PEP06 polypeptide 30 is a novel cluster-dissociating agent inhibiting v integrin/FAK/Src signaling in oral squamous cell carcinoma cells

Collectively migrating tumor cells have been recently implicated in enhanced metastasis of epithelial malignancies. In oral squamous cell carcinoma (OSCC), v integrin is a crucial mediator of multicellular clustering and collective movement ; however, its contribution to metastatic spread remains to be addressed. According to the emerging therapeutic concept, dissociation of tumor clusters into single cells could significantly suppress metastasis-seeding ability of carcinomas. This study aimed to investigate the anti-OSCC potential of novel endostatin-derived polypeptide PEP06 as a cluster-dissociating therapeutic agent . Firstly, we found marked enrichment of v integrin in collectively invading multicellular clusters in human OSCCs. Our study revealed that metastatic progression of OSCC was associated with augmented immunostaining of v integrin in cancerous lesions. Following PEP06 treatment, cell clustering on fibronectin, migration, multicellular aggregation, anchorage-independent survival and colony formation of OSCC were significantly inhibited. Moreover, PEP06 suppressed v integrin/FAK/Src signaling in OSCC cells. PEP06-induced loss of active Src and E-cadherin from cell-cell contacts contributed to diminished collective migration of OSCC . Overall, these results suggest that PEP06 polypeptide 30 inhibiting v integrin/FAK/Src signaling and disrupting E-cadherin-based intercellular junctions possesses anti-metastatic potential in OSCC by acting as a cluster-dissociating therapeutic agent.

Mitochondrial uncoupler BAM15 inhibits artery constriction and potently activates AMPK in vascular smooth muscle cells

Our previous studies found that mitochondrial uncouplers CCCP and niclosamide inhibited artery constriction and the mechanism involved AMPK activation in vascular smooth muscle cells. BAM15 is a novel type of mitochondrial uncoupler. The aim of the present study is to identify the vasoactivity of BAM15 and characterize the BAM15-induced AMPK activation in vascular smooth muscle cells (A10 cells). BAM15 relaxed phenylephrine (PE)-induced constricted rat mesenteric arteries with intact and denuded endothelium. Pretreatment with BAM15 inhibited PE-induced constriction of rat mesenteric arteries with intact and denuded endothelium. BAM15, CCCP, and niclosamide had the comparable IC value of vasorelaxation in PE-induced constriction of rat mesenteric arteries. BAM15 was less cytotoxic in A10 cells compared with CCCP and niclosamide. BAM15 depolarized mitochondrial membrane potential, induced mitochondrial fission, increased mitochondrial ROS production, and increased mitochondrial oxygen consumption rate in A10 cells. BAM15 potently activated AMPK in A10 cells and the efficacy of BAM15 was stronger than that of CCCP, niclosamide, and AMPK positive activators metformin and AICAR. In conclusion, BAM15 activates AMPK in vascular smooth muscle cells with higher potency than that of CCCP, niclosamide and the known AMPK activators metformin and AICAR. The present work indicates that BAM15 is a potent AMPK activator.

Serum vitamin D levels in tuberculosis patients with different blood glucose status / 中华全科医师杂志

