[Correlation between temporomandibular joint osteoarthrosis and coronoid process].

Objective: To investigate the morphological characteristics of coracoid process in different stages of temporomandibular joint osteoarthrosis (TMJOA), and to provide theoretical data for clinical and anatomic study. Methods: A total of 290 patients who were diagnosed with TMJOA in the Department of Temporomandibular Joint, Kunming Medical University School and Hospital of Stomatology from January 2015 to February 2021 were collected, including 69 males and 221 females, with an average age of 35.1±13.7 years (16-69 years old), 64 cases of unilateral lesions (64 sides), and 226 cases of bilateral lesions (452 sides). According to the TMJOA X-ray staging standard put forward by Ma Xuchen in 2005, the affected joints were divided into stage I (227 sides), stage Ⅱ (38 sides), stage Ⅲ (164 sides) and stage Ⅳ (87 sides). Twenty-six patients without clinical and imaging manifestations of temporomandibular disorders in the Department of Radiology, Kunming Medical University School and Hospital of Stomatology from October 2020 to June 2021 were selected as the control group, including 8 males and 18 females. The average age was (34.3±13.9) years (17-60 years). The dicom data of each group were imported into SimplantPro11.04 software to measure the height of coracoid process, anteversion angle and the ratio of coracoid vertex to mandibular corner to condylar vertex to mandibular angle. R 3.6.1 was used to analyze the difference of the morphological characteristics of coracoid process between in the affected side of TMJOA and in the both sides of the control group, in the healthy side and the affected side of unilateral patients and in different stages of TMJOA. Results: The height of the coracoid process [(16.26±2.81 mm)], the ratio of the coracoid process vertex-mandibular angle point and the condyle vertex-mandibular angle point distance [0.96(0.92,1.01)] on the affected side of TMJOA were significantly higher than those in the control group [(15.31±3.03)mm;0.95(0.89、0.99)] (t=2.18, P=0.033; t=2.87, P=0.004). There was no significant difference between the ante-version angle and the control group (t=-1.37, P=0.176). The ratio of the distance between the apex of the coracoid process and the apex of the mandibular angle to the apex of the condyle and the angle of the mandible in the affected side of unilateral patients was significantly greater than that in the healthy side (t=-3.46, P=0.001). There was no significant difference in coracoid height, coracoid anteversion angle and the healthy side (t=-1.85, P=0.069; t=-0.06, P=0.955) in different periods. The intra-group analysis showed that there was no significant difference in the height of the coracoid process in different stages (F=0.37, P=0.774). There was no significant difference in the ante-version angle of the coracoid process: stage I, stage Ⅱ, and stage Ⅲ (P>0.008), but all were significantly smaller than stage Ⅳ (PⅠ-Ⅳ<0.001, PⅡ-Ⅳ=0.009, PⅢ-Ⅳ<0.001). The ratio of the distance between coracoid apex-mandibular angle and condyle apex-mandibular angle: there was no significant difference in stage I, stage Ⅱ, and stage Ⅲ (P>0.008), and stage I and stage Ⅲ were significantly smaller than stage Ⅳ (P<0.001). Conclusions: The coracoid height and the ratio of the coracoid apex-mandibular angle to the condyle apex-mandibular angle distance on the TMJOA side were significantly greater than those without temporomandibular joint disorders. The bone deposition was mainly concentrated in the upper and posterior part of the condyle. TMJOA had a certain correlation with the height of the coracoid process.

[Effects of Porphyromonas gingivalis lipopolysaccharide on the proliferation and migration of human umbilical artery smooth muscle cells under co-culture conditions].

