATERIALS AND METHODS: Bilateral condylectomies with disc preservation were conducted in 12 adult goats. Transport distraction of the mandibular ramus was performed on 1 side and costochondral graft on the other side. Jaw movements, TMJ healing, and condylar remodeling were assessed by clinical, radiologic, and histologic examinations. RESULTS: Transport distraction and costochondral graft established a new condyle separating from the preserved disc with a joint space. Both techniques showed no significant difference in ankylosis scores up to 24 weeks. Signs of calcification were not noted in the transport distraction or costochondral graft side. No significant change of maximal jaw movements was found at any time point from 2 to 24 weeks. CONCLUSIONS: Distraction osteogenesis is an effective reconstruction method for TMJ reconstruction. No significant difference between these 2 techniques in reconstruction of the TMJ was noted.

Efficacy of functional training of the facial muscles for treatment of incomplete peripheral facial nerve injury.

OBJECTIVE: To investigate the efficacy of functional training of facial mimic muscles for patients with incomplete peripheral facial nerve injury. METHODS: Ninety-two patients with 241 injured branches of incomplete peripheral facial nerve injury were divided into a treatment group and a control group. The treatment group consisted of 58 cases that received functional training of facial mimic muscles. The rest of the cases served as controls. Assessment parameters included the House-Brackmann grading system, a quantitative facial nerve function estimating system and electroneurography. According to the three assessments, the facial nerve injury was divided into four grades: normal, minor, moderate and severe. The treatment group started training facial mimic muscle activity 2 weeks after facial nerve injury. After follow-ups of 1 to 4 years, the outcomes were statistically analysed. RESULTS: In the minor facial nerve injury group, there was no significant difference in the time needed for initial recovery (Ti, the time needed for significant recovery of the facial nerve function after injury) and final recovery (Tf, the time point after which no further improvement of facial nerve function was obtained) between the two groups (P > 0.05). No adverse effect was found in these cases. In the moderate facial nerve injury group, the Ti and Tf of the treatment group were shorter than those of the control group (P < 0.05). One case had synkinesis. In the patients with severe facial nerve injury, the recovery rate of facial nerve function in the treatment group was higher than that of the control group and the sequelae were less. CONCLUSION: Functional training of facial mimic muscles cannot shorten the time of recovery for the patients with minor facial nerve injury but it can speed up the recovery and reduce the undesirable sequelae such as synkinesis and hemifacial spasm for the patients with moderate and severe facial nerve injury.

Transport distraction versus costochondral graft for reconstruction of temporomandibular joint ankylosis: which is better?

OBJECTIVE: This study compared the methods of transport distraction and costochondral graft in reconstruction of temporomandibular joint (TMJ) ankylosis. STUDY DESIGN: Bilateral bony TMJ ankylosis was induced in 12 adult goats. Transport distraction of the mandibular ramus was performed on one side, whereas costochondral graft was performed on the other side. Jaw movements, TMJ healing, condylar remodeling, and recurrence of ankylosis were assessed by clinical, radiological, and histological examinations. RESULTS: Both transport distraction and costochondral graft established a neo-condyle separating from the pseudo-disc with a joint space. The ankylosis scores were higher in the distraction side in the first 12 weeks. Both techniques showed no significant difference in the calcification scores and recurrence of ankylosis in the long term. The maximal jaw movements in 3 directions returned to the pre-ankylosis stage by both methods. CONCLUSIONS: Distraction osteogenesis is an effective reconstruction method for TMJ ankylosis, matching the gold standard of costochondral grafting.

[Clinical analysis of 182 cases with facial nerve injury].