Objective To investigate serum vitamin D levels in tuberculosis (TB) patients with different blood glucose status.Methods Two hundred and forty seven TB patients were recruited from tuberculosis clinics in Jilin province and 80 normal subjects who underwent health check up in Beijing Hospital served as controls.Blood samples were collected,fasting blood glucose (FBG) and serum vitamin D [25 (OH)D] levels were measured.Results FBG results showed that there were 95 patients with normal FBG,69 with pre-diabetes (pre-DM) and 83 with diabetes mellitus (DM).Vitamin D measurement showed that 25(OH) D level in TB patients with normal FBG,pre-DM and DM was 16.1 (10.7,26.2) μg/L,12.9 (9.6,20.1) μg/L and 12.4 (10.4,16.9) μg/L,respectively,(x2 =19.608,P ＜ 0.001) and were much lower than that in the normal controls (20.5 μg/L) (x2 =21.701,P ＜ 0.001).Proportion of TB patients with 25 (OH)D sever deficiency(＜ 10.0 μg/L)in patients with normal FBG,pre-DM and DM was 20.0％ (19/95),31.9％ (22/69),and 24.1％ (20/83) respectively (x2 =6.376,P ＜ 0.05);proportion of 25 (OH) D deficiency (10.0-19.9 ng/ml) in three groups was 41.1％ (39/95),40.6％ (28/69),and 57.8 ％ (48/83),respectively (x2 =15.141,P ＜ 0.05);sufficient 25 (OH) D (≥ 30.0 μg/L) was 14.7％ (14/95),7.2％ (5/69),and 1.2％ (1/83),respectively (x2 =19.118;P ＜0.05).While the proportion of TB patient with 25 (OH) D insufficiency (20.0-29.9 ng/ml) was 24.2％ (23/95),20.3％ (14/69),and 16.9％ (14/48) respectively (x2 =0.933,P =0.627).In TB patients with normal FBG,risk factors for 25 (OH) D deficiency were smoking (OR =5.619,95％ CI:1.293-24.424,P =0.021),cold season (OR =14.402,95％CI:4.070-50.965,P ＜ 0.001) and smear negative TB (OR =6.194,95 ％ CI:1.873-20.481,P =0.003).Living in rural area (OR =3.429,95％ CI:1.040-11.299,P =0.043) was the risk factor for 25 (OH) D deficiency in TB patients with pre-DM and age ≥ 60 years (OR =2.474,95％CI:1.086-5.623,P =0.031) was risk factor for 25 (OH) D deficiency in those with DM.Conclusions Vitamin D level is lower in TB patients than that in normal controls.The diabetic TB patients have the lowest 25 (OH) D level and have highest proportion of vitamin D deficiency and sever deficiency,particularly for elderly patients.

Individualized treatment of bilateral carotid body tumor / 中华耳鼻咽喉头颈外科杂志

Objective@#To investigate individualized therapeutic strategy for bilateral carotid body tumors.@*Methods@#Clinical data of 16 patients with bilateral carotid body tumor treated from January 2003 to August 2016 were retrospectively studied. Of the 16 patients, 9 were males and 7 were females; 5 were sporadic and 11 were familial; 8 cases were observed, 1 cases was malignant and treated with chemotherapy, and 7 cases were treated with surgery. The treatment course, perioperative complications and clinical efficacy were assessed. Comprehensive evaluation of bilateral carotid body tumors was performed based on the size of bilateral tumor, clinical manifestations, genetic tests and other indicators. Individual treatment strategies included observation, surgery and observation, bilateral surgery, radiotherapy or chemotherapy. Surgical resection of carotid body tumor was unilateral in 3 cases and bilateral in 3 cases; removal of bilateral carotid body tumors plus unilateral jugular bulb in 1 case; and the internal carotid artery was reconstructed with autologous greater saphenous vein in 1 case.@*Results@#All patients were followed up for 3 months to 12 years. There was no patient death during perioperative period. Superior laryngeal nerve injury occurred in 2 case. Baroreceptor failure syndrome occurred in one patient, but it gradually recoverd with medical treatments.@*Conlusion@#It is important to identify whether bilateral carotid body tumors are hereditary and to make an individualized therapeutic strategy for each patient with bilateral carotid body tumors, focusing on the improvement in the quality of life of patient.

Artificial trachea with pedicle rib cartilage and cilia endothelial wall / 中国耳鼻咽喉头颈外科

[ABSTRACT]OBJECTIVETo explore the method on reconstruction of long and special tracheal defects which can mostly match with natural airway: pedicle rib cartilage with cilia endothelial wall.METHODS8 experimental model of rabbits were trained with cervical double belt blood supply fascia embedding and autologous costal cartilage and nasal septum mucos in the first period, then followed by transplantation in the second period. After the operation, we would assess the physiology, breathing and histopathology index of the rabbits, etc. After the animal experiment, we tried to apply the method to an appropriate clinical case.RESULTS8 cases of experimental rabbits dead after the second period operation with the average survival time of 21.9 days and caused by asphyxia. Histopathological results: rib cartilage and trachea ring up of cartilage cells and fibers have high similarity in histology; cartilage of all cases under the cultivation of the pedicle fascial package has not been absorbed; all cases' nasal septum mucosa in the body and blood supply to differentiation under fascia nutrition. Then we applied the method on a clinical case.CONCLUSIONTrachea ring rib cartilage had a higher similarity to the tracheal cartilage on the histology and biological characteristics that can be used as the preparation of artificial trachea shaping material or cell culture to tissue engineering materials. Package of rib cartilage pedicle fascial can provide adequate blood supply to make up for a free training rib cartilage defect to its absorption. Nasal septum mucosa of pressure in the body and blood supply of the fascia nutrition can simulate the trachea ciliated epithelium, which can play its biological characteristics similar to the inner wall of the trachea.