Objective: To investigate the effects of Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) of type Ⅰand Ⅳ fimA on the proliferation and migration of human umbilical artery smooth muscle cells (HUASMC) under co-culture conditions and to explore the biological basis and possible mechanisms of the relationship between periodontitis and atherosclerosis (As). Methods: Type Ⅰ and Ⅳ fimA Pg were anaerobically cultured and the Pg-LPS was extracted, purified and identified. Human umbilical vein endothelial cells (HUVEC) and HUASMC were cultured in vitro and the HUVEC-HUASMC co-cultured cell model was established using rat tail type Ⅰ collagen. The experiment was divided into three groups: group T1 (co-cultured cells were stimulated with type Ⅰ fimA Pg-LPS at the mass concentrations of 0.5, 1.0, 2.0, 5.0, 10.0 mg/L), group T2 (co-cultured cells were stimulated with type Ⅳ fimA Pg-LPS at the mass concentrations of 0.5, 1.0, 2.0, 5.0, 10.0 mg/L), and negative control group (without LPS). Cell counting kit-8 (CCK-8) was used to detect the proliferation ability of HUASMC and the migration ability of HUASMC was observed by using the Transwell migration chamber. Comparasons of the changes in the proliferation and migration ability of HUASMC after 2, 8, 24 and 48 h of Pg-LPS stimulated co-cultured cells with various mass concentrations of Pg-LPS were conducted. Results: For the co-cultured cells under the action of type Ⅰ and Ⅳ fimA Pg-LPS, at 24 and 48 h, in each mass concentration group of the two types of Pg-LPS (0.5, 1.0, 2.0, 5.0, 10.0 mg/L), HUASMC's A values were significantly up-regulated compared to the negative control group (P<0.05) and for the co-cultured cells after the stimulation of type Ⅳ fimA Pg-LPS at concentrations of 5 and 10 mg/L at 48 h, the A values (1.386±0.044, 1.455±0.058) of HUASMC were significantly higher than that of HUASMC (1.168±0.064, 1.204±0.088) in the same concentrations of type Ⅰ fimA Pg-LPS (P<0.05). In addition, the A values of HUASMC in stimulated co-cultured cells under concentrations of 5.0 and 10.0 mg/L of type Ⅳ fimA Pg-LPS at 48 h were significantly higher than that in stimulated co-culture cells under the concentrations of 0.5 and 1.0 mg/L of type Ⅳ fimA Pg-LPS (1.170±0.082, 1.239±0.089) (P<0.05). The migration results of HUASMC showed that at 8, 24, and 48 h, the numbers of migration of HUASMC in each mass concentration group of type Ⅰ and Ⅳ fimA Pgas-LPS were significantly higher than that of HUASMC under the same Pg-LPS mass concentration at 2 h in the same group (P<0.05). The migration quantities of HUASMC in the other Pg-LPS mass concentration groups at 48 h were significantly higher than that of HUASMC under the same Pg-LPS mass concentration at 24 h in the same group (P<0.05), except for 10.0 mg/L type Ⅳ fimA Pg-LPS. The numbers of HUASMC migration in 2.0 mg/L type Ⅳ fimA Pg-LPS stimulated co-cultured cells at 48 h (204.00±20.98) were significantly higher than that in type Ⅰ fimA Pg-LPS stimulated co-culture cells at the same concentration (141.89±18.28) (P<0.05). Further more, at the same observation time point, the higher the Pg-LPS concentration, the greater the number of HUASMC migration (P<0.05). Conclusions: Both Ⅰ and Ⅳ fimA Pg-LPS could enhance the proliferation and migration ability of HUASMC in the co-culture system, and the virulence of Pg-LPS might be related to the fimA genotype. Ⅳ fimA Pg-LPS showed a more significant stimulating effect and more likely to cause HUASMC dysfunction than Ⅰ fimA Pg-LPS, which provided part of the basis for the progression of As in severe periodontitis.

Identification and validation of novel microenvironment-based immune molecular subgroups of head and neck squamous cell carcinoma: implications for immunotherapy.

Background: Targeting the immune checkpoint pathway has demonstrated antitumor cytotoxicity in treatment-refractory head and neck squamous cell carcinoma (HNSC). To understand the molecular mechanisms underpinning its antitumor response, we characterized the immune landscape of HNSC by their tumor and stromal compartments to identify novel immune molecular subgroups. Patients and methods: A training cohort of 522 HNSC samples from the Cancer Genome Atlas profiled by RNA sequencing was analyzed. We separated gene expression patterns from tumor, stromal, and immune cell gene using a non-negative matrix factorization algorithm. We correlated the expression patterns with a set of immune-related gene signatures, potential immune biomarkers, and clinicopathological features. Six independent datasets containing 838 HNSC samples were used for validation. Results: Approximately 40% of HNSCs in the cohort (211/522) were identified to show enriched inflammatory response, enhanced cytolytic activity, and active interferon-γ signaling (all, P < 0.001). We named this new molecular class of tumors the Immune Class. Then we found it contained two distinct microenvironment-based subtypes, characterized by markers of active or exhausted immune response. The Exhausted Immune Class was characterized by enrichment of activated stroma and anti-inflammatory M2 macrophage signatures, WNT/transforming growth factor-ß signaling pathway activation and poor survival (all, P < 0.05). An enriched proinflammatory M1 macrophage signature, enhanced cytolytic activity, abundant tumor-infiltrating lymphocytes, high human papillomavirus (HPV) infection, and favorable prognosis were associated with Active Immune Class (all, P < 0.05). The robustness of these immune molecular subgroups was verified in the validation cohorts, and Active Immune Class showed potential response to programmed cell death-1 blockade (P = 0.01). Conclusions: This study revealed a novel Immune Class in HNSC; two subclasses characterized by active or exhausted immune responses were also identified. These findings provide new insights into tailoring immunotherapeutic strategies for different HNSC subgroups.