OBJECTIVE: To investigate the recovery patterns and the influencing factors of facial nerve injury in maxillofacial surgery by retrospective analysis of a serial clinical data. METHODS: A total of 182 patients with facial nerve injury were reviewed. The cause of injury, the initial facial nerve function after trauma, the treatment, the initial recovery time of facial nerve function and the total recovery time were recorded. The factors that influenced the outcome of facial nerve function were analyzed. RESULTS: The facial nerve branch injury was common in maxillofacial injury. The injury pattern, location, age and reconstruction time all had effects on the function recovery of the facial nerve. Within 6 months, 45 of 49 (92%) anatomic injured patients completely recovered in 6 months; 53 of 59 patients (90%) began to recover when nerve had been ruptured. In 12 months, 33 of 35 patients (94%) after nerve anastomosed and 5 of 8 patients (62%) with nerve transplantation got complete recovery. CONCLUSIONS: Preserving the facial nerve during surgery is very important. If the facial nerve is injured, reconstructive surgery should be applied as soon as possible.

beta-catenin expression pattern in primary oral squamous cell carcinoma.

BACKGROUND: Beta-catenin, a 92 kDa protein that binds to the cytoplasmic tail of E-cadherin, has an essential role in intercellular adhesion and signal transduction. Aberrant expression of beta-catenin has been associated with progression and metastasis of various human cancers. The aim of this study was to elucidate the expression pattern of beta-catenin in primary oral squamous cell carcinoma and examine the correlation between beta-catenin expression and tumor differentiation, histological grade and lymph node status as well as its clinical significances. METHODS: Seventy-six patients with oral squamous cell carcinoma and sixteen metastatic lymph nodes were studied. The beta-catenin expression was determined by immunohistochemical staining. The correlation with clinical, histological data was analyzed statistically. RESULTS: Normal oral epithelium showed strong beta-catenin expression at the cell membrane, but no cytoplasmic or nuclear expression. Different degrees of reduced expression of beta-catenin at the cell membrane were found in 54 cases with squamous cell carcinoma (71%). Cytoplasmic beta-catenin expression was found in 17 tumors (22.4%). Three cases were found with nuclear beta-catenin expression. In sixteen lymph nodes with metastatic squamous cell carcinoma, negative beta-catenin expression at the cell membrane was seen in 13 tumors (81.2%) and weak expression in 3 tumors (18.8%). Statistical analysis showed that there was an inverse correlation between beta-catenin expression and lymph node status and histological grade of tumors. CONCLUSIONS: Reduced beta-catenin expression at the cell membrane is clearly associated with lymph node metastasis. A reduced expression of beta-catenin may constitute a hallmark of aggressive biological behavior of squamous cell carcinoma.

Surgical induction of temporomandibular joint ankylosis: an animal model.

PURPOSE: The aim of this study was to develop an animal model of temporomandibular joint (TMJ) bony ankylosis for future evaluation of surgical reconstructive methods. MATERIALS AND METHODS: An animal model was selected on the basis of 2 sequential experiments. Four goats and 4 minipigs were used in the first experiment, in which 1 goat and 1 minipig served as control animals. Condylectomy with disc preservation was performed on 1 side of 3 animals in each group. On the contralateral TMJ, condylectomy with discectomy was performed, and the arthroplasty gap was filled with the autogenous bone chips. In the second experiment, TMJ ankylosis was induced bilaterally in 3 additional animals of the species that achieved better ankylosis results in the first experiment. All animals were killed postoperatively at 3 months, and the TMJ complexes were examined by plain radiography, computed tomography, and histological evaluation. RESULTS: In the first experiment, only fibrous ankylosis was observed in the bone-grafted side of 2 goats and 3 minipigs, whereas fibro-osseous ankylosis was achieved in the remaining goat. The extent of ankylosis was found to be more severe in the goats than the minipigs. Hence, goats were selected for bilateral surgery in the second experiment, which achieved consistent bony ankylosis of the TMJ in all animals. CONCLUSIONS: Goats provide a better TMJ bony ankylosis model than minipigs. Consistent bony ankylosis can be induced by bilateral condylectomy, disectomy, and bone grafting of the arthroplasty gap.