Plasmid and stable cell linereconstruction of RET point mutation in medullary thyroid carcinoma pedigree / 中国耳鼻咽喉头颈外科

OBJECTIVEThe plasmid construction and validation of RET point mutation in vitro.METHODS The RET gene exon in the mainland familial medullary thyroid carcinoma family was analized with muon capture two generation sequencing of. In vitro, the RET relative point mutationswere reconstructed in NIH3T3 cells and the expression levels were studied.RESULTSThe corresponding lentiviral plasmids of RET point mutations were successfully obtained after point mutating the wild RET plasmids, it was verified that the target genes were expressed in NIH3T3 cells stably by Western Blot. CONCLUSIONSStable cell lines carrying RET point mutations were reconstructedsuccessfully, which provide a good platform for studying various point mutations.

Clinical efficacy analysis of adult sinonasal rhabdomyosarcoma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the clinical prognostic impact factors of adult sinonasal rhabdomyosarcoma (SNRMS).@*METHOD@#The clinical features, treatment methods, and disease outcome were reviewed retrospectively for twenty-three adult SNRMS between 2006 January and 2014 December. The survival analysis was performed by Kaplan-Meier estimate and the comparison between groups by Log-rank test. Multivariate analysis was carried out by Cox proportional hazard model.@*RESULT@#Patients' ages ranged from 18 to 59 years (median, 23.2 years). With a median follow-up of 20 moths (3-47 moths), 14 cases dead and 9 cases alive, the 1-year and 2-year overall survival (OS) rates were 77.1% and 35.0%, respectively. Within the 1-year and 2-year OS rates,early stage group had a higher overall survival rates than advanced diease group (100.0%, 66.7% and 83.3%, 10.5%, P < 0.05); combined therapy group had a higher overall survival rates than single treatment group (86.7%, 50.0% and 50.8%, 0, P < 0.05). In the non-metastasis group (21 cases), 1-year and 2-year distant metastasis rates were 38.1% and 70.5%, respectively. Multivariate analysis showed that radiotherapy, chemotherapy and tumor diameter less than 5 cm were good prognostic factors (P < 0.05), while the lymph node metastasis, meningeal involvement and orbital involvement were poor prognostic factors (P < 0 05). In the 14 cases of dead patients, 92 8% (13/14) died of distant metastasis.@*CONCLUSION@#Adult RMS had a high advanced rate with poor prognosis. Distant metastasis is the leading cause of death. Controlling distant metastasis is a key to improve the survival rate.

[In situ visual imaging of oral squamous cell carcinoma in mice by using near-infrared quantum dots conjugated with arginine-glycine-aspartic acid peptide fluorescent probes].

OBJECTIVE: To investigate in situ visualization using near-infrared quantum dots (QDs) conjugated with arginine- glycine-aspartic acid (ROD) peptide fluorescent probes in oral squamous cell carcinoma (08CC). METHODS: QDs with emission wavelength of 800 nm (QD800) were conjugated with RGD peptides to produce QD800-RGD fluorescent probes. Human OSCC cell line BcaCD885 was inoculated in nude mice cheeks to establish OSCC mouse models. Frozen BcaCD885 tumor slices were immunofluorescence double stained by using QD800-RGD and CD105 monoclonal antibody and were observed using a laser scanning confocal microscope. QD800-RGD was injected into the OSCC models through the tail veins, and the in situ visualization was analyzed at different time points. The mice were sacrificed 12 h after injection to isolate tumors for the ex vivo analysis of probe localization in the tumors. RESULTS: QD800-RGD specifically targeted the integrin avß3 expressed in the endothelial cells of tumor angiogenic vessels in vitro and in vivo, producing clear tumor fluorescence images after intravenous injection. The most complete tumor images with maximal signal-to-noise ratios were observed 0.5 h to 6 h after injection of the probe and significantly reduced 9 h after the injection. However, the tumor image was still clearly visible at 12 h. CONCLUSION: Using intravenously injected QD800-RGD generates high quality OSCC images when integrin avß3, which is expressed in the endothelial cells of tumor angiogenic vessels, is used as the target. The technique offers great potential in the diagnosis and individual treatment of OSCC.