Can we make an accurate diagnosis of pyriform sinus fistula by fetal MRI?

AIM: To review the fetal magnetic resonance imaging (MRI) features of pyriform sinus fistula (PSF) and to compare them with the postnatal clinical and surgical findings. The relevant medical literature is also discussed. MATERIALS AND METHODS: The location, shape, signal, effects of adjacent structures and in utero changes detected on fetal MRI were reviewed in three cases of PSF. The patient's respiratory status at birth, the application of ex utero intrapartum therapy (EXIT), infectious complications in the neonatal period, and the surgical and pathological findings were also reviewed and discussed. The study consisted of three pregnant women between 18 and 38 years of age, each with a single fetus. Fetal MRI was performed at 26 and 38 weeks of gestation in case 1, at 34 weeks of gestation in case 2, and at 34 and 38 weeks of gestation in case 3. Postnatal clinical follow-up extended from 9 months to 3 years. RESULTS: All lesions were well-circumscribed unilobular cysts with slightly thickened walls at the neck area. Case 1 was irregular in shape, and the other two cases were oval and extended longitudinally along the neck. The fluid contents showed signals consistent with amniotic fluid. Close contact with the thyroid gland was demonstrated to be a fairly characteristic feature that could be recognised on fetal MRI. The in utero follow-up detected slight changes in the signal, wall characteristics, contour, and airway deviation. Case 1 was bilaterally affected, causing it to be initially mistaken as macro-cystic lymphangioma. The diagnosis was corrected with the aid of postnatal imaging; however, a prenatal diagnosis was correctly made in cases 2 and 3. Airway obstruction was not observed in any of the three cases, and thus, EXIT was not applied. The fetal MRI findings were in good agreement with the postnatal clinical and surgical findings. In addition to the higher spatial delineation provided by fetal MRI, as described in the literature, the observation of close contact with the thyroid gland and a flattened deformity of the thyroid gland are other characteristic features. CONCLUSION: Fetal MRI is useful for displaying characteristics of PSF that can enable an accurate prenatal diagnosis.

Expression of transforming growth factor-ß1 in neonatal rats with hyperoxia-induced bronchopulmonary dysplasia and its relationship with lung development.

The aim of this study was to detect the expression of transforming growth factor-ß1 (TGF-ß1) in neonatal rats with hyperoxia-induced bronchopulmonary dysplasia (BPD) and to explore its relationship with lung development. Forty-eight rats (2-3 days old) were randomly divided into a hyperoxia group and a control group (N = 24) which were then fed in ≥95% oxygen atmosphere and air, respectively. On the 1st, 3rd and 7th days of hyperoxia exposure, morphological changes of lung tissues were observed under an optical microscope. TGF-ß1 mRNA and protein levels in lung tissues were detected by real-time quantitative polymerase chain reaction and western blot, respectively. With increasing time of hyperoxia exposure, the hyperoxia group gradually suffered from pathological changes such as poor development of lung tissues, alveolar simplification, decrease in the number of alveoli, and hindered pulmonary microvascular development. On the 7th day of hyperoxia exposure, TGF-ß1 mRNA and protein levels (relative to b-actin) of the hyperoxia group (0.34 ± 0.19 and 0.21 ± 0.09, respectively) were significantly lower than those of the control group (0.83 ± 0.45 and 0.57 ± 0.45, respectively; P < 0.05). TGF-ß1 participates in the pathogenesis of BPD as an important regulatory factor during pulmonary vascular development.

MiR-34c suppresses tumor growth and metastasis in nasopharyngeal carcinoma by targeting MET.

Our previous microarray analysis indicated that miR-34c was downregulated in nasopharyngeal carcinoma (NPC). However, little is known about the function and molecular mechanism of miR-34c in NPC. In this study, miR-34c was found to be significantly downregulated in NPC cell lines and clinical tissues. Ectopic expression of miR-34c suppressed NPC cell viability, colony formation, anchorage-independent growth, cell migration and invasion in vitro, and inhibited xenograft tumor growth and lung metastasis in vivo. MET proto-oncogene (MET) was identified as a direct target of miR-34c using luciferase reporter assays, quantitative RT-PCR, western blotting and immunofluorescent staining. Overexpression of miR-34c markedly reduced MET expression at both the mRNA and protein levels. Knockdown of MET suppressed NPC cell proliferation, migration and invasion, whereas the restoration of MET rescued the suppressive effects of miR-34c. The demethylation agent 5-aza-2'-deoxycytidine (DAC) restored the expression of miR-34c in NPC cell lines. The promoter region of miR-34c was hypermethylated in NPC cells. In conclusion, miR-34c suppresses tumor growth and metastasis in NPC by targeting MET. The newly identified miR-34c/MET pathway provides further insights into the development and progression of NPC, and may represent a novel therapeutic target for NPC treatment.

Mannan-modified adenovirus as a vaccine to induce antitumor immunity.

Tumor vaccine is a useful strategy for cancer therapy. However, priming of the immune system requires the relevant antigen to be presented by antigen-presenting cells (APCs). Here, we employed telomerase reverse transcriptase as a model antigen to explore the feasibility of using mannan-modified adenovirus as a tumor vaccine. We found that tumor immunogene therapy with the vaccine was effective at protective antitumor immunity in mice. The antigen-specific cytotoxic T lymphocytes were found in in vitro cytotoxicity assay. The elevation of the killing activity could be abrogated by anti-CD8 or anti-major histocompatibility complex-I antibodies. Adoptive transfer of purified CD8+ cells, and CD4+ cells to a less extent, was effective at antitumor activity. In vivo antitumor activity could be abrogated by depleting CD4+ T lymphocytes. A possible explanation for the antitumor effects may be the antigen was transferred to APCs in the presence of mannan. These observations provide insights into the design of novel vaccine strategies and might be important for the future application of antigens identified in other diseases.

Systemic inhibition of tumor growth by soluble Flk-1 gene therapy combined with cisplatin.

Soluble Flk-1, a soluble vascular endothelial growth factor (VEGF) receptor, is a potent inhibitor of angiogenesis, which could restrain growth and metastasis of some experimental tumors. However, antiangiogenic agents alone cannot eradicate tumor completely, and should be combined with other therapy to enhance their effects. In this study, we evaluated the antitumor activity of the combination therapy in the immunocompetent BALB/c mice bearing H22 hepatoma and Meth A fibrosarcoma, respectively. Mice were treated with either msFlk-1 i.m. at 100 microg/mouse once every 3 days for four times from day 3 after the tumor cell injection, cisplatin cycled twice (2 mg/kg i.p. on days 4 and 11 after the tumor cell inoculation), or both agents together. Tumor growth and survival time were continually observed. Antiangiogenesis in vivo was determined by CD31 immunohistochemistry. Assessment of apoptotic cells and histological analysis was also conducted in tumor tissues. Our results showed that the combination therapy could evidently improve antitumor efficacy, including tumor growth suppression, mice survival prolongation, tumor cell apoptosis augmentation as well as neovascularization inhibition as compared with controls, without serious adverse effects. Our data suggest that the combination of DDP with msFlk-1 is more effective to suppress tumor growth in mice than either agent alone, and this combination regimen showed its potential for future clinical application.

Immunogene therapy of tumors with vaccine based on Xenopus homologous vascular endothelial growth factor as a model antigen.