In situ visual imaging of oral squamous cell carcinoma in mice by using near-infrared quantum dots conjugated with arginine-glycine-aspartic acid peptide fluorescent probes / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate in situ visualization using near-infrared quantum dots (QDs) conjugated with arginine- glycine-aspartic acid (ROD) peptide fluorescent probes in oral squamous cell carcinoma (08CC).</p><p><b>METHODS</b>QDs with emission wavelength of 800 nm (QD800) were conjugated with RGD peptides to produce QD800-RGD fluorescent probes. Human OSCC cell line BcaCD885 was inoculated in nude mice cheeks to establish OSCC mouse models. Frozen BcaCD885 tumor slices were immunofluorescence double stained by using QD800-RGD and CD105 monoclonal antibody and were observed using a laser scanning confocal microscope. QD800-RGD was injected into the OSCC models through the tail veins, and the in situ visualization was analyzed at different time points. The mice were sacrificed 12 h after injection to isolate tumors for the ex vivo analysis of probe localization in the tumors.</p><p><b>RESULTS</b>QD800-RGD specifically targeted the integrin avβ3 expressed in the endothelial cells of tumor angiogenic vessels in vitro and in vivo, producing clear tumor fluorescence images after intravenous injection. The most complete tumor images with maximal signal-to-noise ratios were observed 0.5 h to 6 h after injection of the probe and significantly reduced 9 h after the injection. However, the tumor image was still clearly visible at 12 h.</p><p><b>CONCLUSION</b>Using intravenously injected QD800-RGD generates high quality OSCC images when integrin avβ3, which is expressed in the endothelial cells of tumor angiogenic vessels, is used as the target. The technique offers great potential in the diagnosis and individual treatment of OSCC.</p>

In situ visual imaging and in vivo distribution of head and neck squamous cell carcinoma in mice by near-infrared fluorescent quantum dots epidermal growth factor receptor monoclonal antibody probe / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the in situ visualization imaging and the in vivo distribution of the near-infrared fluorescent quantum dots (QDs) epidermal growth factor receptor (EGFR) monoclonal antibody (mAb) probe in head and neck squamous cell carcinoma (HNSCC).</p><p><b>METHODS</b>QDs with an emission wavelength of 800 nm (QD800) was conjugated with EGFR mAb to produce QD800-EGFR mAb. QD800-EGFR mAb was co-cultured with BcaCD885 squamous cancer cell line for 30 min and observed with laser scanning confocal microscope (LSCM). QD800-EGFR mAb was injected into HNSCC animal model through the tail vein, and the in situ visualization imaging and the in vivo distribution of QD800-EGFR mAb was analyzed at different time points.</p><p><b>RESULTS</b>BcaCD885 squamous cell carcinoma in the head and neck can be imaged clearly and visually after intravenous injection of QD800-EGFR mAb probe, these fluorescence signals lasted for 24 h. The most complete tumor images with maximal signal-to-noise ratio were observed from 30 min to 6 h after injection of the probe. In vivo tissue distribution studies demonstrated that QD800 aggregated mostly in liver. QD800 aggregation decreased with time in tumors, and QD800 didn't aggregate in heart, brain, intestine, lung and stomach.</p><p><b>CONCLUSION</b>The QD800-EGFR mAb probe can clearly produce visual images in head and neck cancer. It has promising prospects in visualization in vivo imaging and individual treatment of head and neck cancer.</p>