Overcoming immune tolerance of the growth factors associated with tumor growth should be a useful approach to cancer therapy by active immunity. We used vascular endothelial growth factor (VEGF) as a model antigen to explore the feasibility of the immunogene tumor therapy with a vaccine based on a single xenogeneic homologous gene, targeting the growth factors associated with angiogenesis. To test this concept, we constructed a plasmid DNA encoding Xenopus homologous VEGF (XVEGF-p) and control vectors. We found that immunogene tumor therapy with a vaccine based on XVEGF was effective at both protective and therapeutic antitumor immunity in several tumor models in mice. VEGF-specific autoantibodies in sera of mice immunized with XVEGF-p could be found in Western blotting analysis and ELISA assay. The purified immunoglobulins were effective at the inhibition of VEGF-mediated endothelial cell proliferation in vitro, and at antitumor activity and the inhibition of angiogenesis by adoptive transfer in vivo. The elevation of VEGF in the sera of the tumor-bearing mice could be abrogated with XVEGF-p immunization. The antitumor activity and production of VEGF-specific autoantibodies, significantly elevated IgG1 and IgG2b, could be abrogated by the depletion of CD4(+) T lymphocytes. The observations may provide a vaccine strategy for cancer therapy through the induction of autoimmunity against the growth factors associated with tumor growth in a cross reaction with single xenogeneic homologous gene and may be of importance in the further exploration of the applications of other xenogeneic homologous genes identified in human and other animal genome sequence projects in cancer therapy.

Immunotherapy of tumors with xenogeneic endothelial cells as a vaccine.

The breaking of immune tolerance against autologous angiogenic endothelial cells should be a useful approach for cancer therapy. Here we show that immunotherapy of tumors using fixed xenogeneic whole endothelial cells as a vaccine was effective in affording protection from tumor growth, inducing regression of established tumors and prolonging survival of tumor-bearing mice. Furthermore, autoreactive immunity targeting to microvessels in solid tumors was induced and was probably responsible for the anti-tumor activity. These observations may provide a new vaccine strategy for cancer therapy through the induction of an autoimmune response against the tumor endothelium in a cross-reaction.

Enzyme kinetics of thymidine kinase isoenzymes of Ehrlich ascites tumour.

Three isoenzymes of thymidine kinase (TK) identified on Ehrlich ascites tumour corresponding to pl values of 5.3, and 6.9 and 8.3 were studied with respect to their kinetic properties. The isoenzymes were separated by thin-layer isoelectric focusing and measured in the presence of thymidine (TdR) as substrate, adenosinetriphosphate (ATP) as phosphor donor and deoxythymidinetriphosphate (dTTP) as inhibitor. Exponentially growing cells and growth inhibited cells were used with high proportions of the isoenzymes at pl values of 6.9 and 8.3, and 5.3 respectively. The concentrations of the former were further enhanced by the use of S-phase cells isolated by elutriator centrifugation. The isoenzymes were identified as TK1-onc, earlier found in human leukemia cells. In order to confirm the existence of TK1, particular at pl value 5.3, the ability of the isoenzymes to use different phosphor donors (ATP and uridinetriphosphate (UTP)) and substrates (TdR and deoxycytidine (dCdR) was also studied. These results showed that the isoenzymes at pl 6.9 and 8.3 correspond to TK1, while the isoenzyme at pl 5.3 is heterogeneous. Further, high resolution isoelectric focusing confirmed the existence of two isoenzymes at pl values of 5.1 and 5.3, representing mitochondrial (TK2) and cytoplasmic (TK1) isoenzymes.

[Mebendazole-medicated-salt in the control of hookworm infection].

From 1984-1987, mass application of mebendazole-medicated salt was studied for the control of hookworm infection in 5 pilot areas (Wuming, Shanlin, Bobai, Guilin and Rongxian Counties) in Guangxi Zhuang Autonomous Region. The dosages of mebendazole (mixed with salt) administered were 15mg, 25mg, 30mg, 40mg, 50mg, 30mg, 70mg, 80mg, 100mg or 200mg per person per day for 15, 20 or 30 days. The results showed that 40mg/day for 15-20 days could result in stool egg negative conversion rates of 92.3 to 94.4%, while 50mg/day for 30 days resulted in 96.9 to 100%. Concurrently, the hookworm infection rate in pilot areas dropped by 57.9-71.7% in a short time; whereas the stool egg negative conversion rate of Ascaris lumbricoides was as high as 99.1% to 100% at the same dosage, when the dosage of 100mg/day for 30 days was given, the egg negative reversion rate for Trichuris trichiura was 97.6%. Promising result was also recorded concerning the tolerance of the medicated salt, as the side effects in the inhabitants were mild. In conclusion, the authors considered that the application of mebendazole-medicated salt was a simple and effective measure in controlling hookworm infection, especially in the light of solving the problem of incomplete mass detection and selected mass treatment. Furthermore, it is much more economic from the cost-effect point of view.